Extracts from plant and non-plant biomass and uses thereof

[0001] This application claims priority to India application Serial Nos. 653/MUM/2000 and 654/MUM/2000, both filed Jul. 14, 2000 and PCT application PCT/IN01/00132, filed Jul. 04, 2001, International Publication Number WO 02/05830 A2, which are incorporated herein by reference in their entirety.

[0002] Introduction

[0003] The use of plants and herbs for general welfare and effectively treating a variety of conditions and ailments dates back to ancient times. Use of a variety of all other type of materials of biological origin (biomass), both prokaryotes and eukaryotes, has also been known for treating a variety of conditions and ailments. The biomass source materials can be from bacteria, fungi, mushroom, yeast, poultry, birds, marine, amphibian, animal and human products.

[0004] However, using these materials of biological origin can also be dangerous because of endogenous toxins. Effective doses are often achieved when large volumes of these materials are used, thus aggravating toxic effects.

[0005] The plants of different genera and species of the Crassulaceae family have been used to combat inflammation, promote healing, and improve overall well being. The Crassulaceae fix carbon via Crassulacean Acid Metabolism (CAM)in the dark. CAM, so called because originally found in the Crassulaceae family (stonecrops, comprising mostly succulents such as cacti), plants temporally separate the two pathways of carbon fixation, C₃ and C₄, while using both cycles within the same cells. The initial fixation of carbon dioxide, the C₄ pathway, occurs at night (via cytosolic PEP carboxylase), while the C₃ pathway functions during the day. Consequently, the stomata of CAM plants are closed during the day, thus enabling them to withstand brutal environmental conditions, such as drought and low temperatures, and are open at night to take in carbon dioxide. In contrast, C₄ plants have open stomata during the day and closed during night. CAM plants have been reported in at least 23 families of flowering plants, mostly eudicots, including maternity plant, wax plant, snake plant. Less succulent CAM plants include pineapple and Spanish moss. Interestingly, some nonflowering plants also show CAM activity, including the gymnosperm Welwitschia mirabilisi, quillwort (lsoetes), and some ferns (Raven et al., 1999).

[0006] CAM plants are adapted to high stress conditions such as arid zones, including hot and cold deserts, and high altitudes. CAM plants can be found in many genera and are not limited to succulents; these include Kalanchoe, Bryophyllum, Sedum, Sempervium, Rhodiola, Crassulaceae, Aloe, and Cissus sp. CAM plants have been used for many human applications. Most often, plant parts, such as leaves, or plant juices are orally administered. However, the typical dosage is very high, above 100 mg/kg body weight per day (Blazovics et al., 1993; Boikova and Akulova, 1995; Botha et al., 1997; Da Silva et al., 1995; Da Silva et al., 1999; Lans and Brown, 1998; Nadkarni, 1982; Nassis et al., 1992; Obaseiki-Ebor, 1985; Pal et al., 1992; Sendl et al., 1993; Verma et al., 1986; Yoshikawa et al., 1997). Whenever fresh juice or decoction of a particular plant is given internally according to traditional or folk medicine, the useful medicinal agent is believed to be released in the juice from ground leaves, or the decoction of other plant parts; the plant residues remaining after extraction were not added to the composition.

[0007] Kalanchoe is perhaps the most widely known genus in folk medicine. Kalanchoe pinnata (Lam.) pers; Bryophyllum calicinum Salis; Cotyledon pinnata and Bryophullum pinnatum are synomyous, and have been extensively studied. Folk medicine has bestowed nicknames, such as Wonder Plant and descriptions such as Divine, thus illustrating their importance.

[0008] Sedum and Sempervivum are more commonly known in folk medicine of Europe whereas Rhodiola and other genera are known more in China and the Far East. Although the uses of these genera are not as far-reaching as that of Kalanchoe, specific uses have been reported, such as for protecting the liver and lowering lipids for Sempervivum which are not reported so far for Kalanchoe.

[0009] Aloe sp. has been used to promote health worldwide for thousands of years. Aloe vera is the most commonly used species throughout the world. The plant is used both by external topical application and by internal dose. These applications include promotion of general health; specifically, wound and burn healing, surgery recovery, bone growth, immunoprotection against cancer, health in HIV-infected subjects, protection against frostbite, reducing arthritic swelling, bowel inflammation, blood sugar, and protection of superoxide dismutase and glutathione from radiation.

[0010] The effective dosage for Aloe preparations required is typical for many herbal reparations. Dosage is high: an oral dose of 100 mg/kg per day in animal studies or wound healing (Davies et al., 1989) and 150 mg/kg per day has been prescribed to treat arthritis (Davies et al., 1992). For humans, the reported dose of the extract or juice ranges from 2 g/day ( teaspoon) to over 100 g/day. Direct topical application also requires several grams per application. Anthraquinones and other low molecular weight compounds in Aloe are also reported to have cytotoxicity (Avila et al., 1997; Mueller and Stopper, 1999).

[0011] Cissus quadrangularis is the most commonly used species throughout Asia and Africa. It has been used to promote fracture healing both by external topical application and by internal dose. Other uses include treating rheumatic back pain and body pain, irregular menstruation, stomachache and whooping cough.

[0012] Most parts of the plant have been used, including the stem, leaves, and tender shoots. In addition, the entire plant, dried, is used in Arabic countries as a Cure-All medicine. Pastes made from leaves, stem or entire plants have been used for external applications.

[0013] The dosage levels required are typical for many herbal preparations. Oral dose of juice is 10 to 20 grams/day. Typical dosages of dry stem powder are 2 to 4 grams. Topical applications in the form of paste of dried parts is usually applied at least 10 grams or more. However, the usefulness of this plant is diminished: the various previously known compositions are reported to have mutagenic (including clastogenic) activity (Balachandran et al., 1991; Sivaswamy et al., 1991).

[0014] Table 1 summarizes the common uses of CAM plants; for a comprehensive review of Bryophyllum, Kalanchoe, Aloe and Cissus regarding uses, see (Nadkarni, 1982; Gogte, 2000).

TABLE 1
CAM plants and their uses
Plant	Application	Reference
Kalanchoe pinnata (Lam.)	Anti-inflammatory	(Pal et al., 1992)
pers; (also known as	Anti-bacterial	(Obaseiki-Ebor,
Bryophullum calicinum Salis;		1985; Verma et al.,
Cotyledon pinnata and		1985)
Bryophullum pinnatum) and	Neurotoxin (animal	(Botha et al., 1997)
other Kalanchoe sp.	husbandry study)
	Anti-parasite (e.g.	(Da Silva et al.,
	Leishmania)	1995; Da Silva et
		al., 1999)
	Antihistamine (anti-	(Nassis et al., 1992)
	inflammatory)
	Pharmaceutical	(Verma et al., 1986)
	(general)
Sempervivum sp.	Lipid reduction	(Blazovics et al.,
	(circulation)	1993)
Sedum sp.	Anti-inflammatory	(Sendl et al., 1993)
Rhodiula sacra	Antihistamine (anti-	(Yoshikawa et al.,
	inflammatory)	1997)
Aloe sp.	Macrophage	(Djeraba and Quere,
	activation	2000)
	Wound healing,	(Davies et al., 1989;
	circulation	Paturmaj, 2000)
	Immunoregulation	(Qiu et al., 2000)
	General (reviews)	(Reynolds and
		Dweck, 1999;
		Vogler and Ernst,
		1999)
	Inflammation	(Davies et al., 1992)
	Anti-fungal	(Ali et al., 1999)
	Toxicity	(Avila et al., 1997;
		Mueller and
		Stopper, 1999)
Cissus sp.	Animal health	(Barakat et al.,
	(general)	1985)
	Animal health-	(Hifny et al., 1984)
	nervous system
	(brain)
	Fracture healing	(Chopra et al., 1975;
		Chopra et al., 1976)
	Toxicity	(Balachandran et al.,
		1991; Sivaswamy et
		al., 1991)
General Crassulaceae	Ovulation regulation	(Boikova and
references		Akulova, 1995)
	Uses in poultry	(Lans and Brown,
		1998)

[0015] Many other plants front different families and genera have also been used for the treatment of a wide variety of conditions and ailments all over the world. This range includes:

[0016] Angiosperm-Monocotyledon Plants

[0017] Angiosperm-Dicotyledon Plants

[0018] Gymnosperm Plants

[0019] Many parts of the plants have been used, including the stem, leaves, rhizomes, seeds, tender shoots. In addition, the entire plant, dried, is also used in some cases. Pastes made from leaves, stem or entire plants have been used for external applications.

[0020] The dosage levels required are typical for many herbal preparations. Oral dose of juice or decoction of plant parts is 2 to 50 ml/day. Typical dosages of dry powder are 1 to 4 grams. Topical applications in the form of paste of dried parts is usually applied at least 10 grams or more.

[0021] A few illustrative examples are described. For a comprehensive review regarding human dosage and uses, see (Nadkarni, 1982; Gogte, 2000). For typical human dosage, see Tables 1A, 1B and 1C.

[0022] Angiosperm-Monocotyledon Plants

[0023] Cyperous rotundus is well known to folk medicine in the Orient and Africa. Rhizomes or tuberous roots of cyperous are used as anti-inflammatory, anti-pyretic and analgesic particularly for stomach and bowel disorders (Gupta et al., 1971; Seo et al., 2001). Inhibition of NO and superoxide prodution is also reported. Decoctions of tubers or ground powder are given in fever, diarrhoea, dysentery. Paste is applied on the breasts as a galactogouge. (Nadkarni 1982). The human dose of powder is 1 to 3 gm per day. Antimalarial compounds have been isolated (Thebarononth et al., 1995; Weenan et al., 1990). The most active compounds have an IC50 of 5.4 micrograms/ml.

[0024] Andropogon muricatus is known (Nadkarni 1982; Gogte, 2000) as a coolent and anti-pyretic. It is particularly useful to reduce burning sensation. The root is given as powder or in the form of an infusion. It controls diaphoresis.

[0025] Avena sativa or the common cereal, oat, is regarded as a nutritious cereal. Oat bran contains hemicellulose and its intake reduces LDL (Low Density Lipoproteins) and increases HDL (High Density Lipoproteins) levels in the blood.

[0026] Triticum vulgare or the common cereal, wheat, is generally used as a food ingredient. However, general tonic properties of wheat components and preparations are known in folk practice. In wheat grass therapy juice of one week old wheat grass is is ingested as a general tonic. Wheat germ is also used in folk practice as a special tonic for general well-being. Wheat germ agglutinin (0.5-1.0 mg/ml) promoted phagocytosis by human blood PMN cells 1.8 times compared to Zymosan-A control (Stoika et al., 2001). However, at higher concentration (5-10 ng/ml) it strongly inhibited phagocytosis.

[0027] Allium cepa (Onion) and Allium sativum (Garlic) are the two most commonly used substances of the genus Allium. A variety of uses of onion and garlic have been reported in folk medicinal literature (Nadkarni 1982; Gogte, 2000). Garlic is used particularly for stinulating the digestive and circulatory system. Human dose of bulb paste is 1 to 6 gm per day. Onion is useful in arthritis, sciatica, digestion and cardiac debility (Nadkarni 1982; Gogte, 2000). One (1) to three (3) grams powder of the dry onion bulb is used.

[0028] Curcuma genus contains a number of species of medicinal importance such as Curcuma amadal Curcuma longa, Curcuma angustifolia, Curcuma aromatica, Curcuma zedoaria and others. Infusion and paste of the rhizome of Curcuma amada is used (Nadkarni 1982). Internally, the infusion is given for worms and for purification of the blood. Externally, the paste is applied for a variety of skin diseases, often in combination with other medicines. Antifungal activity of the rhizomes has been reported (Ghosh et al., 1980). Human dose of the rhizome powder is 1 to 3 gm per day. Curcuma longa is used (Nadkarni, 1982; Gogte, 2000) as analgesic, anti-inflammatory and chologogouge.

[0029] Zingiber genus contains a number of species of medicinal importance. Zingiber officinale (ginger, the common spice) is the most commonly used species throughout the world. Ginger is taken both internally and applied as a paste externally. Traditional medicine lists many applications of both the juice of fresh rhizome and the powder of dried rhizome. Ginger is used for flatulence, dyspepsia, colic and other painful conditions of the stomach. Other applications of ginger include anti-ulcer, promotion of antioxidant enzymes, stimulation of humoral immunity, lipid lowering, skin tumour protection, rheumatism, gastroprotective and antifilarial.

[0030] The effective dose required is quite high. In animal studies, aqueous decoctions and ethanolic extracts are used. Oral dose of 50 mg/kg per day for anti-ulcer (Agarwal et al. 2000), 100 mg/kg per day for antioxidant enzyme protection (Ahmed et al., 2000), 200 mg/kg per day for lipid lowering (Bhandari et al., 1998), 500 mg/kg per day for gastroprotection (Al-Yahya et al., 1989) and 100 mg/Kg per day for antifilarial (Datta et al., 1987) was used. For humans, the dose of rhizome powder is from 0.75 to 1.5 g/day ( teaspoon juice).

[0031] Crop health applications of Zingiber officinale extracts or active components include IGR, anti-feedant, antifungal and molluscicidal activity.

[0032] The dosage requirements for isolated/derived compounds are quite high. The maximum EC50 activity reported for any compound as IGR is 3.55 mg/ml and as antifungal is 86.46 mg/liter (Agarwal M. et al., 2000).

[0033] Table 1A summarizes the common uses and typical human dosage of a variety of Angiosperm-Monocotyledon plants; for a comprehensive review regarding human dosage and uses, see (Nadkarni, 1982; Gogte, 2000; and the references given below).

TABLE 1A
Other Angiosperm - Monocotyledon plants, their uses
and human dosage
Plant - Angiosperm -
Monocotyledon	Application	Reference
Family - Cyperaceae	Antimalarial	(Thebtaranonth et al.,
Cyperous rotundus Linn.		1995; Weenan et al.,
Human Dose - Root		1990)
Powder 1 to 3 gm per day	Anti-inflammatory,	(Gupta et al., 1971)
	anti-pyretic,
	analgesic
	Inhibition of Nitric	(Seo et al., 2001)
	Oxide and
	Superoxide
	production
Family - Gramineae	Avoids burning	(Nadkarni 1982;
Andropogon muricatus	micturation and	Gogte, 2000)
Retz.	dysurea, Anti-	pyretic
Human Dose - Root powder
3 to 6 gm per day
Family - Gramineae	Lowers LDL and	(Nadkarni 1982;
Avena sativa, Linn.	Increases HDL	Gogte, 2000)
Human Dose - Oat Bran
5 to 20 gm per day
Family - Gramineae	Phagocytosis	(Stoika et al., 2001)
Triticum vulgare	promoter
Human Dose - Seeds
10 to 100 gm per day
Family - Liliaceae,	Stimulant, Anti-	(Nadkarni 1982;
Allium sp. Human Dose -	pyretic,	Gogte, 2000)
Alliun sativumLinn.	Diaphoretic, Skin,
Bud Paste	Anti-inflammatory
1 to 6 gm per day
Allium cepa Linn. - Bulb	Arthritis, Sciatica,	(Nadkarni, 1982;
powder: 1 to 3 gm per day	Digestion, Tonic	Gogte, 2000)
Family - Liliaceae	Galactogouge,	(Nadkarni, 1982;
Asperagus racemosus	Aphrodisiac,	Gogte, 2000)
Willd. Human Dose - Tuber	Alleviates bleeding
powder 3-6 gm per day	disorders
Family - Zingiberaceae	Skin disease,	(Nadkarni, 1982;
Curcuma amada Roxb.	Itching, sprains	Gogte, 2000)
Human Dose - Rhizome	Anti-fungal	(Ghosh et al., 1980)
Powder 1 to 3 gm per day
Family - Zingiberaceae	Anti-inflammatory,	(Nadkarni, 1982;
Curcuma longa Linn.	Analgesic,	Gogte, 2000
Human Dose - Rhizome	Chologouge
Powder 1 to 3 gm per day
Family - Zingiberaceae	Anti-ulcer	(Agarwal et al., 2000)
Zingiber officinale Roscoe.	Antioxidant	(Ahmed et al., 2000)
Human/Animal Health	protection
Human Dose - Rhizome	Humoral Immunity	(Puri et al., 2000)
Powder (Juice): 0.75 to	Lipid Lowering	(Bhandari et al., 1998)
1.5 gm (2-4 ml) per day.	Skin Tumor	(Katiyar et al., 1996)
Zingiber officinale-Crop	Rheumatism	(Srivastava et al.,
Health		1992)
	Gastroprotective	(Al-Yahya et al.,
		1989)
	Antifilarial	(Datta et al., 1987)
	IGR, Antifeedant,	(Agarwal M. et al.,
	Antifungal	2001)
	Fusarium wilt	(Singh R. et al., 2000)
	Molluscicidal	(Singh K. et al., 2000)

[0034] Angiosperm-Dicotyledon Plants

[0035] Momordica charanta juice is used (Nadkarni, 1982; Gogte, 2000) as a galactogouge, and in amenorrhoea and dysmenorrhoea. Juice is also given for sugar control. Extenally, the juce is used for skin disease and for healing of ulcers. The human dose of juice internally is 10 to 30 ml per day.

[0036] The family Apocynaceae has several genera of medicinal importance such as Holarrhena, Rauwolia and Vinca.

[0037] Holarrrhena genus has several medicinally active species. H. floribunda and H. antidysenterica are the most commonly known to traditional medicine. The bark and seed are both used and the applications include antiamoebic, antimicrobial and antitumour. Both are used for toning of vaginal tissues after delivery.

[0038] Total alkaloids from H. antidysenterica showed an MIC of 95 microg/ml against S. aureus (Chakraborty et al., 1999). IC50 values of H. fioribunda for a variety of tumors are 3.4 to 9.8 microg/ml. Human dose of the seed powder is typically 1-2 g/day.

[0039] Rauwolfia serpentine is best known for use on irritated nervous system and as hyotensive agent. Human dose of the root powder is 0.5 to 3 gm per day depending on the severity of symptoms.

[0040] Vinca rosea root powder is used in anti-cancer applications. The human dose of root powder is 1 to 3 gm per day.

[0041] The family Asclepiadaceae has several genera of medicinal importance such as Hemidesmus and Gymnema.

[0042] Hemidesmus indicus is used as a blood purifier, anti-infalmmatory, and as an alterative and tonic. The human dose is 3 to 6 gm of root bark.

[0043] Gymnema sylvestree is used as a hepatic stimulant. Extract of the leaves works on pancreas and adrenal glands and helps regulate sugar in blood and urine. The human dose is 1 to 2 gm of leaf powder.

[0044] Swertia genus has several medicinally active species. Swertia chirata is the most common one. It is used for promoting appetite, as an expectorant and as a blood purifier. It is particularly useful in chronic fevers also malarial fevers.

[0045] Methanolic extracts of whole dried plants are reported to have tyrosinase and PEP inhibitory (Khanom et al., 2000) activity. Methanolic extract at a dose of 100 mg/kg i.p. given to rats was found to be hepatoprotective (Karan et al., 1999). The ethanolic extract given to rats was found to have protective effect against ulcerogenic agents.

[0046] Plyllanthus genus has several plants of medicinal importance. Among these, Phyllanthus emblica Linn. and Phyllanthus amarus (P. niruri Linn.) are the most widely known. Phyllanthus amarus is known to be useful for viral hepatitis. Phyllanthus emblica Linn. is used for disorders of eyes, liver, spleen and lungs. Paste is applied externally for bruning and headache. Juice is added to eyes for eye disorders. The fruit is a rejuvenating agent and is used in cough, asthma, tuberculosis and also as a brain tonic.

[0047] A review of the genus Phyllanthus (Calixto et al., 1998) is available. Hyaluronidase inhibitory activity (Ishizaki et al., 1999) and superoxide scavenging and prolyl endopeptidase inhibitory activities (Khanom et al., 2000) of P. emblica have been reported. Antiviral tannins (Liu et al., 1999) are also reported. Antitumor activity against leukemic cell lines (Sur et al., 1998) for P. emblica has been reported. Being a dietary product, human dosage of P. emblica juice is 12 ml per day and that of fruit powder is 3 to 6 gm per day. Higher doses are not harmful. In case of P. niruri Linn. the total plant is used and the typical dose is 3 to 4 gm powder of total plant per day.

[0048] Linum usitatissimum is a dietary oilseed (Flaxseed or Linseed) rich in omega-3 fatty acids and lignans. The decoction of seeds is useful as an expectorant and is used in cough, pleuritis, pneumonia and whooping cough in children.

[0049] The regular usage of seeds in the diet (25 to 50 gm per day) can lower cholesterol and post-prandial sugar (Cunnane et al., 1993; Harris, 1997). The seeds are also a strong regulator of inflammatory mediators and alleviate conditions such as eczema, psoriasis and arthritis (James et al., 2000).

[0050] Azadiracta indica or the Neem tree has many uses both for human and crop protection applications. Bark, leaves and seed oil are used both for external and internal use. The range of activities and applications include antimicrobial, ant-leprotic, analgesic, wound healing, psoriasis. Human dosage is 1 to 2 gm per day of powder. Cake is generally not given to humans.

[0051] Crop applications of leaves, bark and oil include anti-feedant, IGR, anti-fungal, and anti-nematocidal. Neem cake can be used as a molluscicide (Singh K. et al., 1996). Use of cake mixed with urea and and germinated barley has been suggested as a slow ammonia release (Virk et al., 1989) product. Application of 500 kg/ha of neem cake powder (Rao et al., 1992) caused a striking reduction in the culicine larvae, a vector for Japanese encephalitis virus, in a rice field.

[0052] Terminalia genus has many species with a wide range of medicinal properties. Some of the most widely used species are Terminalia arjuna, Terminalia bellerica and Terminalia chebula.

[0053] T. chebula fruits are used. T. chebula acts as a rejuvenator and is useful in loss of appetite, constipation, ascites, hepatomegaly, splenomegaly, and parasites. It is also useful in disorders of the respiratory system and reproductive system. Application of the fruit or oil extracted from fruit pulp of T. bellerica is useful in painful inflammatory conditions. Oil is used in skin disease, leucorrhoea, and in premature graying of the hair. The rind of the fruit is kept in the mouth acts as a mucolyte. The fruit taken internally is useful in disorders of the respiratory, circulatory and reproductive system.

[0054] Human dosage is 1 gm of fruit powder per day as a general tonic. For purgation, 10 gm dose is used. Mouthwash with a 10% solution of aqueous extract of T. chebula significantly inhibited salivary total streptococcal counts and glycolysis of salivary bacteria post-rinsing. T. chebula increased gastric emptying in rats (Jagtap et al., 1999) at a dose of 100 mg/kg/day given orally for 15 days. Alcoholic extracts of T. chebula and T. bellerica at 200 mg/ml showed interesting activity against a number of pathogenic and opportunistic microorganisms.

[0055] Eugenia jambolana seeds, bark, fruit and leaves are used. Human dose is 12 to 25 nl for juice and 1 to 3 gm for powder. The plant is used for diabetes, diarrhoea and menorrhgia with bleeding, and dysentery.

[0056] Piper genus has several species of wide ranging medeicinal properties. Piper nigrum and Piper longum are the most commonly used species. Both are stimulants of appetite, liver, spleen and nerves. They promote bioabsorption of other drugs and are useful in cough and asthma.

[0057] Embelia ribes has many applications. In rhinitis and migraine, a fine powder is used for nasal adminstration. It is a nervine tonic and is used for abdominal colic, flatulence and particularly against round worm, thread worm and tape worm infestations. Extermally it is used on skin diseases.

[0058] Administration of Embelin from Embelia ribes produces (Chitra et al) a dose-dependant decrease in labeled thymidine uptake, lipid peroxidaation and glutathione levels. Anti-spermatogenic and retinotoxic effects have also been noted.

[0059] Human dosage of fruit powder is 1 to gm per day. For use against worms, a dose of 10 gm is used. Daily subcutaneous administration of embelin (Gupta et al., 1989) at a dose of 20 mg/kg body weight to male albino rats revealed an inhibition of sperm count and other fertility parameters. Chicks fed Embelia ribes at the rate of 0.5 gm/kg per day (Low et al., 1985) showed a dose-dependant degeneration of the retina. Defects were noted above a cumulative dose of 0.25 gm.

[0060] Tinospora cordifolia is used internally for chronic fevers, diabetes, as a restorative, anti-inflammatory and antacid. The human dose of root powder is 1 to 3 gm per day.

[0061] Glycine max (soybean) is used as an oilseed. It is a lactagouge, emenagouge and aphrodisiac. The human dose is 10 to 20 gm seeds per day.

[0062] Glycyrrhiza glabra is used externally as analgesic, helps in hair growth and promotes skin smoothness and complexion. The root powder is used internally in cough, hoarseness of voice, anaemia, bleeding disorders, disurea, pyorrhoea, and for increasing sperm count.

[0063] Oral administration of Lacrinat (Konovalova et al., 2000) containing liquorice root powder for 1 month markedly increased antioxidant activity of the liver. All crude polysaccharide fractions from the shoot and hairy roots of G. glabra induced nitric oxide production by murine peritoneal macrophages in vitro.

[0064] Human dose is 1 to 2 gm of root powder per day. Methanolic extract of G. glabra had a 50% tyrosinase-inhibitory concentration of 21.2 microg/ml (Khanom et al., Sept. 2000). Beta-glycyrrhetinic acid is a potent inhibitor (Kroes et al., 1997) of the classical complement pathway (IC5035 microM).

[0065] Mucuna pruriens roots and seeds are tonic for neurons. Root is used in facial palsey and hemiparesis. Seeds are used in Parkinson's disease. The roots are diuretic and seeds are useful in impotence ans oligospermia.

[0066] Anti-snake venom (Guerranti et al., 2001) properties of M. pruriens extract are a result of its ability to increase precoagulant activity.

[0067] Human dose is 3 to gm of seed powder per day. Higher doses are also used. In a clinical study on Parkinson's disease patients, a concoction in cow's milk containing M. pruriens seed powder was found to give a good response in tremor, bradykinesia, stiffness and cramps (Nagashayana et al., 2000). The daily dose contained 200 mg of L-DOPA. In alloxan-diabetic rabbits (Akhtar et al., 1990) 1 gm/kg per day root powder caused a significant fall in blood glucose levels.

[0068] Phaseolus radiatus and Phaseolus mungo are pulses and are used as a source of high protein food. They are consumed in the range of 5 to 50 gm per day. They are also useful for the health of hair and skin and are anti-inflammatory when applied externally as a paste.

[0069] Pongamia glabra bark, leaves and seeds are antiseptic, antipruritic and analgesic externally. Oil from seeds has antiparasitic, wound healing and analgesic properties. Internally the various parts are used for helminthasis, and as a liver tonic.

[0070] Human dose is 1 to 3 gm of seeds per day. Solvent extracted P. glabra cake (Ravi et al., 2000) can be fed to lambs (20% of feed) without affecting the performance for a period of 98 days. However, expressed Karanj (P. glabra) cake may not be recommended as it adversely affects the intake and digestibility of nutrients. Aqueous extract of deoiled kernels (Sagar et al., 1996) at 100 ppm causes 100% mortality in 4^th in star larvae and pupae of Culex mosquito.

[0071] Trigonella foenum-graecum intake is advised to women in the post natal period to improve excretory and menstrual functions and as a galactogogue. Seed powder is used locally to reduce inflammation and to reduce hair fall. It is also used as a lipolytic agent. Hypoglycaemic and Antiulcer activity has been reported (Zia et al., 201) and Trypsin/Chymotrypsin inhibitor (Weder et al., 1991) activity has also been noted.

[0072] Human dose is 3 to 6 gm of seed powder per day. Methanolic extract given to mice produced antidiabetic effect at a dose of 1 gm/kg per day.

[0073] Santalum album heartwood (2 to 5 gm) and oil (5 to 20 drops) are used for external applications and for internal use. It is used to reduce burning and thirst, dusurea and for the treatment of purulent menstuation. It cures dermatoses and pruritis.

[0074] Ocimum genus has several important medicinal plants. Ocimum sanctum is the most commonly used plant. The juice (10 to 20 ml) and seed (1 to 2 teaspoons) are taken internally. Its main action is on the respiratory system. It is a common remedy for cold, cough and fever. The paste of leaves is also used for ringworm and scabies. Seed is a diuretic and tonic.

[0075] Sesamum indicum seeds and oil are used as food. The intake of seeds is useful for gum and dental health, hair and skin. It is useful in dysmenorrhoea, diabetes and bleeding piles. Oil is used for massage in paralysis, fractures and wounds.

[0076] As a common dietary item, human dose is 10 to 20 gm per day.

[0077] Herpestis monnieri is a valuable plant for brain and the entire nervous system. It is given in mania, epilepsy and retardation. It is also useful in cough and cold. The human dose is 10 ml of leaf juice.

[0078] Withania somnifera is well known for its aphrodisiac property. It is an immunomodulator and also given to reduce pain in rheumatoid arthritis and for abdominal pain. The human dose is 5 gm of the root powder.

[0079] Carum roxburghianum is used as a spice in cooking. It is useful as a carminative, analgesic, anti-inflammatory, and antihelminthic. It is an aphrodisiac and is used for treating amenorrhoea and dysmenorrhoea. The human dose is 5 to 7 gm of seed powder.

[0080] Cuminum cyminum is used as a spice in cooking. It is useful as a carminative, analgesic, anti-inflammatory, and antihelminthic. It also improves lactation. The human dose is 5 to 7 gm of seed powder.

[0081] Ficus genus has several plants of medicinal importance. Ficus bengalensis fruit, bark, aerial roots and latex are all used. Latex is applied on wounds, synovitis, arthritis, toothache, conjunctivitis and piles. Latex, bark and fruit act as antidiabetic. In leucorrhoea and menorrhagia, decoction of bark is used as douche.

[0082] The fruit extracts have antitumour activity in the potato disc bioassay and also antibacterial activity (Mousa et al., 1994). These results support the use in respiratory and certain skin disorders. Leucocyanidin derivatives from the bark have antidiabetic activity (Kumar et al., 1989), and they promote hexokinase and HMGCOA reductase levels in tissues.

[0083] Human dose is 50 to 100 ml decoction of the bark or 3 to 6 gm of powder per day. Leucocyanidin derivates (Kumar et al., 1989) give a significant effect at 100 mg/kg body weight.

[0084] Table 1B summarizes the common uses and typical human doses of a variety of other Angiosperm-Dicotyledon plants; for a comprehensive review regarding human dosage and uses, see (Nadkarni, 1982; Gogte, 2000; and references given below).

TABLE 1B
Other Angiosperm - Dicotyledon plants, their uses and
human dosage
Plant - Angiosperm -
Dicotyledon	Application	Reference
Family - Cucurbitaceae	Diabetes,	(Nadkarni, 1982;
Momordica charantia, Linn.	healing, dysmenorrhoea,	Gogte, 2000)
Human Dosage - Juice of plant or	amenorrhoea, skin
fruit 10 to 30 nl per day	galactogouge
Family - Apocynaceae	Antitumor	(Loukaci A. et al.,
Holarrhena floribunda		2000; Abreu et al.,
Human Dosage - Decoction of		1999)
Seeds	Antileschmania,	(Abreu et al., 1999)
0.5 to 1 gm per day	antimicrobial
Holarrhena	Antibacterial,	(Chakraborty et al.,
antidysenterica, Roxb.	Antimicrobial	1999; Ahmed et al.,
		1998)
	Stimulate phagocytosis	(Atal et al., 1986)
	Hepatotoxicity	(Arseculeratne et al.,
		1981)
Family - Apocynaceae	Sedative, Lowering of	(Nadkarni, 1982;
Rauwolfia serpentina, Benth Ez	B.P., Promotes Uterine	Gogte, 2000)
Kurazaa Human Dosage - Root	contractions,
0.5 to 3 gm per day
Fanily - Apocynaceae	Anti-cancer	(Nadkarni, 1982;
Vinca rosea, Linn.		Gogte 2000)
Human Dosage - Roots
1 to 3 gm per day
Family - Asclepiadaceae	Blood purifier, Tonic,	(Nadkarni, 1982;
Hemidesmus indicus, R. Br.	Alterative,	Gogte, 2000)
Human Dosage - Root Bark paste	Anti-inflammatory
3 to 6 gm
Family - Asclepiadaceae	Hepatic stimulant,	(Nadkarni, 1982;
Gymnema sylvestree, R. Br.	Anti-diabetic	Gogte, 2000)
Human Dosage - Leaf powder
1 to 2 gm
Family - Gentianaceae	Liver Toxicity	(Karan et al., 1999;
Swertia chirata, Ham.		Reen et al., 2001)
Human Dosage - Whole Plant	Tyrosinase and Prolyl	(Khanom et al.,
2 to 6 gm per day	Endopeptidase	April and Sept.
	Inhibitor	2000)
	Gastric Ulcer	(Rafatullah et al.,
	Protection	1993)
Family - Euphorbiaceae	General Review	(Calixto et al., 1998)
Phyllanthu niruri, Linn.	Superoxide scavenging	(Khanom et al.,
Human Dosage: Total Plant	and PEP Inhibition	April 2000)
Powder	Hyaluronidase	(Ishizaki et al.,
3 to 6 gm per day	Inhibition	1999)
Phyllanthus emblica, Linn.	Anti-tumor	(Sur et al., 1998)
Human Dosage - Fruit	Anti-viral	(Liu et al., 1999)
Powder: 3 to 6 gm per day
Juice: 12 ml per day
Family - Linaceae	Cholesterol	(Harris, 1997)
Linum usitatssiimum, Linn.	Post prandial blood	(Cunnane et al.,
Human Dosage - seeds	glucose	1993)
5 to 50 gm per day	Inflammatory Mediator	(James et al., 2000)
	Production
Family - Meliaceae	Slow ammonia release	(Virk et al., 1989)
Azadiracta indica, A. Juss.	Molluscicidal	(Singh K. et al.,
Human Dosage - Bark Powder: 1		1996)
to 2 gm per day. Leaf Juice: 12 ml	Culex Mosquito Larvae	(Rao et al., 1992)
per day	in Rice Fields(vector
Cake is not used for humans.	for encephalitis virus)
Family - Conbretaceae	Antimicrobial	(Ahmad et al., 1998)
Terminalia sp.	Gastric Emptying	(Tamhane et al.,
Terminalia bellericaa, Roxb.		1997)
Erminalia chebula, Retz.	Anticaries agent	(Jagtap et al., 1999)
Human Dosage - Fruit Powder	HIV Reverse	(el-Mekkawy et al.,
1 gm per day	Transcriptase	1995)
3 to 6 gm for purgation
Family - Myrtaceae	Diabetes,	(Nadkarni, 1982;
Eugenia jambolana, Lam.	Liver Funcction	Gogte, 2000)
Human Dosage - Fruit, Seed,	Diarrhoea, Dysentry,
Bark, Leaves:	Menorrhagia with
Juice 12 to 25 ml, Powder 1 to 3 gm	bleeding
Family - Piperaceae	Stimulant for appetite,	(Nadkarni, 1982;
Piper nigrum, Linn. and	liver, spleen, nerves	Gogte, 2000)
Piper longum, Linn.	Promote bioabsorption
Human Dosage - Fruit	of other drugs
0.25 to 0.5 gm for P. nigrum	cough and asthma
5 to 10 gm for P. longum
Family - Myrsinaceae	Anti-neoplastic	(Chitra et al.)
Embelia ribes, Burm.	Anti-spermatogenic	(Gupta et al., 1989)
Human Dosage - Fruit	Retinotoxic	(Low et al., 1985)
1 to 2 gm per day. 10 gm for
worms.
Family - Menispermaceae	Chronic fevrs,	(Nadkarni, 1982;
Tinospora cordifolia, Willd	Diabetes, Anti-	Gogte, 2000)
Meirs.	inflammatory, Antacid,
Human Dosage - Bark powder	Restorative
1 to 3 gm
Family - Leguminosae	Lactagouge and	(Nadkarni, 1982;
Glycine max, Merr.	emengouge,	Gogte, 2000)
Human Dosage - Seeds	aphrodisiac
10 to 20 gm per day
Family - Leguminosae	Tyrosinase Inhibitor	(Khanom et al.,
Glycyrrhiza glabra, Linn.		Sept. 2000)
Human Dosage - Root Powder	Macrophage Activation	(Nose et al., 1998)
1 to 2 gm per day	Anti-inflammatory	(Kroes et al., 1997)
	Antioxidant	(Konovalova et al.,
		2000)
Family - Legyminosae	Anti-snake venom	(Guerranti et al.,
Mucuna pruriens, Bak.		2001)
Human Dosage -	Parkinson's Disease	(Nagashayana et al.,
Seed Powder: 3 to 6 gm per day		2000)
Root Extract: 50 to 100 ml per	Antidiabetic	(Akhtar et al., 1990)
day
Family - Leguminosae	Anti-inflammatory	(Nadkarni, 1982;
Phaseolus radiatus	Complexion Promoter	Gogte, 2000)
Human Dosage - Seeds
5 to 50 gm per day
Family - Leguminosae	Anti-mosquito	(Sagar et al., 1996)
Pongamia glabra, Vent.	Solvent Extracted Cake	(Ravi et al., 2000)
Human Dosage -	As Feed Supplement to
Juice of Bark or Leaves: 6-12 ml	Lambs
per day
Seeds: 1 to 3 gm per day
Family - Leguminosae	Hypoglycaemic	(Zia et al., 2001)
Trigonella foenum - graeceum,	Trypsin/Chymotrypsin	(Weder et al., 1991)
Linn.	Inhibitor
Human Dosage - Seed Powder
2 to 6 gm per day.
Family - Santalaceae	Burning sensation and	(Nadkarni, 1982;
Santalum slba Linn.	thirst, Dermatoses and	Gogte, 2000)
Human Dosage - Heartwood, Oil	Pruritis, purulent
Powder 2 to 5 gm, Oil 5 to 20	menstruation
drops
Family - Labiatae	Expectorant, Fever,	(Nadkarni, 1982;
Ocimum sanctum Linn.	Cough, Cold,	Gogte, 2000)
Human Dosage - Juice 10 to ml	Diaphoretic, Ringworm
Seeds - 1 to 2 teaspoon
Family - Pedaliaceae	Lactagouge and	(Nadkarni, 1982;
Sesamum indicum	emenagouge,	Gogte, 2000)
Human Dosage - Seeds	aphrodisiac
10 to 20 gm per day
Family - Scrophulariaceae	Epilepsy, Depresion,	(Nadkarni, 1982;
Herpestis monnieri, H. B.K.	Retardation,	Gogte, 2000)
Human Dosage - Leaf Juice	Cough and Cold
2 teaspoon (10 ml)
Family - Solanaceae	Aphrodisiac,	(Nadkarni, 1982;
Withania somnifera, Dunal.	Immune modulator,	Gogte, 2000)
Human Dosage - Root Powder	Rhematic Arthritis,
5 gm per day	Abdominal Pain
Family - Umbelliferae	Anti-inflammatory,	(Nadkarni, 1982;
Carum roxburghianum, Benth	Analgesic, Carminative,	Gogte, 2000)
Human Dosage - Seeds	Antihelminthic,
	Amenorrhoea,
	Dysmenorrhoea
Family - Umbelliferae	Anti-inflammatory,	(Nadkarni, 1982;
Cuminum cyminum, Linn.	Analgesic, Carminative,	Gogte, 2000)
Human Dosage - Seeds	Antihelminthic,
3 to 6 gm per day	Galactogouge
Family - Moraceae	Antitumor,	(Mousa et al., 1994)
Ficus bengalensis, Linn.	Antibacterial
Human Dosage -	Antidiabetic	(Kumar et al., 1989)
Decoction of Bark: 50 to 100 ml
per day.
Powder: 3 to 6 gm per day.

[0085] Gymnosperm Plants

[0086] There are three species known by the name Talispatra. They are: Abies webbiana Lindle, Taxus baccata and Rheododendron anthropogon. One of them is highly poisonous, Taxus baccata a species from which taxol, an anticancer drug, has been isolated. Leaves are used as an appetizer and in cough, asthma, general debility and pthisis. The dose of leaf powder is 0.5 to 1 gm.

[0087] Pinus deodar Roxb. Sergent is used widely. Leaf powder, oil and latex are all used. Externally the latex is used on abscesses. The oil (turpentine oil) is useful for pulmonary edema, arthritis, flatulence. Internally, the plant is used as hepatostimulant, on acute and chronic bronchitis and a variety of skin diseases.

[0088] Human dosage powder and oil is 1-3 gm and 1-2 gm, respectively. Overdose causes, diarrhoea, vomiting, paralysis and sensory loss.

[0089] Table 1C summarizes the common uses and typical human doses of a variety of Gymnosperm plants; for a comprehensive review regarding human dosage and uses, see (Nadkarni, 1982; Gogte, 2000; and references given below).

TABLE 1C
Gymnosperm plants, their uses and human dosage
Plant - Gymnosperm	Application	Reference
Order - Conifereae	Anti-tumor	(Mantle et al., 2001)
Taxus baccata(also Abies	Toxicity	(Kite et al., 2000)
webbiana Lindle)
Human Dosage - Leaf powder
0.5 to 1.0 gm per day

[0090] Non-Plant Biomass Sources

[0091] A wide range of many other biomass sources have also been used for the treatment of a wide range of ailments all over the world. This range of biomass includes:

[0092] Microbial, Mycological, Marine, Poultry, Animal and Human.

[0093] The preparations start with many different parts of biomass such as blood, mik, urine, organ meat, cartilage and chitin, skin. Total cell mass may also be used. As many of these are food materials, typical human dosage levels required on the basis of fresh part is 10 to 50 gm per day. In the case of dry powders of mushroom, yeast or lactobacilli, the dosage may be 1 to 10 gm.

[0094] The total list of non-plant biomass sources used includes the entire set of life forms and is therefore too exhaustive to be cited here. General use and nutritional applications of meat, milk and eggs from poultry, seafood and animals are also well known and are not repeated here. Only a few illustrative examples of non-plant biologicals are described.

[0095] Mushrooms have been used medicinally for centuries, particularly in traditional Chinese and Japanese medicine (Lombardi R. M. 2002). They are considered as promoters of health and vitality and are adaptogens. The constituents show, immunomodulatory, amtobacterial, antiviral, antitumour, antiparasitic, cardiovascular hypercholesteromiac properties. Active substances include beta-glucans and polysaccharide-protein complexes Button mushroom (Agaricus bisporus) is a very common type of mushroom, and is used as a source of high protein food ingredient. It is also known to promote digestion and help lower blood pressure.

[0096] Beta-glucans are also isolated from the cell walls of yeast (Bacon et al. 1969) and oat and barley bran. Beta-glucans activate the anti-infection and antitumour activity of macrophages. Hence, yeast and oat and barley bran also have immune modulating activities. Beta-glucan dose is typically 2 mg/kg per day.

[0097] Shrimp(Prawn) outer shell and shark fin are used as a source of glucosamine and chondroitin in commercial preparations and are given internally for rebuilding cartilage in joints. No specific medicinal activity has been claimed for them.

[0098] Table 1D summarizes the common uses and typical human dose of a variety of non-plant biomass.

TABLE 1D
Non-plant Biomass Sources, their uses and human dosage
Biomass Source	Application	Reference
Yeast	Digestion,	Bacon et al. 1969
Active dried Baker's Yeast	Immunomodulation
Saccharomyces cerevisiae
Human Dosage - dry powder 2
to 6 gm
Beta-glucan - 200 mg
Mushroom	Immunomodulation,	Lombardi, 2001
Agaricus bispores (Button	Digestion, Blood
Mushroom)	Pressure Lowering
Human Dosage - dry powder 2
to 6 gm
Beta-glucan 200 mg
Prawn (Shrimp)	Source for
	glucosamine
Shark cartillage	Source for
	chondroitin

[0099] Toxicity

[0100] Because of the relatively high effective doses and the traditional methods of preparation, toxins from CAM plants that might be otherwise negligible can adversely affect a subject. For example, Crassulaceae juices and aqueous extracts from various plants have cytotoxic substances (Avila et al., 1997; Balachandran et al., 1991; Botha et al., 1997; Mueller and Stopper, 1999; Sivaswamy et al., 1991). High therapeutic doses of leaf juice for internal use (more than 100 mg/kg bbody weight daily) coupled with high mammalian toxicity close to the habitual level of use (LD₅₀ in mice is 230 mg/kg and in rat is 560 mg/kg, respectively (Verma et al., 1986)) render these compositions less useful.

[0101] Toxicity issues rendering compositions less useful is not restricted to CAM plants. Many other plants and other biologicals also have toxic substances and their toxic effects are known. As an example, Embelia ribes used for the treatment of worms is known to be retinotoxic (Low et al., 1985). Holarrhena antydysenterica used for amoebic dysentery is known to have hepatotoxicity (Arsecularatne, 1981).

[0102] Shelf Life and Potency

[0103] The traditional methods of preparation, juice extraction and ground leaf, suffer from poor shelf life, especially fresh plant juice, which ferments readily if not sterilized or stored properly. Even when dry leaf or stem powder is used, the shelf life of such products is 6 months to one year. Because preparation methods have not been optimized, potency varies by preparation, and thus each preparation may have different effective doses.

[0104] Previous Oil Extraction Methods

[0105] Medicated oils using herbal materials are known in Indian traditional medicine. The base oils used for such preparations are sesame oil and ghee (clarified butter). In South Indian practice coconut oil may replace sesame oil. Sharangdharsamhita, an ancient treatise by Sharangdharacharya (1961), a standard reference treatise of traditional medicine describes a standard method of preparing such medicated oils.

[0106] In traditional practice, such extracts are made by boiling together a mixture of kalka (ground paste or homogenate), oil and other liquid substances. The recommended ratio of kalka:oil:liquid substances changes with the nature of liquid substance used. Water, plant derived liquids and juices are the three types of liquid substance described. Plant derived liquids used for making oil extracts are decoctions of plant parts in water. The decoctions are the filtrates prepared by boiling plant part in water, and filtering to obtain a clear liquid or decoction. The ratios of kalka:oil:liquid substance for these three cases are 1:4:16; 1:6:24; and 1:8:32, respectively. Therefore, in these three cases the overall ratio of water to kalka, plant derived liquids to kalka and juice to kalka are recommended to be 16:1, 24:1 and 32:1.

[0107] Juice based or decoction based preparation is the commonly used preparation in practice for fresh or dried succulents. Thus, the standard preparations for succulents call for a very high ratio of juice to ground paste (32:1) or decoction to ground paste (24:1) in making the medicated oil extracts. Traditionally, the effective substances were thought to be present only in the fresh juice or decoction; the bulky residue from ground paste was considered unimportant. Furthermore, the typical recommended dose of such medicated oils is as high as 4 tola (1 tola11.4 g). This standard procedure is practiced in Ayurveda, the traditional medicine of India. These oil preparations are thus characterized by predominant use of juice or decoction and a high dose. Classical treatises and other references specify particular plants for such methods since the belief is that the various healing substances are liberated from the plants in very specific ways (Nanal, 1995).

[0108] However, Nanal (1995), in reviewing the use of Kalanchoe in the context of theory and practice, remarks that Parnabeeja (Kalanchoe) is not mentioned in any Ayurvedic texts. Nanal mentions several different preparations from Kalanchoe that includes oils, both in sesame and in clarified butter, but does not specify the usefulness of such preparations, and he does not recommend dosages. Such Kalanchoe preparations are usually prepared from the juice of the plant; with only a small amount of leafy residue. These juice-based preparations are seldom used because of serious toxic side effects at very low doses, and when used, are only topically (as opposed to internally) administered.

[0109] Aloe extracts are used commercially in hair oils, often as part of multi-herb medicated oils. Such preparations use extracts prepared from fresh Aloe juice or decoctions of dried Aloe pulp. Aloe oils, by themselves, are not generally recommended for topical or internal use; instead, Aloe liquids, gels or pulp are used.

[0110] The use of oil extracts of Cissus is unknown.

[0111] In a more general way, oil or ghee (clarified butter) extracts of plants are more commonly used in multi-herb compositions used for both external and internal applications. Specific detailed description for individual plants is difficult to come by. The generalized procedures described above are therefore the guidelines to go by.

[0112] Oil extract preparations for non-herbal biomass are not described in traditional medicine.

[0113] The use of paste or residue of total plant or plant parts as the predominant component compared to the use of juice or decoction as the dominant component in extractions is contrary to the teachings of traditional medicine. However, surprisingly, the methods of the invention allow for the preparation of compositions that have an enormous potential to improve health by mining the beneficial effects and minimizing toxicity of plants. These methods also produce compositions of high potency at very low doses, thus further reducing any potential for toxicity.

[0114] The methods of this invention also produce compositions of high potency and low toxicity from all non-herbal biomass sources including microbial, mycological, veterinary, human and aquatic.

[0115] The invention circumvents the problems of toxicity and shelf life by providing a general way to make compositions that incorporate oil extracts of a wide variety of plants and other non-plant biomass. These compositions have a very high useful activity (on the basis of biomass material dose) which allows extremely low dosage and as a result, the toxicity is considerably reduced. These compositions bring out the activity characteristic of each biomass. As a result, these compositions are useful for a wide variety of applications, including human, veterinary and plant applications, for both known and novel uses. These applications include broad general effects such as disease resistance, stress resistance, general promotion in health and growth, delaying senescence and special effects such as wound healing, skin repair, stimulation of hair growth, bone repair and lipid lowering.

[0116] This invention relates to compositions comprising biomass extracts to be used in human, veterinary and agricultural aquatic, mycological and microbial applications.

[0117] The novel compositions are prepared from biomass by taking the entire biomass or appropriate parts, washing them with water, soaking them (in case of hard seeds or dry material), pounding them (in case of hard materials such as seeds), or cutting them into pieces (in case of soft plant material), mixing them with water, homogenizing the mixture, and filtering the homogenate to obtain two fractions: juice (J) fraction (as the filtrate) and the leafy residue (or stem, biomass part, etc.; LR) fraction. The fractions may be mixed together, or kept separate as J or LR fractions. The total homogenate or any form and any proportion of the fractions may be mixed with oil or fat, adding water, bringing the mixture to boil, stirring the mixture to provide good contact of biomass with both water and oil and to minimize sticking of biomass to the vessel bottom, removing the water by boiling, cooling the mixture, and filtering the mixture to separate the oil extract from the residue, which are referred to as the first oil extract and first residue, respectively. A second extract from the particular fraction may be obtained by washing the corresponding first residue with oil and filtering to obtain a second extract. Subsequently, the first and second extracts may be combined. The composition can be used to treat a variety of human and animal ailments, and has manifold applications in agriculture, using exceptionally low doses and without toxic side effects. These uses will become apparent as the various embodiments of the invention are discussed.

[0118] The novel herbal compositions of the invention, is prepared by a method wherein the total biomass or one or more biomass parts are first reduced in size by soaking, pounding and cutting as required, and are then homogenized, adding water as required. The total homogenate (kalka) is filtered to separate the juice fraction (J) from the concentrated stem/leaf/plant parts residue (LR). The total homogenate may be added or the two fractions may be added separately (mixed in any proportion) to oil with additional water as required; the water is then removed by boiling. Stirring is provided to promote good contact between the biomass, oil and water.

[0119] Total biomass or any part or parts of the biomass can be used to prepare a range of extracts. The concentrated seed/rhizome/stem/leaf/biomass parts residue may be used to prepare LR fraction; or, only the juice fraction may be used to obtain J fraction. Thus, the proportion of the biomass residue fraction to the juice fraction may be 1:0 or 0:1. The combined extract comprises both the J and LR fractions. By varying the starting biomass materials (including plant, non-plant materials and their parts, etc.), the admixing of J and LR fractions, and filtration provides the preparation of extracts with a variable biological activity that are suitable for specific applications (see Examples). Because various factors can be adjusted during the preparation of the compositions of the invention (biomass parts, ratio of leaf, stem, biomass parts, J fraction, LR fraction, oil, etc.), the drawbacks of traditional methods of preparation, such as cytotoxicity and excessively high doses, are circumvented.

[0120] In addition, the compositions of the invention have unexpected and useful results, including high potency coupled with low toxicity, an exceptionally long shelf life, and a wide range of usefulness.

[0121] The two important features of this invention in the context of plant extracts are the ratio of oil to homogenate or kalka of biomass/biomass parts and the ratio of liquid substances to homogenate or kalka of biomass/biomass parts taken for boiling.

[0122] The traditional methods recommend an oil to kalka ratio of 4:1, 6:1 and 8:1 for use depending upon the use of water, plant decoction or juice as the liquid substance used. This invention uses the lowest possible ratio consistent with ease of processing. Thus, with succulent leaves and fruit as the plant part, the typical ratio of oil to starting plant part is 1 to 2 in the presence of water. A ratio of homogenate to the oil of 0.1 to 1.5 can also be used. In the case of dried material and particularly where the material forms a pasty mass after homogenization, oil to starting plant part ratio of 4:1 may be used in the presence of water.

[0123] The traditional methods recommend the overall ratio of water to kalka, plant derived liquids to kalka and juice to kalka to be 16:1, 24:1 and 32:1, respectively.

[0124] This invention uses ratios which are significantly different. In general, present invention uses overall ratio of water to kalka, plant derived liquigds to kalka and juice to kalka less than 16:1, less than 24:1 and less than 32:1, respectively. This is best understood in the context of one of three cases.

[0125] The first case is where the total homogenate is used for boiling without separating the LR and J fractions. In this case, unlike the traditional method, there is no additional decoction or juice is used at all. Thus, it is equivalent to using a ratio of decoction or juice to kalka or homogenate of zero instead of 16:1 and 24:1 and 32:1.

[0126] The second case is where the LR fraction is taken further for boiling. In this case, not only no additional decoction or juice is used but most of the juice inherently present in the succulent starting material is also washed away. Thus, this method is equivalent to having a negative ratio of juice to initial plant material.

[0127] In both these cases, some water inevitably gets added for homogenization and also to control the time of boiling. The total water to initial plant material is typically in the ratio of 1:1 to 8:1. The low ratio is likely to be used with succulents where total homogenate is being used. The higher ratio is required when starting with dry materials which tend to form a thick paste. Even the higher ratio is much lower than the 16:1 ratio recommended in traditional literature.

[0128] Thus, for total homogenate or for the LR fraction, considerably lower total liquid quantities are used. This drastically cuts down the processing time, avoiding loss of activity caused by excessive heating for a long time. At the same time, this still provides an ability to adjust the ratio of water and helps in controlling and optimizing the boiling time to the desired level as required for each specific extract.

[0129] This invention also incorporates another novel feature as an Extraction Concept. Traditionally, plants are typically extracted in one solvent. The filtrate is then concentrated and evaporated to dryness to obtain the active concentrate. U.S. Pat. No. 5,529,778 (1996) describes a composition made by such a process where the plant material is extracted in distilled water and the filtrate is evaporated to dryness. Sometimes, the residue from the first solvent extraction may be further extracted by a series of different solvents, all the filtrates combined together and then evaporated to obtain the active material. In some other cases, the plant is extracted in one solvent and then the filtrate is back-extracted into another solvent.

[0130] However, the concept of fractionating the total homogenate into LR and J fractions and separately processing them in oil to produce two separate compositions with widely different biological activity is novel and forms a special feature of this invention. This can be explained in the following manner. The step of homogenization brings the plant material with intimate contact with a solvent (e.g. cold water). After filtration, the two fractions, LR and J, contain a selective fractionation of plant components in them. Subsequent oil extraction, therefore, yields two separate extracts with widely differing biological activity in many cases. In some cases one fraction gets enhanced activity and reduced toxicity than the total homogenate for a particular application. In some other cases, one of the fractions becomes stronger in one activity while the other fraction also develops strong activity for a different application. Thus, this scheme of fractionation/extraction provides a general method to greatly enhance the utility of many extracts and therefore the utility of many plants as per this invention. In the oil extract of the present invention, the ratio of the first residue to the oil may be from 0.05:0.5. Alternatively, in the oil extract of the present invention, the ratio of the biomass juice to the oil may be 0.5 to 10.

[0131] The key features, methods and the Extraction Concept outlined above are also applicable to making compositions from all non-plant biomass materials as per this invention.

[0132] Potency

[0133] Doses of less than 1 mg/kg body weight/day on the basis of total fresh leaf or stem or plant parts or non-plant biomass weight for human (and mammalian) internal use is sufficient to produce significant therapeutic effects compared to greater than 50 mg/kg body weight therapeutic dosage traditionally used. A 5 to 50 mg plant or other biological material equivalent is sufficient for topical applications, compared to the traditional use of 5 to 10 g of juice or homogenate. A dose level of less than 1 mg/kg body weight per day of plant equivalent is effective in poultry applications, compared to approximately 100 mg/kg body weight per day as traditionally used.

[0134] Low Toxicity

[0135] Oil extracts of Kalanchoe pinnata (Lam.), when prepared according to the methods of the invention, are not toxic when given in doses of 50 mg/kg/day for 6 months to Sprague-Dawley rats. Even at doses of 500 mg/kg/day, changes in mortality rates or histopathology are not observed. The compositions are not cytotoxic in vitro when administered to 60 different tumor cell lines at doses up to 250 ppm. Thus, compared to the toxicity levels reported for the traditionally prepared compositions, the toxicity of the compositions of the present invention is negligible, even at high doses.

[0136] Similar enhancement in the efficacy/toxicity ratio is expected for other plants and non-plant biologicals as well.

[0137] Shelf Life

[0138] Fresh juice or extracts prepared by traditional methods ferments rapidly. However, the compositions of the invention remain potent much longer. In case of Crassulaceae, the extracts have remained potent even after at least 7 years.

[0139] A. Human

[0140] The compositions of this invention can be made from a wide variety of plants and non-plant biomass. For each biomass, the key activities as reported in the literature can be incorporated in the compositions of this invention. Hence, the compositions of this invention have a wide variety of human applications. An illustrative summary of examples of the many embodiments for selected CAM plants is given in Table 2.

[0141] Thus, in the context of CAM plants alone, the compositions of the invention may be used to treat respiratory disorders and skin conditions, modulate the immune system, lower blood lipid levels, improve digestion, promote healing, regulate menstruation and ovulation, and may be used as an anti-inflammatory agent. Dosages are unexpectedly low when compared to traditional applications, from 100 to 1000 times less.

[0142] The compositions may also be used prophylactically.

TABLE 2
Human embodiments of uses for the compositions of
the invention
General
embodiment	Specific embodiments
Respiratory	Treating coughs, colds and congestion
	Treating asthma, including allergy and stress-induced
Circulatory	Promoting circulation in feet
	Lowering low density lipoproteins (LDL)/cholesterol
	Lowering triglycerides
Digestive	Treating ulcers from Diabetes
	Reducing stomach acidity
	Reducing stomach upsets
	Promoting appetite
Growth	Promoting weight gain
	Promoting height growth in children
Healing/Wound	Promoting healing of bruises and cuts
repair	Promoting healing of ulcers from leprosy
	Promoting healing of bedsores
	Promoting healing of burns
	Promoting healing of piles (hemorrhoidal tumors)
	Treating fistulas
Stress and energy	Promoting sound sleep
levels	Promoting lowered stress and tension
	Promoting higher energy level in elderly
Inflammation	Reducing general pain and swelling
	Treating spondylitis (inflammation of the vertebrae)
	Treating arthritis
	Treating gingivitis
	Treating toothaches
Reproduction	Treating oligospermia
	Promoting sperm motility
	Regulating ovulation
	Regulating menstruation
	Managing menstruation pain
	Treating irregular, especially prolonged (menorrhagia),
	menses
Dermatology	Treating pimples
	Treating sunburn and tan
	Treating lichenplanus
	Treating hyperpigmentation
	Treating eczema/dermatitis
	Treating psoriasis
	Preventing hair loss
	Promoting hair growth
Vision	Promoting vision recover after macular surgery
	Treating dry cornea
	Treating styes

[0143] Utility of the compositions of this invention are not restricted to the embodiments listed in Table 2. Innumerable other embodiments for particular plant and non-plant biomass are possible as the invention is applicable to a wide variety of plant and non-plant biomass. The range of embodiments can be further increased by using appropriate mixtures of biomass. A few illustrative examples of such applications are given below:

[0144] reduce sciatica pain,

[0145] improve handgrip post paralytic stroke,

[0146] restore Foot Drop condition post viral polyneuritis attack,

[0147] restore concentration and memory post brain haemorrage,

[0148] alleviate colitis,

[0149] reduce hyperpigmentation,

[0150] regulate sugar om blood and

[0151] alleviate headache.

[0152] B. Veterinary

[0153] The compositions of the invention may also be used to improve livestock productivity, treat animals for a variety of conditions, and improve animal health. Additionally, other benefits may be realized, such as an early onset of maturity, improvement in the shelf life of buffalo milk, an improvement in feed conversion efficiency (more production for less feed), and a decrease in mortality. Table 3 summarizes examples of embodiments with CAM plants in which the compositions of the invention may be used on animals. Utility of the compositions of this invention are not restricted to the embodiments listed in Table 3. Innumerable other embodiments for particular biomass are possible as the invention is applicable to a wide variety of plant and non-plant biomass.

TABLE 3
Veterinary embodiments of uses for the compositions of
the invention
	General embodiment	Specific embodiments
	Growth	Increasing weight gain
		Increasing growth rate
		Decreasing mortality (overall
		improving health)
		Hastening maturity
	Productivity	Increasing egg laying with less feed
		(egg-laying birds)
		Improving quality of milk (buffalo)

[0154] C. Agricultural

[0155] The utility of the compositions, of the present invention extends to all areas of the Plant Kingdom. For example, the compositions of the invention have beneficial effects on vegetables, ornamentals, flowers, fruits, trees, cereals, legumes, herbs and medicinal plants. Table 4 summarizes examples of embodiments in which the compositions of the invention may be used in plants. The utility of the compositions of this invention are not restricted to the embodiments listed in Table 4. Innumerable other embodiments for particular biomass are possible as the invention is applicable to a wide variety of plants.

TABLE 4
Embodiments of uses for the compositions of the
invention
	General embodiment	Specific embodiments
	Germination	Promoting vigorous rooting and
		shooting and germination vigour
	Vegetative growth	Promoting branching
		Promoting growth (especially height)
	Leaf production	Promoting increased chlorophyll levels
		Promoting larger leaves and more leaf
		area per plant
		Promoting higher carbohydrate content
		Promoting higher number of leaf active
		days (LAD)
		Extending leaf life
		Delaying senescence
	Flowering	Promoting early onset
		Reducing flower drop
		Promoting larger bloom size
		Promoting uniform bloom size
		Increasing production
	Fruit	Reducing fruit drop
		Promoting larger sized fruits
		Promoting fruit appearance (e.g.
		shine)
		Promoting production
	Productivity	Promoting increased yields, whether
		fruit, flower, or vegetable
		Increasing primary metabolites (e.g.,
		sugars, proteins, and oil content)
		Increasing secondary metabolites (e.g.,
		anti-oxidants, aromatics, and medicinal
		substances)
	Herbicide	Eliminating unwanted plants/grasses
		Controlling growth of plants/grasses
		Acting as a synergist with pre-emergent
		herbicides
	Shelf life	Promoting shelf life of fruit and flowers
	Pest defenses	Promoting higher levels of defense
		chemicals (e.g., polyphenols and
		alkaloids)
		Reducing damage by pests (e.g., white
		fly, aphid, jassid, fruit fly, fruit borer,
		mite, stem borer, millibug)
		Reducing incidence of viral attacks
		Reducing incidence of fungal damage
	Environmental stress	Promoting frost resistance
		Promoting drought tolerance
		Increasing osmolyte levels (e.g.,
		proline)
		Allowing co-existence of insects while
		decreasing insect damage
	Qualitative	Decreasing thorny habits
		Promoting natural plant colors (e.g.,
		ornamentals) and shiny leaves/fruit

[0156] Other embodiments of the invention will be apparent to those of skill in the art.

[0157] Crassulacean Acid Metabolism (CAM)

[0158] CAM involves the use of both the C₃ and C₄ pathways of carbon fixation. However, unlike C₄ plants, CAM plants temporally separate, as opposed to spatially separate, the C₃ and C₄ cycles.

[0159] The C₃ cycle (Calvin cycle) takes place in the stroma of the chloroplasts, starts and ends with the five carbon sugar, ribulose 1,5-bisphosphate (RuBP). The Calvin cycle occurs in three stages. (1) Carbon dioxide enters the cycle and is enzymatically combined (fixed) to RuBP. The resultant six-carbon compound, an unstable enzyme-bound intermediate, is immediately hydrolyzed to generate two molecules of 3-phosphoglycerate or 3-phosphoglyceric acid (PGA). Each PGA molecule contains three carbon atoms. RuBP carboxylase/oxygenase (Rubisco) catalyzes this reaction. (2) In the second stage, 3-phosphoglycerate is reduced to glyceraldehydes 3-phosphate, or 3-phosphoglyceraldhyde (PGAL), requiring NADPH as the nucleotide cofactor for reduction. (3) In the third stage, five of the six molecules of clyceraldehyde 3-phosphate are used to regenerate three molecules of ribulose 1,5-bisphosphate. Many plants use only the C₃ cycle.

[0160] The C₄ cycle (Hatch-Slack pathway) involves a first step of fixing carbon dioxide to phosphoenolpyruvate (PEP) by the enzyme PEP carboxylase. PEP carboxylase uses the hydrated form of carbon dioxide, bicarbonate ion. Depending on the species, the resulting oxaloacetate is either reduced to malate or transaminated to aspartate through the addition of an amino group. The malate or aspartate then releases the carbon dioxide for use in the Calvin cycle. Plants that are C₄ spatially separate the different steps of carbon fixation: oxaloacetate and malate (or aspartate) are produced in the mesophyll cells, but then the malate (or aspartate) moves to bundle-sheath cells, where decarboxylation occurs and the Calvin cycle. Hence, C₄ plants spatially separate the C₃ and C₄ cycles. Kranz leaf anatomy clearly identifies most C₄ plants, wherein mesophyll cells are orderly arranged around a layer of large bundle-sheath cells, so that together, the two form concentric layers around the vascular bundle.

[0161] CAM plants are distinguished by their ability to fix carbon dioxide in the dark through the activity of PEP carboxylase in the cytosol. The initial carboxylation product is oxaloacetate, which is immediately reduced to malate. The malate is stored as malic acid in the vacuole. During the following light period, the malic acid is recovered from the vacuole, decarboxylated, and the carbon dioxide transferred to RuBP of the Calvin cycle within the same cells. Structurally, CAM plants have cells with large vacuoles (for aqueous storage of malic acid), and chloroplasts, where the carbon dioxide obtained from the malic acid can be transformed into carbohydrates.

[0162] CAM plants are largely dependent upon nighttime accumulation of carbon dioxide for their photosynthesis because their stomata are closed during the day to retard water loss. In general, CAM plants, while able to survive harsh environmental conditions, grow more slowly and if forced to compete with C₃ and C₄ species (in favorable environments), will compete poorly (Raven et al., 1999).

[0163] Examples of CAM plants include Crassula sp., Faucaria sp., Lithops sp. Rhodia sp., Cactaceae, Euphorbiaceae, Agave sp., Spanish moss, epiphytic bromeliads, pineapple, and vanilla orchids. Other examples are given in Table 5.

TABLE 5
Examples of CAM plants
	Family	Genera
	Agavaceae	Agave, Yucca
	Aizoaceae	Aptenia, Bergeranthus, Carpobrotus,
		Conophytum, Drossanthenum,
		Faucaria, Lithops,
		Mesembryanthemum, Tetragonia,
		Titanopsis, Trichodeadema
	Asclepiadaceae	Caralluma, Hoya, Stapelia
	Asteraceae	Aster, Kleinia, Notonia, Senecio
	Bromeliaceae	Acanthostachys, Aechmia,
		Ananas, Araeocassus, Billbergia,
		Bromelia, Canistrum, Dyckia,
		Guzmania, Hoplophytum,
		Neoregelia, Nidularium, Orthophytum,
		Puya, Quesnelia,
		Tillandsia,
	Cactaceae	Bergerocactus, Carnegiea, Cereus,
		Cephalocereus, Echinocereus,
		Echinopsis, Eulychnia, Ferocactus,
		Lobivia, Lophocereus,
		Machaerocereus, Mammillaria,
		Metacactus, Myrtillocactus, Neichilena,
		Nopalea, Notocactus,
		Opuntia, Pachycereus, Phyllocactus,
		Pilocopiapoe, Trichocereus, Zygocactus
	Crassulaceae	Aeonium, Bryophyllum, Cotyledon,
		Crassula, Dudleya, Echeveria,
		Kalanchoe, Rochea, Sedum,
		Sempervivum,
	Cucurbitaceae	Xerosicyos
	Didiereaceae	Alluaudia, Didieria
	Euphorbiaceae	Euphorbia, Monodenium, Synadenimum
	Geraniaceae	Geranium, Pelargonium
	Labiateae	Plectranthus
	Lilliaceae	Aloe, Gasteria, Haworthia,
		Sanservieria
	Orchidaceae	Arachnis, Aranda, Aranthera,
		Brassovora, Brassolaeliocattleya
		Bulbophyllum, Cattleya, Dendrobium,
		Encyclia, Epidendrum Laelia, Lanium,
		Oncidium, Phalaenopsis, Pleurothris,
		Schomburgkia, Sophrontis, Vanilla
	Oxalidaceae	Oxalis
	Piperaceae	Peperomia
	Polypodiaceae	Drymoglossum, Pyrrosia
	Portulacaceae	Portulacaria, Calandrinia
	Vitaceae	Cissus
	Welwitschiaceae	Wewitschia

[0164] A more general complete list of plants can be considered to be in three categories: Angiosperm-Monocotyledon, Angiosperm-Dicotyledon, and Gymnosperm. Table 5A, 5B, and 5C below give just some illustrative examples of each category of plants.

TABLE 5A
Examples of Angiosperm - Monocotyledon Plants
	Order	Family	Genera
	Graminales	Cyperaceae	Cyperus
	Graminales	Gramineae	Triticum, Andropogon,
			Avena, Cynodon
	Liliales	Liliaceae	Allium, Asperagus
	Schitaminales	Zingiberaceae	Curcuma
	Schitaminales	Zingiberaceae	Zingiber

[0165]

TABLE 5B
Examples of Angiosperm - Dicotyledon Plants
	Order	Family	Genera
	Campanulales	Cucurbitaceae	Momordica
	Gentianales	Apocynaceae	Holarrhena, Rauwolfia,
			Vinca
	Gentianales	Asclepiadaceae	Hemidesmus, Gymnema
	Gentianales	Gentianaceae	Swertia
	Geraniales	Euphorbiaceae	Phyllanthus
	Geraniales	Linaceae	Linum
	Geraniales	Meliaceae	Azadiracta
	Myrtales	Combretaceae	Terminalia
	Myrtales	Myrtaceae	Eugenia
	Piperales	Piperaceae	Piper
	Primulales	Myrsinaceae	Embelia
	Ranales	Menispermaceae	Tinospora
	Rosales	Leguminosae	Glycine,Glycyrrhiza,
			Mucuna, Phaseolus,
			Pongamia, Trigonella
	Santales	Santalaceae	Santalum
	Tubiflorales	Labiatae	Ocimum
	Tubiflorales	Pedaliaceae	Sesamum
	Tubiflorales	Scrophulariaceae	Bacopa
	Tubiflorales	Solanaceae	Withania
	Umbellales	Umbelliferae	Carum, Cuminum
	Urticales	Moraceae	Ficus

[0166]

TABLE 5C
Examples of Gymnosperm Plants
	Order	Family	Genera
	Conifereae		Pinus, Taxus

[0167] An illustrative list of non-plant biomass includes mushroom, yeast, shrimp, shark fin, milk, organ meat and human hair.

[0168] Extract

[0169] An extract is most simply a preparation that is in a different form than its source. A cell extract may be as simple as mechanically lysed cells. Such preparations may be clarified by centrifugation or filtration to remove insoluble debris.

[0170] Extracts also comprise those preparations that involve the use of a solvent. Examples of solvents are water, a detergent, an oil or an organic compound. Extracts may be concentrated, removing most of the solvent and/or water; and may also be fractionated, using any method common to those of skill in the art (such as a second extraction, filtration, size fractionation by gel filtration or gradient centrifugation, etc.). In addition, extracts may also contain substances added to the mixture to preserve some components, such as the case with protease inhibitors to prolong protein life, or sodium azide to prevent microbial contamination.

[0171] When oils are used as a solvent, generally all oils that are appropriate for the application can be used. Examples include vegetable (corn, hempnut, mustard, rapeseed, safflower, sesame, sunflower, flaxseed, canola, soybean, olive, grape seed, walnut, peanut, anise, balm, bay, bergamont, borage, cajeput, castor (including Turkey Red (sulfated castor)), cedarwood, cinnamon, clove, coconut, cottonseed, evening primrose, jojoba bean, linseed (boiled or not), macadeamia, orignaum (thyme), Tea Tree, wheat germ, Neem (Azadirachta indica), Karanj (Pongamia glabra) and almond), animal (lard, fish, and butterfat from milk from various species), and those produced by the extraction industries (mineral, immersion and halocarbon). Purified oil components (lipids) may also be used. While all combinations of such oils and fats can be used, it is preferred to avoid those oils and oil combinations that polymerize or form gum during the extraction procedure that would interfere with extraction and fractionation.

[0172] Often, cell or tissue extracts are made to isolate a component from the intact source; for example, growth factors, surface proteins, nucleic acids, lipids, polysaccharides, etc., or even different cellular compartments, including Golgi vesicles, lysosomes, nuclei, mitochondria and chloroplasts may be extracted from cells.

[0173] A biomass extract may be made from any part of, or the entire, biomass. Plant parts include leaves, stems, flowers, inflorescences, shoots, cotyledons, etc. Non-plant biomass parts include milk, organ meat, blood, fruiting bodies, mycelium, hair, horn etc. The various parts may be dehydrated or used fresh. Often, the biomass parts are washed before processing. Fractionation with organic solvents may be desired to separate out organic-soluble components, such as chlorophyll.

[0174] The term biomass extract in the context of the current invention refers to any extract, made from a plant or non-plant biomass, that has at least one activity of the biomass extracts and compositions of the invention. A biomass extract activity is one that is evident throughout the description of the invention, including, but not limited to, Tables 1, 1A, 1B, 1C, 1D and Tables 2, 3, and 4.

[0175] Vigor

[0176] Vigor refers to the active, healthy, and well-balanced growth of plants or animals. For example, a vigorous plant has a fast growth rate coupled with non-etiolated habit and copious reproduction (seed or spore). A vigorous animal also has a fast growth rate coupled with adequate body strength.

[0177] Resistance

[0178] Resistance is of two types. A plant or animal may resist pests or opportunistic infections.

[0179] A plant or animal may also show resistance or tolerance to environmental stresses, such as heat, drought, frost, osmotic stresses and sudden fluctuations in the environment.

[0180] Production, Yield, and Feed Conversion

[0181] Production refers to the aspect of a plant or animal that is used for human purposes. For example, tomato plants are grown for their tomatoes; a tomato variety that produces many fruits per plant is more productive than one that produces few fruit but many leaves. On the other hand, a lettuce plant with many leaves is more productive than one that bolts early.

[0182] Yield refers the actual production per unit, unit referring to an organism such as a plant or animal.

[0183] Feed conversion is tied into production and yield. Feed conversion refers to the ability of an animal to efficiently produce per amount of feed.

[0184] Quality

[0185] Quality refers to subjective criteria that are used commercially to distinguish goods. For example, a high quality apple is one of a certain weight, certain, shape, free of blemishes, ripened and has a desired coloration, flavor, and texture. Qualitative assessments are well known to those of skill in the various arts.

[0186] Longevity

[0187] Longevity refers to criteria that define delaying of senescence such as a longer green life of a leaf or longer shelf life of flower or fruit.

[0188] A. Extraction

[0189] The following describes the preparation of extract from Angiosperm-monocotyledon, Angiosperm-dicotyledon and Gymnosperm plants and from non-plant biomass. General extraction methods are defined both for fresh, soft, easily homogenizable biomass and biomass parts and for dry, hard, difficult to homogenize biomass and biomass parts.

[0190] A1: Fresh, Soft, Easy to Homogenize Biomass and Biomass Parts:

[0191] The following describes the preparation of an extract prepared from Kalanchoe pinnata (Lam.). Also see Examples. It will be apparent to one of skill in the art that many variations of the following procedure may yield extracts with similar activities. In general, any extract produced from Kalanchoe pinnata (Lam.) or any other whole plant or non-plant biomass or parts of plant or non-plant biomass with soft, easy to homogenize material that has at least one of the activities of the extract (see examples) is contemplated by the inventors.

[0192] However, any extract comprising regeneration and other specific activities can be similarly prepared from any CAM plant, such as Aloe vera or Cissue quadrangularis or from any other whole plant or parts of plant or with any other whole non-plant biomass or parts of non-plant biomass with fresh, soft, easy to homogenize material. Such extracts will have at least one activity of the compositions of the invention (see Examples). Such a procedure can be used for a wide variety of fresh, soft plant and non-plant materials including but not restricted to Allium cepa, Allium sativum, Bacopa monnieri, Fresh Ocimum santum, Phyllanthus indica (Fresh Fruit), and Zingiber officinalis (Fresh Rhizome), baker's yeast, fresh mushroom, ground prawn paste, milk, bacterial and fungal cell mass, and organ meat.

[0193] A mixture of small, medium and large leaves (1205 g) of Kalanchoe pinnata (Lam.) is plucked. After washing in water, the leaves are blended in a household blender, adding water to the mixture to allow the blades of the blender to contact the leaves such that the leaves are reduced to a pulp. Generally, water equal to half the weight of fresh leaves suffices. Next, 1205 g of sesame oil is heated to 100-120 C., but well below the smoke point of the oil in a stainless steel pot. The leaf mixture is charged to the pot and brought to boil. Thus, the ratio of oil to total homogenate for this soft material was 1:1. Stirring is provided to bring about an intimate contact of the plant material with oil and water and to minimize sticking on the walls of the vessel or charring. Boiling is continued until only fine bubbles or fine foam is formed, and bubbling nearly ceases. When the oil just starts to smoke, the extract is sufficiently free of water and is ready for filtration. The boiling time may be anywhere from 15 minutes to over 6 hours, depending on a variety of variables, including the starting material, volumes of water, etc. Heating is then stopped, the mixture cooled and filtered through cheesecloth to separate the first extract from the leafy residue. The leafy residue is mixed with sesame oil, 0 to 1 times the weight of the filtrate and filtered through a double layer of cheesecloth to obtain a second extract. The two extracts are combined, and additional sesame oil is added to adjust the total weight to 1205 g to obtain a final oil strength of 100, i.e., R-100. The composition is based on 100 g of leaf equivalent per 100 g of total final extract.

[0194] A2: Dry, Hard, Difficult to Homogenize Biomass and Biomass Parts:

[0195] The following describes the preparation of an extract prepared from Curcuma amada Roxb. Also see Examples. It will be apparent to one of skill in the art that many variations of the following procedure may yield extracts with similar activities. In general, any extract produced from Curcuma amada or any other whole plant or parts of plant or any other whole non-plant biomass or parts of non-plant biomass with dry, hard, difficult to homogenize material that has at least one of the activities of the extract (see examples) is contemplated by the inventors.

[0196] However, any extract comprising regeneration and other specific activities can be similarly prepared from any other whole plant or parts of plant with dry, hard, difficult to homogenize material including bark, rhizomes and seeds, bones, cartilage. Such extracts will have at least one activity of the compositions of the invention (see Examples). Such a procedure can be used for a variety of plants with hard, difficult to homogenize material including but not restricted to Azadirachta indica cake, Carum copticum, Cuminum cyminum, Curcuma longa, Ficus bengalensis, Embelia ribes, Eugenia jambolana, Hemidesmus indicus, Gymnema sylvestree, Glycine max, Glycyrrhiza glabra, Holarrhena antidysenterica, Momordica charantia, Phaeolus radiatus, Piper longum, Piper nigrum, Pongamia glabra cake, Rauwolfia serpentina, Santalum alba, Trigonella foecum-graecum, Terminalia chebula, Terminalia bellerica, Taxus baccata, Tinospora cordifolia, Mucuna pruriens, Sesamum indicum, Triticum vulgare, Swertia chirata, Cyperous rotundus, Vinca rosea, Withania somnifera.

[0197] Dry rhizomes (500 g) of Curcuma amada Roxb were obtained from the market. After washing in water, the rhizomes were pounded in a mortar to break them up into small pieces below 3 mm in diameter. These pieces were then soaked in 2 litres of water for 2 hours. All of the material, including the soak water, was blended by adding some more water to the mixture to allow the blades of the blender to contact the pieces such that the pieces are reduced to a very fine grind. Generally, for such dry, hard materials, water equal to 4 to 8 times the weight of dry starting material suffices. The homogenate was filtered to obtain 1180 g of wet residue. Next, 1840 g of sesame oil is heated to 100-120 C., but well below the smoke point of the oil in a stainless steel pot. With very hard biomass from seeds and dry bark/roots, about 4 kg oil per kg biomass is needed to keep the mass stirrable to the end. The wet residue is charged to the pot along with an additional 800 ml water and brought to boil. Stirring is provided to bring about an intimate contact of the plant material with oil and water and to minimize sticking on the walls of the vessel or charring. Boiling is continued until only fine bubbles or fine foam is formed, and bubbling nearly ceases. When the oil just starts to smoke, the extract is sufficiently free of water and is ready for filtration. The boiling time may be anywhere from 25 minutes to over 6 hours, depending on a variety of variables, including the starting material, volumes of water, etc. Heating is then stopped, the mixture cooled and filtered through cheesecloth to separate the first extract from the grind residue. The grind residue is mixed with sesame oil, 0 to 1 times the weight of the filtrate and filtered through a double layer of cheesecloth to obtain a second extract. The two extracts are combined, and additional sesame oil is added to adjust the total weight to 2000 g. The composition is based on 25 g of rhizome equivalent per 100 g of total final extract.

[0198] In both of above general procedures, several variables can be adjusted to achieve extracts of desired potency. For example, the starting material may consist of leaves, stems, shoots, seeds, bark, rhizome or the entire plant. Alternatively, juice that has been manually extracted, or expressed, from the plant or plant parts may also be used. Pounding can be done in a mortar or with any other device that can reduce the size of the starting material. Instead of a blender to homogenize the plant tissues, a mortar and pestle, or any other device or method that can destroy the integrity of the plant tissue, may be used. Boiling time may range from 25 minutes to 6 hours without losing efficacy. The oil may be any known in the art, including coconut, sesame, mineral and butterfat. It will be apparent to one of skill in the art to adjust other variables as appropriate, as, for example, when large-scale preparations are desired.

[0199] The compositions thus made may also be further diluted with oils to achieve extracts of different strengths that are suitable for various applications. Dilution serves important functions, including reducing any irritants and providing convenient doses. General penetrants and absorption aides such as Isopropyl myristate or MCT (Medium Chain Tryglyceride fraction) oil may be admixed.

[0200] B. Pharmaceutical Compositions

[0201] The compositions of the invention can be incorporated into pharmaceutical compositions. Such compositions typically comprise the plant extracts of the invention.

[0202] A pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration (Gennaro, 2000). Preferred examples of such carriers or diluents include, but are not limited to, water, saline, Finger's solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used. Except when a conventional media or agent is incompatible with an active compound, use of these compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.

[0203] The pharmaceutical compositions for the administration of the active compounds, such as those of any of the plant extracts, may conveniently be presented in dosage unit form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing the active compound or plant extracts into association with the carrier that constitutes one or more accessory ingredients. In general, the pharmaceutical compositions are prepared by uniformly and intimately bringing the active compound into association with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product into the desired formulation. In the pharmaceutical composition the active compound is included in an amount sufficient to produce the desired effect upon the process or condition of diseases.

[0204] General Considerations

[0205] A pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration, including intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and agents for the adjustment of tonicity such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0206] Injectable Formulations

[0207] Pharmaceutical compositions suitable for injection include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, paraffin oils such as CREMOPHOR EL (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid so as to be administered using a syringe. Such compositions should be stable during manufacture and storage and must be preserved against contamination from microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (such as glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures. Proper fluidity can be maintained, for example, by using a coating such as lecithin, by maintaining the required particle size in the case of dispersion and by using surfactants. Various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, and thimerosal, can contain microorganism contamination. Isotonic agents, for example, sugars, polyalcohols such as manitol, sorbitol, and sodium chloride can be included in the composition. Compositions that can delay absorption include agents such as aluminum monostearate and gelatin.

[0208] Sterile injectable solutions can be prepared by incorporating the active compound or composition, such as plant extracts, in the required amount in an appropriate solvent with one or a combination of ingredients as required, followed by sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium, and the other required ingredients as discussed. Sterile powders for the preparation of sterile injectable solutions, methods of preparation include vacuum drying and freeze-drying that yield a powder containing the active ingredient and any desired ingredient from a sterile solution.

[0209] Oral Compositions

[0210] Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included. Tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, PRIMOGEL, or corn starch; a lubricant such as magnesium stearate or STEROTES; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

[0211] Compositions for Inhalation

[0212] For administration by inhalation, the compounds are delivered as an aerosol spray from a nebulizer or a pressurized container that contains a suitable propellant, e.g., a gas such as carbon dioxide.

[0213] Systemic Administration, Including Patches

[0214] Systemic administration can also be transmucosal or transdermal. For transmucosal or transdermal administration, penetrants that can permeate the target barrier(s) are selected. Transmucosal penetrants include: detergents, bile salts, and fusidic acid derivatives. Nasal sprays or suppositories can be used for transmucosal administration. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams.

[0215] Creams are useful for a variety of external applications such as on chapped lips, cracked feet, heat rash, face cream, pimples, hand and body lotion to restore darkened skin after sun exposure, etc.

[0216] The compounds can also be prepared in the form of suppositories (e.g. with bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

[0217] Carriers

[0218] In one embodiment, plant extracts are prepared with carriers that protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Such materials can be obtained commercially from ALZA Corporation (Mountain View, Calif.) and NOVA Pharmaceuticals, Inc. (Lake Elsinore, Calif.), or prepared by one of skill in the art. Liposomal suspensions can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, such as in (Eppstein et al., U.S. Pat. No. 4,522,811, 1985).

[0219] Unit Dosage

[0220] Oral formulations or parenteral compositions in unit dosage form can be created to facilitate administration and dosage uniformity. Unit dosage form refers to physically discrete units suited as single dosages for the subject to be treated, containing a therapeutically effective quantity of active compound in association with the required pharmaceutical carrier. The specification for the unit dosage forms of the invention are dictated by, and directly dependent on, the unique characteristics of the active compound and the particular desired therapeutic effect, and the inherent limitations of compounding the active compound.

[0221] Dosage

[0222] The pharmaceutical composition and method of the present invention may further comprise other therapeutically active compounds, such as plant compositions, as noted herein that are usually applied in the treatment of wounds or other associated pathological conditions.

[0223] In the treatment of human conditions which require the compositions of the invention, an appropriate dosage level will generally be about 0.01 to 10 mg per kg patient body weight per day which can be administered in single or multiple doses. Preferably, the dosage level will be about 0.01 to about 10 mg/kg per day; more preferably about 0.01 to about 2.0 mg/kg per day, and most preferably 0.01 to about 0.4 mg/kg per day. A suitable dosage level may be about 0.001 to 10 mg/kg per day, about 0.01 to 2 mg/kg per day, or about 0.01 to 50 mg/kg per day. Within this range the dosage may be 0.05 to 0.5, 0.5 to 5 or 5 to 50 mg/kg per day.

[0224] For oral administration, the compositions are preferably provided in the form of tablets containing 0.1 to 10 milligrams of the active ingredient, particularly 0.1, 0.2, 0.5, 1.0, 1.5, 2.0, 2.5, 5.0, 7.5 and 10.0 milligrams of the active ingredient. The compounds may be administered 1 to 4 times per day, preferably once or twice per day.

[0225] For topical applications, the composition may have a dosage of about 0.001% to 50%, more preferably 0.01% to 10%, delivering 0.1 mg to 100 mg per 1 g application. The compositions may be administered 1 to 8 times per day, preferably once or twice per day. Alternatively, pads and other materials may be impregnated with such compositions and held in contact to the surface of the subject for chronic application.

[0226] The dosages outlined above are also suitable for veterinary applications. It will be understood, however, that the specific dose level and frequency of dosage for any particular subject may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy. In addition, the site of delivery will also impact dosage and frequency. Also understood, however, is that dosage for livestock may also differ. A skilled artisan will know how to adjust the unit dosage.

[0227] Kits for Pharmaceutical Compositions

[0228] The pharmaceutical compositions can be included in a kit, container, pack, or dispenser together with instructions for administration. When the invention is supplied as a kit, the different components of the composition may be packaged in separate containers and admixed immediately before use. Such packaging of the components separately may permit long-term storage without losing the active components' functions.

[0229] Containers or Vessels

[0230] The reagents included in the kits can be supplied in containers of any sort such that the life of the different components are preserved, and are not adsorbed or altered by the materials of the container. For example, sealed glass ampoules may contain lyophilized plant extracts or buffer that have been packaged under a neutral, non-reacting gas, such as nitrogen. Ampoules may consist of any suitable material, such as glass, organic polymers, such as polycarbonate, polystyrene, etc., ceramic, metal or any other material typically employed to hold reagents. Other examples of suitable containers include simple bottles that may be fabricated from similar substances as ampoules, and envelopes, that may consist of foil-lined interiors, such as aluminum or an alloy. Other containers include test tubes, vials, flasks, bottles, syringes, or the like. Containers may have a sterile access port, such as a bottle having a stopper that can be pierced by a hypodermic injection needle. Other containers may have two compartments that are separated by a readily removable membrane that upon removal permits the components to mix. Removable membranes may be glass, plastic, rubber, etc.

[0231] Instructional Materials

[0232] Kits may also be supplied with instructional materials. Instructions may be printed on paper or other substrate, and/or may be supplied as an electronic-readable medium, such as a floppy disc, CD-ROM, DVD-ROM, Zip disc, videotape, audiotape, etc. Detailed instructions may not be physically associated with the kit; instead, a user may be directed to an internet web site specified by the manufacturer or distributor of the kit, or supplied as electronic mail.

[0233] Delivery Methods

[0234] Interstitial Delivery

[0235] The composition of the invention, such as plant extracts, may be delivered to the interstitial space of tissues of the animal body, including those of muscle, skin, brain, lung, liver, spleen, bone marrow, thymus, heart, lymph, blood, bone, cartilage, pancreas, kidney, gall bladder, stomach, intestine, testis, ovary, uterus, rectum, nervous system, eye, gland, and connective tissue. Interstitial space of the tissues comprises the intercellular, fluid, mucopolysaccharide matrix among the reticular fibers of organ tissues, elastic fibers in the walls of vessels or chambers, collagen fibers of fibrous tissues, or that same matrix within connective tissue ensheathing muscle cells or in the lacunae of bone. It is similarly the space occupied by the plasma of the circulation and the lymph fluid of the lymphatic channels. They may be conveniently delivered by injection into the tissues comprising these cells. They are preferably delivered to sites of injury, preferably to live cells and extracellular matrices directly adjacent to dead and dying tissue.

[0236] Any apparatus known to the skilled artisan in the medical arts may be used to deliver the compositions of the invention to the site of injury interstitially. These include, but are not limited to, syringes, stents and catheters.

[0237] Systemic Delivery

[0238] Any apparatus known to the skilled artisan in the medical arts may be used to deliver the compositions of the invention to the circulation system. These include, but are not limited to, syringes, stents and catheters. One convenient method is delivery via intravenous drip. Another approach would comprise implants, such as transdermal patches, that deliver the compositions of the invention over prolonged periods of time. Such implants may or may not be absorbed by the subject over time.

[0239] Surgical Delivery

[0240] The compositions of the invention may be delivered in a way that is appropriate for the surgery, including by bathing the area under surgery, implantable drug delivery systems, and matrices (absorbed by the body over time) impregnated with the compositions of the invention.

[0241] Superficial Delivery

[0242] Direct application of the compositions of the invention, such as plant extracts, may be used. For example, gauze impregnated with plant extracts or active components may be directly applied to the site of damage, and may be held in place, such as by a bandage or other wrapping. Alternatively, the compositions of the invention may be applied in salves, creams, or other pharmaceutical compositions known in the art meant for topical application.

[0243] C. Agricultural/Horticultural Compositions

[0244] Compositions Suitable for Application to Plants

[0245] In its simplest form, plant extract compositions that are suitable for agricultural compositions are simply diluted in water. Oil, powder and tablets of the CAM plant extract compositions may be used.

[0246] It is also possible to prepare combinations with other pesticidally active substances, fertilizers and/or growth regulators, for example in the form of a ready mix or a tank mix. These can be thought of us to be carriers for the plant extracts.

[0247] Wettable powders are preparations which are uniformly dispersible in water and which, besides the active substance, also comprise ionic and/or nonionic surfactants (wetting agents, dispersants), for example polyoxyethylated alkylphenols, polyoxyethylated fatty alcohols, polyoxyethylated fatty amines, fatty alcohol polyglycol ether sulfates, alkanesulfonates, alkylbenzenesulfonates, sodium lignosulfonate, sodium 2,2-dinaphthylmethane 6,6-disulfonate, sodium dibutylnaphthalene-sulfonate, or else sodium oleoylmethyltaurinate, in addition to a diluent or inert substance.

[0248] Emulsifiable concentrates are prepared by dissolving the plant extracts in an organic solvent, for example butanol, cyclohexanone, dimethylformamide, xylene, or else higher-boiling aromatics or hydrocarbons, or mixtures of the organic solvents with the addition of one or more ionic and/or nonionic surfactants (emulsifiers). Examples of substances which can be used as emulsifiers are: calcium alkylarylsulfonates such as calcium dodecylbenzenesulfonate, or nonionic emuslifiers such as fatty acid polyglycol esters, alkylaryl polyglycol ethers, fatty alcohol polyglycol ethers, propylene oxide/ethylene oxide condensates, alkyl polyethers, sorbitan esters, for example sorbitan fatty acid esters, or polyoxyethylene sorbitan esters, for example polyoxyethylene sorbitan fatty acid esters.

[0249] Dusts are obtained by grinding or mixing the plant extracts with finely distributed solid substances, for example, talc, natural clays such as kaolin, bentonite and pyrophyllite, or diatomaceous earth.

[0250] Suspension concentrates can be water-based or oil-based. They can be prepared, for example, by wet grinding using commercially available bead mills with or without an addition of surfactants, for example those that have already been mentioned above in the case of the other formulation types.

[0251] Emulsions, for example oil-in-water emulsions (EW), can be prepared, for example, by means of stirrers, colloid mills and/or static mixers using aqueous organic solvents in the presence or absence of surfactants, for example, in the case of the other formulation types.

[0252] Granules can be prepared either by spraying the active substance onto adsorptive, granulated inert material or by applying active substance concentrates to the surface of carriers such as sand, kaolinites or granulated inert material with the aid of binders, for example polyvinyl alcohol, sodium polyacrylate or else mineral oils. Suitable active substances can also be granulated in the manner that is conventional for the preparation of fertilizer granules, if desired as a mixture with fertilizers.

[0253] As a rule, water-dispersible granules are prepared by the customary processes such as spray drying, fluidized-bed granulation, disk granulation, mixing with high-speed mixers, and extrusion without solid inert material.

[0254] For the preparation of disk, fluidized-bed, extruder and spray granules see, for example (1973; 1979).

[0255] In wettable powders, the concentration of active substance is, for example, approximately 0.01% to 90% by weight, more preferably 0.01% to 0.5%, the remainder to 100% by weight being composed of customary formulation components. In the case of emulsifiable concentrates, the concentration, of active substance may be approximately 0.01% to 90%, preferably 0.01% to 0.5% by weight. Formulations in the form of dusts comprise 0.01% to 30% by weight of active substance, in most cases preferably 0.01% to 0.5% by weight of active substance; sprayable solutions comprise approximately 0.01% to 80%, preferably 0.01% to 0.5% by weight of active substance. In the case of water-dispersible granules, the active substance content depends partly on whether the active compound is in liquid or solid form and on which granulation auxiliaries, fibers and the like are being used. The active substance content of the water-dispersible granules is, for example, between 0.01% and 95% by weight, preferably between 0.01% and 0.5% by weight.

[0256] Alternatively, the rate of application of an active biomass extract is 2 to 100 g per hectare per year, applied in 4 to 20 sprays per year (or 2-5 sprays per season). More preferably, 3 to 30 g per hectare per year is applied. For herbicidal effects or for control of excessive growth mediated by biomass extracts, the extract concentration is increased to 25 to 500 g per hectare per year.

[0257] Besides, the above mentioned formulations of active substances may comprise, if appropriate, the adhesives, wetting agents, dispersants, emulsifiers, penetrants, preservatives, antifreeze agents, solvents, fillers, carriers, colorants, antifoams, evaporation inhibitors and pH and viscosity regulators which are customary in each case.

[0258] For use, the formulations that are in commercially available form are, if desired, diluted in the customary manner, for example using water in the case of wettable powders, emulsifiable concentrates, dispersions and water-dispersible granules. Preparations in the form of dusts, granules and sprayable solutions are usually not diluted any further with other inert substances prior to use. The necessary rate of application of the safeners varies with the external conditions such as temperature and humidity.

[0259] Components that can also be present in biomass extract compositions suitable for plant (agricultural) application, such as fertilizers or pesticides, include natural enzymes, growth hormones such as the gibberellins (gibberellic acid and gibberellin plant growth hormones), and control agents including pesticides such as acaracides and molluskicides, insecticides, fungicides, nematocides, and the like, depending of course on their compatibility with particular plant extracts. Examples of control agents that can be used in the compositions of the invention, depending on particular biomass extract compatibility, include inorganic compounds such as elementary sulfur and inorganic sulfur compounds, e.g., calcium polysulfide and sodium thiosulfate, which are effective fungicides, copper, zinc, and other metal in organics such as copper carbonate copper oxychloride, copper sulfate, and copper zinc sulfate. Organometallic compounds such as iron and tin compounds, e.g., triphenyl tin hydroxide exhibit both insecticidal and pesticidal activity. Saturated higher alkyl alcohols, either straight or branched chain, such as nonyl and decyl alcohol, can be present as insecticides. Aldehydes such as metaldehyde, are effective molluskicides, e.g., useful against snails. Carbonic acid derivatives, especially their mixed esters, are potent acaracides and fungicides; when sulfur is also present, e.g., mixed esters of thio- and di-thiocarbonic acids, activity is enhanced. 6-methylquinoxaline-2,3-dithiocyclocarbonate is an effective acaricide, fungicide, and insecticide. Carbamic acid derivatives such as aryl esters of N-methylcarbamnic acid, e.g., 1-naphthyl-N-methylcarbamate can also be used. Halogen substituted aliphatic monobasic and dibasic carboxylic acids are effective pesticides. Natural pyrethrins and their synthetic analogs are also effective pesticides. Salicylanilide is effective against leaf mold and tomato brown spot. Hetercyclic compounds possessing insecticidal and/or fungicidal activity can also be used. Halogen derivatives of benzene, such as paradichlorobenzene, are effective pesticides, often used against the sugarbeet weevil. Chitin-containing products are effective menatocides. Other compounds that can be used include aliphatic mercaptans having four or fewer carbon atoms, organic sulfides and thioacetals, nitro compounds such as chloropicrin dichloronitroethane, and chloronitropropane, copper and zinc inorganic and organic compounds, e.g., copper linoleate, copper naphthenate, etc., organophosphorous compounds of which there are well over a hundred, e.g., DDVP, tris-(2,4-diphenoxyethyl) phosphite, derivatives of mono- and dithiophosphoric acids, such as 0,0-diethyl S (2-ethylthio)-ethyl)phosphorodithioate, phosphoric acid derivatives, pyrophosphoric acid derivatives and phosphonic acid derivatives, quinones, sulfonic acid derivatives, thiocyanates and isocyanates, phytoalexins, insect killing soaps such as potassium fatty acid salts, and antiallatotropins such as 7-methoxy-2,2-dimethylchromene and the 6,7-dimethoxy analog. Diatomaceous earth can be used, which kills crawling insects.

[0260] These components can comprise from 0.001 to 10% or more by weight of the biomass extract compositions suitable for plant application. Also, alkalizing agents such as ground limestone and acidifying agents such as inorganic acids or acid salts can be added as needed or desired.

[0261] The biomass extract compositions suitable for plant application can be in solid form or in the form of an aqueous solution. Solid forms include powders and larger particulate forms, e.g., from 20 to 200 mesh. Where the biomass extract compositions are in solid form and biomass extracts are sensitive to light, air, or compounds in the composition, or to optional added components, the biomass extract compositions can be separately encapsulated in water soluble coatings, e.g., dyed or undyed gelatin spheres or capsules, or by micro-encapsulation to a free flowing powder using one or more of gelatin, polyvinyl alcohol, ethylcellulose, cellulose acetate phthalate, or styrene maleic anhydride. The separately encapsulated biomass extracts can then be mixed with the powder or larger particulates of another unencapsulated component and any optional components.

[0262] The presence of biomass extracts in the compositions suitable for agricultural use provides further enhancement of plant growth, and where applicable, crop production, i.e., by further enhancement is meant benefits in plant growth and crop production in addition to the benefits provided by the components other than plant extracts, and/or provides control of pest damage and resistance to stress. Biomass extracts also improve the effectiveness of beneficial microorganisms, and promote nutrient absorption and assimilation.

[0263] Particular biomass extracts may be added to herbicides, known in the art, to increase their effectiveness; as such, biomass extracts can also be used to control unwanted proliferation of weeds and other vegetative growth.

[0264] The following examples are included to demonstrate preferred embodiments of the present invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent techniques discovered by the inventors to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments that are disclosed and still obtain a like or similar result without departing form the spirit and scope of the invention. For example, heating could be provided with steam or hot heat transfer mediaum to reduce degradation due to wall hot spots. Use of nitrogen or other inert gas blanketing may also be used to reduce degradation. The extraction process can also be carried out in a continuous manner by using the well established engineering methods of continuously introducing biomass, oil and water at one end and continuously withdrawing the liquid slurry at the other end of a suitably designed vessel.

[0265] The following examples illustrate biomass extractions; however, one of skill in the art will know how to vary the various variables to obtain extracts with the activity of the biomass extracts of the invention. Table A summarizes the designations for the various extracts used throughout the following examples.

[0266] In several of the following examples of biomass extraction, final extract weight is the same as the starting fresh biomass weight; hence all extracts are equivalent on fresh weight basis and have equivalent potencies. This, however, is not essential. As illustrated in Table A, any proportion of biomass weight to final extract can be used and suitabley designated as explained herein.

[0267] Using Kalanchoe pinnata

[0268] A mixture of small, medium and large leaves (1205 g) of Kalanchoe pinnata (Lam.) pers. was harvested. The leaves were washed with water, and blended in a household blender by addition of water, approximately half the weight of plant material, 600 g (or 600 ml). Separately, an equal weight of sesame oil was heated in a stainless steel pot. The blended mixture of leaves and water was charged to the pot and boiled for about 2 hours and 45 minutes until a very fine foam appeared. Heating was stopped, the mixture cooled and filtered through a once-folded (double) layer of cheesecloth to separate the first extract from leafy residue. The leafy residue was stirred with sesame oil equal to half the weight of the wet residue and filtered through a double layer of cheesecloth to obtain a second extract. The two extracts were combined and sesame oil was added to adjust the total weight of the final extract to 1205 g. This composition is based on 100 g of leaf equivalent per 100 g of total extract. The final extract was named R-100.

[0269] Large, thick leaves (380 g) of Kalanchoe pinnata (Lam.) pers. were harvested and processed as in Example 1, except for a boiling time of 25 minutes. The final extract weight was adjusted with sesame oil to 380 g. This extract was named R-100.

[0270] Leaves of Kalanchoe pinnata (Lam) pers. (2000 g) were harvested. Procedure as outlined in Example 1 was followed, but with a boiling time of 6 hours. The final extract weight was adjusted with sesame oil to 2000 g. This extract was named R-100

[0271] Coconut Oil

[0272] Leaves (1380 g) of Kalanchoe pinnata (Lam.) pers. were harvested and washed with water. Leaves were blended as in Example 1. Separately, 2000 g of refined coconut oil was heated in a stainless steel pot. The total leaf homogenate was charged to the pot and brought to boil. After boiling for 4 hours and 45 minutes, until a very fine foam formed and started to subside, heating was stopped. The mixture was cooled and filtered through a double layer of cheesecloth to separate the first extract from leafy residue. The extract was adjusted to 1380 g by adding coconut oil. This composition, based on 100 g of leaf equivalent per 100 g of total final extract, was named R-100.

[0273] Safflower Oil; also Demonstrating Different Homogenization

[0274] Leaves (600 g) of Kalanchoe pinnata (Lam.) pers. were harvested and washed with water. Leaves were then ground in a pestle and mortar, adding water as in Example 1 to produce a leaf homogenate. Separately, 400 g of safflower oil was heated in a stainless steel pot. The total leaf homogenate was charged to the pot and boiled for 45 minutes until a fine foam formed and subsided. The mixture was cooled and filtered a double layer of cheesecloth to separate the first oil extract. The leafy residue was stirred with an equal amount by weight of safflower oil. The two extracts were combined, and additional safflower oil was added to a final total weight of 600 g. This composition, based on 100 g of leaf equivalent per 100 g of total final extract, was named R-100.

[0275] Using Aloe vera

[0276] 800 g of Aloe vera, Linn. leaves were plucked and washed with water. Leaves were blended in a household blender by addition of 200 ml water as in Example 1 to produce a total homogenate. This homogenate was filtered over a cloth to separate the juice (J fraction) from the leafy residue concentrate (LR fraction). Separately, sesame oil was heated in two separate stainless steel pots.

[0277] The LR fraction was charged to one of the pots containing 800 g of sesame oil; 400 ml of water was then added, and the mixture was boiled for 45 minutes until a fine foam formed and subsided. The mixture was cooled and filtered through double layer of cheesecloth to separate the first extract. The residue was stirred with an equal weight of sesame oil. The two extracts were combined and additional sesame oil was added to a final total weight of 800 g. This composition from the LR fraction based on 100 g of initial leaf equivalent per 100 g of total final extract was named A-100 PLUS.

[0278] The J fraction was charged to a second stainless steel pot containing 800 g of sesame oil and the mixture was boiled for 1 hour and 20 minutes until the foam subsided. This extract was filtered through a double layer of cheesecloth and additional sesame oil added to a final weight of 800 g. This composition from the J fraction based on 100 g of initial total leaf equivalent per 100 g of final extract is designated as A-100 MINUS.

[0279] Using Cissus quadrangularis

[0280] The stem portion of Cissus quadrangularis (650 g) was harvested and washed with water. The stems were then blended in a household blender by addition of water as in Example 1 to produce a total homogenate. The homogenate was filtered over a double layer of cheesecloth to separate the juice (J fraction) from the fibrous stem residue concentrate (LR fraction). Separately, sesame oil was heated in two separate stainless steel pots.

[0281] The LR fraction was charged to one of the pots containing 650 g of sesame oil. 600 ml of water was then added, and the mixture boiled for 30 minutes until a fine foam formed and subsided. The mixture was cooled and filtered a double layer of cheesecloth to separate the extract. Additional sesame oil was added to a final total weight of 650 g. This composition from the LR fraction, based on 100 g of initial stem equivalent per 100 g of total final extract, was named C-100 PLUS.

[0282] The J fraction was charged to a second stainless steel pot containing 640 g of sesame oil, and the mixture was boiled for 1 hour and 20 minutes until the foam subsided. This extract was filtered, and additional sesame oil added to a final weight of 650 g. This composition from the J fraction, based on 100 g of initial total stem equivalent per 100 g of final extract, is designated as C-100 MINUS.

[0283] Leaves of Kalanchoe pinnata were harvested and washed with water. Leaves were blended in a household blender by adding water as in Example 1 to produce a leaf homogenate. Separately, 2000 g of butterfat (ghee) was heated in a stainless steel pot. The total leaf homogenate was charged to the pot and brought to boil. Boiling was continued for 1 hour and 30 minutes to drive off the water. The mixture was cooled and filtered through a double layer of cheesecloth while warm to recover butterfat. The final composition, weighing 1600 g and based on 50 g of leaf equivalent per 100 g of total final extract was named R-50.

[0284] In all of these extract preparations, stirring was provided while boiling in oil or fat.

[0285] Extracts by using the general procedures outlined in Examples 1 to 7 can also be made from all other CAM plants. The same general procedures can also be used to make extracts from other soft, fresh plants and their parts. These can be any plants from the Angiosperm-monocotyledon, Angiosperm-dicotyledon and Gymnosperm plants including but not limited to fruit pulp of Phyllanthus emblica Linn., and rhizome of Zingiber officinalis Roscoe.

[0286] The following examples 1A to 4A, illustrate extractions of dry, hard biomass and their parts.

[0287] The bark portion of Ficus bengalensis Linn. (500 g) was procured and washed with water. The bark pieces were then soaked in 1 litre water for 2 hours. The soaked pieces were then blended in a household blender by addition of another 500 ml water to produce a total homogenate. The homogenate was filtered over a double layer of cheesecloth to separate the juice (J fraction) from the fibrous bark residue concentrate (LR fraction). Separately, sesame oil was heated in two separate stainless steel pots.

[0288] The LR fraction was charged to one of the pots containing 1840 g of sesame oil. Then, 200 ml of water was added, and the mixture boiled with continuous stirring for 50 minutes until a fine foam formed and subsided. The mixture was cooled and filtered a double layer of cheesecloth to separate the extract. The oil wet residue was stirred with 940 g of sesame oil and filtered again to obtain the second oil extract. The two oil extracts were combined to have a final total extract of 1708 g. This composition from the LR fraction, based on 29.3 g of initial bark equivalent per 100 g of total final extract, was named FEBNG-29.3 PLUS.

[0289] The J fraction was charged to a second stainless steel pot containing 485 g of sesame oil, and the mixture was boiled for 35 minutes until the foam subsided. This extract was filtered, and additional sesame oil added to a final weight of 500 g. This composition from the J fraction, based on 100 g of initial bark equivalent per 100 g of final extract, is designated as FBENG-100 MINUS.

[0290] Seeds of Taxus baccata Linn. were procured (500 g) and washed with water. They were blended in a household blender by adding water. The homogenate becomes very viscous and thick. Hence, 3 litres of water was used to produce a homogenate. Separately, 1840 g of sesame oil was heated in a stainless steel pot. The total homogenate was charged to the pot and brought to boil. Boiling with continuous stirring was continued for 55 minutes to drive off the water. The mixture was cooled and filtered through a double layer of cheesecloth to separate the first oil extract from the oil wet cake. The oil wet cake was stirred with 920 g sesame oil, and filtered to obtain a second oil extract. The two extracts were combined to produce a final composition, weighing 1858 g and based on 27 g of seed equivalent per 100 g of total final extract was named TABA-27.

[0291] Dry, tuberous roots of Cyperous rotundus Linn. (500 g) were procured and washed with water. These roots do not easily get pulverized in a mortar and pestle. They were soaked in 2 litres of water for 4 hours. The soaked material was blended in a household blender by using an additional 1.2 litres of water to produce a homogenate. The homogenate was filtered over a double layer of cheesecloth to separate the juice (J fraction) from the fibrous residue concentrate (LR fraction). Separately, sesame oil was heated in two separate stainless steel pots.

[0292] The LR fraction (947 g water wet residue) was charged to one of the pots containing 1840 g of sesame oil. Then, 400 ml of water was added and the mixture boiled with continuous stirring for 45 minutes until a fine foam formed and subsided. The mixture was cooled and filtered a double layer of cheesecloth to separate the first oil extract from oil wet residue. The oil wet residue was stirred with 464 g of sesame oil and filtered again to obtain the second extract. The two oil extracts were combined to have a final total extract of 1773 g. This composition from the LR fraction, based on 28.2 g of initial tuberous root equivalent per 100 g of total final extract, was named CROT-28.2 PLUS.

[0293] The J fraction was charged to a second stainless steel pot containing 920 g of sesame oil, and the mixture was boiled for 50 minutes until the foam subsided. This extract was filtered to obtain 798 g of extract. This composition from the J fraction, based on 62.7 g of initial tuberous root equivalent per 100 g of final extract, is designated as CROT-62.7 MINUS.

[0294] Seeds of Mucuna pruriens Bak were procured (500 g) and washed with water. The seeds were then pounded in a mortar and pestle to produce fines and pieces smaller than 5 mm. The larger pieces from the pounded mass were then soaked in 400 ml water for 4 hours. The soaked mass was blended in a household blender you produce a very thick paste. Separately, 1840 g of sesame oil was heated in a stainless steel pot. The thick paste and fines were charged to the pot along with an additional 800 ml water and brought to boil. Boiling with continuous stirring was continued for 55 minutes to drive off the water. The mixture was cooled and filtered through a double layer of cheesecloth to separate the first oil extract from the oil wet cake. The oil wet cake was stirred with 467 g sesame oil, and filtered to obtain a second oil extract. The two extracts were combined to produce a final composition, weighing 1843 g and based on 27.1 g of seed equivalent per 100 g of total final extract was named MPRU-27.1.

[0295] Extracts by using the general procedures outlined in Examples 1A to 4A can also be made from all other dry, hard plants and plant parts. These can be any plants from Angiosperm-monocotyledon, Angiosperm-dicotyledon and Gymnosperm including but not restricted to Azadirachta indica cake, Ficus bengalensis, Embelia ribes, Glycyrrhiza glabra, Holarrhena antidysenterica, Pongamia glabra cake, Trigonella foecum-graecum, Terminalia chebula, Terminalia bellerica, Taxus baccata, Mucuna pruriens, Sesamum indicum, Triticum vulgare, Swertia chirata, Cyperous rotundus.

[0296] Table A illustrates the method of designationg extracts from each plant. Each plant extract is designated by the following:

[0297] A number of alphabets which indicate the plant name, e.g. C stands for Cissus quadrangularis, CROT stands for Cyperous rotundus etc.

[0298] A further set of alphabets which may designate the particular part.

[0299] A number from 0 to 100 which designates the potency of the extract. Potency is designated on the basis of starting biomass material per 100 g of final extract.

[0300] The words, PLUS or MINUS which indicate use of LR and J fraction, respectively.

[0301] Thus, as illustrative examples,

[0302] TBEL-R-35 means an extract of the seed outer rind of Terminalia bellerica prepared such that 35 g of rind ends up as 100 g of final extract.

[0303] PE-100 PLUS means an extract of the LR fraction of the fruit of Phyllanthus emblica prepared such that 100 g of fruit ends up as 100 g of final extract.

TABLE A
Designations of the various extracts as illustrative
examples
	Total
Plant source	homogenate	LR fraction	J fraction
Kalanchoe	R-100	R-100 PLUS	R-100 MINUS
pinnata
LEAF
Kalanchoe	RS-10	RS-10 PLUS	RS-10 MINUS
pinnata
LEAF STEM
Kalanchoe	KPMS-100	KPMS-100 PLUS	KPMS-100
pinnata			MINUS
MAIN STEM
Aloe vera	A-100	A-100 PLUS	A-100 MINUS
LEAF
Cissus	C-100	C-100 PLUS	C-100 MINUS
quadrangularis
STEM
Cyperous	CROT-25	CROT-25 PLUS	CROT-100
rotundus ROOT			MINUS
Triticum vulgare	TVUL-25	TVUL-25 PLUS	TVUL-100
SEED			MINUS
Allium cepa	ACEP-76	ACEP-76 PLUS	ACEP-76 MINUS
BULB
Allium sativum	ASAT-100	ASAT-100 PLUS	ASAT-100
BULB			MINUS
Asperagus	ASRA-50	ASRA-50 PLUS	ASRA-50
racemosus -			MINUS
ROOT
Curcuma amada	CAMA-25	CAMA-25 PLUS	CAMA-100
RHIZOME			MINUS
Curcuma longa	CLNG-I-34	CLNG-I-34	CLNG-I-34
		PLUS
RHIZOME	CLNG-II-28	CLNG-II-28	MINUS
		PLUS	CLNG-II-28
			MINUS
Zingiber officinale	ZOFF-50	ZOFF-50 PLUS	ZOFF-100
RHIZOME			MINUS
Momordica	MOCH-35	MOCH-35 PLUS	MOCH-35
charantia			MINUS
FRUIT
Holarrhena	HA-35	HA-35 PLUS	HA-100 MINUS
antidysenterica
SEED
Rauwolfia	RSER-75	RSER-75 PLUS	RSER-75 MINUS
serpentina
ROOT
Vinca rosea	VR-37	VR-37 PLUS	VR-37 MINUS
ROOT
Gymnema	GYSY-26.4	GYSY-25 PLUS	GYSY-25
sylvestree			MINUS
LEAF
Swertia chirata	SCHIR-35	SCHIR-35 PLUS	SCHIR-100
TOTAL PLANT			MINUS
Phyllanthus	PE-100	PE-100 PLUS	PE-100 MINUS
emblica FRUIT
Linum	LUST-35	LUST-35 PLUS	LUST-100
usitatisimum			MINUS
SEEDS
Azadiricta indica	AZIN-C-35	AZIN-C-35	AZIN-C-100
		PLUS
CAKE	N-50	N-50 PLUS	MINUS
LEAF			N-50 MINUS
Terminalia	TCHEB-35	TCHEB-35 PLUS	TCHEB-100
chebula FRUIT			MINUS
Terminalia	TBEL-35	TBEL-35-PLUS	TBEL-100
bellerica	TBEL(R)-35	TBEL(R)-35	MINUS
T. bellerica RIND	TBEL(P)-35	PLUS	TBEL(R)-35
T. bellerica PULP		TBEL(P)-35	MINUS
		PLUS	TBEL(P)-35
			MINUS
Eugenia	EJAM-28	EJAM-28 PLUS	EJAM-28
jambolana			MINUS
SEED
Piper longum	PILO-29	PILO-29 PLUS	PILO-29 MINUS
FRUIT
Piper nigrum	PINI-32	PINI-32 PLUS	PINI-32 MINUS
FRUIT
Embelia ribes	ER-35	ER-35 PLUS	ET-100 MINUS
FRUIT
Tinospora	TICO-34	TICO-34 PLUS	TICO-34 MINUS
cordifolia
STEM
Glycine max	GLMX-54	GLMX-54 PLUS	GLMX-54
SEED			MINUS
Glycyrrhiza	GGLAB-35	GGLAB-35	GGLAB-35
glabra ROOT		PLUS	MINUS
Mucuna pruriens	MPRU-35	MPRU-35 PLUS	MPRU-100
SEED			MINUS
Phaseolus	PRAD-28	PRAD-28 PLUS	PRAD-29
radiatus			MINUS
SEED
Pongamia glabra	PGL-C-35	PGL-C-35 PLUS	PGL-C-100
CAKE			MINUS
Trigonella	TRIGF-35	TRIGF-35 PLUS	TRIG-35 MINUS
foecum-graecum
SEED
Santalum alba	SAAL-65	SAAL-65 PLUS	SAAL-65
HEARTWOOD			MINUS
Ocimum sanctum	OSAT-36	OSAT-36 PLUS	OSAT-36
TOTAL PLANT			MINUS
Sesamum	SIND-35	SIND-35 PLUS	SIND-100
indicaum SEEDS			MINUS
Bacopa monnieri	BAMO-37	BAMO-37 PLUS	BAMO-37
LEAF			MINUS
Withania somifera	WISO-43	WISO-43 PLUS	WISO-43
STEM			MINUS
Carum copticum	CACO-42	CACO-42 PLUS	CACO-42
SEED			MINUS
Cuminum	CUCY-29	CUCY-29 PLUS	CUCY-29
cyminum			MINUS
SEED
Ficus bengalensis	FBENG-35	FBENG-35 PLUS	FBENG-100
BARK			MINUS
Baker's Yeast -	YBD-45	YBD-45 PLUS	YBD-45 MINUS
granulated
Button Mushroom	MUSH-A-67	MUSH-A-57	MUSH-A-
Agaricus bisporus		PLUS	57 MINUS
Prawn - ground -	PRWN-54	PRWN-54 PLUS	PRWN-54
whole			MINUS

[0304] The following examples illustrate compositions of plant extracts from CAM plants for a variety of applications; however, they are not meant to be limiting. The same principles and methods can be used to prepare useful compositions from any Angiosperm-monocotyledon, Angiosperm-dicotyledon and Gymnosperm plant and from any non-plant biomass. It will be apparent to one of skill in the art how to modify the various preparations for specific applications.

[0305] Extracts made by the general procedure described in Examples 1-7 were further diluted with sesame oil to make extracts of different strengths. For example, extract made by mixing one part of R-100 with 9 parts of sesame oil was named R-10. Similarly, one part of R-100 with 19 parts of sesame oil, and one part of R-100 with 49 parts of sesame oil were designated R-5 and R-2, respectively. Thus, one can make diluted oil extracts of any desired strength such as R-1, R-2, R-5, R-10, etc. as convenient and stable dosage forms.

[0306] R-5 can be used effectively on simple burns and open wounds. However, applications in more sensitive skin application require a much lower concentration such as R-1 or one may have to go even lower and use skin lotions as described in subsequent examples. R-5 to R-1 range can also be conveniently given in the form of one or more drops as such or in drinking water.

[0307] In veterinary applications, R-10 to R-1 constitute more convenient dosage forms for addition to feed or drinking water.

[0308] In crop health applications, too, R-5 to R-1 can be conveniently added to the root zone or added to the water for spraying purposes.

[0309] Extracts from other plants and from J or LR fractions were also diluted to different strengths and designated by the degree of dilution and the fraction used, such as A-5, A-5 PLUS, A-5 MINUS, C-2, C-2 PLUS, C-2 MINUS.

[0310] R-100 extract made by the general procedure described in Example 1 is further diluted by mixing it with carriers such as sucrose, lactose, or other sugars. Alternatively, R-100 can be absorbed on porous supports such as precipitated calcium carbonate, talc, precipitated silica, etc. Powders made by mixing one part of R-10 with 9 parts of solids were named R-10(P). Similarly, one part of R-100 with 19 parts of solids and one part of R-100 with 49 parts of solids are named R-5(P) and R-2(P), respectively. Thus, one can make these diluted powders of any desired strength such as R-1(P), R-2(P), R-5(P), R-10(P), etc. as convenient and stable dosage forms.

[0311] R-1(P) and lower strengths up to R-0.1(P) can be used effectively in talcum powder formulations, dental preparations or other powder formulations for dusting applications on skin.

[0312] For internal veterinary applications, the quantities required are so small that R-10(P) to R-1(P) dilutions can be conveniently used directly for feed mixing in the conventional mixing equipment.

[0313] In crop health applications, too, R-5(P) to R-1(P) can be conveniently added to the root zone or added to the water for spraying purposes.

[0314] Tablets of different strength can be made as convenient and stable dosage forms for a variety of applications. A few typical recipes for tablet making are given in the examples below. However, a variety of other excipients may also be used, with or without other adjuvants, for tablet making.

[0315] Tablets of any desired strength of mg of R-100 can be made. In the following examples, they were named accordingly as T-1, T-2, T-5, T-10, etc. Tablet making also allows convenient dose metering. Thus, for internal human consumption, T-1 represents one of the convenient forms as the typical human dose is one T-1 per day for an adult.

[0316] In veterinary applications also, for the same reason, T-10 to T-1 constitute a more convenient dosage form for addition to feed.

[0317] In crop health applications, one T-5 in 5 liters water is a very convenient spray dose per 100 m² field area. Also, one T-5 at the root zone is the typical dose for a new tree sapling. Larger trees need root zone application dose in multiples of T-5.

[0318] T-1 and multiples can be conveniently used at the root zone for small potted plants and one T-1 per liter is a useful spray solution.

[0319] 150 mg Tablets

[0320] 100 g of R-100 was mixed with 900 g sucrose and homogenized in a pestle and mortar to make R-10(P). R-10(P) was then mixed with other components, processing aids and binding agents in the proportion of 1 kg R-10(P), 0.3 kg gum Arabic, 0.3 kg gelatine, 1.15 kg magnesium stearate, 0.3 kg talc, and 11.95 kg sucrose (for a total of 15 kg) and made into tablets weighing 150 mg in a tablet making machine.

[0321] 250 mg Tablets

[0322] 100 g of R-100 is mixed with 900 g sucrose and homogenized in a pestle and mortar to make R-10(P). R-10(P) was then mixed with other components, processing aids and binding agents in the proportion of 1.0 kg R-10(P), 0.1 kg gum Arabic, 0.1 kg gelatine, 0.35 kg magnesium stearate, 0.1 kg talc, and 3.35 kg potassium chloride (for a total of 5.0 kg) and made into tablets weighing 250 mg in a standard tablet making machine.

[0323] The notations to describe the strength of oil, powder and tablet in terms of leaf equivalent are also applicable to creams.

[0324] All Purpose Cream

[0325] Two mixtures were prepared. Mixture A consisted of 3% stearic acid, 40% mineral oil (70 viscosity), 7% lanolin, 10% petrolatum (USP), 2% cetyl alcohol, 2% microcrystalline wax and 0.10 R-100. Mixture B consisted of 5% MgAl silicate (as a 5% dispersion), 1.78% triethanoloamine, and 29.22% water.

[0326] Mixtures A and B were heated separately to 70 C. Mixture B was then added to mixture A and stirred continuously. Then, the mixtures was cooled to 35-40 C. A negligeable amount of fragrance (such as lavendar) and preservatives (such as parabens) was then added, and the mixture mixed until dispersion was complete. Evaporation loss was replaced with water.

[0327] Hand and Body Lotion (Oil in Water Emulsion)

[0328] Two mixtures were prepared. Mixture A consisted of 2.5% stearic acid, 2% mineral oil (70 viscosity), 1% glyceryl monostearate, 2% isopropyl palmitate, 1% petrolatum (USP), 1% cetyl alcohol, 0.25% PEG 40 stearate wax and 0.10% R-100. Mixture B consisted of 7% Carbomer 934 (as a 2% dispersion), 5% glycerine, 1% triethanolamine (as 99% solution), and 77% deionized water.

[0329] Mixtures A and B were heated separately to 70 C. Mixture B was then added to mixture A and then agitated. Then, the mixtures were mixed to 35 C. A negligible amount of lavender and parabens were added for fragrance and stability, respectively, and the mixture mixed until dispersion was complete.

[0330] Methods

[0331] Oil, powder, or tablets made as described in the preceding examples were used in all cases for internal human administration. Oils, creams or lotions were made as described in Examples 8-11 and were used in all topical applications. These formulations were based on oil extracts made according to Examples 1-7 and 1A-4A. Multi-herb combinations were made by: 1) mixing the oil extracts made as per the preceding examples and 2) further adding medium chain fatty acid tryglycerides and sesame oil as per the final stength desired. The examples given below illustrate the use of individual extract compositions as also the use of multi-herb compositions.

[0332] Results

[0333] In all of these reported cases in Table 6, the dose administered internally was 1 to 2 drops of R-5 oil or between 1 to 4 tablets per day each containing 1 mg of leaf equivalent per day, i.e. 1 to 4 mg of R-100 per day, and in a vast majority of cases, 1 tablet per day or 1 drop of R-5 oil per day. The topical application (1 to 2 times a day) using oil or body lotion was also less than or equal to 5 mg of R-100 or leaf equivalent per day.

TABLE 6
Results of human applications of CAM plant oil
extracts
Indication	% Positive Respondents	Number
Respiratory disorders (prevent/cure)		78
Cough/cold/congestion	70	66
Asthma: Allergic or Stress	80	12
Induced
Stress/energy level	90	26
Sound sleep		3
Lowering of stress/tension		12
Higher energy level in elderly		9
Improved circulation in feet		2
Digestive system	80	13
Reduced acidity		4
Reduced stomach upsets		7
Improved appetite		2
Healthy growth	80	28
Gain in height/weight		28
Blood lipid levels	80	10
Lowering of LDL cholesterol		8
Lowering of triglycerides		8
Healing response	90	52
General bruises/cuts		4
Diabetic ulcers		13
Leprotic ulcers		7
Varicose ulcers		1
Bedsores		6
Burns		12
Piles		2
Fistula		1
Anti-inflammatory		35
General pain/swelling	90	12
Spondulitis	50	6
Arthritis	50	8
Gingivitis	80	5
Toothache	80	4
Reproductive system		11
Oligospermia/sperm motility		3
Ovulation		2
Menstual discomfort/pain		4
Menorrhagia		2
Skin/hair		26
(inflammation/repair/recovery)
Pimples		4
Sunburn	100	2
Lichenplanus		1
Eczema/dermatitis		5
Psoriasis		6
Hair loss prevention	100	5
Hair vigorous growth	100	3
Eyes		5
Vision recovery post-macular surgery		3
Dry cornea		1
Sty		1

[0334] As an example, none of the persons whose LDL and triglyceride levels have responded to the use of these compositions made any special changes in their diet or their lifestyle during the trial period (Table 6). At a dose level of 2 to 4 T-1 tablets/day, or at a dose level of 2 to 4 mg leaf equivalent/day, levels of serum LDL and/or serum triglyceride were reduced considerably within a few months (Table 7).

TABLE 7
Case Summary: Lipid-lowering effect of the
compositions of the invention
CASE SUMMARY - LIPID PROFILE
		mg/dl	mg/dl	mg/dl	mg/dl	T-1/day
CASE NO./Dates	Sex/Age(yrs.)	Cholesterol	HDL	LDL	Trigly	Dose/Remarks
1.
26/05/93	Male(50)	170	58	96	80	Base Data
09/12/93		168	55	98	77	Base Data
16/11/94		138	62	67	49	2 taken for two
						months bef. test
2.
10/07/95	Male(40)	219	44		179	2 dose start
23/09/95		177	40	125	123	2
27/02/96		185	43	116	127	1-2
10/08/96		185	41	118	127
3.
23/09/95	Fem(12)	204	48	138	90	1 dose start
19/06/97		151	46	90	78	Taken a total
						150 tabs on/off
4.
09/10/95	Male(45)	268	37	176	273	1 dose start
30/1/96		247	38	151	247
5.
21/09/96	Male(60)	230	47	120	313	1 started from
26/02/97		225	41	130	269	4th may 1996
26/03/98		228	53		194	Intermittently
6.
27/08/93	Male(40)	258	53	151	270	1 dose started
01/03/97		245	40	169	180	taken a total 150
						tabs from 5/5/96
7..
15/11/97	Male(35)	240				4 dose started
06/02/98		148				from 8/12/97
8.
06/10/98	Fem(50)	220	60	137	115	2 dose started
18/12/98		170	60	89	105
9.
06/10/98	Fem(24)	245	65	166	70	2 dose started
18/12/98		195	70	115	50
10.
06/11/98	Male(80)	310
01/03/99		174				2 Taken for two
						months before test

[0335] Notes on Table 6:

[0336] Stress Resistance/Energy Level

[0337] As reported in Table 6, a number of persons experienced effects such as reduction in stress-induced asthma, increase in sound sleep, etc. These effects were obtained by a daily intake of one T-1 tablet for 2 to 4 weeks. A number of elderly persons (over 70 years) found an enhanced sense of well-being, higher energy levels, a general reduction in stomach upsets and a reduction in seasonal coughs and colds by daily intake of one T-1 tablet.

[0338] Healthy GrowthHeight/Weight Gain in Children

[0339] As reported in Table 6, several children who were otherwise had a lack of appetite, routine headaches, low hemoglobin, fatigue, etc, responded positively to the intake of one T-1 tablet per day and started registering healthy height and weight gain with alleviation of these other symptoms. Significant improvement was noted after treatment for more than one month.

[0340] Healing Response

[0341] Diabetic, leprotic, varicose ulcers; bedsores and burns were treated successfully with the daily application of one to four drops of R-5 oil to the ulcer (1 to 6 mg of leaf equivalent). In case of deep leprotic ulcers, the whiteness near the top of the wound changes to a healthy pink color by topical application of R-5 oil, indicating local promotion of angiogenesis. Faster growth of a tougher collagen layer in healing of diabetic ulcer was also observed. Infected diabetic wounds were cleared by topical application.

[0342] Skin Inflammation and Repair

[0343] In cases of eczema and psoriasis, topical application gave relief from the inflammatory process. In case of lichenplanus, the lesions healed readily. Tan caused by sunburn on exposed arms was eliminated by topical application of a 0.1% R-100 body lotion. Inflammation of pimples reduced by facial application of a 0.1% R-100 body lotion. In a few cases, inflammation and wound due to piles were also controlled.

[0344] Hair health

[0345] As reported in Table 6, several persons losing hair on account of ill health or poor hair health, the loss was arrested and vigorous re-growth of hair started by the intake of one T-1 tablet per day. Observable effects were noted within two weeks of starting the intake. Application of hair oil containing 0.1% by weight of R-100 to the scalp produced the same effect.

[0346] Eyes

[0347] In a few cases, rapid vision recovery post-macular surgery by oral intake of T-1 tablets was noted. Recovery from sty infection and dry corneas were achieved by topical application.

[0348] Reproductive System

[0349] A few cases of oligospermia/sperm motility were corrected by oral intake of T-1 tablets for three months.

[0350] The examples below illustrate that the compositions of the present invention produce preventive and curative therapeutic effects in many different human embodiments.

[0351] The cases described below also illustrate how a variety of compositions using combinations of biomass extracts can be used effectively in a variety of human embodiments. In a majority of cases these effects are expressions of effects expected fror the same biomass as described in the available literature. However, there are also some novel therapeutic effects which are not described in the literature. All combination extracts also contain 20 gm of MCT Oil (a 70:30 mixture of capryllic/capric tryglycerides obtained from Subhash Chemical lndustriesd, Pune)/100 gm.

[0352] The examples also illustrate the key aspect of this invention in that these effects are produced at an extremely small dose in terms of mg per day of the starting biomass.

[0353] In all of these reported cases below the dose administered internally was 1 to 10 drops of apprpriate final oil extract composition containing 2-50 mg of total plant equivalent per day. In a majority of cases, 1-2 drop of appropriate oil extract per day was adequate. The topical application (1 to 2 times a day) using oil or body lotion was also less than or equal to 10 mg of plant equivalent per day. The designations of extracts are as defined hereinabove and also as defined herein.

[0354] Digestive System/General Health

[0355] A mixture of AMAVAT-20 oil was prepared. This oil is based on the extract of 20 gms of total herbal material/100 gm of oilextract. The herbal mixture contains equal parts of: 1) an equal part mixture of Zingiber officinale, Carum copticum, Cuminum cyminum, Piper longum, 2) a mixture of Terminalia Bellerica (1 part), Terminalia chebula (3 parts), Phyllanthus emblica (6 parts), Aloe indica (10 parts), Glycyrrhiza glabra (10 parts), 3) an equal part mixture of Trigonella, Linum usitatisum, Phaseolus radiatus, Triticum vulagare and 4)an equal part mixture of Holarrhena antidysenterica, Embelia ribes, Tinospora cordifolia, Swertia chirata. Each drop of this AMAVAT-20 oil contains 8 mg equivalent of all the above herbs combined. One drop of this oil was given twice a day to 6 persons suffering from chronic stomachache and tendancy for loose bowels. All of them reported considerable reduction in stomach discomfort within 3 to 4 days. The tendancy for loose bowels also stopped.

[0356] Another case (female, 60 years age) suffering from recurrent indigestion, diarrhea, urticaria has responded well to two drops per day of AMAVATA-20.

[0357] Menstrual Health

[0358] A 13 year old girl suffering from pain during menstruation, frequent colds and poor concentration in studies was given 2 drops per day of AMAVATA-20 and 1 tab/day of 1 mg equivalent of R-100. Over a period of 2 months, pain during menstuation has reduced considerably as also the frequency of colds. There is also an improvement in concentration in studies.

[0359] Two teenage girls suffering from delayed menstruation (35 to 40 day cycle) were given 2 drops per day of A-50(Aloe). Over a period of 2-3 months (two cycles) the menstruation cycle was restored to normalcy. The toal herbal dose was equivalent to 40 mg per day.

[0360] Arthritis, Backache and Joint Pain

[0361] A person (58 years, male) was suffering from lower back joint inflammatory pain, which was aggravated by vigourous walking. He started taking one drop per day of a composition designated as RCPE-10 PLUS and consisting of a mixture of R-5 PLUS (2.5 parts), C-5 PLUS (2.5 parts), and PE-5 PLUS (5 parts). The total initial plant material per drop is 4 mg. After one week, there was a considerable reduction in the joint inflammation and pain.

[0362] A former jet fighter pilot (58 years old) was suffereing from chronic lower back pain and stiffness, particularly in the morning for several years. He started taking one drop per day of a composition designated ARTH-10. The total starting plant material per drop is 4 mg. ARTH-10 contains equal parts of A-100 PLUS, R-100 PLUS, C-100 PLUS, ZOFF-55.4 PLUS, CAMA-26.2 PLUS and GGLAB-7.4 PLUS. After one week, he has found a considerable relief from his nagging back pain and stiffness.

[0363] Cough/Throat Irritation/Cold

[0364] A male (58 years old) developed a cold and early stage of a cough with throat irritation. One drop each of 1 R-5 PLUS, TABA-5 and GGLAB-5 in a cup of hot water taken three times a day alleviated the symptoms within two days. Thus, the total daily dose was 6 mg of each of three plants.

[0365] In case of some other subjects, a drop of COUGH-5 was given thrice a day at the onset of cough, throat pain and cold. The throat pain and cough disappeared within two days as also the symptoms of cold. COUGH-10 is a mixture of: 2.4 parts of: a mixture of Zingiber officinale, Carum copticum, Cuminum cyminum, Piper longum, 0.6 parts of Glycyrrhiza glabra; and 2 parts of Piper nigrum. The total daily herbal dose in this case was 6 mg.

[0366] In cae of four persons suffering from hinitis, runy nose and headache due to congestion, one drop of OSAT-48 was applied with a cotton bud inside the nostrils. Within 15 minutes, there was a complete drainage of mucous with considerable relief from the symptoms of cold and headache.

[0367] Sciatica

[0368] A 78 year old female was suffering from throbbing pain due to sciatica. A mixture of 1 drop each of MPRU-5 and PE-5 along with 1 drop of Medium Chain Fatty Acid Tryglycerides was applied externally to the sore spot once a day. The throbbing pain completely stopped within 4 days. The total daily dose in this case was 2 mg each of Mucuna pruriens and Phyllanthus emblica.

[0369] An 80 year old male suffering from sciatica in both the legs was given a mixture of SHUKRA-5 (a mixture of Kalanchoe pinnata, Withania somnifera, Asperagas racemosus and Mucuna pruriens) and R-100 PLUS in the ratio of 3:2. Application of one drop per day to the tender spot on each leg and 1 drop internally has resulted in a considerable relief in the pain.

[0370] Neurology-Muscle Spasm

[0371] A person (60 years, female) has been suffering from hemi-facial spasm for a long time. The spasm starts from above the left eye, travels down the nose and cheeks up to lips. The spasm lasts for several hours and causes face distortion and pain. She started applying one drop of an equal part mixture of PE-5 PLUS and MPRU-5. When the drop is applied, the spasm subsides immediately and does not recur at least for 12 hours. Application of one drop also prevents the initiation of the spasm. The total herbal dose is just 2 mg per day.

[0372] A person (58 yeares, male) has been suffering from a chronic muscle spasm and painful area on the upper part of right foot between the fingers and ankle. Application of 2 drops of MPRU-5 (4 mg of herbal starting material) has eliminated the spasm and pain within two days.

[0373] NeurologyPost-Paralysis Recovery of Muscle Activity and Mental Concentration.

[0374] A person (58 years, male) had a stroke (left frontal MCA infarc) in 1996 with almost complete loss of speech. He was not able to concentrate on reading for more than 15 minutes at a time. Also, his handgrip was poor. Thus, recovery had been very very slow. From 1^st Sep., 2001, the person was given one drop per day each of PE-5 PLUS, MPRU-4.8 and GGLAB-5. The total plant material equivalent is 6 mg per day. By mid-November, 2002 (2.5 months) there was a considerable inprovement in ability to concentrate. He can now read for 2 hours at a time (compared to 15 minutes at a time before treatment) and the retention has improved. There is also a considerable improvement in the hand grip.

[0375] Another case is of a 20 year man. He had suffered a brain hemorrhage and consequently his ability to learn was severely constrained. He started taking NEURO-35 at the rate of two drops per day (an equal mixture of Phylanthus indica, Mucuna pruriens and Bacopa monnieri) equivalent to a total herbal dose of 45 mg per day. After one and a half month, there is now a noticeable improvement in his ability to learn a new language (Russian).

[0376] NeurologyViral Polyneuritis

[0377] A person (male) was unable to lift his foot after an attack of viral polyneuritis. He was given one drop twice a day of NEURO-35 (an equal mixture of Phylanthus indica, Mucuna prurients and Bacopa monnieri) equivalent to a total herbal dose of 28 mg per day. After a few weeks, the ability of the muscles revived and he is able to lift his foot. The circumferance near the ankle also increased by 1 cm indicating a restoration of the wasted muscle.

[0378] NeurologySpeech Centres

[0379] A person suffering from heavy speech slurring for 15 years (triggered because of pesticidal neuritoxicity) was given NEURO-10 at the dose of 1 drop, twice a day equivalent to a total herbal dose of 8 mg per day. His speech clarity has improved within a few weeks.

[0380] In another case, a 50 year old person had serious speech coordination problems as a result of occupational stress particularly if he tried to speak fast. By taking two drops per day of NEURO-35 equivalent to a total herbal daily dose of 28 mg per day, his speech clarity has also improved within 2 weeks.

[0381] Reproductive System

[0382] A 13 year old girl was suffering from dysmenorrhea and did not menstruate for 3 months. She was given A-10 PLUS at the rate of 4 drops per day. This is equivalent to 16 mg/day of total plant material. Within 15 days she began to menstruate and a regular cycle was established.

[0383] PRAWN

[0384] Application of two drops of prawn extract (PRWN-5) alleviated knee pain for 1 hour. A mixture of one drop each of PRWN-5 and R-5 alleviated the pain for several hours.

[0385] MUSHROOM

[0386] A few persons suffering from a lack of appetite were given 1 drop of the button mushroom extract (MUSH-A-10) twice a day. Within four days, there was an improvement in the appetite. The total daily dose in this case was 8 mg per day of mushroom equivalent.

[0387] Agricultural Applications

[0388] Methods

[0389] In the following examples using Kalanchoe pinnata, R-5 or R-2 oil, R-10(P) powder and tablets made as per examples above were used. In the case of extracts of other plants, different potency oils were used as described in particular examples.

[0390] Plants were either grown in hydroponic or soil media.

[0391] Administration was accomplished by a variety of means, including direct application to the root zone, foliar spray, application of a solution at the root zone after dissolving/dispersing tablet/oil in water, injection in to the trunks or stems, application to terminal buds, addition to tissue culture medium, etc.

[0392] In the following examples, the typical dosage of extract for field crops was 0.5 to 1 g of R-100 oil or leaf equivalent per hectare per spray. The number of sprays can be typically at a frequency of once every one to three weeks. The dose for tree crops varied from 5 mg to 50 mg per tree of R-100 per year, depending on the size of the sapling/tree.

[0393] Multiple high dose sprays of particular preparations such as R-100 MINUS (5 to 25 g per spray per hectare) act to reduce the flower set, total seeds produced and the size of seeds. This effect can be used to control the propagation of hardy weeds such as Congresss Grass (Parthium sp.), Lantana sp., Cyperus sp. and others.

[0394] Toxicity

[0395] The Onion Root Tip Assay was used to study genotoxicity profile of CAM plant extracts. The results of this test can be usefully related to the expected cytotological profile in animal cells or human lymphocytes (Meenakumari, 1995; Mercykutty, 1980).

[0396] Bulbs of Allium cepa, L. cv N-2-4-1 (2n16) were used. Selected bulbs were washed and the root systems of the bulbs were kept in 100 ml of aqueous solutions in cavity blocks containing different amounts of herbal oil extract. Roots were treated for 48 hours.

[0397] At the end of 48 hours, the root tips were recovered and fixed in acetic acid-alcohol (1:3). For cytological preparations, root tips were hydrolyzed in 1 N HCl and squashed in 1% acetocarmine. Slides were examined under a microscpoe. Cells were observed and scored (Table 8), and the status of cells with respect to mitosis and various other physiological (clumping, stray and lagging chromosomes) and clastogenic (anaphase, fragments, binucleate) aberrations was recored. The total number of roots and the average length of the roots were also measured and sprouting from the tip of the bulb was also noted.

TABLE 8
Effect of R-100 PLUS vs. R-100 MINUS on Onion Root Tip
Concentration,	No. cell	Mitotic Index	No. cells showing aberrations	Root no./
l/liter	scored	- % dividing -	Physiological	Clastogenic	(length, cm)
0 (CONTROL)	1120	36.25	0	0	42 (4.3)
R-100 PLUS
10	1105	39.12	0	0	50 (4.6)
30	1121	42.58	0	0	52 (4.7)
100	1085	40.12	8	4	44 (3.9)
300	1048	30.23	58	8	42 (3.6)
R-100
10	1012	38.56	0	0	48 (4.5)
30	1025	40.12	12	0	45 (4.1)
100	1045	25.26	87	7	38 (3.8)
300	1005	18.23	126	14	36 (1.6)
R-100 MINUS
10	1052	38.36	8	12	43 (4.2)
30	930	25.25	49	31	38 (3.2)
100	936	8.25	487	45	26 (1.2)
300	856	1.22	671	69	12 (0.9)

[0398] Roots that formed at higher concentrations of R-100 MINUS were short, yellowish and had curved tips. There was excellent sprouting on top of the bulb in case of control and 10 l/liter of R-100 PLUS. Sprouting was moderate with 10 l/liter of R-100 and 30 l/liter of R-100 PLUS. There was no sprouting at all in any of the other sets.

[0399] In the above data, major aberrations were physiological and mainly clumping of chromosomes. However, the onset of aberrations was shown by R-100 MINUS-treated root tips at 10 l/liter whereas R-100 PLUS-treated root tips began to show aberrations at 100 l/liter, a tenfold higher concentration. The mitotic index, and the number and average root length also confirmed this observation.

[0400] R-100 PLUS, at least up to 30 l liter promoted cell division, rooting and sprouting. In contrast, R-100 MINUS began to act as a mitogen and root system inhibitor beyond 10 l/liter.

[0401] Experiments were carried out using A-100 PLUS, A-100 MINUS, C-100 PLUS, C-100 MINUS, R-100 PLUS, R-100 MINUS and sesame oil at the solution concentrations given in Table 9.

[0402] Bulbs of Allium cepa, L. cv N-2-4-1(2n16) were used. Selected bulbs were washed and the root systems of the bulbs were kept in 100 ml aqueous solution in cavity blocks containing different amount of herbal oil extract. Roots were treated for 12 hours and then recovered in 10% glucose for another 12 hours.

[0403] At the end of recovery, the root tips were recovered and fixed in acetic acid-alcohol (1:3). For cytological preparations, root tips were hydrolyzed in 1 N HCl and squashed in 1% acetocarmine. Slides were examined under a microscope. Cells were observed and scored (Table 9), and the status of cells with respect to mitosis and various other physiological (clumping, stray and lagging chromosomes) and clastogenic (anaphase, fragments, binucleate) aberrations was recorded. The results are summarized in Table 9.

TABLE 9
Effect of CAM Plant Fractions on Onion Root Tip
Concentration,	No. cell	Mitotic Index	% cells showing aberrations
l/liter	scored	-% dividing-	Physiological	Clastogenic
Sesame oil
0	1552	30.12	0	0
100	1230	31.23	0.08	0
300	1452	32.59	0.34	0
R-100 PLUS
30	1530	32.24	0	0
100	1547	33.56	0.52	0
300	1531	18.63	1.31	0.2
R-100 MINUS
30	1498	28.27	0.13	0
100	1521	14.45	1.78	0.26
300	1530	9.63	3.01	0.92
A-100 PLUS
30	1521	32.21	0	0
100	1511	33.52	0	0
300	1505	31.89	0.8	0
A-100 MINUS
30	1563	31.25	0	0
100	1524	32.68	0.2	0
300	1541	28.30	0.97	0
C-100 PLUS
30	1621	32.33	0.06	0
100	1563	26.23	1.09	0
300	1518	11.14	1.98	0.4
C-100 MINUS
30	1546	31.28	0.45	0
100	1543	12.65	1.94	0.32
300	1532	8.25	3.26	0.91

[0404] In all cases, the PLUS fraction (LR) appeared to be an excellent promoter of mitosis or cell proliferation compared to sesame oil controls. In some cases 100 PLUS and A-100 PLUS), this activity was retained up to 100 l/liter concentration in this assay. The PLUS fractions also had lower toxicity than the corresponding MINUS (J) fractions in terms of mitogenic activity inhibition and genotoxicity.

[0405] The use of the PLUS fraction and exclusion of the MINUS fraction for medicated oil preparations is contradicts the teachings of traditional medicine. Surprisingly, the method of the invention improves the overall potential of health promotion and broadens the safe operating range. This method also allows compositions of high potency at low dose; thus further reducing the toxicity potential.

[0406] These positive effects, particularly in case of Kalanchoe and Cissus, have greatly extended their safe operating range, considerably improved their efficacy/toxicity ratio and therefore extended their utility in applications that hitherto were restricted due to the toxicity of the use of juice with a high dosage.

[0407] The higher toxicity of the juice-based extract can be used in applications to eliminate unwanted vegetation or control plant growth.

[0408] Experiments were carried out using TABA-28, CROT-62.66 MINUS, TVUL-32 PLUS, SCHIR-55 MINUS, HA-29.7, PE-100 PLUS, PE-100 MINUS, and sesame oil at the solution concentrations given in Table 9 A. R-100 PLUS was also used as comparison.

[0409] Bulbs of Allium cepa, L. cv N-2-4-1(2n16) were used. Selected bulbs were washed and the root systems of the bulbs were kept in 100 ml aqueous solution in cavity blocks containing different amount of herbal oil extract. Roots were treated for 48 hours.

[0410] At the end of 48 hours, the root tips were recovered and fixed in acetic acid-alcohol (1:3). For cytological preparations, root tips were hydrolyzed in 1 N HCl and squashed in 1% acetocarmine. Slides were examined under a microscope. Cells were observed and scored (Table 9 A ), and the status of cells with respect to mitosis and various other physiological (clumping, stray and lagging chromosomes) and clastogenic (anaphase, fragments, binucleate) aberrations was recorded. The results are summarized in Table 9A.

TABLE 9A
Effect of Non-CAM Plant Fractions on Onion Root Tip
		Number
		of	Mitotic	Physiological	Clstogenic	Total
	Concentration	Cells	Index, %	Aberration,	Aberration,	Aberration,
Extract	l/liter	Scored	Dividing	% Cells	% Cells	% Cells
Control	0	1236	23.14	0.0	0.0	0.0
Sesame	3	1235	24.17	0.0	0.0
Oil	10	1230	23.17	0.16	0.0	0.16
	30	1452	21.49	0.34	0.0	0.34
R-100	3	1250	26.00	0.08	0.0	0.08
PLUS	10	1230	23.50	1.06	0.0	1.06
	30	1520	16.84	1.84	0.26	2.11
TABA-	3	1698	22.50	0.59	0.0	0.59
28	10	1615	20.12	1.67	0.31	1.98
	30	1602	17.85	2.56	0.69	3.25
CROT-	3	1629	23.51	0.74	0.0	0.74
62.66	10	1622	16.09	1.97	0.31	2.28
MINUS	30	1628	11.36	3.28	0.92	4.30
TVUL-	3	1521	25.97	0.13	0.0	0.13
32	10	1566	24.39	0.38	0.0	0.38
PLUS	30	1590	22.39	1.45	0.0	1.45
SCHIR-	3	1611	22.66	0.62	0.06	0.68
55	10	1598	20.03	1.94	0.38	2.32
MINUS	30	1596	12.84	3.13	0.69	3.82
HA-	3	1562	24.71	0.26	0.0	0.26
29.7	10	1624	19.77	1.35	0.12	1.47
	30	1615	14.98	2.85	0.74	3.59
PE-	3	1595	24.20	0.69	0.0	0.69
100	10	1622	17.82	2.10	0.43	2.53
PLUS	30	1659	12.18	3.01	0.66	3.68
PE-		1652	21.55	0.85	0.0	0.85
100		1623	17.51	2.45	0.67	3.12
MINUS		1623	11.58	3.94	0.99	4.93

[0411] Fractions from Kalanchoe (R-100 PLUS), wheat (TVUL-32 PLUS), Holarrhena antidysenterica (HA-29.7) and Phyllanthus emblica (PE-100 PLUS) show mitogenic index increase or cell division promotion at/upto 3 l/liter of the oil extract. At higher concentration, these and other extracts all show a decline in mitigentic index or inhibition of cell division. However, in all these cases, the % of clastogenic aberrations is rather low. Thus, these extracts, at higher concentrations can be used for contolling cell proliferation without causing significant mutagenic effects.

[0412] Applications to Dicotyledonous Plants

[0413] Germination Promotion

[0414] Using R-10 (P) (batch 881128) in Phaseolus radiatus, a legume, the seeds were soaked in a solution of R-10 (P) and observations were taken at 24, 48 and 120 hours after soaking. The results are summarized in Table 10.

TABLE 10
Germination promotion
	Concentration,
	ppm,	ROOT LENGTH (cm), RANGE
	R-10(P)	24 Hrs	48 Hrs	120 Hrs
	0(Control)	0.2-1.0	3.5-6.0	4.2-8.6
	1	0.2-2.0	4.0-6.2	3.5-12.5
	5	0.2-2.5	4.0-6.8	5.7-12.1
	20	2.0-2.8	4.5-7.1	13.9-17.8
	500	0.5-2.5	0.5-4.7	3.6-11.2

[0415] Thus, germination promotion is observed with increasing concentration up to 20 ppm of R-10(P). At 20 ppm of R-10(P) or 2 ppm of R-100 equivalent, there is a particularly strong promotion of germination.

[0416] Germination experiments were carried out with R-100 PLUS vs. R-100 MINUS and also with the standard R-100 extract. Twenty-five (25) seeds of Phaeolus mungo were placed in a plate with 5 ml of distilled water containing various concentrations of the R-100 extracts. On the b 7^th day after initiation of experiment, mean values of 11 seedlings were taken and reported. The results are summarized below in Table 11.

TABLE 11
Germination in Phaeolus mungo; comparison of
different forms of extract
			R-100
Concentration	R-100 PLUS	R-100	MINUS	Sesame oil
Root Length (cm)
.0	4.62(0.26)	4.62(0.26)	4.62(0.26)	4.62(0.26)
0.33	5.22(0.08)	5.18(0.40)	4.76(0.11)	4.52(0.29)
1.0	5.38(0.08)	5.30((0.10)	4.86(0.39)	4.92(0.18)
3.0	5.62(0.08)	5.56(0.11)	4.94(0.11)	4.96(0.19)
30.0	5.30(0.07)	5.12(0.09)	4.68(0.11)	5.28(0.22)
Shoot Length (cm)
0.0	9.08(0.29)	9.08(0.29)	9.08(0.29)	9.08(0.29)
0.33	11.48(0.24)	11.48(0.53)	13.64(0.38)	10.56(0.23)
1.0	12.44(0.23)	13.50(0.07)	15.02(0.22)	10.96(0.18)
3.0	13.70(0.37)	13.56(0.23)	15.70(0.20)	11.84(0.17)
30.0	13.46(0.15)	11.4(0.16)	15.12(0.49)	12.54(0.11)
Dry weight, g (10 seedlings) - mean of two observations
0.0	0.33	0.33	0.33	0.33
0.33	0.35	0.34	0.32	0.33
1.0	0.42	0.41	0.34	0.33
3.	0.40	0.38	0.31	0.34
30	0.35	0.32	0.27	0.35
Note:
Concentration in the medium is in l/50 ml distilled water.
Values in parenthesis are standard deviation.

[0417] All fractions showed significant biological activity at very low doses. Both R-100 and R-100 PLUS show significant promotion of root and shoot growth and biomass weight at the end of 7 days compared to sesame oil controls at 1 and 3 mg levels. The PLUS fraction showed the best overall promotional effect.

[0418] R-100 MINUS did show shoot growth compared to sesame oil control. However, there was no root growth and no increase in dry biomass weight up to 3 mg. At the higher dose level, R-100 also showed a sharper drop in root length and biomass retention.

[0419] Thus, this data corroborates the contrasting behavior of LR and the J fractions-based compositions from Kalanchoe described in Example 14 above.

[0420] Twenty-five (25) seeds of Phaseolus mungo were placed in a plate with 5 ml of distilled water, containing various concentrations of oil extracts or plain base oil. On the 7^th day after initiation of experiment, mean values of 10 seedlings were taken and reported in Table 12.

TABLE 12
Effect of CAM plant fractions on seed germination activity
Sesame Oil	A-100	A-100	C-100	C-100
Concentration Plain	PLUS	MINUS	PLUS	MINUS
Root Length(cm)
0.0	5.09(0.065)	5.12(0.065)	5.12(0.065)	5.12(0.065)	5.12(0.065)
1.0	5.20(0.048)	6.33(0.068)	6.12(0.087)	6.75(0.055)	5.42(0.063)
3.0	5.30(0.052)	6.51(0.052)	6.39(0.047)	4.33(0.065)	5.23(0.071)
10.0	5.67(0.061)	6.69(0.061)	6.41(0.045)	4.67(0.062)	4.51(0.042)
Shoot Length(cm)
0.0	10.35(0.058)	10.35(0.058)	10.35(0.058)	10.35(0.058)	10.35(0.058)
1.0	12.08(0.062)	12.12(0.054)	11.86(0.062)	13.35(0.064)	13.62(0.048)
3.0	12.98(0.047)	13.12(0.062)	13.02(0.068)	12.33(0.56)	12.65(0.052)
10.0	13.56(0.054)	13.56(0.057)	13.12(0.077)	11.38(0.054)	12.17(0.051)
Dry Weight(gm/10 seedlings) mean of two observations. Numbers
in parenthesis indicate range of the two observations.
0.0	0.332(0.0023)	0.332(0.0023)	0.332(0.0023)	0.332(0.0023)	0.332(0.0023
1.0	0.351(0.0023)	0.371(0.0022)	0.384(0.0022)	0.352(0.0022)	0.364(0.0018)
3.0	0.363(0.0018)	0.408(0.0023)	0.397(0.0021)	0.382(0.0023)	0.377(0.0019)
10.0	0.362(0.0019)	0.427(0.0019)	0.415(0.0023)	0.336(0.0021)	0.322(0.0023)
Note:
Concentration in the medium is in l of oil/50 ml distilled water.
Values in parenthesis indicate standard deviation.

[0421] PLUS and MINUS fractions of both CAM plants promoted auxin-like (rooting promotion), gibberellin-like (shooting promotion) and cytokine-like (biomass preservation/growth) activity up to 1 l/50 ml DW. A-100 PLUS and MINUS both promoted rooting and a higher dry biomass at the end of 7 days, even at the higher concentration of 10 l/50ml distilled water. Thus, A-100 PLUS and MINUS promote a wide range of endogenous hormones at a low concentration and this promotional effects continue up to a high concentration. However, C-100 PLUS and MINUS had a different activity profile. They showed auxin, gibberellin and cytokine-like activities at 1 l/50 ml DW as well. However, at higher concentrations (10 l/50 ml distilled water), a reversal in all activities was observed. Thus, this process of inhibition of activity started earlier with C-100 than the other extracts.

[0422] Agar (0.8 wt. % agar in distilled water) was digested in water bath to get transperant medium and 100 ml of this solution (base medium) was poured in 300 ml culture bottles. Different doses of oil extracts of various Angiosperm-monocotyledon, Angiosperm-dicotyledon and Gymnosperm plants, non-plant biomass or plain base sesame oil were added to each bottle. Bottles were sterilized at 15 lbs for 20 mnutes in an autoclave. Surface sterilized 10 seeds of Phaseolus radiatus were added to each bottle amd germinated in dark for 5 days. On the 5^th day after initiation of experiment, values of shoot lenth, root lenth, and dry weight of biomass (dried in an oven at 70-80 deg. C. for constant dry weight) were measured for 10 seedlings. Mean values are reported in Table 12A-12K, 12M, 12O, 12Q, 12S, 12U, 12W. Several extracts of plants and their fractions and non-plant biomass promoted rooting, shooting and biomass mobilization at low doses.

TABLE 12A
Effect of plant extracts on P. radiatus seed germination activity
Extract/		Sesame	PE 100	PE 100	TBEL	TBEL	TBEL
Bottle		Oil	PLUS	MINUS	R 50	P 50	27.7
0	Shoot	23.5
Control	(cm)	(1.09)
	Root	8.3
	(cm)	(0.74)
	Total	31.3
	(cm)	(1.38)
1	Shoot	24.7	25.8	25.1	24.9	24.6	26.0
	(cm)	(1.03)	(1.04)	(1.01)	(0.70)	(0.83)	(1.04)
	Root	9.4	11.3	12.1	10.8	9.9	9.6
	(cm)	(0.51)	(0.95)	(0.61)	(0.61)	(0.62)	(0.40)
	Total	34.1	37.1	37.2	35.6	34.5	35.6
	(cm)	(0.97)	(1.50)	(0.90)	(1.09)	(0.82)	(1.08)
3	Shoot	25.2	26.3	26.8	25.5	25.3	25.7
	(cm)	(0.99)	(1.02)	(0.86)	(0.71)	(0.69)	(0.80)
	Root	10.8	12.8	12.3	12.82	11.4	11.0
	(cm)	(0.40)	(1.06)	(0.96)	(0.82)	(0.93)	(0.92)
	Total	36.1	39.1	39.1	38.3	36.7	36.6
	(cm)	(1.02)	(1.5)	(1.51)	(1.16)	(0.90)	(1.13)
10	Shoot	25.0	26.8	21.7	24.7	26.0	26.5
	(cm)	(1.01)	(1.36)	(1.05)	(1.21)	(1.27)	(0.92)
	Root	10.0	8.8	12.4	11.4	10.7	12.4
	(cm)	(0.55	(0.74)	(0.76)	(0.70)	(0.83)	(1.32)
	Total	35.1	35.7	34.1	36.1	36.7	38.8
	(cm)	(1.39)	(1.95)	(1.23)	(1.46)	(1.20)	(1.44)
Note:
Extract/Bottle in the medium is in l of oil/100 ml base medium.
Values in parenthesis are standard deviation of the estimate.

[0423] Dry biomass weight per 10 seedlings at the end of 5 days is reported in Table 12B below.

TABLE 12B
Effect of plant extracts on P. radiatus germination activity
Extract/		Sesame	PE 100	PE 100	TBEL	TBEL	TBEL
Bottle		Oil	PLUS	MINUS	R 50	P 50	27.7
0	E.A.	230
	(mg)
1	E.A.	228	294	302	220	240	240
	(mg)
3	E.A.	226	268	306	246	210	232
	(mg)
10	E.A.	224	240	238	260	270	266
	(mg)
Note:
Extract/Bottle in the medium is in l of oil/100 ml base medium.
E.A.Embryo Axis (Root Shoot)

[0424] Phyllanthus emblica (PE 100 PLUS and MINUS) fractions show strong promotion, particularly of rooting and biomass growth (E.A. value) up to 3 l of oil/100 ml base medium. Rooting promotion is shown by the TBEL R 50 fraction (Terminalia bellerica Rind) at 3 l of oil/100 ml base medium. Both the Rind (R) and Pulp (P) fraction of TBEL also show biomass growth at the higher concentration.

TABLE 12C
Effect of plant extracts on P. radiatus seed germination activity
			FBENG	FBENG		CAMA	ZOFF
Extract/		Sesame	29	100	TCHEB	98.4	100
Bottle		Oil	PLUS	MINUS	27.8	MINUS	MINUS
0	Shoot	22.8
Control	(cm)	(0.92)
	Root	8.1
	(cm)	(0.71)
	Total	30.9
	(cm)	(0.95)
1	Shoot	23.7	23.8	24.2	23.7	24.0	23.8
	(cm)	(1.11)	(1.1)	(0.62)	(1.17)	(0.90)	(0.87)
	Root	8.3	9.6	9.3	8.9	8.3	8.6
	(cm)	(0.82)	(0.84)	(0.61)	(0.46)	(0.64)	(0.72)
	Total	32.1	33.4	33.5	32.7	32.2	32.4
	(cm)	(1.38)	(1.66)	(0.90)	(1.32)	(0.88)	(1.06)
3	Shoot	24.2	24.0	28.3	23.6	24.5	24.4
	(cm)	(0.98)	(0.91)	(0.84)	(0.99)	(0.74)	(0.69)
	Root	8.6	10.1	9.1	10.0	9.8	8.8
	(cm)	(1.12)	(0.83)	(0.72)	(0.79)	(0.34)	(0.69)
	Total	32.8	34.1	37.3	33.5	34.3	33.2
	(cm)	(1.68)	(0.96)	(1.28)	(1.21)	(0.64)	(0.97)
10	Shoot	24.0	23.8	25.1	23.4	25.8	24.0
	(cm)	(1.22)	(1.06)	(0.83)	(0.72)	(0.82)	(1.21)
	Root	8.9	8.1	9.4	8.3	7.9	8.5
	(cm)	(0.92)	(0.64)	(0.76)	(0.82)	(1.07)	(0.53)
	Total	32.9	32.0	34.5	31.7	33.7	32.5
	(cm)	(1.65)	(1.31)	(1.16)	(1.17)	(0.86)	(1.32)
Note:
Extract/Bottle in the medium is in l of oil/100 ml base medium.
Values in parenthesis indicate standard deviation.

[0425] Dry biomass weight/10 seedlings at the end of 5 days is reported in Table 12D below.

TABLE 12D
Effect of plant extracts on P. radiatus germination activity
			FEBG	FBENG		CAMA	ZPFF
Extract/		Sesame	29	100	TCHEB	98.4	100
Bottle		Oil	PLUS	MINUS	27.8	MINUS	MINUS
0	E.A.	322
	(mg)
1	E.A.	318	314	288	296	292	298
	(mg)
3	E.A.	256	300	320	270	308	278
	(mg)
10	E.A.	290	288	312	294	322	284
	(mg)
Note:
Extract/Bottle in the medium is in l of oil/50 ml base medium.
E.A.Embryo Axis (Root Shoot)

[0426] Ficus bengalensis fractions (FBENG 29 PLUS and FBENG 100 MINUS show promotion of growth (rootshoot) upto 3 l of oil/100 ml base medium. Moderate rooting promotion is shown by Terminalia chebula (TCHEB 27.8) up to 3 l of oil/100 ml base medium. However, promotion is not seen in biomass growth on a consistent basis in these cases.

TABLE 12E
Effect of plant extracts on P. radiatus seed germination activity
			SCHIR	SCHIR	ER	ER
Extract/		Sesame	27.8	55	28.8	100	TABA	MPRU	HA
Bottle		Oil	PLUS	MINUS	PLUS	MINUS	28	27.1	29.7
0	Shoot	24.0
Control	(cm)	(0.97)
	Root	8.0
	(cm)	(0.81)
	Total	32.0
	(cm)	(1.44)
1	Shoot		24.2	25.0	23.4	26.5	28.3	26.6	22.8
	(cm)		(1.19)	(0.98)	(1.12)	(1.03)	(0.81)	(1.09)	(0.78)
	Root		12.1	11.9	10.3	10.0	10.4	9.3	10.3
	(cm)		(1.34)	(1.03)	(1.0)	(0.67)	(0.55)	(0.59)	(0.65)
	Total		36.3	36.9	33.8	36.6	38.7	35.9	33.1
	(cm)		(2.04)	(1.32)	(0.99)	(1.31)	(1.16)	(1.37)	(0.84)
3	Shoot	24.2	25.1	26.5	23.3	27.1	28.5	27.9	29.1
	(cm)	(0.98)	(0.80)	(0.97)	(1.04)	(0.96)	(1.01)	(0.51)	(0.87)
	Root	10.5	12.7	10.3	11.3	11.2	11.9	11.5	12.1
	(cm)	(0.87)	(0.70)	(1.01)	(0.93)	(0.86)	(0.74)	(0.63)	(0.85)
	Total	34.7	37.8	36.8	34.7	38.3	40.3	39.3	41.3
	(cm)	(1.23)	(1.08)	(0.99)	(0.87)	(1.59)	(0.90)	(0.76)	(1.49)
10	Shoot		28.5	24.6	28.7	23.9	20.3	24.4	26.2
	(cm)		(1.20)	(1.18)	(1.11)	(1.05)	(0.91)	(0.77)	(1.05)
	Root		9.2	7.4	11.0	10.8	5.0	8.2	9.7
	(cm)		(0.73)	(.79)	(1.05)	(0.82)	(0.54)	(0.72)	(0.69)
	Total		37.7	32.0	29.7	34.7	25.3	32.6	35.9
	(cm)		(1.61)	(1.35)	(1.95)	(1.33)	(1.15)	(1.09)	(1.27)
Note:
Extract/Bottle in the medium is in l of oil/100 ml base medium.
Values in parenthesis indicate standard deviation.

[0427] Dry biomass weight/10 seedlings at the end of 5 days is reported in Table 12F below.

TABLE 12F
Effect of plant extracts on P. radiatus germination activity
			SCHIR	SCHIR	ER	ER
Extract/		Sesame	27.8	55	28.8	100	TABA	MPRU	HA
Bottle		Oil	PLUS	MINUS	PLUS	MINUS	28	27.1	29.7
0	E.A.(mg)	200
1	E.A.(mg)		289	276		253		300	197
3	E.A.(mg)	190	266	287	259	283	244	331	250
10	E.A.(mg)		222	296	250	260	232	193	244
Note:
Extract/Bottle in the medium is in l of oil/100 ml base medium.
E.A. Embryo Axis (Root Shoot)

[0428] All fractions in the above Table have shown a maximum biomass growth promotion (E.A.) at 3 l of oil/100 ml base medium. Several fractions have also shown a maximu rootshoot growth promotion at 3 l of oil/100 ml base medium. In most cases, this is a result of a strong shoot growth promotion. In several cases, there is a sharp reduction in rootshoot growth at 10 l of oil/100 ml base medium.

TABLE 12G
Effect of plant extracts on P. radiatus seed
germination activity
Extract/		Sesame	AZIN-	PGL-	SIND-	TRIGF-
Bottle		Oil	C-29	C-33	28.8	29.62
0	Shoot	22.1
Control	(cm)	(0.75)
	Root	8.2
	(cm)	(0.87)
	Total	30.3
	(cm)	(1.05)
1	Shoot	23.1	22.3	23.2	23.7	22.4	( )
	(cm)	(0.82)	(0.81)	(0.80)	(0.66)	(0.86)
	Root	8.6	8.6	9.0	8.8	8.3	( )
	(cm)	0.49)	(0.75)	(0.79)	(1.01)	(0.94)
	Total	31.7	30.9	32.3	32.6	30.7	( )(
	(cm)	(1.10)	(1.02)	(1.19)	(1.2)	(1.43)
3	Shoot	24.0	19.1	21.0	24.1	24.1	( )
	(cm)	(0.84)	(0.59)	(0.71)	(0.72)	(0.52)
	Root	8.8	7.2	8.8	9.1	8.8	( )
	(cm)	(1.07)	(0.85)	(0.56)	(0.92)	(0.46)
	Total	32.9	26.4	29.3	33.3	32.9	( )
	(cm)	(1.33)	(1.33)	(0.79)	(1.55)	(0.88)
10	Shoot	24.1	*13.3	19.0	24.8	25.9	( )
	(cm)	(0.70)	(0.88)	(1.09)	(0.84)	(1.02)
	Root	9.0	*6.3	7.6	9.8	9.2	( )
	(cm)	(0.95)	(0.57)	(0.84)	(0.74)	(0.72)
	Total	33.1	*19.6	26.5	34.6	35.16	( )
	(cm)	(1.34)	(0.60)	(1.1)	(1.35)	(1.26)
Note:
Extract/Bottle in the medium is in l of oil/100 ml base medium.
Values in parenthesis indicate standard deviation.

[0429] In this set, there was a complete liqufaction of the base medium. As result, only five seedlings grew and the others sank to the bottom. Hence, the readings are an average of 5 seedlings.

[0430] Dry biomass weight per 10 seedlings at the end of 5 days is reported in Table 12H below.

TABLE 12H
Effect of plant extracts on P. radiatus germination
activity
Extract/		Sesame	AZIN-	PGL-	SIND-	TRIGF-
Bottle		Oil	C-29	C-33	28	29.62
0	E.A.(mg)	280
Control	Root(mg)	62
	Shoot(mg)	218
1	E.A.(mg)	364	377	380	349	228
	Root(mg)	70	78	62	60	63
	Shoot(mg)	294	299	318	289	165
3	E.A.(mg)	350	324	336	389	322
	Root(mg)	68	80	70	75	68
	Shoot(mg)	282	244	266	314	254
10	E.A.(mg)	314	*287	312	334	307
	Root(mg)	64	*73	58	72	59
	Shoot(mg)	250	*214	254	262	260
Note:
Extract/Bottle in the medium is in l of oil/100 ml base medium.
E.A.Embryo Axis (Root Shoot)

[0431] In this set, there was a complete liqufaction of the base medium. As result, only five seedlings grew and the others sank to the bottom. Hence, the weight reading for 5 seedlings per set has been multipied by 2 and recorded for comparison.

[0432] In this set, the Pongamia glabra cake (PGL-C-33) and particularly the Azadirachta indica cake (AZIN-C-29) extracts have shown a strong inhibitory activity for rooting and shooting above 3 l of oil/100 ml base medium. This is confirmed by low biomass growth (E.A.) in case of Azadiracta indica cake. However, biomass growth in case of Pongamia glabra cake has not gone down. In case of Sesamum indicum (SIND-28) there is no significant increase in root and shoot length. However, there is a significant in crease in the biomass at 1 and 3 l of oil/100 ml base medium. Thus, a wide range of activity has been observed.

TABLE 12I
Effect of plant extracts on P. radiatus seed
germination activity
			CROT	CROT	TVUL	TVUL	ZOFF	CAMA
Extract/		Sesame	28.2	62.66	32.83	66.84	55.4	26.2	GGLAB
Bottle		Oil	PLUS	MINUS	PLUS	MINUS	PLUS	PLUS	27.43
0	Shoot	20.0
Control	(cm)	(0.82)
	Root	9.0
	(cm)	(0.70)
	Total	29.1
	(cm)	(1.17)
1	Shoot	20.5	21.9	20.7	20.7	20.2	24.1	22.4	21.9
	(cm)	(0.72)	(0.90)	(1.33)	(0.57)	(0.25)	0.50)	(0.67)	(0.97)
	Root	9.8	10.3	10.7	10.4	10.3	11.5	10.7	10.6
	(cm)	(0.51)	(0.41)	(0.72)	(0.67)	(0.63)	(0.65)	(0.31)	(0.39)
	Total	30.3	32.2	31.4	31.1	30.5	35.6	33.1	32.5
	(cm)	(0.83)	(0.92)	(1.58)	(0.77)	(0.57)	(0.82)	(0.72)	(1.06)
3	Shoot	20.6	22.4	20.5	23.3	20.4	23.3	23.1	23.7
	(cm)	(0.80)	(0.59)	(0.88)	(0.53)	(1.04)	(0.47)	(0.42)	(0.39)
	Root	10.1	11.1	10.2	11.7	11.2	10.9	9.8	111.4
	(cm)	(0.49)	(0.52)	(0.42)	(0.67)	(0.50)	(0.51)	(0.55)	(0.72)
	Total	30.8	33.5	30.5	35.0	31.7	32.2	33.0	35.1
	(cm)	(0.95)	(0.60)	(0.75)	(0.73)	(0.95)	(0.48)	(0.72)	(0.87)
10	Shoot	22.9	20.72	19.5	20.4	23.5	20.6	22.5	21.1
	(cm)	(0.48)	(0.55)	(0.98)	(1.14)	(0.64)	(1.15)	(0.37)	(0.58)
	Root	11.0	11.0	8.0	11.0	12.3	9.7	8.7	11.0
	(cm)	(0.54)	(0.41)	(0.64)	(0.46)	(0.43)	(0.42)	(0.71)	(0.48)
	Total	34.0	31.7	27.5	31.4	35.8	30.3	31.2	32.0
	(cm)	(0.81)	(0.95)	(1.27)	(1.29)	(0.93)	(1.16)	(0.92)	(0.63)
Note:
Concentration in the medium is in l of oil/100 ml base medium.
Values in parenthesis indicate standard deviation.

[0433] Dry biomass weight/10 seedlings at the end of 5 days is reported in Table 12J below.

TABLE 12J
Effect of plant extracts on P. radiatus germination
activity
			CROT	CROT	TVUL	TVUL	ZOFF	CAMA
Extract/		Sesame	28.2	62.66	32.83	66.84	55.4	26.2	GGLAB
Bottle		Oil	PLUS	MINUS	PLUS	MINUS	PLUS	PLUS	27.43
0	E.A.	313
Control	(mg)
	Root	71
	(mg)
	Shoot	242
	(mg)
1	E.A.	320	291	317	294	272	303	281	284
	(mg)
	Root	74	73	86	74	62	67	61	62
	(mg)
	Shoot	246	218	231	220	204	236	220	222
	(mg)
3	E.A.	290	279	316	298	307	311	297	244
	(mg)
	Root	72	67	82	80	67	73	75	72
	(mg)
	Shoot	218	212	234	218	240	238	222	172
	(mg)
10	E.A.	314	297	324	327	314	298	308	254
	(mg)
	Root	77	83	82	84	82	78	73	75
	(mg)
	Shoot	237	214	242	243	232	220	235	179
	(mg)
Note:
Concentration in the medium is in l of oil/100 ml base medium.
E.A.Embryo Axis (Root Shoot)

[0434] Cyperous rotundus fractions (CROT 28.2 PLUS and CROT 62.66 MINUS) show differential activity. The PLUS fraction appears to promote shootroot length at the lower dose; however, this isnot reflected in the biomass growth. The minus fraction does not siginificantly promote shootroot growth and starts showing inhibitory effects at 10 l of oil/100 ml base medium. Triticum vulagare fractions (TVUL 32.83 PLUS and TVUL 66.84 MINUS) promote shoot growth at 3 l of oil/100 ml base medium and 10 l of oil/100 ml base medium, respectively. This, in terms of starting wheat seeds, is approx. at 1 mg of wheat equivalent/100 ml base medium for the PLUS fraction and 6.7 mg of wheat equivalent/100 ml base medium for the minus fraction. Curcuma amamada (CAMA 26.2 PLUS) promotes particularly shoot growth at the lower concentrations of 1 and 3 l of oil/100 ml base medium. Glycyrrhiza glabra (GGLAB 27.43) also promotes rootshoot growth at 1 and 3 l of oil/100 ml base medium. However, this growth is accompanied by a reduction in biomass growth.

TABLE 12K
Effect of plant extracts on P. radiatus seed
germination activity
Extract/		Sesame	A-100	A-100	B-100	B-100	C-100	C-100
Bottle		Oil	PLUS	MINUS	PLUS	MINUS	PLUS	MINUS
0	Shoot	16.4
Control	(cm)	(0.69)
	Root	8.13
	(cm)	(0.55)
	Total	24.53
	(cm)	(0.86)
1	Shoot	16.78	14.98	18.53	17.23	16.6	15.56	17.02
	(cm)	(0.49)	(0.46)	(0.62)	(0.54)	(0.66)	(0.35)	(0.48)
	Root	7.88	7.73	9.29	11.67	8.9	9.59	8.41
	(cm)	(0.33)	(0.64)	(0.67)	(0.31	(0.44)	(0.40)	(0.55)
	Total	25.18	22.71	27.82	28.9	25.5	25.15	25.42
	(cm)	(0.55)	(0.58)	(1.01)	(0.69)	(0.71)	(0.63)	(0.91)
3	Shoot	17.51	16.84	17.32	16.23	17.25	16.84	16.29
	(cm)	(0.57)	(0.16)	(0.34)	(0.42)	(0.40)	(0.54)	(0.51)
	Root	8.4	7.43	8.79	10.92	9.21	9.12	7.69
	(cm)	(0.39)	(0.39)	(0.64)	(0.47)	(0.45)	(0.42)	(0.43)
	Total	25.91	24.27	26.11	27.15	26.46	25.96	23.88
	(cm)	(0.62)	(0.48)	(0.99)	(0.83)	(0.75)	(0.58)	(0.43)
10	Shoot	17.99	16.89	16.48	16.33	19.34	19.69	15.5
	(cm)	(0.51)	(0.64)	(0.27)	(0.43)	(0.36)	(0.67)	(0.32)
	Root	8.55	7.08	7.04	9.84	11.29	10.2	6.88
	(cm)	(0.79)	(0.50)	(0.43)	(0.65)	(0.44)	(0.45)	(0.53)
	Total	26.54	23.97	23.52	26.17	30.63	29.89	22.19
	(cm)	(0.87)	(0.84)	(0.40)	(0.74)	(0.50)	(0.95)	(0.70)
Note:
Extract/Bottle in the medium is in l of oil/100 ml base medium.
Values in parenthesis indicate standard deviation.

[0435] In this set, different compositions show differential activity. The A-100 MINUS and B-100 PLUS both show a strong root and shoot promotional activity at the 1 l of oil/100 ml level and the activity declines at higher concentrations. The B-100 MINUS and C-100 PLUS show strong promotional activity at the 10 l of oil/100 ml level.

TABLE 12M
Effect of plant extracts on P. radiatus seed
germination activity
				RS-
				10			R-25	R-25	R-25
Extract/		Sesame	R-	Leaf	R-100	R-100	30 Min	120 Min	240 Min
Bottle		Oil	100	Stem	(J L/4)	(4J L/8)	boiling	boiling	boiling
0	Shoot	16.4
Control	(cm)	(0.69)
	Root	8.13
	(cm)	(0.55)
	Total	24.53
	(cm)	(0.86)
1	Shoot	16.78	16.4	18.84	15.76	16.24	16.71	12.93	15.51
	(cm)	(0.49)	(0.41)	(0.51)	(0.61)	(0.37)	(0.44)	(0.46)	(0.30)
	Root	7.88	8.63	10.07	5.32	5.96	8.53	4.98	8.73
	(cm)	(0.33)	(0.27)	(0.76)	(0.53)	(0.64)	(0.39)	(0.52)	(0.31)
	Total	25.18	25.08	28.91	21.08	22.2	25.24	17.91	24.24
	(cm)	(0.55)	(0.32)	(0.47)	(0.65)	(0.81)	(0.48)	(0.54)	(0.44)
3	Shoot	17.51	17.03	17.51	16.31	16.16	16.65	18.18	16.27
	(cm)	(0.57)	(0.35)	(0.42)	(0.36)	(0.30)	(0.51)	(0.48)	(0.43)
	Root	8.4	10.07	8.77	10.67	6.59	9.26	8.36	11.04
	(cm)	(0.39)	(0.52)	(0.66)	(0.47)	(0.54)	(0.46)	(0.37)	(0.30)
	Total	25.91	27.1	26.28	26.98	22.75	25.91	26.54	27.31
	(cm)	(0.62)	(0.61)	(1.0)	(0.60)	(0.58)	(0.62)	(0.63)	(0.56)
10	Shoot	17.99	18.27	12.57	15.34	16.11	18.51	16.5	19.23
	(cm)	(0.51)	(0.64)	(0.34)	(0.29)	(0.21)	(0.82)	(0.50)	(0.36)
	Root	8.55	11.2	7.41	8.62	5.48	10.87	8.61	11.39
	(cm)	(0.79)	(0.37)	(0.33)	(0.47)	(0.53)	(0.33)	(0.36)	(0.35)
	Total	26.54	29.47	19.98	23.96	21.69	29.38	25.11	30.62
	(cm)	(0.87)	(0.77)	(0.55)	(0.69)	(0.67)	(0.66)	(0.51)	(0.52)
Note:
Extract/Bottle in the medium is in l of oil/100 ml base medium.
Values in parenthesis indicate standard deviation.

[0436] In this set, R-100, made as per a typical recipe of this invention is compared with R-100(4JL/8) made using a typical traditional recipe deacribed in Sharangdharsamhita. The (4JL/8) indicates that the juice taken for boiling was 4 times the oil and the kalka taken for boiling was ^th the weight of oil. Although R-100 shows promotion, R-100(4JL/8) shows inhibition when compared to both the plain control and sesame oil alone.

[0437] RS-10 made just from the leaf stem also shows a strong promotion. The last three sets of columns show that over a boiling time range of 30 minutes to 240 minutes, the compositions made as per the present invention show promotion compared to control.

TABLE 12O
Effect of plant extracts on P. radiatus seed
germination activity
				CLNG
Extract/		Sesame	CLNG I	II	EJAM	RSER	VR
Bottle		Oil	33.75	26.47	34.16	29	25
0	Shoot	19.71
Control	(cm)	(1.06)
	Root	8.3
	(cm)	(0.48)
	Total	28.01
	(cm)	(1.32)
1	Shoot	22.37	13.42	20.53	17.53	19.96	19.96
	(cm)	(0.55)	(0.63)	(1.43)	(1.24)	(0.39)	(0.45)
	Root	7.9	4.29	8.26	5.93	8.12	7.54
	(cm)	(0.76)	(0.70)	(0.46)	(1.14)	(0.55)	(0.81)
	Total	30.32	17.17	28.79	23.4	27.45	24.5
	(cm)	(0.78)	(0.91)	(1.51)	(1.83)	(1.5)	(1.13)
3	Shoot	22.22	17.0	22.83	20.12	22.25	20.54
	(cm)	(1.71)	(0.71)	(0.54)	(1.36)	(0.37)	(0.96)
	Root	9.31	7.9	10.14	10.03	11.42	10.29
	(cm)	(0.50)	(0.27)	(0.62)	(0.61)	(0.56)	(0.51)
	Total	31.53	24.90	32.71	30.20	33.7	30.83
	(cm)	(1.9)	(0.75)	(1.05)	(2.0)	(0.93)	(1.22)
10	Shoot	22.33	21.92	23.1	21.63	18.52	20.99
	(cm)	(1.19)	(0.30)	(0.56)	(2.1)	(1.38)	(0.57)
	Root	10.61	6.53	10.5	10.13	7.73	8.48
	(cm)	(0.55)	(0.38)	(0.52)	(0.34)	(0.50)	(1.37)
	Total	32.94	28.45	33.60	31.76	26.25	29.47
	(cm)	(1.42)	(0.60)	(0.74)	(1.47)	(1.78)	(1.44)
Note:
Extract/Bottle in the medium is in l of oil/100 ml base medium.
Values in parenthesis indicate standard deviation.