FIRST REPORT ON FIBRINOLYTIC AND THROMBOLYTIC ACTIVITY OF EUTYPHOEUS GAMMIEI AN EARTHWORM SPECIES COLLECTED FROM TRIPURA, NORTHEAST INDIA

Objective: The present investigation for the first time evaluated the in vitro fibrinolytic and thrombolytic activities of crude extracts from Eutyphoeus gammiei, native, large size earthworm of Tripura, Northeast, India. The present study was designed to evaluate the therapeutic use of the organism E. gammiei as a source of fibrinolytic and thrombolytic agent(s).Methods: The fibrinolytic activity was studied using by fibrin plate and zymography assays. Thrombolytic assay was carried out according to Prasad et al. (2006) using whole blood.Results: The results obtained clearly indicated E. gammiei as a potential source of fibrinolytic and thrombolytic agents. Both in fibrin plate assay and thrombolytic assay with whole blood, E. gammiei crude homogenate showed similar and close results in respect to that of streptokinase. Fibrin zymography also showed antifibrinolytic activity with producing clear bands. Dose and time dependency also is evident from the results.Conclusion: The results of the present study conclude that the studied earthworm species E. gammiei possessed profound fibrinolytic and thrombolytic activity on human blood and E. gammiei might prove to be useful alternative source for the development of new drugs for treatments involving blood coagulation and fibrinolysis

REACTIVE OXYGEN SPECIES AS POSSIBLE MEDIATOR OF ANTIBACTERIAL ACTIVITY OF PARKIA JAVANICA, AGAINST BACTERIAL SPECIES PREDOMINANTLY FOUND IN CHRONIC WOUND.

The crude methanol extract of Parkia javanica was screened for antibacterial activity. against bacterial species predominantly found in chronic wound, by serial dilution technique. Growth kinetics study was performed and percentage of ROS production was measured by NBT reduction assay. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were obtained with a range of IC100 5-40 mg/ml in case of standard bacterial strains. The lag phase of all extract treated bacteria is extended compared to untreated cells. The normalized % of ROS is increased in presence of crude extract. This study suggests that the crude methanol extract of Parkia javanica possesses promising antimicrobial substances which are having activity against Standard ATCC bacterial species and ROS induced DNA damage could be the possible mediator of its antimicrobial activity. Keywords: Parkia javanica, antibacterial activity, standard ATCC bacterial strains, growth curve, ROS, DNA damag

A Rare HBV Subgenotype D4 with Unique Genomic Signatures Identified in North-Eastern India –An Emerging Clinical Challenge?

BACKGROUND/AIMS: HBV has been classified into ten genotypes (A-J) and multiple subgenotypes, some of which strongly influence disease outcome and their distribution also correlate with human migration. HBV infection is highly prevalent in India and its diverse population provides an excellent opportunity to study the distinctiveness of HBV, its evolution and disease biology in variegated ethnic groups. The North-East India, having international frontiers on three sides, is one of the most ethnically and linguistically diverse region of the country. Given the paucity of information on molecular epidemiology of HBV in this region, the study aimed to carry out an in-depth genetic characterization of HBV prevailing in North-East state of Tripura. METHODS: From sera of chronically HBV infected patients biochemical/serological tests, HBV DNA quantification, PCR-amplification, sequencing of PreS/S or full-length HBV genomes were done. HBV genotype/subgenotype determination and sequence variability were assessed by MEGA5-software. The evolutionary divergence times of different HBV subgenotypes were estimated by DNAMLK/PHYLIP program while jpHMM method was used to detect any recombination event in HBV genomes. RESULTS: HBV genotypes D (89.5%), C (6.6%) and A (3.9%) were detected among chronic carriers. While all HBV/A and HBV/C isolates belonged to subgenotype-A1 and C1 respectively, five subgenotypes of HBV/D (D1-D5) were identified including the first detection of rare D4. These non-recombinant Indian D4 (IndD4) formed a distinct phylogenetic clade, had 2.7% nucleotide divergence and recent evolutionary radiation than other global D4. Ten unique amino acids and 9 novel nucleotide substitutions were identified as IndD4 signatures. All IndD4 carried T120 and R129 in ORF-S that may cause immune/vaccine/diagnostic escape and N128 in ORF-P, implicated as compensatory Lamivudine resistance mutation. CONCLUSIONS: IndD4 has potential to undermine vaccination programs or anti-viral therapy and its introduction to North-East India is believed to be linked with the settlement of ancient Tibeto-Burman migrants from East-Asia

Determination of virulence factors and biofilm formation among isolates of vulvovaginal candidiasis

Context: Under morphogenesis-inducing conditions, Candida spp. begins to undergo yeast-to-hypha switch. This shift from commensal to pathogenic state is dependent on several virulence factors. Aim: To find out whether the isolated Candida spp. were pathogens causing vulvovaginal candidiasis or mere bystanders. Settings and Design: Cross-sectional observational study conducted on 275 symptomatic hospital patients in Tripura between August 2012 and April 2015. Subjects and Methods: Discharge was collected from patients and identified by Grams staining and wet mount test. Culturing was done in Sabouraud dextrose agar followed by speciation. To test for virulence factors, assays for adherence, plasma coagulase, phospholipase, lipase, protease, hemolysin, and biofilm formation were carried out. Statistical Analysis Used: Significance between two groups was compared using one-way analysis of variance along with Tukey test, and Chi-square 2 × 2 contingency table at 95% confidence interval. Results: Fifty-six Candida spp. could be isolated in the study which was used for further virulence tests. One hundred percent of isolates expressed adherence. Among other virulence factors, maximum virulence 25 (45%) was shown through protease production. Hemolysin production and biofilm formation were the second most 22 (39%) expressed virulence factors. In a comparison of virulence factors between biofilm-forming isolates and planktonic cells, significant difference was seen for plasma coagulase and hemolysin production. Conclusions: All the isolates expressed one or more virulence factors. Adherence was expressed in all isolates but highest number was observed for Candida albicans. Furthermore, C. albicans strain number was highest for protease, hemolysin and coagulase expression and biofilm formation. Candida krusei isolates were the least in number for expressing any of the virulence factors. Significantly higher number of biofilm forming isolates produced hemolysin and coagulase in comparison to planktonic cells

ACTIVITY OF THE MEDICINAL PLANT PARKIA JAVANICA AGAINST NEISSERIA GONORRHOEAE MULTI DRUG RESISTANT AND OTHER CLINICAL ISOLATES: Running title: Anti-gonococcal activity of Parkia javanica

Objective: The objective of this study was to look into the in vitro activity of Parkia javanica against isolates of Neisseria gonorrhoeae. Methods: Methanolic extract of P. javanica bark (MEPJ) and organic fractionation were tested against one standard strain and 10 clinical isolates including one multidrug-resistant (MDR) isolate of N. gonorrhoeae through minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) tests. Results: The MDR isolate, standard strain, as well as all the clinical isolates were inhibited by MEPJ as well as all the fractions except water fraction. Chloroform fraction showed the best activity with MIC and MBC values, both being 0.2 mg/ml. Ethyl acetate fraction also showed MIC value at 0.2 mg/ml; however, MBC value was at 0.3 mg/ml. MIC and MBC values of MEPJ were both 0.3 mg/ml. Conclusion: Chloroform fraction, ethyl acetate fraction, and MEPJ bark showed the lowest MIC values and can be considered as prospective candidates for the development of antigonococcal topical drugs

Oroxylin A: A Promising Flavonoid for Prevention and Treatment of Chronic Diseases

There have been magnificent advancements in the understanding of molecular mechanisms of chronic diseases over the past several years, but these diseases continue to be a considerable cause of death worldwide. Most of the approved medications available for the prevention and treatment of these diseases target only a single gene/protein/pathway and are known to cause severe side effects and are less effective than they are anticipated. Consequently, the development of finer therapeutics that outshine the existing ones is far-reaching. Natural compounds have enormous applications in curbing several disastrous and fatal diseases. Oroxylin A (OA) is a flavonoid obtained from the plants Oroxylum indicum, Scutellaria baicalensis, and S. lateriflora, which have distinctive pharmacological properties. OA modulates the important signaling pathways, including NF-κB, MAPK, ERK1/2, Wnt/β-catenin, PTEN/PI3K/Akt, and signaling molecules, such as TNF-α, TGF-β, MMPs, VEGF, interleukins, Bcl-2, caspases, HIF-1α, EMT proteins, Nrf-2, etc., which play a pivotal role in the molecular mechanism of chronic diseases. Overwhelming pieces of evidence expound on the anti-inflammatory, anti-bacterial, anti-viral, and anti-cancer potentials of this flavonoid, which makes it an engrossing compound for research. Numerous preclinical and clinical studies also displayed the promising potential of OA against cancer, cardiovascular diseases, inflammation, neurological disorders, rheumatoid arthritis, osteoarthritis, etc. Therefore, the current review focuses on delineating the role of OA in combating different chronic diseases and highlighting the intrinsic molecular mechanisms of its action

Estimates of Evolutionary Divergence over Sequence Pairs between HBV/D subgenotypes (D1–D9) and Indian HBV/D4 (IndD4).

<p>Genetic distances of Indian HBV/D4 (IndD4) with other subgenotypes of D are indicated in bold and the genetic distance between IndD4 and reported D4 is enclosed within a box.</p><p>Estimates of Evolutionary Divergence over Sequence Pairs between HBV/D subgenotypes (D1–D9) and Indian HBV/D4 (IndD4).</p