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Table of Contents
July 2016
Volume 12 | Issue 47 (Supplement)
Page Nos. 389-496
Online since Friday, September 30, 2016
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ORIGINAL ARTICLES
Human cytochrome P450 enzyme modulation by
Gymnema sylvestre
: A Predictive Safety evaluation by LC-MS/MS
p. 389
Bera Rammohan, Karmakar Samit, Das Chinmoy, Saha Arup, Kundu Amit, Sarkar Ratul, Karmakar Sanmoy, Adhikari Dipan, Sen Tuhinadri
DOI
:10.4103/0973-1296.191441
PMID
:27761064
Background:
Traditionally GS is used to treat diabetes mellitus. Drug-herb interaction of GS via cytochrome P450 enzyme system by substrate cocktail method using HLM has not been reported.
Objective:
To evaluate the
in-vitro
modulatory effects of GS extracts (aqueous, methanol, ethyl acetate, chloroform and
n
-hexane) and deacylgymnemic acid (DGA) on human CYP1A2, 2C8, 2C9, 2D6 and 3A4 activities in HLM.
Material and Methods:
Probe substrate-based LCMS/MS method was established for all CYPs. The metabolite formations were examined after incubation of probe substrates with HLM in the presence or absence of extracts and DGA. The inhibitory effects of GS extracts and DGA were characterized with kinetic parameters IC50 and Ki values.
Results:
GS extracts showed differential effect on CYP activities in the following order of inhibitory potency: ethyl acetate > Chloroform > methanol >
n
-hexane > aqueous > DGA. This differential effect was observed against CYP1A2, 2C9 and less on CYP3A4 and 2C8 but all CYPs were unaffected by aqueous extract and DGA. The ethyl acetate and chloroform extract exhibited moderate inhibition towards CYP1A2 and 3A4. The aqueous extract and DGA however showed negligible inhibition towards all five major human CYPs with very high IC50 values (>90μg/ml).
Conclusion:
The results of our study revealed that phytoconstituents contained in GS, particularly in ethyl acetate and chloroform extracts, were able to inhibit CYP1A2, 3A4 and 2C9. The presence of relatively small, lipophillic yet slightly polar compounds within the GS extracts may be attributed for inhibition activities. These suggest that the herb or its extracts should be examined for potential pharmacokinetic drug interactions
in vivo
.
Abbreviations used:
GS:
Gymnema sylvestre
, GSE:
Gymnema sylvestre
extract, DGA: deacyl gymnemic acid, CYP: cytochrome P450, DMSO: dimethylsulphoxide, HLM: human liver microsomes, LC-MS/MS: liquid chromatography tandem mass spectroscopy, NADPH: reduced nicotinamide adeninedinucleotide phosphate, NRS: nicotinamide adeninedinucleotide phosphate regenerating system, CHE: chloroform extract, EAE: ethyl acetate extract, NHE-
n
-hexane extract, AE: aqueous extract, ME: methanol extract,
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Evaluation of the genotoxicity and cytotoxicity of semipurified fractions from the Mediterranean brown algae,
Dictyopteris membranacea
p. 395
Najoua Akremi, Davie Cappoen, Roel Anthonissen, Abderrahman Bouraoui, Luc Verschaeve
DOI
:10.4103/0973-1296.188318
PMID
:27761065
Dictyopteris membranacea
, a species of Mediterranean brown algae,is believed to have potential pharmacological and nutritional applications. However, such potentials only make sense when devoid of any adverse health consequences. The present study should be seen in this context. It aimed at evaluating the genotoxicity and cytoxicity of its organic extract (F0) and semi purified fractions (F
4
, F
5
, and F
6
).Extracts were tested using the bacterial Vitotox
®
test and micronucleus assay in different concentrations (from 1.25 μg/mL up to 100 μg/mL, depending on the test and the extract). Applied concentrations were based on a preliminary dose-finding test with the neutral red uptake assay. The results show that all extracts were not genotoxic in the presence or absence of a rat metabolic enzyme fraction (S9). This is encouraging and justifies further investigations on the therapeutic and other values of this algae.
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In vitro
accumulation of polyphenols in tea callus derived from anther
p. 400
Naga Pavan Kumar Chevala, Naga Thirumalesh Chevala, Kirubakaran Dhanakodi, Rama Rao Nadendla, Chandrashekara Krishnappa Nagarathna
DOI
:10.4103/0973-1296.191442
PMID
:27761066
Background:
Tea is an economic important crop with high medicinal value due to rich polyphenols content. In the present research we studied the accumulation of polyphenols of
in vitro
regenerated callus from anthers.
Objective:
Callus induction of tea anthers and
in vitro
accumulation of phenolic compounds from the anther-derived callus.
Materials and Methods:
Standardization of callus induction for tea anthers.
In vitro
generated callus was screened for
in vivo
accumulation of catechins and its isomers were screened by FC reagent staining technique. The methanol extract of dry and green callus obtained were estimated qualitatively by Fourier transform infrared spectroscopy (FTIR)-alternative total reflection (ATR) and quantitatively by HPLC method.
Results:
Anthers inoculated on half strength MS media fortified with 2,4-dichloro acetic acid (2 mg/L), Kn (1 mg/L), and BAP (1 mg/L) induced callus under photoperiod of 9:15 h light. The
in vivo
histochemical studies revealed the accumulation of polyphenols in the callus. The
in vitro
generated fresh and dry callus were used for extraction and screened for accumulated polyphenols [galic acid, (+)-catechin (C), (−)-epicatechin, (−)-epigallocatechin, (−)-epigallocatechin gallate, (−)-gallocatechins, (−)-epicatechin gallate] were estimated qualitatively by FTIR-ATR method and quantitatively by HPLC method.
Conclusion:
The FC staining technique used here helps in localization of polyphenol compounds accumulation in the tissues by instant microscopic studies. The study have scope in large-scale isolation of various medicinally important flavonol by using anther culture.
Abbreviations used:
HPLC: high pressure liquid chromatography; FTIR: Fourier transform infrared spectroscopy; 2,4-D: 2,4-dichloro acetic acid; BAP: N
6
-benzyl amino purine; kn: kinetin
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Genetic and chemical profiling of
Gymnema sylvestre
accessions from central India: Its implication for quality control and therapeutic potential of plant
p. 407
Ashutosh Kumar Verma, Sunita Singh Dhawan, Seema Singh, Kumar Avinash Bharati, Jyotsana
DOI
:10.4103/0973-1296.191443
PMID
:27761067
Background:
Gymnema sylvestre
, a vulnerable plant species, is mentioned in Indian Pharmacopeia as an antidiabetic drug
Objective:
Study of genetic and chemical diversity and its implications in accessions of
G. sylvestre
Materials and Methods:
Fourteen accessions of
G. sylvestre
collected from Central India and assessment of their genetic and chemical diversity were carried out using ISSR (inter simple sequence repeat) and HPLC (high performance liquid chromatography) fingerprinting methods
Results:
Among the screened 40 ISSR primers, 15 were found polymorphic and collectively produced nine unique accession-specific bands. The maximum and minimum numbers of amplicones were noted for ISSR-15 and ISSR-11, respectively. The ISSR -11 and ISSR-13 revealed 100% polymorphism. HPLC chromatograms showed that accessions possess the secondary metabolites of mid-polarity with considerable variability. Unknown peaks with retention time 2.63, 3.41, 23.83, 24.50, and 44.67 were found universal type. Comparative hierarchical clustering analysis based on foresaid fingerprints indicates that both techniques have equal potential to discriminate accessions according to percentage gymnemic acid in their leaf tissue. Second approach was noted more efficiently for separation of accessions according to their agro-climatic/collection site
Conclusion:
Highly polymorphic ISSRs could be utilized as molecular probes for further selection of high gymnemic acid yielding accessions. Observed accession specific bands may be used as a descriptor for plant accessions protection and converted into sequence tagged sites markers. Identified five universal type peaks could be helpful in identification of
G. sylvestre
-based various herbal preparations.
Abbreviations used:
HPLC: High Performance Liquid Chromatography, ISSR: Inter Simple Sequence Repeats, CTAB: Cetyl Trimethylammonium Bromide, DNTP: Deoxynucleotide Triphosphates
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Potential therapeutic agents for the treatment of fatty degeneration of liver and atheromatous plaques: An experimental study in rats
p. 414
Sibghatullah Muhammad Ali Sangi
DOI
:10.4103/0973-1296.191444
PMID
:27761068
Background:
Since long high fat diet (HFD) is being blamed for causing fatty degeneration of liver and formation of atheromatous plaques. At present, no proper pharmacotherapy is available for both the conditions. In this study, different substances containing monounsaturated fatty acids were used to observe their protective effects in the HFD induced damage to liver and coronary vessels.
Objectives:
To discover effective therapeutic agents for HFD induced fatty degeneration of liver and atheromatous plaques.
Materials and Methods:
The study was conducted from September 2015 to April 2016. In this study, rats were divided into nine groups according to dietary regimen. Each group comprised six rats. Saturated fat was given in the form of butter, and unsaturated fat was given in the form of corn oil, olive oil,
Nigella sativa
oil, and crushed garlic. Serum samples were taken to estimate lipid profile, liver functions, cardiac functions, and kidney functions. Visceras were removed after animal sacrifice, and histopathological examination was done.
Results and Conclusion:
During the study period, the weight of animals changed significantly in some groups. Those animals which were given crushed garlic along with high saturated fat diet, showed protection against accumulation of lipids in the hepatocytes. Olive oil and
Nigella sativa
oil were comparatively less effective.
Abbreviations used:
HFD: High Fat Diet; NS: Nigella Sativa; TQ: Thymoquinone; KFMRC: King Fahad Medical Research Center; BUN: Blood Urea Nitrogen; BNF: Buffered Neutral Formalin; G: Group
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Influence of extracting solvent on pharmacological activity and cytotoxicity of
Polygonum minus
, a commonly consumed herb in Southeast Asia
p. 424
Parayil Varghese Christapher, Subramani Parasuraman, Palanimuthu Vasanth Raj, Sultan Ayesh Mohammed Saghir, Mohd. Zaini Asmawi, Murugaiyah Vikneswaran
DOI
:10.4103/0973-1296.191451
PMID
:27761069
Objective:
To investigate the antihyperlipidemic, antioxidant, and cytotoxic effect of aqueous and methanol extract of leaves of
Polygonum minus
.
Materials and Methods:
Acute antihyperlipidemic effect was studied on chemically induced hyperlipidemic rat model. Treated groups received aqueous and methanol extract of leaves of
P. minus
respectively (1000 mg/kg; oral) whereas standard treated group received atorvastatin (60 mg/kg; oral) for 3 consecutive days. Blood samples were collected at fixed intervals for lipid profile analysis. Antioxidant effects were studied using 1,1-diphenyl-2-picrylhydrazyl, 2,2-azinobis 3-ethylbenzothiazoline 6-sulfonate, and ferric reducing antioxidant power assays. The total flavonoids content and total phenolic contents were also estimated. Cytotoxicity of both extracts was studied on one normal and three cancer cell lines using 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay method.
Results:
The methanol extract showed significant reduction in total cholesterol (
P
< 0.001), triglycerides (
P
< 0.01), LDL (
P
< 0.05), VLDL (
P
< 0.01), atherogenic index (
P
< 0.001), and elevation of HDL (
P
< 0.05) levels than the aqueous extract. Similarly, the antioxidant investigations also demonstrated that the methanol extract had higher antioxidant capacity than aqueous extract. Both extracts were not toxic to normal (EA.hy926) as well as to cancer (HCT116, HT29, and HeLa) cells. Significant correlation was demonstrated between total phenolic and total flavonoids contents with the antioxidant activity but not with the antihyperlipidemic effect, suggesting other groups of chemical constituents may be mainly responsible for the antihyperlipidemic effect of this plant.
Conclusion:
The study demonstrated that the presence and extent of bioactivities are influenced by solvents used for extraction. This study confirmed the antihyperlipidemic effect of leaves of
P. minus
in acute hyperlipidemic rat model.
Abbreviations used:
CVDs: Cardiovascular diseases, LDL: Low-density lipoprotein, DDPH: 1 ,1-Diphenyl-2-picrylhydrazyl radical, TPTZ
:
2,4,6,-tris(1-pyridyl)-5-triazine, ABTS
:
2,2?-Azino-di-[3-ethylbenzthiazoline Sulfonate], HDL
:
High-density lipoprotein,
VLDL:
Very low-density lipoprotein,
TC:
Total cholesterol,
TG:
Triglycerides,
EC
50
:
Half maximal effective concentration,
LD
50
:
Median lethal dose.
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Phytochemicals and cytotoxicity of
Launaea procumbens
on human cancer cell lines
p. 431
Preeti Rawat, Lokesh M Saroj, Anil Kumar, Tryambak D Singh, SK Tewari, Mahesh Pal
DOI
:10.4103/0973-1296.191452
PMID
:27761070
Background:
The plant
Launaea procumbens
belongs to the family Asteraceae and traditionally used in the treatment rheumatism, kidney, liver dysfunctions and eye diseases. In the present study Phytochemical analysis and fractions of methanolic extract of
L. procumbens
leaves were tested
in vitro
for their cytotoxicity.
Objectives:
Phytochemical analysis and cytotoxic activity of methanolic extract and fractions of
Launaea procumbens
against four cancer cell lines K562, HeLa, MIA-Pa-Ca-2 and MCF-2 by SRB assay.
Materials and Methods:
Powdered leaves of
Launaea procumbens
were extracted sequentially with hexane, ethyl acetate, butanol and water by cold extraction. Phytochemical analysis and cytotoxicity assay were carried out for these fractions using SRB assay against four human cancer cell lines, namely leukemia (K562), cervix (HeLa), pancreatic (MIA-Pa-Ca-2) and breast (MCF-7).
Results:
Ethyl acetate extract exerts potent cytotoxicity against human leukemia (K562), cervix (HeLa) and breast (MCF-7) cell lines IC
50
value of 25.30±0.50, 19.80±0.10 and 36.90±4.90 μg/ ml respectively. Moderately cytotoxic effect found in hexane extract IC
50
value of 41±8 and 48.20±0.50 μg/ ml against leukemia (K562), and breast (MCF-7) cancer cell line respectively. The Chemical composition analyzed by GC-MS showed considerable differences in solvent fractions of
Launaea procumbens
.
Conclusion:
This study revealed the cytotoxic potential of ethyl acetate and hexane fractions of
L.procumbens
leaves on different cancer cell lines.
Abbreviations used:
SRB: Sulforhodamine B assay, MW: Molecular weight
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Inhibitory and cytotoxic activities of chrysin on human breast adenocarcinoma cells by induction of apoptosis
p. 436
Saeed Samarghandian, Mohsen Azimi-Nezhad, Abasalt Borji, Malihe Hasanzadeh, Farahzad Jabbari, Tahereh Farkhondeh, Mohammad Samini
DOI
:10.4103/0973-1296.191453
PMID
:27761071
Objectives:
Chrysin, an active natural bioflavonoid found in honey and many plant extracts, was first known for its antioxidant and anti-inflammatory effects. The fact that antioxidants have several inhibitory effects against different diseases, such as cancer, led to search for food rich in antioxidants. In this study, we investigated the antiproliferative and apoptotic effects of chrysin on the cultured human breast cancer cells (MCF-7).
Materials and Methods:
Cells were cultured in Roswell Park Memorial Institute medium and treated with different chrysin concentrations for three consecutive days. Cell viability was quantitated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The percentage of apoptotic cells was determined by flow cytometry using Annexin V-fluorescein isothiocyanate.
Results:
The MTT assay showed that chrysin had an antiproliferative effect on MCF-7 cells in a dose- and time-dependent manner. The 50% cell growth inhibition values for chrysin against MCF-7 cells were 19.5 and 9.2 μM after 48 and 72 h, respectively. Chrysin induced apoptosis in MCF-7 cells as determined by flow cytometry. Chrysin inhibits the growth of the breast cancer cells by inducing cancer cell apoptosis which may, in part, explain its anticancer activity.
Conclusion:
This study shows that chrysin could also be considered as a promising chemotherapeutic agent and anticancer activity in treatment of the breast cancer cells in future.
Abbreviations used:
Human breast cancer cells (MCF-7), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), phosphate-buffered saline (PBS), normal fi broblast mouse (L929).
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Sambar, an Indian dish prevents the development of dimethyl hydrazine-induced colon cancer: A preclinical study
p. 441
Vutturu Ganga Prasad, Neetinkumar Reddy, Albi Francis, Pawan G Nayak, Anoop Kishore, Krishnadas Nandakumar, Mallikarjuna C Rao, Rekha Shenoy
DOI
:10.4103/0973-1296.191454
PMID
:27761072
Background:
Colon cancer (CC) is the third commonly diagnosed cancer and the second leading cause of mortality in the US when compared to India where prevalence is less. Possible reason could be the vegetarian diet comprising spices used in curry powders. Researchers believe that 70% of the cases are associated with diet. Spices have inherited a rich tradition for their flavor and medicinal properties. Researchers have been oriented towards spices present in food items for their antitumorigenic properties.
Objective:
We investigated the effects of sambar as a preventive measure for 1,2-dimethyl hydrazine (DMH)-induced CC in Wistar albino rats.
Materials and Methods:
The animals were divided into three groups (
n
= 6) namely control, DMH, and sambar. At the end of the experimental period, the animals were killed using anesthesia and the colons and livers were examined.
Results:
All the treatment groups exhibited a significant change in the number of aberrant crypt foci (ACF). Sambar group showed a significant change in the colonic GSH when compared to both normal and DMH groups. A significant reduction in the liver GSH was noted in the sambar group. Only sambar group showed a significant change in the liver catalase levels when compared to DMH. There was a significant reduction in the colonic nitrite in the sambar-treated group; 2.94 ± 0.29 when compared to DMH control at 8.09 ± 1.32. On the contrary, a significant rise in the liver nitrite levels was observed in the sambar-treated rats.
Conclusion:
Sambar may prevent the risk of CC when consumed in dietary proportions.
Abbreviations used:
ACF: aberrant crypt foci, CC: colon cancer, DMH: 1,2-dimethyl hydrazine, GSH: glutathione, IL-6: Interleukin-6, TNF-α: Tumor necrosis factor-alpha.
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Antihepatotoxic effect and metabolite profiling of
Panicum turgidum
extract via UPLC-qTOF-MS
p. 446
Mohamed A Farag, Ahlam M El Fishawy, Sayed A El-Toumy, Khadiga F Amer, Ahmed M Mansour, Hala E Taha
DOI
:10.4103/0973-1296.191455
PMID
:27761073
Background:
Panicum turgidum,
desert grass, has not reported any detailed phytochemical or biological study as yet
Objective:
To establish
P. turgidum
secondary metabolite profile and to assess its antihepatotoxic effect
Materials and Methods:
Ultra-performance liquid chromatography (UPLC) coupled to quadrupole high-resolution time of flight mass spectrometry (qTOF-MS) was used for large-scale secondary metabolites profiling in
P. turgidum
extract, alongside assessing median lethal dose
(LD
50
)
and hepatoprotective effect against carbon tetrachloride (CCl
4
) intoxication
Results:
A total of 39 metabolites were identified with flavonoids as the major class present as
O/C
-glycosides of luteolin, apigenin, isorhamnetin and naringenin, most of which are first time to be reported in
Panicum
sp. Antihepatotoxic effect of
P. turgidum
crude extract was revealed via improving several biochemical marker levels and mitigation against oxidative stress in the serum and liver tissues, compared with CCl4 intoxicated group and further confirmed by histopathological examination.
Conclusion:
This study reveals that
P. turgidum
, enriched in
C
-flavonoids, presents a novel source of safe antihepatotoxic agents and further demonstrates the efficacy of UPLC-MS metabolomics in the field of natural products drug discovery.
Abbreviations used:
UPLC:
Ultra-performance liquid chromatography (UPLC), LD50: median lethal dose,
MDA:
malondialdehyde,
GSH:
glutathione reductase,
CAT:
catalase,
SOD:
superoxide dismutase,
ALT:
alanine aminotransferase,
AST:
aspartate aminotransferase,
ALP:
alkaline phosphatase,
TG:
triglycerides.
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Catha edulis
(khat) induces apoptosis in Madin-Darby bovine kidney cell line
p. 454
Hussein M Ageely, Ahmed E Agag, Syam Mohan, Atef Shehata
DOI
:10.4103/0973-1296.191456
PMID
:27761074
Background:
Khat (
Catha edulis
) is a controversial plant having a euphoretic effect, at the same time part of culture in many countries such as Africa and Arabian Peninsula. The presence of amphetamine-like substance, cathinone and cathine make this plant banned in many countries. Many neurological and other system related studies have been carried out in this plant, but the lack of toxicity studies are there especially the mechanism.
Objective:
In this study, Madin-Darby Bovine Kidney cell line was used as an
in vitro
model to study the cell death mechanism. Crude extract of fresh Khat plant leaves were prepared and exposed to cells.
Materials and Methods:
Trypan blue assay, phase-contrast microscopy, fluorescent microscopy, clonogenic assay, annexin-V assay, and hematoxylin and eosin (H and E) staining were employed to check the objectives.
Results:
Reductions in cellular viability were observed at concentrations above 1.25 mg/ml while using Trypan blue assay. The results of the clonogenic assay had shown that the untreated control with the highest number of colonies (100% survival) and the 0.1562 concentration could not prevent the colony formation significantly. The high concentrations reduced the colony formation at concentration dependent manner 27.4% and 24.9%, for 0.625 mg/ml and 1.25 mg/ml concentrations, respectively. The acridine orange/ethidium bromide experiment had observed the cells were intact with round nucleus while the apoptosis features such as blebbing and nuclear chromatin condensation were clearly observed in treatment. The shrinkage of cells was clearly observed in H and E staining.
Conclusion:
In addition, annexin-V binding confirmed the presence of apoptosis significantly on Khat treatment.
Abbreviation used:
PS: Phosphatidylserine (PS); MDBK: Madin-Darby Bovine Kidney; DMEM: Dulbecco's modified Eagle's medium; PI: propidium iodide; EB: ethidium bromide; PBS: Phosphate Buffer saline; FITC: fluorescein isothiocyante; TUNEL: Terminal deoxynucleotidyl transferase dUTP nick end labeling.
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Induction, subculture cycle, and regeneration of callus in Safed musli (
Chlorophytum borivilianum
) using different types of phytohormones
p. 460
Jaafar Juju Nakasha, Uma Rani Sinniah, Nurashikin Kemat, Kumara Swamy Mallappa
DOI
:10.4103/0973-1296.191457
PMID
:27761075
Background:
Chlorophytum borivilianum
is an industrially valued medicinal crop. Propagation through seeds is not feasible because of low germination percentage and long dormancy period. Therefore, callus culture and plant regeneration can be an alternative to improve this crop production. Also, callus can serve as an alternative source of bioactive compounds.
Objective:
To evaluate the effect of different phytohormones on callus induction, subculture cycle, and regeneration studies of callus in
C. borivilianum
.
Materials and Methods:
Young shoot buds of
C. borivilianum
were inoculated on Murashige and Skoog medium fortified with 3% sucrose and different concentrations (0, 1, 5, 10, and 15 mg/L) of either naphthalene acetic acid or 2,4-dichlorophenoxyacetic acid or indole-3-acetic acid and callus induction was evaluated up to four subcultures cycles. Shoot regeneration from callus was studied on Murashige and Skoog media fortified with 6-benzylaminopurine andkinetin or thidiazuron at varied levels (0, 0.5, 1, 2, and 3 mg/L). Microshoots were rooted on Murashige and Skoog media supplemented with 1.0 mg/L indole-3-butyric acid and plantlets were acclimatized before transferred to the natural conditions.
Results:
Callus induction was better evidenced on Murashige and Skoog media containing 5 mg/L 2,4-dichlorophenoxyacetic acid up to fourth subculture. Callus differentiated into shoots on Murashige and Skoog media fortified with 6-benzylaminopurine or kinetin, whereas thidiazuron completely failed to regenerate shoots. Furthermore, microshoots rooted on 1.0 mg/L indole-3-butyric acid containing Murashige and Skoog media. The rooted plantlets were successfully acclimatized and established in soil with 88.3% survivability.
Conclusion:
The type of auxins played an important role in inducing callus tissue from shoot bud explants of Safed musli. In future, this
in vitro
protocol could benefit in crop improvement programs and serve as a new source of bioactive compounds from Safed musli callus tissue for various therapeutic applications.
Abbreviations used:
MS: Murashige and Skoog, NAA: naphthalene acetic acid, 2,4-D: 2,4-dichlorophenoxyacetic acid, IAA: indole-3-acetic acid, BAP: 6-benzylaminopurine, Kn: Kinetin, TDZ: thidiazuron, IBA: indole-3-butyric acid, RCBD: Randomized Complete Block Design, DMRT: Duncan's Multiple Range Test
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Elicitation of phenolics from the micropropagated endangered medicinal plant
Calligonum polygonoides
L. (Polygonoaceae)
p. 465
Asmaa I Owis, Nada S Abdelwahab, Adel A Abul-Soad
DOI
:10.4103/0973-1296.191458
PMID
:27761076
Background:
Calligonum
polygonoides
L. subsp.
comosum
(L'Hér.) Sosk. is a plant species belonging to family Polygonaceae. Susceptibility to threaten, presence of various chemical constituents, and many medicinal effects reported for this plant in addition to rareness of
in vitro
culture studies have fuelled the need for its micropropagation and phytochemical investigations of the produced cultures.
Objectives:
To employ
in vitro
culture technique for
ex
situ
conservation of
C.
polygonoides
, using the fruit as an explant; establish callus and cell suspension cultures from
in vitro
germinated plantlets; investigate the production of phenolics through callus, redifferentiated shoot, and cell suspension cultures; attempt to enhance cell capacity to accumulate phenolics using salicylic acid and yeast extract and provide a brief demonstration of biosynthetic pathway leading to phenolic production.
Materials and Methods:
Modified Murashige and Skoog media supplemented with growth hormones such as kinetin, 1-naphthaleneacetic acid, 6-benzylaminopurine, and indole-3-acetic acid were used to establish callus, redifferentiated shoots, and cell suspension cultures. Elicitation of cell suspension culture was performed using salicylic acid and yeast extracts. A reversed phase-high performance liquid chromatography method for determination of phenolic content in the aforementioned cultures was developed.
Results:
The unorganized callus and cell suspension cultures contained fewer amounts of phenolic compounds than redifferentiated shoots. Elicitation produced massive quantitative reprogramming of phenolic content.
Conclusion:
The present study offers an alternative and renewable source for this valuable natural plant, provide a chance to improve secondary metabolite yield and serve as a useful tool for studying the biosynthesis of these compounds and its regulation in plant cells.
Abbreviation used:
H
2
O
2
: Hydrogen peroxide, Kin: Kinetin, NAA: Naphthaleneacetic acid, BAP: 6-benzylaminopurine, IAA: Indole-3-acetic acid, HPLC: High-performance liquid chromatography.
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Assessment of
In vitro
antibacterial activity and cytotoxicity effect of
Nigella sativa
oil
p. 471
Ayse Ruveyda Ugur, Hatice Turk Dagi, Bahadir Ozturk, Gulsum Tekin, Duygu Findik
DOI
:10.4103/0973-1296.191459
PMID
:27761077
Background:
Methicillin resistance is a serious health concern since it has spread among
Staphylococcus aureus
and coagulase-negative
Staphylococci
(CoNS) that are frequent community and nosocomial pathogens worldwide. Methicillin-resistant strains are often resistant to other classes of antibiotics, making their treatment difficult.
Nigella
sativa
oil is known to be active against Gram-positive cocci, yet its
in vitro
cytotoxicity is rarely investigated, is a proper and powerful candidate for treatment of methicillin-resistant isolates.
Objectives:
The aim of this study is to evaluate the
in vitro
antibacterial activity and cytotoxicity effect of
N.
sativa
oil.
Materials and Methods:
The minimal inhibitory concentrations (MICs) of
N.
sativa
oil were determined by broth microdilution method against four different American Type Culture Collection strains, 45 clinical isolates of methicillin-resistant
S.
aureus
(MRSA), and 77 methicillin-resistant CoNS (MRCoNS). The effects of different dilutions (0.25 μg/mL, 0.5 μg/mL, and 1 μg/mL) of
N.
sativa
oil on the proliferation of gingival fibroblasts were evaluated.
Results:
The MIC values of
N.
sativa
oil against clinical isolates of
Staphylococci
were between <0.25 μg/mL and 1.0 μg/mL. Compared to the control group, there was no cytotoxic effect on the proliferation of the gingival fibroblasts.
Conclusion:
In the present study, the oil of
N.
sativa
was very active against MRSA and MRCoNS and had no
in vitro
cytotoxicity at relevant concentrations. These findings emphasize that there is a requirement for further clinical trials on
N.
sativa
oil for "safe" medical management of infections caused by methicillin-resistant
Staphylococci
.
Abbreviation used:
ATCC: American Type Culture Collection; CLSI: Clinical and Laboratory Standards Institute; CoNS: Coagulase-negative Staphylococci; DMEM: Dulbecco's modified Eagle's medium; DMSO: Dimethyl sulfoxide; FBS: Fetal bovine serum; HGF: Human gingival fi broblast; MIC: Minimal inhibitory concentration; MRCoNS: Methicillin-resistant CoNS;MRSA: Methicillin-resistant S. aureus
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Postprandial antihyperglycemic and antioxidant activities of
Acalypha indica
Linn stem extract: An
in-vivo
study
p. 475
Charles Lekhya Priya, KV Bhaskara Rao
DOI
:10.4103/0973-1296.191461
PMID
:27761078
Background:
α-glucosidase inhibitors controls postprandial hyperglycemia (PPHG) by lowering sharp rise in blood glucose levels after ingestion of carbohydrate rich meal in type 2 diabetic (T2D) individuals.
Acalypha indica
commonly known as Indian copper leaf is used in traditional medicinal system to treat various diseases. In our previous
in-vitro
investigation, methanolic extract of
A. indica
stems (AIS) proved to be an effective a-glucosidase inhibitor, antioxidant, and well tolerated in acute and subchronic toxicity studies in albino wistar rats
Objective:
In this perspective, this study was designed to evaluate postprandial antihyperglycemic potential of AIS in maltose, sucrose, and glucose loaded streptozotocin (STZ)-induced normal and diabetic rats. As, the acute hyperglycemia at postprandial period has more triggering effect on oxidative stress, study was also aimed to evaluate the antioxidant potential of AIS on STZ-induced Albino-Wistar rats.
Materials and Methods:
Rats were treated with AIS (300-600 mg/kg b.w.) to investigate effect of AIS in controling PPHG after carbohydrate loading. Hepatoprotective activity of AIS is evaluated in diabetic rats by treating them at the dosages 300-600 mg/kg b.w.
Results:
Studies revealed 69.10 and 80.35% blood glucose-lowering effect of AIS in maltose and sucrose loaded diabetic rats in comparison with the diabetic control group. AIS recovered the liver damage caused by streptozotocin
Conclusion:
The present study confirmed high potential of AIS in controling PPHG by inhibiting a-glucosidase enzyme in maltose and sucrose loaded diabetic rats. AIS also exhibited hepatoprotective activity in STZ-induced diabetic rats. Thus, AIS could be used as a nutraceutical supplement to treat T2D effectively.
Abbreviations used:
AIS: Acalypha indica Stems, ALP: Alkaline Phosphatase, b/w: Body Weight, PPHG: Postprandial hyperglycemia, SE: Standard Error, SGOT: Serum glutamate oxaloacetate transaminase, SGPT: Serum glutamate pyruvate transaminase, SOD: Superoxide dismutase, STZ: Streptozotocin, TB: Total Bilirubin, T2D: Type 2 diabetes
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Elucidation of molecular mechanism(s) of cognition enhancing activity of Bacomind
®
: A standardized extract of bacopa monnieri
p. 482
Shekhar Dethe, M Deepak, Amit Agarwal
DOI
:10.4103/0973-1296.191464
PMID
:27761079
Background:
Bacopa monnieri
(L.) Wettst., commonly known as Brahmi, is renowned in Indian traditional system for its potent memory enhancing activity, which has been validated by various scientific studies.
Objective:
The objective of this study was to understand the molecular mechanism of memory enhancing activity of BacoMind
®
(BM), a standardized extract of
B. monnieri.
Materials and Methods:
BM was screened
in vitro
in a panel of cell-free and receptor-transfected cell assays. The purified enzymes/membrane homogenates/cells were incubated with substrate/standard ligand in the absence or presence of the test compound. The IC
50
values and EC
50
values were determined by nonlinear regression analysis of the concentration-response curves generated with mean replicate values using Hill equation curve fitting.
Results:
BM was found to inhibit three enzymes; Catechol-O-methyl transferase (COMT), Prolyl endopeptidase (PEP), and Poly (ADP-ribose) polymerase (PARP). It also had an antagonistic effect on serotonin 6 and 2A (5-HT
6
and 5-HT
2A
) receptors
,
known to influence the different neurological pathways, associated with memory and learning disorders, age-associated memory impairment.
Conclusion:
BM was found to inhibit three enzymes namely, Catechol-O-methyl transferase (COMT), Prolyl endopeptidase (PEP), and Poly (ADP-ribose) polymerase (PARP). It also exhibited an antagonistic effect on 5-HT
6
and 5-HT
2A
receptors.
Abbreviations used:
HTRF: Homogenous time resolved fluorescence, cAMP: Cyclic adenosine monophosphate,
CHO
: Chinese hamster ovary,
RFU
: Relative fluorescence unit, pNP: Para nitro phenol, AMC: 7-amino-4-methylcoumarin, ELISA: Enzyme linked immunosorbent assay, Z-Pro-Pro-CHO: Z-prolyl-prolinal, HEK: Human embryonic kidney, TE: Trolox equivalent.
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Free radical scavenging, α-glucosidase inhibitory and anti-inflammatory constituents from Indian sedges,
Cyperus scariosus
R.Br and
Cyperus rotundus
L.
p. 488
Lavanya Kakarla, Suresh Babu Katragadda, Ashok K Tiwari, K Srigiridhar Kotamraju, K Madhusudana, D Anand Kumar, Mahendran Botlagunta
DOI
:10.4103/0973-1296.191467
PMID
:27761080
Background:
Cyperus scariosus
R. Br and
Cyperus rotundus
L are widely used in ayurvedic preparation for the treatment of diabetes and other diseases. The early literature ,so far, does not indicate the presence of any bioactive principle isolated from these plants.
Objective:
To identify free radical scavenging, anti-diabetic and anti- inflammatory principles from these two species.
Materials and Methods: The
bioassay guided fractionation and isolation of active constituents was done by chromatographic techniques. They also evaluated their anti-oxidant activity by DPPH and ABTS. The anti-diabetic activity was screened by α- glucosidase and α- amylase assays.Also, the further evaluation of
in vitro
anti-inflammatory activity using THP-1 monocytic cells and
in vivo
anti- inflammatory activity, was confirmed by carrageenan induced rat paw edema as model.
Results:
The activity guided isolation led to isolation of twelve compounds Which are: Stigmasterol
[1]
, β- sitosterol
[2]
, Lupeol
[3]
, Gallic acid
[4]
, Quercetin
[5]
, β- amyrin
[6]
, Oleanolic acid
[7]
, β- amyrin acetate
[8]
, 4- hydroxyl butyl cinnamate
[9]
, 4- hydroxyl cinnamic acid
[10]
, Caffeic acid,
[11]
and Kaempferol
[12]
respectively. Among the isolates, the compounds 4 and 5 displayed potent radical scavenging activity with an IC
50
values of 0.43 and 0.067 ΅g/ml. The compounds 4, 5 and 10 showed significant anti-diabetic activities. while lupeol
[3]
showed potent IL-1 β activity inhibition in THP-1 monocytic cells and also displayed significant (p<0.0025)
in vivo
anti-inflammatory activity.
Conclusion:
Inbrief, we isolated twelve compounds from both the species. Collectively, our results suggested that aromatic compounds showed good anti-oxidant and anti-diabetic activities.
Abbreviations used:
DPPH: 2,2- Diphenyl-1-1-picryl hydrazyl, ABTS: 2,2- Azinobis-3-ethylbenzo thiazoline-6-sulfonic acid, THP-1: Human leukaemia monocytic cell line, IL-1
β
: Interleukin-1
β
, IC
50
- Inhibitory concentration 50%.
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