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   2016| January-June  | Volume 7 | Issue 1  
    Online since April 13, 2016

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Cichorium intybus: A concise report on its ethnomedicinal, botanical, and phytopharmacological aspects
Sneha Das, Neeru Vasudeva, Sunil Sharma
January-June 2016, 7(1):1-12
Cichorium intybus L. (chicory) is a Mediterranean plant species belonging to the Asteraceae family. Chicory is gaining interests because of its culinary features, nutritional values and medicinal characteristics. C. intybus has been implemented in folk medicine from North Africa to South Asia for several 100 years. In Indian medicine, it has been used to treat fever, diarrhea, spleen enlargement, jaundice, liver enlargement, gout, and rheumatism. In China, it is valued for its tonic effects upon the liver and digestive tract. In Germany, chicory has been used as a folk medicine for everyday ailments. Thus, C. intybus is a plant of great economic potential due to high concentrations of fructooligosacharide, known as inulin, in its roots, used as a replacement ingredient for sugar and fat. The other various phytoconstituents reported in chicory are sucrose, cellulose, proteins, caffeic acid derivatives, flavonoids, polyphenols, carotenoids, anthocyanins, tannins, coumarins, sesquiterpene lactones, fatty acids, pectin, cholins, benzo-isochromenes, alkaloids, vitamins, amino acids, and minerals. The therapeutic investigations reveal that C. intybus is useful for maintaining normal health and has nematicidal, antihepatotoxic, antidiabetic, cardioprotective, antiallergic, antihyperlipidemic, anti-inflammatory, antineoplastic, calcium homeostater, bulking agent, immunostimulatory, prebiotic, protective against pancreatitis, antimicrobial, and antioxidant effects. This review encompasses botany, ethnomedicinal uses, phytoconstituents, pharmacological uses, and toxicity studies of C. intybus L.
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Strain improvement of Phaffia rhodozyma for astaxanthin production
Hina Nangia, Mojeer Hasan, Mohd. Azhar , Prakash Chandra Bhatt, Bibhu Prasad Panda
January-June 2016, 7(1):63-68
Objective: The aim of the present study is to evaluate and optimize different carbon sources at different temperatures in shake flask for cell growth and astaxanthin accumulation in ultraviolet (UV) mutant Phaffia rhodozyma. The focus is to make the process cost-effective and fruitful with higher productivity. Materials and Methods: Adaptive strain development was carried out so that the P. rhodozyma can give the best astaxanthin at a higher temperature (35°C), increasing the process economy. P. rhodozyma was cultivated with three broad spectrum antibacterial drugs - streptomycin, gentamycin, and ampicillin (200 μL) and with antifungal drug - fluconazole (200 μL) to determine the effect on yeast growth and astaxanthin production. Rice, cane juice, and sucrose were selected as carbon source and compared with dextrose at different fermentation temperatures - 22°C, 30°C, 33°C, and 35°C for astaxanthin production. Results: P. rhodozyma was resistant to antibiotic drugs inhibiting bacterial and fungal contaminants. Rice being more economical was selected as carbon source, but the strain was not able to utilize starch-rich substrate leading to its rejection. When P. rhodozyma was grown in cane juice, biomass is highest (OD 2.77) at 30°C and 610 nm wavelength, whereas astaxanthin productivity is highest (OD 2.64) in dextrose media at 30°C and 450 nm. Comparative metabolic and growth results for UV mutants showed significantly higher biomass and astaxanthin productivity in comparison to wild strain. Conclusion: The most efficient carbon source in terms of penny-pinching and astaxanthin productivity was found to be dextrose containing media at 30°C.
  3,709 489 1
In vitro α-glucosidase and α-amylase inhibition by aqueous, hydroalcoholic, and alcoholic extract of Euphorbia hirta L.
Manjur Ali Sheliya, Rayhana Begum, Kolappa Krishna Pillai, Vidhu Aeri, Showkat Rasool Mir, Abuzer Ali, Manju Sharma
January-June 2016, 7(1):26-30
Background: Euphorbia hirta L. (Euphorbiaceae), commonly known as Dudhani, is distributed in the warm region of India and China. Traditionally, it is used in respiratory and gastrointestinal disorders. In addition, the antidiabetic property of the plant was also reported in the literature. This study was designed to evaluate the effect of aqueous, hydroalcoholic, and methanolic extracts (MEHs) of E. hirta on α-glucosidase and α-amylase in vitro. Materials and Methods: Aqueous, hydroalcoholic, and MEHs of E. hirta were prepared as per application program interface. In α-glucosidase activity, α-glucosidase (0.1 μ/mL) and substrate, 2.5 mM p-nitrophenyl-α-D-glucopyranoside was used; absorbance was recorded at 405 nm. In α-amylase activity, α-amylase solution (1.0 μ/mL) and substrate, 0.25% starch were used, and absorbance was measured at 540 nm. The IC50values were calculated by linear regression. Results: All the extracts showed α-glucosidase inhibitory activity comparable to acarbose with MEH having highest inhibitory activity among tested extracts. The observed IC50values were 213.63, 146.9, 78.88, and 8.07 μg/mL for aqueous, hydroalcoholic, MEH, and acarbose, respectively. All the extracts have shown mild α-amylase inhibitory activity compared to acarbose. Lineweaver–Burk plot has shown that the MEH is a mixed noncompetitive inhibitor for α-glucosidase enzyme. Conclusion: The results from this in vitro study clearly indicated that MEH of E. hirta had strong inhibitory activity against α-glucosidase and mild inhibitory activity against α-amylase. It can be used for management of postprandial hyperglycemia with lesser side effects, and provide a strong rationale for further animal and clinical studies.
  3,305 445 6
Quality control analysis of Khamira Abresham Hakim Arshad Wala and Khamira Marwareed
Masood Shah Khan, Washim Khan, Wasim Ahmad, Mhaveer Singh, Sayeed Ahmad
January-June 2016, 7(1):47-50
Background: Unani system of medicine is based on the principles proposed by Galen, a Greek practitioner. Since then, many Arab and Persian scholars have contributed to the system. Among them, Ibn-e-Sina, an Arab philosopher and Physicist, who wrote “Kitab-al-shifa” are worth mentioning. This system has an extensive and inspiring record in India. Khamira Abresham Hakim Arshad Wala (KAHAW) and Khamira Marwareed (KM) are semisolid sugar-based preparations with potent cardiac tonic and well-known antioxidant properties. It has been studied that therapeutic intervention of KAHAW may be used to prevent or to decrease the deterioration of cognitive function and neurobehavioral activities, often associated with the generation of a free radical. However, no considerable attempt has been made for quality evaluation of KAHAW. Materials and Methods: In the present investigations, KAHAW and KM were evaluated for their phytochemical screening, physicochemical standards, presence of minerals, contaminants such as aflatoxins, pesticide residues, thin layer chromatography, and high performance thin layer chromatography (HPTLC) as per WHO and AYUSH guidelines for development of quality standards, which can be used by Unani industries. Results: Physicochemical parameters of the prepared compound formulation KAHAW and KM were studied such as total ash, acid insoluble ash, water soluble ash, solubility matter in alcohol and water, and loss on drying at 105°C, pH as per the methods described in WHO guidelines. The heavy metal, aflatoxins, and pesticide residues were also determined for safety evaluation. The total phenolic and flavonoid contents were estimated using catechin and quercetin as standard after preparing calibration plot, respectively. HPTLC fingerprinting of hydrolyzed chloroform extracts of KAHAW and KM was carried out to determined a number of spots present in them. Conclusions: This study on pharmacognostical standardization of KAHAW and KM will provide useful information for its identification and quality control and can be applied by different manufacturers of these formulations.
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Doxorubicin prodrug for cytoplasmic and nuclear delivery in breast cancer cells
Animikh Ray, Kishore Cholkar, Ravinder Earla, Ashim K Mitra
January-June 2016, 7(1):13-19
Purpose: The objective of this study was to develop a novel peptide prodrug of doxorubicin which can evade over-expressed efflux pumps in breast cancer cells. This approach may lead to increased uptake and higher drug accumulation in nuclei of cancer cells. Materials and Methods: L-val-L-val doxorubicin prodrug was synthesized following standard f-moc chemistry. The prodrug was analyzed for stability, cellular and nuclear uptake and interaction with efflux and peptide transporters. Breast cancer cells (T47D) were grown on polystyrene 12-well plates. Result: The prodrug Val-Val-doxorubicin was found to be very stable in breast cancer cell homogenate. It was able to evade efflux pumps. The prodrug penetrated cytoplasm and nucleus of cancer cells by interacting with peptide transporters.These transporters (pepT1 and pepT2) are expressed both on plasma and nuclear membrane of breast cancer cells. Uptake of prodrug was found to be 10 times more than parent drug. Conclusion: Peptide prodrug derivatization of doxorubicin has potential to evade efflux pumps and increase availability and nuclear accumulation of doxorubcin in breast cancer cells.
  2,480 364 3
Quality control analysis of Safoof-e-Pathar phori: Antiurolithiatic formulation
Wasim Ahmad, Washim Khan, Masood S Khan, Mohd Mujeeb , SM Arif Zaidi, Sayeed Ahmad
January-June 2016, 7(1):20-25
Aim: Safoof-e-Pathar phori (SPP), a Unani polyherbomineral formulation used for antilithiatic activity. The present study involves standardization of SPP to assess the quality. SPP were subjected to pharmacognostic studies, physiochemical properties, phytochemical analysis, high-performance thin layer chromatography (HPTLC), high-performance liquid chromatography (HPLC), and gas chromatography-mass spectrometry (GC-MS) fingerprint profile to set the standards, which can be used as reference of quality by herbal industries for its preparation and human use. Materials and Methods: The quality control of SPP has been carried out as per the AYUSH and WHO guidelines. The HPTLC fingerprinting has been done using chloroform: ethyl acetate (9:2, v/v) for petroleum ether extract, chloroform: toluene: ethyl acetate (7:2:4, v/v/v) for chloroform extract and for methanol extract petroleum ether: ethyl acetate (9:2, v/v) was used as mobile phase. HPLC was carried out using mobile phase consisted of acetonitrile and water in the ratio of 50:50 (v/v) for the methanolic extract. GC-MS fingerprinting profile has been carried out using hexane extract. Result: SPP was subjected to qualitative estimation of phytochemicals using standard methods, which revealed the presence of various bioactive components such as anthraquinone glycosides, carbohydrates, resins, proteins, flavonoids, phenolics, tannins, and terpenoids. The quantitative estimation of total phenolics and flavonoid content showed 0.44 mg/g and 1.02 mg/g, respectively. The HPTLC fingerprint showed presence of number of compounds for extracts at different Rfvalues. However, HPLC fingerprinting showed presence of 23 well-separated compounds and GC-MS showed presence of 22 compounds. Conclusion: The quality control parameters in present study reveal complete standardization profile of SPP for the 1st time, which would be of immense value in checking quality of developed formulation for human use.
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Antimicrobial activity of sugar-based semisolid polyherbal ayurvedic formulation: Chyawanprash
Mohammad Ahmed Khan, Satyajyoti Kanjilal, Arun Gupta, Sayeed Ahmad
January-June 2016, 7(1):31-33
Objective: This study evaluated the antimicrobial activity of chloroform (CHCl3) extracts of chyawanprash (Dabur India Ltd., Uttar Pradesh, India) and hydrolyzed chyawanprash against Escherichia coli and Staphylococcus aureus on nutrient agar media using cup plate method. Materials and Methods: CHCl3extracts of chyawanprash were prepared directly as well as by previously hydrolyzing with dilute HCl. Three test groups of 10, 20, and 50 mg/ml were prepared for each CHCl3extract, whereas ampicillin 20 mg/ml was used as standard group. Further, six agar plates were prepared for each group, i.e., three for E. coli and three for S. aureus. Cups were bored on each plate with the help of steel cup borer and cups on each plate were marked for identification. Extracts of different concentrations (i.e., 10, 20, and 50 mg/ml) were poured into cups and all plates were then incubated at 37°C for 48 h. After incubation, plates were observed for bacterial growth and zone of inhibition were measured. Results: Results showed that both CHCl3as well as hydrolyzed CHCl3extract of chyawanprash showed concentration-dependent antimicrobial activity. The largest zone of inhibition was observed at the concentration of 50 mg/ml of both CHCl3extracts of chyawanprash (12.7 ± 1.5 mm for E. coli and 15.0 ± 1.0 mm for S. aureus) and hydrolyzed chyawanprash (14.3 ± 0.6 mm for E. coli and 16.3 ± 0.6 mm for S. aureus). Conclusion: Chyawanprash possesses promising potential for use as an antimicrobial agent.
  2,615 176 1
In vitro anticancer potential of Semecarpus anacardium Linn
Md. Nasar Mallick, Washim Khan, Mhaveer Singh, Mohd. Zeeshan Najm, Mohammad Kashif, Sayeed Ahmad, Syed Akhtar Husain
January-June 2016, 7(1):55-58
Background: Keeping in view the toxicity of Semecarpus anacardium Linn. as reported in the traditional literature, the present study was carried out to evaluate the in vitro cytotoxic activity of ethanolic extract of Semecarpus on two different cell lines. Materials and Methods: The ethanolic extract of Semecarpus was prepared using cold extraction method. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay of ethanolic extract was carried out on HeLa and SiHa cell lines for determination of cytotoxicity. Results: The IC50values of ethanolic extract of S. anacardium in HeLa and SiHa cell lines were 44.0 μg/ml and 57.0 μg/ml, respectively. The extract was standardized by thin-layer chromatography and Gas chromatography-mass spectrometry. Conclusion: The results showed good cytotoxic activity in the ethanolic extract of S. anacardium in both the cell lines may be due to the presence of toxic flavones.
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Simultaneous determination of 18α-glycyrrhetinic acid and 18β-glycyrrhetinic acid in Glycyrrhiza glabra root by reversed phase high-performance liquid chromatography
Ambika Chamoli, Makhmur Ahmad, Mojeer Hasan, Bibhu Prasad Panda
January-June 2016, 7(1):59-62
Background: The aim of the present research work is to develop a high-performance liquid chromatography (HPLC) method for simultaneous analysis of 18α-glycyrrhetinic acid (18α-GA) and 18β-GA (18β-GA) of Glycyrrhiza glabra. Materials and Methods: About 20 μL aliquots of each 18α-GA and 18β-GA were analyzed using reversed-phase C-18 column. The mobile phase was acetonitrile:tetrahydrofuran:water (10:80:10, v/v/v). The run time was 10 min at flow rate of 1 ml/min. Ultraviolet detection was carried out at 254 nm. Results: 18α-GA and 18β-GA were well resolved in reversed phase C-18 column using mobile phase acetonitrile: tetrahydrofuran: water (10:80:10, v/v/v, pH 7.9). The Rtof 18α-GA and 18β-GA was detected at 2.091 and 2.377 min, respectively. Conclusion: The developed chromatography method could be extended for potential quantification or simultaneous determination of these markers in plant as well as in herbal formulation.
  2,204 233 1
Screening of two new herbal formulations in rodent model of urolithiasis
Mohammad Ahmed Khan, Satyendra Kumar, Arun Gupta, Sayeed Ahmad
January-June 2016, 7(1):34-38
Background: Kidney stone formation or urolithiasis is a complex process that is a consequence of an imbalance between promoters and inhibitors in the kidneys. The recurrence of urolithiasis also represents a serious problem in patients. Not all standard pharmaceutical drugs used to prevent urolithiasis are effective in all patients, and many have adverse effects. The present study was undertaken to evaluate the antiurolithiatic potential of two new herbal formulations DRDC/AY/8080 (tablet) and DRDC/AY/8081 (syrup) against 28-day ethylene glycol (EG)-induced urolithiasis model in Wistar rats. Materials and Methods: Animals were divided into five groups (n = 6). The control group was given normal saline, and the toxicant group was given 0.75% EG with 1% w/v of ammonium chloride (AC) for 10 days followed by 0.75% w/v EG for next 18 days in drinking water. Treatment groups received respective oral co-treatment with DRDC/AY/8080 (265 mg/kg), DRDC/AY/8081 (2.65 ml/kg), and standard (2.65 ml/kg) for 28 days along with EG and AC as given in toxicant group. After 28th day urine, blood and kidney tissue were collected. Ca2+, Mg2+, Na+, and K+ levels were estimated in urine, creatinine, and urea levels were estimated in serum whereas the extent of lipid peroxidation was measured in kidney tissue. Further, crystalluria and histopathological evaluation were carried out in urine and kidney tissue, respectively. Results: Toxicant group showed significant elevation (P < 0.001 vs. control) in serum creatinine, blood urea, tissue lipid peroxide, and urinary Mg2+ levels and significant reduction in (P < 0.001 vs. control) urinary Na+ and Ca2+ levels. Histopathology of the toxicant group showed damaged proximal tubules with deposits of refractile crystals and loss of tubular epithelium. Both tablet and syrup treated groups showed nephroprotective activity as evident from lower serum creatinine, blood urea, and lipid peroxide levels. Treatment with tablet and syrup formulations also showed significant (P < 0.001 vs. toxicant) elevation in urinary Na+, Ca2+, and reduction in Mg2+ levels. Histologically, both tablet/AY/8080) and syrup treatment showed protected against urolithiasis and nephrotoxicity. Conclusion: It can be concluded that the two herbal formulations DRDC/AY/8080 and DRDC/AY/8081 possess significant potential in the management of renal calculi.
  2,160 194 -
First in human trials: Are there any benefits of including placebo treatments?
Akosua Adom Agyeman, Richard Ofori-Asenso
January-June 2016, 7(1):73-74
  2,109 228 -
Standardization of Unani polyherbal formulation, Qurse-e-Hummaz: A comprehensive approach
YT Kamal, Mhaveer Singh, Shahana Salam, Sayeed Ahamd
January-June 2016, 7(1):39-42
Background: An increase in the awareness about the advantages of the traditional system of medicines has led to the commercialization of the formulations used for the treatments. Manufacture of these medicines to meet this increasing demand has resulted in a decline in their quality, primarily due to a lack of adequate regulations pertaining to this sector of medicine. Hence, it is necessary to come up with a systematic approach to develop well-designed methodologies for the standardization of polyherbal formulations which are used in traditional systems of medicine. Materials and Methods: Qurs-e-Hummaz formulations were prepared by a qualified “Hakim” (Unani medical practitioner) of Faculty of Unani Medicine, Hamdard University, as per the formula and instruction given in National Formulary of Unani Medicine. Results: Various quality control parameters such as organoleptic evaluations (color, odor, taste, and consistency), physicochemical evaluations (loss on drying, disintegration time, moisture content, total ash, acid insoluble ash, water soluble ash, pH of 1 and 10% solution, extractive values, water soluble matter, alcohol-soluble matter, and total phenolic content) and thin layer chromatography fingerprint profiling have been carried out in triplicate. The evaluation of contaminants such as heavy metals, aflatoxins, pesticide residues, and microbial contamination has also been carried out in the formulation. Conclusion: Help in maintaining the quality and batch to batch consistency of many important polyherbal formulations.
  2,098 172 -
Quantification of vasaka alkaloids in in vitro cultures and in natural leaves from Indian subcontinents by reversed phase- high performance liquid chromatography
Madhukar Garg, Ennus Tajuddin Tamboli, YT Kamal, SH Ansari, Sayeed Ahmad
January-June 2016, 7(1):51-54
Background: The present study was designed to develop an efficient protocol for studying the enhancement of in vitro production of secondary metabolites in Adhatoda vasica leaves collected from the varied geographical locations of Indian sub-continents by a validated simultaneous high-performance liquid chromatography gradient method for the quantification of vasicine and vasicinone in the developed calli and crude samples. Materials and Methods: The analysis was carried out on a reverse phase C18column using 0.1% of orthophosphoric acid: Acetonitrile in gradient manner and carried out the detection at 280 nm wavelength keeping the flow rate of 1.0 mL/min. Results: The simultaneous method was found linear with regression coefficient r2 = 0.991 in a wide range (100–500 μg/mL) precise, accurate, and robust for quinazoline alkaloids in samples. Results clearly showed a significant increase in the concentration of alkaloids in in vitro cultures as compared to natural ones. The proposed method was validated as per the International Conference on Harmonization guidelines for accuracy, precision, robustness, limit of detection, and limit of quantitation. Conclusion: The developed method was found suitable for quality control of A. vasica and for the analysis of vasicine and vasicinone in any type of sample.
  2,050 200 2
Quality control of an antipsoriatic ayurvedic herbal Formulation: Lajjalu Keram
MT Athar, SM Musthaba, AK Al-Asmari, S Baboota, J Ali, Sayeed Ahmad
January-June 2016, 7(1):43-46
Background: Psoriasis is an autoimmune disorder, which affects a large group of human population of world (3%). Till date, there is no treatment for psoriasis except some herbal drugs and its constituents. Since Ayurveda is the main traditional system of medicine in India, here, we have selected one ayurvedic formulation - Lajjalu Keram, which has been used since long for their quality control. Methods: Total microbial load of formulations were carried out for total fungal count and total bacterial count. Lajjalu Keram was also tested by high-performance liquid chromatographic (HPLC) for aflatoxins (B1, B2, G1, and G2), which showed its presence below the permissible limit; similarly, pesticides residues were analyzed using gas chromatography/mass spectrometry for organophosphates and organochlorides, which showed that pesticides were below detection limit (0.1 ppb). The content of heavy metals was analyzed using AAS, which demonstrated the presence of cadmium, lead, and arsenic below permissible limit, whereas mercury was found absent. Results: The result of quality control analysis showed the presence of alkaloids, tannins, carbohydrate, saponins, proteins and amino acids, lipid/fats, phenolic compounds, and flavonoids in formulation. The dermal toxicity (LD50) of Lajjalu Keram in Wistar rats was found more than 2000 mg/kg (safe for the management of psoriasis). Formulation was also analyzed for their composition of fatty acids. It was found to have 13 fatty acids, out of which, seven were saturated fatty acids (95.2%) and the rest were unsaturated fatty acids (3.27%). A rapid HPLC method for quantification of mimosine (an unusual amino acid present in formulation) has been developed and validated. The mimosine content in Lajjalu Keram was found to be 0.0070% w/w with % relative standard deviation of 0.41. Conclusion: The formulation afforded significant and better protection of carrageenan-induced rat paw edema (72.11% inhibition) as compared to control. The shelf life studies on antipsoriatic herbal formulations were carried out for 6 months at different time intervals. No significant variation in analysis parameters was observed on the storage of formulations up to 6 months. The assay of constituents using mean curve of sigma plot showed 44.2 month of shelf life for Lajjalu Keram.
  2,051 176 -
Screening and bioconversion of glycyrrhizin of Glycyrrhiza glabra root extract to 18β-glycyrrhetinic acid by different microbial strains
Makhmur Ahmad, Jalaluddin , Mohammad Ali, Bibhu Prasad Panda
January-June 2016, 7(1):69-72
Objective: The objective of the present study is to perform screening of different microorganisms (7 bacteria and 14 fungi) for conversion of glycyrrhizin (GL) to 18β-glycyrrhetinic acid (GA). Penicillium chrysogenum produced the highest concentration of β-glucuronidase enzyme (61 U/mL) and produced GA of 52 μg/mL while E. coli produced the highest β-glucuronidase of 376 U/mL with GA concentration of 2.1 μg/mL. Materials and Methods: Submerged and solid state biotransformation of GL was carried out. To 9.0 mL of bacterial supernatant, 1.0 mL 0.2% w/v of aqueous Glycyrrhiza glabra root extract was added and incubated at 37°C for 24 h. β-glucuronidase activity was measured and high-performance liquid chromatography analysis was carried out. Results and Discussion: Induced-Escherichia coli produces 2.1 μg/mL of GA with an enzyme activity of 376 U/mL which shows that the enzyme has a potential biotransformation capability. Rhizopus oryzae and P. chrysogenum have the potential ability to biotransform GL to GA with 2.6 μg/mL and 61 μg/mL of GA with enzyme activity of 569 U/mL and 61 U/mL, respectively. Conclusions: G. glabra roots containing GL can be hydrolyzed by microbial β-glucuronidase enzyme under sub-merged fermentation (SmF). β-glucuronidase, an enzyme of E. coli, was found to be the best microbial source of enzyme which biocatalyzed the reaction than fungal strain under SmF.
  2,008 193 -