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1.  Fate of β-asarone in Ayurvedic Sodhana process of Vacha 
Calamus (Acorus calamus Linn., Araceae) rhizome synonymously called sweet flag or Vacha is an aromatic herb indigenous to Central Asia and Eastern Europe. It has been used by the Ayurvedic practitioners since time immemorial for diseases ranging from weakness of memory to being used as an anthelminthic. Reports of its use have been found in books like Charak Samhita, Sushruta Samhita, etc., The major constituent of the oil of Vacha is a phenyl propanoid called β-asarone, which is reported to show carcinogenic properties. Due to the toxic effects of β-asarone, sodhana prakriya (detoxification process) has been prescribed for Vacha before its inclusion in the Ayurvedic medicines. Shodhanaprakriya (S. prakriya) of Vacha has been mentioned in the Ayurvedic texts.
This study was undertaken with an aim to find out the mechanism involved in the S. prakriya of Vacha and also to suggest an alternate method for the conventional one.
Materials and Methods:
The conventional method was studied in the laboratory and equivalent alternate methods were designed based on the mechanism involved. Vacha samples were subjected to the conventional method as well as the alternate methods and the content of β-asarone in the different samples was monitored using Gas Chromatography technique.
Various alternate methods have been devised based on the mechanism involved in the S. prakriya which have given results comparable with those of the conventional method.
The scientific mechanism involved in the S. prakriya of Vacha has been established and alternate methods have been proposed.
PMCID: PMC3667428  PMID: 23741157
Acorus calamus; β-asarone; Sodhana; Vacha
2.  Experimental evaluation of antidepressant effect of Vacha (Acorus calamus) in animal models of depression 
Ayu  2010;31(2):153-158.
Depression is a common diagnosis throughout India. It is one of the major sequelae of modern lifestyle which is full of stress. Several drugs and therapies have been tried but a safe and effective treatment of depressive illness is yet not fully established. The main objective of this experimental study on animal models is to evaluate the antidepressant action of plant drug Vacha (Acorus calamus). The behavioral study was conducted and at the same time 5-HT receptor involvement was evaluated. The experimental study was done in rats to evaluate their Open Field Behavior (OFB), High Plus Maze (HPM) activity and 5-hydroxytryptamine (5-HT) receptor syndrome, before and after feeding Vacha. Concurrent Vacha administration in the depression model prevented the development of behavioral deficit in ambulation and rearing due to stress. Similarly, in High Plus Maze Test (HPMT), exploratory activity of rat was restored with Vacha administration. In adopted model of depression, when the animal was subjected to Vacha administration, the behavioural deficit was prevented very well as compared to stressed group. While eliciting the 5-HT syndrome, only two components out of five were influenced by Vacha, indicating that Vacha does not sensitize postsynaptic 5-HT1A receptors, which explains the behavioral deficit prevention in stressed rat group. Vacha definitely has antidepressant effects in animal model of depression.
PMCID: PMC3215357  PMID: 22131703
5-hydroxytryptamine receptors; depression; Vacha
3.  Preliminary physico-chemical profile of Brahmi Ghrita 
Ayu  2013;34(3):294-296.
Brahmi Ghrita was processed as per the process of Snehapaka procedure described in classics. It contained Brahmi (Bacopa monneri), Vacha (Acorus calamus), Kushtha (Sassurea lappa), Shankhapushpi (Convolvulos pluricalis), and Purana Ghrita. In the preparation of Brahmi Ghrita, Brahmi Swarasa, Kalka Dravya of Brahmi, Vacha, Kushtha, and Shankhapushpi were mixed in Purana Ghrita and heated for three hours at 110°C every day for three days. On the third day Ghrita was filtered to obtain the finished product. In this manner, three samples of Brahmi Ghrita were prepared. To understand the changes that occurred during the preparation, Brahmi Ghrita and Purana Ghrita were analyzed by using modern parameters such as Acid value, Saponification value, and so on. After the analysis, it was found that the Acid values of Sample A, B, and C of Brahmi Ghrita were 4.26, 4.03, and 4.03; the Saponification values of Samples A, B, and C of Brahmi Ghrita were 227.2, 230.01, and 230.01, and the Iodine values of Samples A, B, and C were 34.75, 35.88, and 35.88, respectively, and the Acid value, Saponification value, and Iodine value of Purana Ghrita were 1.57, 199.15, and 31.04, respectively. The present study revealed that, there was no significant variation in the analytical values among all three samples of Brahmi Ghrita.
PMCID: PMC3902597  PMID: 24501526
Brahmi Ghrita; acid value; saponification value; iodine value
4.  Clinical study of the Immunoglobululin Enhancing effect of “Bala compound” on Infants 
Ancient Science of Life  2009;28(3):18-22.
Kaumarbhritya a branch of Asthanga Ayurveda deals with neonatal, infant and child health care. Multicentric studies conducted in various developed and developing countries have indicated that Infant Mortality Rate (I.M.R.) is very high in developing countries, and infection has been observed as the major cause. Immune system in neonates is not yet fully functional. Bala compound having the ingredients of Atibala (Abutilon indicum Linn), Amalaki (Emblica officinalis Linn), Vidanga (Emblica ribes burn), Guduchi (Tinospora cordifolia Welld Miers), Pippali (Piperlongum linn), Yashtimadhu (Glycyrrhiza glabra Linn), Shankhapuspi (Convolvulus pluricaulis Chois ), Vacha (Acorus calamus Linn), Musta (Cyperus rotundus Linn) and Ativisha (Aconitum heterophyllum wall) are Medhya as well as Rasayana drugs mention in Ayurvedic classics. ‘Bala compound” was tried in infants in the form of oral drops for a period of six months and result was assessed for serum immuoglobulins IgG, IgM, IgA for three months of interval of two follow ups (i.e., third and six month of infant). There is significant increase of immunoglobulins observed after six months administration of ‘Bala compoumd”
PMCID: PMC3336317  PMID: 22557316
5.  Usage trends for memory and vitality-enhancing medicines: A pharmacoepidemiological study involving pharmacists of the Gujarat region 
The aim of the study was to explore the trends and rationale of use of memory and vitality-enhancing medicines (MVEM) in the Gujarat region.
Materials and Methods:
A prospective pharmacoepidemiological study involving pharmacists of Gujarat region was carried out in the year 2005. Pharmacists (n = 351) working in general and Ayurvedic medical stores were selected from 12 districts of Gujarat region. The pharmacists were explained about the objective of the study and were given a pretested, validated questionnaire.
Outcome Measures:
The questionnaire included the questions regarding herbal MVEM used most commonly, percentage sale of herbal MVEM – sold with or without prescriptions – age group of patients and professional groups who used these drugs most commonly.
The number of individuals using MVEM was highest in the age group of 11–20 years (17.54%), followed by the 21–40 years group (17.12%), supporting the results that the professional group of students (17.29%) and the persons of business or service class (15.29%) are the highest users of these medicines. Evaluation of various constituents in the marketed polyherbal MVEM revealed that Brahmi (Bacopa monniera), Shankhpushpi (Evolvulus alsinoides), Ashwangandha (Withania somnifera), Jatamansi (Nardostychos jatamansi), Vacha (Acorus calamus) and Amla (Phyllanthus emblica) were the common ingredients in the polyherbal preparations.
This study highlights commonly used Ayurvedic medicines that can be explored for safely enhancing memory and vitality performance. Hence, detailed and scientifically designed research on these drugs would help to identify safe and effective drugs for enhancing the same.
PMCID: PMC2996570  PMID: 21170204
Memory-enhancing medicines; polyherbal formulations; use of herbal medicines; vitality-enhancing medicines
6.  Immunomodulatory activity of Vachadhatryadi Avaleha in albino rats 
Ayu  2011;32(2):275-278.
The present study is carried out to evaluate the immuno-modulatory activity of Vacha Dhatryadi Avaleha in albino rats. Vacha Dhatryadi Avaleha was prepared by classical method and evaluated for humoral antibody formation and cell-medicated immunity in established experimental models. Test formulation was administered at the dose of 900 mg/kg and parameters like hemagglutination titer, ponderal changes, histopathology of immunological organs and immunological paw edema were recorded. Vacha Dhatryadi Avaleha significantly enhanced antibody formation and moderately suppressed the immunological edema. The present study concludes that Vachadhatryadi Avaleha has immunopotentiating activity.
PMCID: PMC3296354  PMID: 22408316
Cell mediated immunity; hemagglutination titer; Vachadhatryadi
7.  Protective role of Ashwagandharishta and flax seed oil against maximal electroshock induced seizures in albino rats 
Ayu  2012;33(1):114-118.
Ashwagandharishta, an Ayurvedic classical formulation, is the remedy for Apasmara (epilepsy), Murchha (syncope), Unmada (psychosis), etc. Recent studies in animal models have shown that n-3 PUFAs can raise the threshold of epileptic seizures. The indigenous medicinal plant, called Atasi (Linum usitatissimum Linn.) in Ayurveda, or flax seed, is the best plant source of omega-3 fatty acids. The present study is designed to investigate whether Ashwagandharishta and Atasi taila (flax seed oil) protect against maximal electroshock (MES) seizures in albino rats. Further, a possible protective role of flax seed oil as an adjuvant to Ashwagandharishta in its anticonvulsant activity has also been evaluated in the study. MES seizures were induced for rats and seizure severity was assessed by the duration of hind limb extensor phase. Phenytoin was used as the standard antiepileptic drug for comparison. Both flax seed oil and Ashwagandharishta significantly decreased convulsion phase. Pre-treatment with flax seed oil exhibited significant anticonvulsant activity by decreasing the duration of tonic extensor phase. Contrary to the expectations, pre-treatment with flax seed oil as an adjuvant to Ashwagandharishta failed to decrease the tonic extensor phase; however, it significantly decreased the flexion phase (P < 0.001) and duration of the convulsions (P < 0.05). Both the drugs exhibited an excellent anti-post-ictal depression effect and complete protection against mortality.
PMCID: PMC3456848  PMID: 23049195
Antiepileptic; Ashwagandharishta; flax seed oil; maximal electroshock seizure; omega-3 fatty acid
8.  PA01.33. Mode of action of medhya drugs: A review 
Ancient Science of Life  2012;32(Suppl 1):S83.
Memory is combination of power of acquisition (Grahan), retention (Dharan) and recollection (Smaran). Vata, pitta, and kapha are the three psychobiological dimensions (energy) or biological rhythms regulating the entire functioning of the human body. Pitta is responsible for understanding and attaining knowledge due to its Satva, Aashukari and Tikshna property. Vata is responsible for association of ideas. Kapha provides stability which is necessary for retention of memory. Several Central Nervous System disorders (CNS) are often associated with impairment in cognitive functions. Disorders of memory can range from mild to severe. It is major health problem of 21st century. Main objective of this work is to understand the ayurvedic aspect of learning and memory mode of action of some medhya drugs W.S.R. Vacha, Shankhpushpi, Endri, Jyotishmati and brahmi.
A review on available scientific information in terms of references in classical texts, and Nighantus, chemical constitute, pharmacological activity is prepared with a view to understand mode of action of these Medhya dravya.
Pitta is responsible for understanding and attaining knowledge due to its Satva, Aashukari and Tikshna property. Vata is responsible for association of ideas. Kapha provides stability which is necessary for retention of memory. Sheetvirya and improves retention power and ushna virya drugs enhance power of acquiring new information and data interpretation.
Ushna virya and sheet virya drugs acts through different mechanism and enhance cognition. It may provide parameters for selection of medhya drug in different memory disorder.
PMCID: PMC3800965
9.  Effect of Purificatory Measures Through Cow's Urine and Milk on Strychnine and Brucine Content of Kupeelu (Strychnos Nuxvomica Linn.) Seeds 
Strychnos nux vomica Linn.(Loganaceae) commonly known as Nux vomica (Kupeelu), is a poisonous plant and its seeds are used widely in Ayurvedic system of medicine since time immemorial. Ayurveda advocates that nux vomica seeds are to be administered in therapeutics only after going through certain purificatory measures (Shodhana). There are more than six media: cow's urine (Go mutra), cow's milk (Go dugdha), cow's ghee (Go ghrita), Kanji (thin gruel), castor oil (Eranda taila) and fresh ginger juice (Ardraka swarasa) etc., which have been reported in different classical texts of Ayurveda for proper processing of nux vomica seeds. In this study, an attempt has been made to purify the seeds by using three different methods as described in ancient treatise by using cow's urine and cow's milk as media alone and together. This study revealed that all the methods studied reduced the toxicity of strychnine and brucine contents in comparison to the raw seeds as determined by HPTLC. Out of these three methods maximum reduction in strychnine and brucine contents was found when the seeds were purified by keeping them in cow's urine for seven days followed by boiling in cow's milk for three hrs.
PMCID: PMC3746529  PMID: 23983327
Kupeelu; Strychnos nuxvomica; Shodhana; strychnine; Ayurveda; brucine; Cow's milk; Cow's urine
10.  Urushiol-induced contact dermatitis caused during Shodhana (purificatory measures) of Bhallataka (Semecarpus anacardium Linn.) fruit 
Ayu  2012;33(2):270-273.
Bhallataka (Semecarpus anacardium Linn.; Ancardiaceae) is mentioned under Upavisha group in Ayurvedic classics and it is described as a poisonous medicinal plant in Drugs and Cosmetics Act (India), 1940. Fruit of Bhallataka is used either as a single drug or as an ingredient in many compound formulations of Indian systems of medicine to cure many diseases. Tarry oil present in the pericarp of the fruit causes blisters on contact. The major constituent of the tarry oil is anacardic acid and bhilawanol, a mixture of 3-n-pentadec(en)yl catechols. Bhilawanol A and B are known as Urushiols, and also, anacardic acid is closely related to Urushiol. Urushiol-induced contact dermatitis is the medical name given to allergic rashes produced by the oil Urushiol. This paper deals with five case reports of contact dermatitis caused during different stages of Shodhana (purificatory measures) of Bhallataka fruit due to improper handling of the utensils and disposal of media used in Shodhana procedure and their Ayurvedic management. To combat these clinical conditions, the affected persons were advised external application with pounded Nimba (Azadirachta indica A. Juss.) leaves on the affected parts and internal administration of Sarivadyasava 30 ml thrice daily after food and Triphala Churna 5 g before food twice daily. Reduction of itching and burning sensation was observed after topical application.
PMCID: PMC3611655  PMID: 23559802
Bhallataka; bhilawanol; blisters; dermatitis; Semecarpus anacardium Linn.; urushiol
11.  Acute and Subchronic Toxicity Study of Tamra Bhasma (Incinerated Copper) prepared from Ashodhita (Unpurified) and Shodhita (Purified) Tamra in Rats 
The use of metals in traditional medicines is very often seen as matter of concern these days, especially the Bhasma preparations which are always under stringent observations for containing highly reactive inorganic elements such as lead, mercury, arsenic and others. One of the Bhasma extensively used in routine Ayurvedic practice is Tamra (copper) bhasma. If it is not prepared properly or Shodhana procedure is not done properly, it acts as a poison. To indicate its toxic potential, Ashtamahadoshas (eight major ill effects) have been quoted in classics and due emphasis have been given to its Shodhana procedure. In the present study, Tamra bhasma prepared from Shodhita and Ashodhita Tamra was subjected to oral toxicity study to ascertain the role of Shodhana process on safety profile of Tamra bhasma on subchronic administration to albino rats. Both the samples were administered to rats for 45 consecutive days at the doses of 5.5, 27.5, and 55 mg/kg. Animals were sacrificed on 46th day and parameters like hematological, serum biochemical, and histopathology of various organs were studied. Results showed that Tamra bhasma prepared from Ashodhita Tamra has pathological implications on different hematological, serum biochemical and cytoarchitecture of different organs even at therapeutic dose level (5.5 mg/kg). Whereas, Tamra bhasma prepared from Shodhita Tamra is safe even at five-fold to therapeutic equivalent doses (27.5 mg/kg). These observations emphasize the role of Shodhana and importance of dose in expression of toxicity of the medicinal preparations.
PMCID: PMC3783753  PMID: 24082351
Amrutikarana; copper; herbomineral formulation; LD50; Marana; Shodhana
12.  Efficacy of Acorus calamus on collagen maturation on full thickness cutaneous wounds in rats 
Pharmacognosy Magazine  2014;10(Suppl 2):S299-S305.
The rhizomes of Acorus calamus and their essential oil are widely used in the flavoring industry and production of alcoholic beverages in Europe. Recent reports have confirmed the presence of several pharmacological components in the rhizomes of A. calamus.
The objective of this study was to find out the efficacy of topical administration of ethanolic extract of A. calamus on dermal wound healing in rats. Wound healing is a natural process occurring in living organisms, which results in a complete or partial remodeling of injured tissue and ultimately progresses to the formation of a fibrous scar. Several natural products have been reported to augment the wound healing process.
Materials and Methods:
An ethanolic extract of A. calamus was prepared and its wound-healing efficacy was studied. An excision wound was made on the back of the rat and 200 μL (40 mg/kg body weight) of the A. calamus extract was applied topically once daily for the treated wounds. The control wounds were treated with 200 μL of phosphate buffered saline.
The granulation tissues formed were removed at 4, 8 and 12 days and biochemical parameters such as deoxyribonucleic acid, total protein, total collagen, hexosamine and uronic acids were measured. The amount of type I/III collagen formed in control and treated wound tissues was evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The epithelialization time, tensile strength and histological examination of the wounds were also studied. Biochemical analyses of the granulation tissues revealed a significant increase in collagen, hexosamine and uronic acid when compared with the control. The tensile strength of extract treated wounds was found to increase by 112%. A significant reduction in lipid peroxide levels suggested that A. calamus possesses antioxidant components.
The results strongly confirm the beneficial effects of A. calamus in augmenting the wound healing process.
PMCID: PMC4078342  PMID: 24991107
Acorus calamus; collagen; epithelialization; sodium dodecyl sulfate-polyacrylamide gel electrophoresis; tensile strength; wound healing
13.  Marker based standardization of polyherbal formulation (SJT-DI-02) by high performance thin layer chromatography method 
Preparation of highly standardized herbal products with respect to chemical composition and biological activity is considered to be a valuable approach in this field. SJT-DI-02 polyherbal formulation was successfully developed at our institute and filed for patent at Mumbai patent office.
The present work was marker based standardization of patented, novel and efficacious polyherbal formulation namely SJT-DI-02 for the treatment of diabetes. The SJT-DI-02 was comprised of dried extracts of rhizomes of Acorus calamus, leaves of Aegle marmelose, fruits of Benincasa hispida, roots of Chlorophytum arendinaceum, seeds of Eugenia jambolana, leaves of Ocimum sanctum, pericarp of Punica granatum, seeds of Tamarindus indica. Selected plants were collected, dried and extracted with suitable solvents. The formulation was prepared by mixing different fractions of extracts.
Materials and Methods:
For successful and best standardization, first of all selection and procurement was carried out. Selection is done on the basis of therapeutic efficacy and amount of the marker present in the particular plant part. At the time of procurement side by side phytochemical screening and estimation of phytoconstituents was carried out. After completion of preliminary screening using characterized markers, we tried to develop best TLC systems using selected solvent composition. Finally well-developed TLC systems were applied in HPTLC. In the present study polyherbal formulation was standardized by using different four markers. TLC Densitometric methods were developed using HPTLC for the quantification of these marker compounds. Solvent systems were optimized to achieve best resolution of the marker compounds from other components of the sample extract. The identity of the bands in the sample extracts were confirmed by comparing the Rf and the absorption spectra by overlaying their UV absorption spectra with those of their respective standards. The purity of the bands due to marker compounds in the sample extracts were confirmed by overlaying the absorption spectra recorded at start, middle and end position of the band in the sample tracks. After conforming all these things fingerprints were developed for all three formulations which will be act as authentification and quality control tool.
% w/w of asarones is 3.61, % w/w of marmelosin is 4.60, % w/w of gallic acid is 10.80 and % w/w of lupeol is 4.13. The method was validated in terms of linearity, precision, repeatability, limit of detection, limit of quantification and accuracy. In well-developed mobile phase system linearity was found to be in the range of 0.983-0.995, % recovery was found to be in the range of 97.48-99.63, % RSD for intraday and interday was found to be 0.13- 0.70 and 0.32 -1.41 and LOD and LOQ was found to be in the range of 0.15- 0.61 and 0.45 -1.83 microgram per ml.
Thus High performance thin layer chromatography (HPTLC) methods were developed and validated in terms of linearity, precision, repeatability, limit of detection, limit of quantification and accuracy. The methods were rapid, sensitive, reproducible and economical. It does not suffer any positive or negative interference due to common other component present in the formulation and would also serve as a tool for authentication of herbal products containing marmelosin, gallic acid, lupeol and asarones. Thus this work provides standardized and therapeutically active polyherbal formulations for the different ailments.
PMCID: PMC4097936  PMID: 25035642
Asarones; gallic acid; HPTLC; lupeol; marmelosin; SJT-DI-02
14.  A comparative anti-inflammatory activity of raw and processed Kupeelu (Strychnos nux-vomica Linn.) seeds on albino rats 
Ancient Science of Life  2011;31(2):73-75.
Seeds of Kupeelu (Strychnos nux-vomica Linn.), a known poisonous drug, is used extensively in various Ayurvedic formulations with great therapeutic significance. Ayurveda recommends the administration of Kupeelu only after passing through specific purificatory procedures in different media like cow's urine (Go mutra), cow's milk (Go dugdha), cow's ghee (Go ghrita), Kanji (thin gruel) etc. Strychnos nux vomica seeds are extensively advocated for nervous debility, paralysis, and weakness of limbs, sexual weakness, dyspepsia, and dysentery and in rheumatism where it can be assumed that besides other properties, Kupeelu may have some sort of anti-inflammatory activity too. In the present study, the powder of raw and processed Kupeelu seeds (processed / purified with Kanji i.e sour gruel) as test drugs were assessed for anti-inflammatory activity by employing Carrageenan and Formaldehyde induced hind paw oedema in Wistar strain albino rats at a dose of 22.5 mg/kg body weight orally. This study reveals that both raw and purified Kupeelu showed presence of highly significant anti-inflammatory activity against formaldehyde induced hind paw oedema, but did not have similar activity against Carrageenan induced hind paw oedema.
PMCID: PMC3530271  PMID: 23284209
Kupeelu; Kanji; anti-inflammatory; purification; Shodhana; purificatory procedure
15.  Comparative physico-chemical profile of Gunja (Abrus precatorius Linn.) seeds processed through water and Nimbu Swarasa (lemon juice) 
Ayu  2013;34(4):411-416.
Gunja (Abrus precatorius Linn.), known as Indian liquorice, is reputed as one of the world's most deadly but most beautiful seed belonging to the family Fabaceae, characterised under the Upavisha (semi-poisonous drugs) and used extensively in various Ayurvedic formulations with great therapeutic significance. Ayurveda recommended the administration of Gunja only after proper Shodhana (purification procedures) in different media such as Godugdha (cow's milk), Kanji (sour gruel), etc., Apart from the classical methods, some traditional practitioners use Nimbu Swarasa for the Shodhana of Gunja seeds. In this study, an attempt has been made to carry out Shodhana of Gunja seeds using Nimbu Swarasa and water. This study revealed differences in physico-chemical parameters of purified samples, in comparison to raw drugs.
PMCID: PMC3968706  PMID: 24696580
Abrin; Abrus precatorius; Gunja; Nimbu Swarasa; Shodhana
16.  Effects of WIN 55,212-2 (a non-selective cannabinoid CB1 and CB2 receptor agonist) on the protective action of various classical antiepileptic drugs in the mouse 6 Hz psychomotor seizure model 
Journal of Neural Transmission  2014;121(7):707-715.
The aim of this study was to characterize the influence of WIN 55,212-2 (WIN—a non-selective cannabinoid CB1 and CB2 receptor agonist) on the anticonvulsant effects of various classical antiepileptic drugs (clobazam, clonazepam, phenobarbital and valproate) in the mouse 6 Hz-induced psychomotor seizure model. Limbic (psychomotor) seizure activity was evoked in albino Swiss mice by a current (32 mA, 6 Hz, 3 s stimulus duration) delivered via ocular electrodes. Drug-related adverse effects were ascertained by use of the chimney test (evaluating motor performance), step-through passive avoidance task (assessing learning) and grip-strength test (evaluating skeletal muscular strength). Total brain concentrations of antiepileptic drugs were measured by fluorescence polarization immunoassay to ascertain any pharmacokinetic contribution to the observed antiseizure effect. Results indicate that WIN (5 mg/kg, administered intraperitoneally) significantly enhanced the anticonvulsant action of clonazepam (P < 0.001), phenobarbital (P < 0.05) and valproate (P < 0.05), but not that of clobazam in the mouse 6 Hz model. Moreover, WIN (2.5 mg/kg) significantly potentiated the anticonvulsant action of clonazepam (P < 0.01), but not that of clobazam, phenobarbital or valproate in the 6 Hz test in mice. None of the investigated combinations of WIN with antiepileptic drugs was associated with any concurrent adverse effects with regard to motor performance, learning or muscular strength. Pharmacokinetic experiments revealed that WIN had no impact on total brain concentrations of antiepileptic drugs in mice. These preclinical data would suggest that WIN in combination with clonazepam, phenobarbital and valproate is associated with beneficial anticonvulsant pharmacodynamic interactions in the mouse 6 Hz-induced psychomotor seizure test.
PMCID: PMC4065376  PMID: 24549572
6 Hz psychomotor seizure model; Antiepileptic drugs; Cannabinoids; Drug interactions; WIN 55,212-2
17.  Anticonvulsant potential of ethanol extracts and their solvent partitioned fractions from Flemingia strobilifera root 
Pharmacognosy Research  2013;5(4):265-270.
Flemingia strobilifera (FS) R.Br. (Fabaceae) is an important medicinal plant. In wealth of India it has been reported that roots of FS are used by santals in epilepsy, hysteria, insomnia, and to relieve pain. In Burma also the roots of F. strobilifera are used to treat epilepsy.
To investigate anticonvulsant potential of 95% ethanol extract and four subsequent fractions (petroleum ether, chloroform, ethyl acetate, and aqueous fractions of the roots of FS against pentylenetetrazole (PTZ) and maximal electroshock (MES) induced convulsions.
Material and Methods:
All the fractions and crude ethanol extract were administered (i.e., 200, 400, 600 mg/kg, p.o.) for 7 days and at the end of the treatment convulsions were induced experimentally using pentylenetetrazole and Maximal electroshock Test. Diazepam and phenytoin (4 mg/kg, i.p. and 20 mg/kg, i.p., respectively) were used as reference anticonvulsant drugs against experimentally induced convulsions. The latency of tonic convulsions and the numbers of animals protected from tonic convulsions were noted.
High doses (200 and 300 mg/kg, p.o.) of ethyl acetate fraction and 95% ethanol crude extract (400 and 600 mg/kg, p.o.) significantly reduced the duration of seizure induced by maximal electroshock (MES). The same dose also protected from pentylenetetrzole-induced tonic seizures and significantly delayed the onset of tonic seizures. However, pet, ether, chloroform, and aqueous fraction at any of the doses used (i.e., 100, 200, 300 mg/kg, p.o.) did not show any significant effect on PTZ and MES induced convulsions. The treatment with crude ethanolic extract and ethyl acetate fraction caused signs of central nervous system depressant action in the locomotor activity test, confirmed by the potentiation of sodium pentobarbital sleeping time. Both did not cause disturbance in motor coordination assessed by rotarod test.
The data suggest that crude ethanol extract and ethyl acetate fraction of roots of Flemingia strobilifera have a central nervous system depressant action and behave as a potential anticonvulsant. It may produce its anticonvulsant effect via non-specific mechanism since it reduced the duration of seizures produced by maximal electroshock as well as delayed the latency of seizures produced by pentylenetetrazole.
PMCID: PMC3807991  PMID: 24174820
Antiepileptic; Flemingia strobilifera;  maximal electroshock; pentylenetetrazole; seizures
18.  Seizure recurrence and remission after switching antiepileptic drugs 
Epilepsia  2012;54(1):187-193.
Studies of seizure outcome in patients undergoing serial antiepileptic drug trials have all been uncontrolled, with no account made for the spontaneous changes in disease state that could confound the elucidation of drug effects. In addition, no study has ever looked at outcome following antiepileptic drug switch in seizure-free patients, despite the fact that this is done routinely in clinical practice. We aimed to address both of these issues using a matched case-cohort design.
We followed patients taking phenytoin or carbamazepine in monotherapy for focal epilepsy who were being crossed over to a newer agent as part of studies on the metabolic effects of anticonvulsant therapy. Many had been seizure-free but were being switched nonetheless due to side effects or concerns about long-term adverse consequences. Each patient was matched with two controls of the same seizure status who were on anticonvulsant monotherapy and whose drug was not switched. Seizure freedom over the ensuing 6 months was the primary endpoint.
Key Findings
There were 43 cases and 86 matched controls. Twenty-three case patients had been seizure-free on their old drug; 5 (21.7%) had seizure recurrence after drug switch compared to 2/46 matched controls (4.3%). Twenty case patients were having seizures on their old drug; 6 (30%) entered remission after drug switch, compared to 8/40 matched controls (20%). The two groups differed at baseline in number of anticonvulsants previously failed, which was the most important factor for prognosis. After statistical adjustment to account for this, seizure-free patients had 6.53 times higher odds of seizure recurrence if switched to a new drug (95% CI 1.02 – 61.19; p=0.06). Non-seizure-free patients had 1.66 times higher odds of remission if they remained on the same drug compared to switching, though this was not significant (95% CI 0.36 – 8.42; p=0.532). Neither dose changes, nor drug mechanism, nor duration of seizure-freedom had any bearing upon the results.
While the large majority of seizure-free patients remain so when switched to another agent, about one-sixth have a recurrence attributable to the change. Conversely, our study design provides the first evidence to suggest that most improvements in drug-resistant patients are likely due to spontaneous remissions, not new drug introductions. These findings have conflicting implications for two competing models of comparative antiepileptic drug efficacy, which will require further study to elaborate.
PMCID: PMC3519976  PMID: 22931161
antiepileptic drugs; seizure recurrence; seizure remission; crossover
19.  Retrospective Analysis of Levetiracetam Compared to Phenytoin for Seizure Prophylaxis in Adults with Traumatic Brain Injury 
Hospital Pharmacy  2013;48(9):757-761.
Phenytoin is standard of care for seizure prophylaxis following traumatic brain injury (TBI). Levetiracetam, an alternative antiepileptic drug, is utilized for seizure prophylaxis despite limited data supporting its use.
Our primary outcome was post-TBI seizure activity measured by electroencephalogram (EEG) for levetiracetam versus phenytoin. Secondary outcomes were length of intensive care unit (ICU) stay, requirement for additional antiepileptic drugs (AED), and drug and monitoring costs.
A retrospective review was performed of patients admitted to neurosurgical or surgical trauma ICU. Adult patients with at least 1 day of EEG monitoring were included. Patients were excluded if they had history of epilepsy, prior TBI, less than 48 hours of AED therapy, or additional AED prior to EEG monitoring.
A total 90 patients met inclusion criteria, with 18 receiving levetiracetam and 72 receiving phenytoin. Prevalence of EEG-confirmed seizure activity was similar between the levetiracetam and phenytoin groups (28% vs 29%; P = .99). ICU length of stay (13 vs 18 days; P = .28), time to EEG-confirmed seizure activity (4 vs 6 days; P = .24), and duration of seizure prophylaxis (9 vs 14 days; P = .18) were also similar. The median daily cost of levetiracetam therapy was $43 compared to $55 for phenytoin therapy and monitoring (P = .08). When all anticonvulsant therapy and monitoring were included, costs were lower for the levetiracetam group ($45 vs $83; P = .02).
Levetiracetam may provide an alternative treatment option for seizure prevention in TBI patients in the ICU. Total antiepileptic drug and monitoring costs were lower for levetiracetam patients.
PMCID: PMC3857128  PMID: 24421550
head injuries; intensive care units; levetiracetam; phenytoin; seizures
20.  Anticonvulsant and neurotoxicity profile of the rhizome of Smilax china Linn. in mice 
Indian Journal of Pharmacology  2011;43(1):27-30.
To study the anticonvulsant activity and neurotoxicity of ethanolic extract and ethyl acetate fraction of the rhizome of Smilax china (EESC and EAF, respectively) in mice.
Materials and Methods:
The anticonvulsant activities of EESC and EAF were studied against maximal electroshock (MES) and pentylenetetrazole (PTZ)-induced seizures in mice and neurotoxicity was determined using rotarod test.
The duration of hindleg extension in MES test was reduced significantly (P < 0.001) by EESC at a dose level of 400 mg/kg and EAF at both higher dose levels (200 and 400 mg/kg). In PTZ model, the seizure latency was prolonged by all the test groups.
The EESC and EAF may help to control petit mal and grand mal seizures.
PMCID: PMC3062115  PMID: 21455417
Epilepsy; China root; Smilax china; neurotoxicity
21.  PA01.12. Evaluations of role of vishesha shodhana of metals with specific reference to tamra shodhana 
Ancient Science of Life  2012;32(Suppl 1):S61.
In this research work, an effort was made to observe effects of Samanya Shodhanottara Vishesha Shodhana and only Vishesha Shodhana upon Tamra.
Methods described in Rasatangini were followed to perform Samanya as well as Vishesha Shodhana. The study was carried out in two parts. In first part, actual process of Shodhana by various methods was performed and six samples were prepared. In second part, analysis of Ashodhita and Shodhita Taamra (Samanya Shodhanottara Vishesha Shodhit as well as only Vishesh Shodhita) was carried out. For this purpose, following parameters of analysis were considered- Change in physical characteristics of copper foils pre and post Shodhana, Change in weight of copper foils pre and post Shodhana, Pattern of XRPD for each sample, Percentage of copper in each sample by ICP AES, Change in electrical conductivity of each sample.
After Samanya Shodhana, the previous shine of copper was totally lost and it had crumpled appearance. Overall, Changes in tenacity of metal were also observed. The hardness went on decreasing and brittleness went on increasing. Steady fall in weight of Tamra was observed after each Shodhana. The XRPD samples of all the samples of Shodhita Tamra showed presence of copper and copper oxides in them. However, the peak positions and intensities of them were varying. There was marked decline in electrical conductivity post Shodhana.
It appeared from this study that Vishesha Shodhana of metals if preceded by their Samanya Shodhana produces better results. This time tested exercise certainly brings about significant changes in metallic properties of metals. Performing only Vishesha Shodhana or only Samanya Shodhana is not adequate.
PMCID: PMC3800942
22.  Degradative actions of microbial xylanolytic activities on hemicelluloses from rhizome of Arundo donax 
AMB Express  2014;4:55.
Polysaccharidases from extremophiles are remarkable for specific action, resistance to different reaction conditions and other biotechnologically interesting features. In this article the action of crude extracts of thermophilic microorganisms (Thermotoga neapolitana, Geobacillus thermantarcticus and Thermoanaerobacterium thermostercoris) is studied using as substrate hemicellulose from one of the most interesting biomass crops, the giant reed (Arundo donax L.). This biomass can be cultivated without competition and a huge amount of rhizomes remains in the soil at the end of cropping cycle (10–15 years) representing a further source of useful molecules. Optimization of the procedure for preparation of the hemicellulose fraction from rhizomes of Arundo donax, is studied. Polysaccharidases from crude extracts of thermophilic microorganisms revealed to be suitable for total degradative action and/or production of small useful oligosaccharides from hemicelluloses from A. donax. Xylobiose and interesting tetra- and pentasaccharide are obtained by enzymatic action in different conditions. Convenient amount of raw material was processed per mg of crude enzymes. Raw hemicelluloses and pretreated material show antioxidant activity unlike isolated tetra- and pentasaccharide. The body of results suggest that rhizomes represent a useful raw material for the production of valuable industrial products, thus allowing to increase the economic efficiency of A. donax cultivation.
PMCID: PMC4086442  PMID: 25024928
Biorefinery; Xylan; Thermophilic enzymes; Xylanases; Xylooligosaccharides; Arundo donax
Ancient Science of Life  1995;14(4):276-280.
Standardization of ayurvedic drugs has become necessary as in all other walks of life. This paper deals with the detection of Acorus calamus in Ayurvedic preparations. It is one of the most commonly used ingredients in ayurvedic preparations. This layer chromatographic technique was used for detection. Two solvent systems were developed and the spots were visualized in iodine vapour. The spot of Rf 0.85 in I and 0.70 in (ii) obtained for Acorus calamus was present in all the finished products, taken thus showing the present of Acorus calamus in the finished products. Co. T.L.C. was done to confirm the observation. This was further confirmed by testing o market samples containing Acorus calamus.
PMCID: PMC3331246  PMID: 22556710
24.  A Review of the Effect of Anticonvulsant Medications on Bone Mineral Density and Fracture Risk 
Osteoporosis and seizure disorders are common diagnoses in older adults and often occur concomitantly.
The goal of this review was to discuss the current hypothesis for the pathogenesis of anticonvulsant-induced bone density loss and the evidence regarding the risk for osteoporosis and fractures in older individuals.
A review of the literature was performed, searching in MEDLINE and CINAHL for articles published between 1990 and October 2009 with the following search terms: anticonvulsant OR antiepileptic; AND osteoporosis OR bone density OR fracture OR absorptiometry, photon. Studies within the pediatric population, cross-sectional studies, and studies whose results were published in a language other than English were excluded.
A search of the published literature yielded >300 results, of which 24 met the inclusion and exclusion criteria and were included in this review. Hepatic enzyme induction by certain anticonvulsant medications appears to contribute to increased metabolism of 25-hydroxyvitamin D to inactive metabolites, which results in metabolic bone disease. There is increasing evidence that anticonvulsant use is associated with a higher risk of osteoporosis and clinical fractures, especially among older agents such as phenobarbital, carbamazepine, phenytoin, and valproate. Several observational studies suggest a class effect among anticonvulsant agents, associated with clinically significant reductions in bone mineral density and fracture risk. The use of anticonvulsant medications increases the odds of fracture by 1.2 to 2.4 times. However, only 2 large-scale observational studies have specifically examined the risk among those aged >65 years. This review also identified a randomized controlled trial whose results suggest that supplementation with high-dose vitamin D may be associated with increased bone mineral density in patients taking anticonvulsant medications. However, no randomized controlled trials investigating therapeutic agents to prevent fracture in this population were identified. Consequently, there are no formal practice guidelines for the monitoring, prevention, and management of bone disease among those taking anticonvulsants.
Observational studies suggest an association between use of anticonvulsant medications, reduced bone mineral density, and increased fracture risk. Randomized clinical trials are needed to guide the management of bone disease among those who use anticonvulsants.
PMCID: PMC3570810  PMID: 20226391
anticonvulsants; bone density; osteoporosis; fractures
25.  Comparative powder microscopy of Alpinia calcarata Roscoe and Alpinia galanga (Linn.) Willd 
Ayu  2012;33(3):441-443.
Medicinal plant materials are being adulterated in commerce due to many reasons such as similar morphological features, same name as written in classical text, presence of similar active principles in the substituted plant etc., that may badly affect the therapeutic activity of the finished products. Therefore, systematic identification is becoming essential in order to produce standardized finished herbal products. The present study includes two medicinal plant rhizomes; Alpinia calcarata Roscoe (Heenaratta) and A. galanga (Linn.) Willd (Aratta) whose microscopical build up was different from each other and was assessed by standard Pharmacognostical methods. Diagnostic identification characters of A. calcarata were compound starch grins, triangular-shaped starch grains, and plenty of simple starch grains in one parenchyma cell compared to that of A. galanga. Diamond-shaped silica crystals were found only on A. galanga rhizome powder. Present study has revealed an easy technique to identify two similar medicinal plant materials microscopically and this method can also be employed to detect the degree of adulteration in powdered raw medicinal plant materials as well.
PMCID: PMC3665095  PMID: 23723656
Adulteration; Alpinia galanga; Alpinia calcarata; microscopy

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