Monthly Publishing Peer Reviewed Journal

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1. Acute and Subacute Toxicity Evaluation of Alkaloids of Peganum harmala L. in Experimental Mice
Hassina Guergour, Rima Allouni, Nadia Mahdeb, Abdelouahab Bouzidi
The objective of the study was to evaluate the acute and subacute toxicity of total alkaloids seeds of Peganum harmala in female mice. All morphological, biochemical, hematological, histopathological changes, in addition to mortality and bodyweight changes were recorded. After acute intraperitoneal administration of dose118 mg/kg, there were no any remarkable changes in general appearance and mortality. After 24h, a significant increase in relative weights of heart and brain with no change in hematological, biochemical parameter compared to the normal group. For 5 days a significant reduction in the relative weights of the kidneys and increase of brain with significant change in hematological (MCHC, MCV) and biochemical parameter (AST, ALP and Urea). Subacute study of dose (18 mg/kg) for 28 days showed no remarkable changes in general appearance and deaths occurred in experimental group. A significant increase in relative weight of brain compared to the normal group was observed. In biochemical parameters, a significant increase was seen in both ALT and AST enzyme activities. There was no significant change in hematological parameters. Histopathological examination revealed a ground glass appearance of hepatocytes and the vascular congestion. Alkaloids seeds of P.harmala showed significant toxicity in female mice.

2. A Review on Cucurbita pepo
Neel Ratnam, Vandana, Md Najibullah, Md Ibrahim

Cucurbita pepo is widely used as food and herbal remedies around the world. It contains several phyto-constituents belonging to the categories of alkaloids, flavonoids, and palmitic, oleic and linoleic acids. Many pharmacological studies have demonstrated hepatoprotection, inhibit benign prostatic hyperplasia, hypoglycemic agent, antioxidant, anticancer, antimicrobial, anti-inflammatory, anti-diabetic, and antiulcer activities supporting its traditional uses. This aims a comprehensive study of the chemical constituents, pharmacological and clinical uses.

3. Impact of Virucide; Carrageenan in Suppression of Potato Virus Y in Potato Plants
Gomaa Hanaa H A
Background: Potato plants are usually subjected to numerous diseases caused by viruses. Potato virus Y belongs to a group of potyvirus that causes serious diseases to potato plants and is mainly responsible for the majority of losses caused in potato production.  Objective: The present study aims to evaluate the potential effect of carrageenan (virucide) which was extracted from red algae (Acanthophora specifira) in an induction of systemic acquired immunity for bio-control of PVY infection in potato plants. Methods: Potato tubers (Solanium tuberoisum cv. spounta) were obtained. The virus isolate was obtained and maintained on Datura metel plant. Specimens of the red alga Acanthophora specifira were collected from El Shoaiba coast, the eastern part of Saudi Arabia; Red Sea. The tested material carrageenan was extracted from the selected seaweeds species as antiviral agents. The infectious sap was prepared and inoculated on the primary and free potato leaves. The tested plants were sub-classified into 5 groups; 2 healthy groups (control group and virucide-treated group), 3 PVY infected plants groups (one without virucide, PVY infected plants either before or after virucide). Reduction in the disease severity was recorded and inhibition in PVY infectivity as a result of periodical treatment of foliar potato plants with virucide (carrageenan) was assessed in pre and post-treatment groups. Results:  PVY infection resulted in significant decrease in shoot length, leave an area and fresh and dry weight of shoots. Improvement of morphological characters of periodically-treated foliar with virucide (carrageenan) was observed. Chlorophyll and carotenoids contents were increased in PVY infected foliar potato plants treated with virucide (carrageenan). Biochemical markers as indicators for systemic acquired resistance were detected via significantly increased total proteins, phenol compounds, free proline (PO), polyphenol oxidase (PPO) and superoxide dismatase (SOD) enzyme activities. As well as expressed proteins of induced potato plants were determined using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The results indicated that newly expressed proteins were produced and a difference in number and density of bands in foliar treatment plants with virucide (carrageenan) were compared with healthy ones. Conclusion: It has been suggested that the induction of systemic acquired resistance (SAR) was successfully achieved; it also protected potato plants against PVY infection.

4. Alpha Mangostin and Xanthone from Mangosteen (Garcinia mangostana l.) Role on Glucose Tolerance and Glucose Transporter-4 in Diabetes Mellitus
Ratwita W, Sukandar E Y, Adnyana I K, Kurniati N F
Objective: This research elaborated role of alpha mangostin and xanthone on glucose tolerance and Glucose Transporter (GLUT)-4 by measuring blood glucose level and GLUT-4 expression on cardiac cell musce and adipocyte cell culture. Methods: Glucose tolerance test were conducted using male wistar rat divided into 9 groups, which were normal, control (D-Glucose induced only), glibenclamide, various doses of a-mangostin and xanthone (5, 10, 20 mg/kgbw). All group induced by D-glucose 3 g/kg orally 30 minutes later. Blood glucose levels changes were observed at 90th and 150th minute. GLUT-4 study conducted for 3 weeks on mice that divided into 10 groups which were normal, diabetic control mice (alloxan induced only), metformin, glibenclamide, various doses of a-mangostin and xanthone (5, 10, 20 mg/kgbw). GLUT-4 expression than observed on cardiac cell muscle. While other study observed GLUT-4 expression on adipocyte cell culture that treated with a-mangostin/xanthone/pioglitazone. Results: Normal group (non-diabetic) responds slightly to the administration of glucose in glucose tolerance test.  Blood glucose level in every group in the 90th up to 150th minute decreased significantly when compared to the positive control group (p <0.05). This shows that glucose tolerance does not occur in all treated groups  althought they were treated with high glucose consentration. GLUT-4 expressions in mice cardiac-musce cells that treated with a-mangostin/xanthone/ glibenclamide/metformin significantly increased when compared to the positive control group, except in group treated with xanthone 5 mg/kgbw. GLUT-4 expressions also increase in adipocytes that treated with 3.125 mM; 6.25 mM and 25 mM 𝛼–mangostin, equivalent to pioglitazone. All treatment group results significantly different when compared with control. The effect of 𝛼–mangostin on GLUT-4 expression better than xanthone’s. Conclusion: Alpha mangostin and xanthone are two substances that showed protective effect to glucose tolerance and also have potential effect to improve insulin resistance by increasing GLUT-4 on cardiac muscle and adipocyte.

5. Traditional and Ethnobotanical uses Premna barbata Wall. Ex Schauer in Kumaun and Garhwal Regions of Uttarakhand, India & Other Western Himalayan Countries- A Review
Joshi Amit Kumar, Juyal Divya
The Himalayas are a repository of tremendous wealth of medicinal plants. Kumaun and Garhwal are characterized by a rich diversity of ethno-medicinal plants as well as a rich heritage site in traditional medicine system in the western Himalayan region. Present study deals with the status, identification and to explore the traditional knowledge of plant Premna Barbata Wall. Ex Schauer about its uses in local medicine by village people in Kumaun, Garhwal region in India and other western Himalayan Countries. The genus Premna contains 200 species under the family Verbenaceae all over the world and out of that approximately 30 species are present in India and are native mainly to the tropical and subtropical Asia, Africa, Australia and the Pacific islands. Premna Barbata Wall. Ex Schauer plant is belonging to family Verbenaceae. It is distributed in the forest area of the Eastern & northern parts of India, Pakistan, Nepal, and Myanmar and nearby regions.The present paper is a review on ethno-medicinal uses of Premna Barbata Wall. Ex Schauer, which is widely used by different tribes and medicinal practitioners for treatment of various diseases. Different parts of the plant (bark, wood and leaves) are variably used in treating wide range of diseases such as joints and back pain, dropsy, ulcer, diarrhea, wound healing, throat infection, antibacterial, fever, herpes complex disease, arthritic pain. With this review it is concluded that there is a need for conducting further investigation and studies for pharmacognostic characteristics and the isolation of individual active constituents form Premna Barbata Wall. Ex Schauer plant. Also the study is expected to provide basic data for further studies aimed at conservation of traditional medicine.

6. Macrofilaricidal Activity of Leaf Extracts of Rauvolfia tetraphylla L. Against Bovine Filarial Parasite Setaria cervi.
Behera D R, Bhatnagar Sunita
Leaf extracts of Rauvolfia tetraphylla were evaluated for antifilarial potential using Setaria cervi as target parasite. Four extracts were tested these were hexane, chloroform, acetone and methanol.    The activity was assessed by means of worm motility assay, MTT reduction and GST enzyme inhibition assay using female worms of Setaria cervi. DEC was used as a standard drug.    Methanol extract exhibited significant activity among all the extracts with percentage of reduction 89.28% at 10mg/ml concentration. DEC was comparitively less active than methanolic extract with 79.22% activity at the same dose. The IC50 value for methanolic extract was found to be 0.03mg/ml, which was better than the standard DEC i.e. 2.84mg/ml. The methanol extract significantly inhibited the GST enzyme activity of Setaria worms as compared to untreated worms with percentage of inhibition value of 56.41% at 10mg/ml. The most active methanol  extract was finally explored for cell viability study by using trypan blue dye exclusion test. The methanolic extract significantly killed the oocytes of Setaria worms as all the oocytes stained blue in treated worms and unstained in control oocytes. The findings indicate that the methanolic extract has potent macrofilaricidal ativity.


7. Effect of Euphorbia Prostrata on the Wound Healing in Excisional Wound Model in Rats
T R Patil, R P Limaye
Background-Euphorbia prostrata possesses many actions one of which is wound healing. Skin wound healing is a significant health care problem which  clinician faces still. Hence this study was planned to evaluate effect of euphorbia prostrata on skin wound healing. Material and methods-This study was conducted among randomly selected wistar rats (n=6 per group). Group A and Group B had Cream and 1% euphorbia prostrate cream applied topically on the excisional wound respectively. Wound healing and epithelisation was assessed on Days 4,8,12,16,20. Results:  Group A showed an average wound size of 348±13.0 on day 4, 278±10.5 on day 8, 119±9.8 on day 12, 86±9.2 on day 16 and 100% wound healing was observed day 20 onwards. Group B showed an average wound size of 296±11.0 on day 4, 147±9.5 on day 8 and complete wound healing was observed day 12 onwards in the test group. Conclusion: Euphorbia prostrata holds promise as effective drug therapy for wound healing.

8. Ginger: A Potential Neutraceutical, An Updated Review
Singh Rudra Pratap, Gangadharappa H V, Mruthunjaya K
Ginger (rhizome of Zingiber officinale) belongs to the Zingiberaceae family, is widely used and is most popular as a culinary spice and in Traditional medicines to add flavor for more than thousands of years. It is also used in pharmaceuticals, nutraceuticals and in cosmetics. Ginger contains a fusion of an aroma oils both volatile (zingiberene) and non-volatile (oleoresin) oils and phenolic compounds (gingerol and shogaol, zingerone and paradol). In ginger rhizomes, also contains starch, saccharides, proteins, colouring matter and trace minerals that plays a huge role as a spice ingredient. In ginger, starch comprises of 40 – 60 % w/w, protein is 6.2 – 19.8 %, wax or lipid is 5.7 – 14.5 % and crude fiber is 1.1 – 7.0 % and oleoresin approximately 4 – 7.5 %. Ginger is available in fresh, dried, pickled, preserved, crystallized, candied and powdered or ground form. It is unstable due to acidic environment or as a result of the increase in temperature; presence of light, air, heat and long term storage but it can be enhanced by Nano emulsion formulation. Ginger has shown various pharmacological effects such antioxidant, anti – inflammatory, gastro protective, anti-bacterial, anti-diabetic etc.

9. Qualitative and Quantitative Analytical Studies on Poora parpam– A Siddha Medicine
Kabilan N, Murugesan M, Balasubramanian T, Geethalakshmi S
Siddha system of medicine is a potent and unique indigenous system of medicine, which deals with the diseases of human being efficiently with the knowledge of both subtle and also the gross material body. This study was to prepare the Poora Parpam a Siddha medicine which contains herbo-mineral compounds responsible for the therapeutic activity and find out the compound by qualitative and quantitative analysis by different methods. Literature review reveals many herbo–mineral formulations available in market which is useful in anemia, diabetes, cancer, liver diseases, skin diseases etc. Therefore our aim of the study was to determine the compound by qualitative and quantitative analysis of ICP-MS, FTIR, MS, SEM and TEM. The study found some compound by above studies which were responsible for different activity.

10. Chromatography Analysis of Fatty Acids, Volatile Compounds and Alkaloids of Ephedra alata Growing Wild in Southern Tunisia and Evaluation of their Antioxidant Activity
Mighri H, Bennour N, Eljeni H, Neffati M, Akrout A
In order to provide more information about phytochemical composition of Tunisian Ephedra alata known for their medicinal uses, fatty acids, volatile compounds and alkaloids have been analyzed in fresh or dry plant materials whether powdered or not. Extraction was performed with hexane and ethanol in soxhlet extractor and all extracts were analyzed by GC-MS. Phosphomolybdenum, DPPH and FRAP methods were used to evaluate their antioxidant activities. The dry matter present a more diversified fatty acids composition dominated by palmitic acid, oleic acid and linolelaidic acid. Hydrocarbons represent the largest group of volatile compounds, followed by alcohols. The most known detected ephedrine-type alkaloid characteristic of the Ephedra genus are dominated by pseudoephedrine, norephedrine, ethylephedrine. Some other compounds are mentioned for the first time in E. alata alkaloids such as azetidine, 1,2-dimethyl-3-phenylaziridine, N-ethyl benzamide and N-methyl-mandelamide. Lower total antioxidant and anti-radicular effects were shown with hexane extracts and alkaloid fractions instead the ferric reducing antioxidant power test showed the highest results with obtained alkaloid from powdered fresh and dry plant materials. Lipids, volatile compounds and alkaloids in E. alata that have been assessed could provide possible roles in case of human breast cancer and health benefits and therefore a research on this plant might be of value in drug industry.

11. Toxicity Studies of Aqueous Extract of Prismatomeris glabra
Razali M S, Mizaton H H, Hazilawati H, Aishah A
This study was conducted to estimate levels for human consumption of an aqueous extract of Prismatomeris glabra (PG) roots. PG extract was prepared by boiling powdered roots for 10 minutes before drying in spray dryer. Cytotoxicity studies were conducted using cell lines from embryonic human liver (WRL68), liver cancer (HEP G2), colon cancer (HCT116), leukemic cells (K562) and mouse Leydig cells (CRL1714). Established method of comet assay was used to determine genotoxicity. Toxicity studies in mice were conducted for acute, subacute and subchronic effects. In all experiments, age-matched control mice were given normal saline. Gross necropsy biochemistry analyses were conducted following killing. PG showed no cytotoxic effects on WRL68, HEP-G2, K562 and CRL1714 cell lines. IC50 values in these cell lines were 2818, 2291, 1778 and 2400 µg/ml, respectively. PG also showed no effect on DNA. In vivo, mice were able to tolerate PG to a maximum single dose of 3 g/kg, p.o., 500 mg/kg/d, p.o., daily for 14 days, and at 100 mg/kg/d, p.o., daily for 3 months, respectively, without showing signs of toxicity or abnormal biochemical markers. In conclusion, based on toxicity data, PG root aqueous extract is generally safe for consumption.

12. Mussaenin A Isolated From Mussaenda glabrata Induces Apoptosis in the Liver Cancer Cells Via Mitochondrial Pathway
Lipin Dev Mundur Sahadevan, Darsan Balakrishnan Menon
Plants belonging to Rubiaceae family have been used in Chinese folk medicine and Ayurveda. The aim of the study was to determine the effect of Mussaenin A isolated from root of Mussaenda glabrata on liver cancer cell line, Hep G2. MTT assay was performed to check the ability of Mussaenin A to induce death in cancerous Hep G2 cells and normal NIH3T3 cells. Acridine Orange/Ethidium Bromide staining, Hoechst staining and DNA fragmentation assays were used to confirm the apoptosis inducing ability of MA on Hep G2 cells. RT-PCR and western blotting was performed to check the expression of pro- and anti-apoptotic factors. Calorimetric assay was done to check caspase-3 and caspase-9 activities. Mussaenin A at lower concentrations was found to induce cell death selectively in the liver cancer cells (IC50 = 11.38 µg/mL). The transcriptional expression studies of the pro-apoptotic Bax and anti-apoptotic Bcl-2 and Cox-2; and the western blot analysis of pro-apoptotic BAK and BAD showed that MA upregulated the expression of pro-apoptotic factors and down regulated the expression of anti-apoptotic factors in Hep G2 cells. Caspase-9 and 3 activities were found to be upregulated in the calorimetric studies. The down regulation of anti-apoptotic factors and upregulation of pro-apoptotic factors show that the Mussaenin A induced apoptosis in the liver cancer cells via the intrinsic or the mitochondrial pathway.

13. Determination of Plant Constituents in Fractions of Cordia obliqua Willd. Leaf Methanol Extract using GC-MS Analysis
R Gupta, G D Gupta
Cordia obliqua Willd. is an important medicinal plant of family Boraginaceae. It is commonly known as Clammy Cherry. It is a deciduous tree and found throughout Himalayan region in India. Traditionally, it possesses a number of medicinal activities like diuretic, expectorant, antipyretic, anthelmintic and maturant. The current work is aimed to investigate chemical constituents present in fractions of leaf methanol extract of Cordia obliqua plant using GC-MS analysis. Two fractions of methanol extract (CO1 & CO3) were studied and about 30 components were detected in each sample. Highest peak area was observed for moncyclic sesquiterpene alcohol α-Bisabolol oxide b at RT 20.84 in CO1 and for o-methyl-d-glucose at RT 16.53 in CO3. The important constituents detected were Amino acids, Oleic acid, Oleanolic acid, Sesquiterpene lactone, trietrpenoids, various steroids, galactopyranoside, Hexadecanoic acid, Phthalic acid, Azafrin, Decatrienoic acid, Docasenamide, Morphinan and carotenoids. Among these Morphinan is an opoid alkaloid and good psychoactive drug. Other constituents are also medicinally important and many have reported biological activities. Thus GC-MS analysis revealed the existence of various types of constituents in Cordia obliqua leaf methanol extract fraction which confirm the application of this plant for a number of medicinal activities.

14. Isolation and Characterization of β-Sitosterol from Justicia gendarussa burm. F.-An Anti-Inflammatory Compound
N D Phatangare, K K Deshmukh, V D Murade, P H Naikwadi, D P Hase, M J Chavhan, H E Velis
Justicia gendarussa Burm.f. has become important source of β-sitosterol which is associated with other phenolic,terpenoids,alkaloids and steroids. Plant sterols show anti-inflammatory activity. β-sitosterol is one of phytosterol, in a mouse model of acute inflammation, and β-sitosterol effect on leukocyte recruitment, cytokines levels, and oxidative stress. The anti-inflammatory activities of β-sitosterol were assessed by measuring paw edema induced by different inflammatory agents. It separated from Justicia gendarussa burm.f. and characterization of β-sitosterol carried out by IR, NMR, and mass spectrometry. β-sitosterol shows potent as Anti-inflammatory activity by releasing histamine (30.07%), serotin and bradykinin (52.25%), and prostaglandin (69.43%) as compared to standard (Dicolfenac 5mg/kg).  Objectives:To isolate, separate and characterization of β-sitosterol and to evaluate the anti-inflammatory activity of β-sitosterol extracted from Chloroform extract of Justicia Gendarussa Burm.f.

International Journal of Pharmacognosy and Phytochemical Research