Antidiabetic Property of Symplocos cochinchinensis Is Mediated by Inhibition of Alpha Glucosidase and Enhanced Insulin Sensitivity

The study is designed to find out the biochemical basis of antidiabetic property of Symplocos cochinchinensis (SC), the main ingredient of ‘Nisakathakadi’ an Ayurvedic decoction for diabetes. Since diabetes is a multifactorial disease, ethanolic extract of the bark (SCE) and its fractions (hexane, dichloromethane, ethyl acetate and 90% ethanol) were evaluated by in vitro methods against multiple targets relevant to diabetes such as the alpha glucosidase inhibition, glucose uptake, adipogenic potential, oxidative stress, pancreatic beta cell proliferation, inhibition of protein glycation, protein tyrosine phosphatase-1B (PTP-1B) and dipeptidyl peptidase-IV (DPP-IV). Among the extracts, SCE exhibited comparatively better activity like alpha glucosidase inhibition (IC₅₀ value-82.07±2.10 µg/mL), insulin dependent glucose uptake (3 fold increase) in L6 myotubes, pancreatic beta cell regeneration in RIN-m5F (3.5 fold increase) and reduced triglyceride accumulation (22% decrease) in 3T3L1 cells, protection from hyperglycemia induced generation of reactive oxygen species in HepG2 cells (59.57% decrease) with moderate antiglycation and PTP-1B inhibition. Chemical characterization by HPLC revealed the superiority of SCE over other extracts due to presence and quantity of bioactives (beta-sitosterol, phloretin 2′glucoside, oleanolic acid) in addition to minerals like magnesium, calcium, potassium, sodium, zinc and manganese. So SCE has been subjected to oral sucrose tolerance test to evaluate its antihyperglycemic property in mild diabetic and diabetic animal models. SCE showed significant antihyperglycemic activity in in vivo diabetic models. We conclude that SC mediates the antidiabetic activity mainly via alpha glucosidase inhibition, improved insulin sensitivity, with moderate antiglycation and antioxidant activity.     

Effects of diallyl disulfide (DADS) on expression of apoptosis associated proteins in androgen independent human prostate cancer cells (PC-3)

Prostate cancer is a leading cause of death among the aging men. Surgical or radiotherapy is effective when the cancer is confined to the prostate gland but once the cancer spreads beyond the pelvis even chemotherapy and hormonal ablation therapy fails in curing this disease. Our previous studies have shown that diallyl disulfide (DADS) induces cell cycle arrest and also induces apoptosis in PC-3 cells. And now the present study is focused to see whether there is an activation of caspase cascade pathway. Hence, in the present study the apoptotic effect of DADS is studied by Western blot analysis of caspase-3, -9, -10 and Bcl-2, Bad, and Bax protein. The Apoptotic cells were assessed by Hoechst 33342 staining with 25 and 40 microM concentrations of DADS for 24 h. The results have shown that DADS at 25 and 40 microM concentrations has induced the activation of caspases. There is a significant increase in the expression of caspases (3, 9, and 10). The proapoptotic protein Bax has significantly increased at 40 microM of DADS treatment and there is significant increase of Bad protein at both the concentration. Bcl-2 protein has significantly decreased in DADS treated cells. Therefore, the present investigation serves as evidence that DADS may be a therapeutic drug in the treatment of prostate cancer.     

Scanning Electron Microscopy of the Infection Process of Cercospora henningsii on Cassava Leaves

Germination, penetration and sporulation of Cercospora henningsii (Allesch.) on cassava leaves were studied by scanning electron microscopy. Conidia started to germinate 9 h postinoculation producing one to two germ tubes. The germ tubes entered the leaf tissue through the abaxial surface by direct penetration of the epidermis without forming appressoria. The cassava leaf is characterized by its papillose epidermis on the abaxial surface. The penetrations occurred at smooth areas of the leaf epidermis between the papillae. The germ tubes did not enter stomata even when they passed over stomatal openings. Leaf spots started to appear 9 days after the inoculation (dpi), and the emergence of conidia occurred 14 dpi. The symptoms appeared first on the abaxial leaf surface, followed 2 days later on the adaxial. Conidia emerged in clusters through ruptured epidermis on both sides of the leaves. Conidia emerged also through the epidermal papillae and the leaf veins. Even though small groups of conidia emerged through stomata also, emergence through stomata appeared to be random rather than a preferred route. Each conidium was born on a short conidiophore with a swollen base.     

Widespread Emergence of Multidrug Resistant <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> Isolated from CSF Samples

Bacterial infections of the central nervous system, especially acute infections such as bacterial meningitis require immediate, invariably empiric antibiotic therapy due to the widespread emergence of resistance among bacterial species. Nosocomial infections by Pseudomonas aeruginosa have been described with an increasing trend towards multidrug resistance. P. aeruginosa isolates n = 53 (66%) isolated from the cerebrospinal fluid (CSF) were used for this study. Antibiotic resistance in 53 P. aeruginosa clinical isolates from 80 CSF samples were evaluated. Of these, n = 42 (80%) of the isolates showed multidrug resistance to more than eight antibiotics and n = 17 (32%) isolates were found to be imipenem resistant P. aeruginosa (IMPR-Pa). Genotypical examination by ERIC based PCR revealed minor genetic variations. Polymicrobial infections are common in the CSF samples. However, high prevalence of P. aeruginosa as an opportunistic pathogen has been developing with increased resistance to antimicrobial agents and thus becoming a significant threat.     

High-Level Oxacillin and Gentamycin Resistance with Reduced Susceptibility to Vancomycin in <Emphasis Type="Italic">Staphylococcus aureus-</Emphasis>Carrying <Emphasis Type="Italic">mecA</Emphasis> and <Emphasis Type="Italic">femA</Emphasis> Gene Complex

Staphylococcus aureus oxiva 10 and oxiva 14 strains clinically isolated from diabetic patients were resistant to gentamycin and oxacillin The minimal inhibitory concentrations (MICs) of oxacillin and gentamycin were 720 and >2048 μg/mL, respectively, for oxiva 10 and 680 and 400 μg/mL. respectively, for oxiva 14; both strains carry mecA and femA genetic determinants in their genomes. In addition, both are vancomycin-intermediate Staphylococcus aureus (VISA) isolates. The addition of vancomycin led to significant decreases in oxacillin resistance of both oxiva 10 and oxiva 14 strains, whereas the addition of vancomycin to gentamycin plates showed a decrease in gentamycin resistance of non-high-level gentamycin-resistant (non-HLGR) oxiva 14 and indifference in gentamycin resistance in HLGR oxiva 10. Transmission electron microscopy of representative strains unveils a remarkable increase in the thickness of the cell wall, indicating that thickening of the cell wall is a common phenotype associated with vancomycin resistance in VISA isolates. The present study reports that the rate of synergism and synergistic effect in the combination vancomycin–gentamycin vary according to the MICs of gentamycin.     

Increase in cell size and acid tolerance response in a stepwise-adapted methicillin resistant <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> mutant

A methicillin-susceptible strain of Staphylococcus aureus (MSSA) showed stepwise adaptation when grown in increasing concentrations of oxacillin, eventually reaching a maximum of 35 μg/ml. The resultant oxacillin resistant mutant strain was stable and did not revert to susceptibility on frequent subculturing. The response of the cells to different concentrations of oxacillin was examined by scanning electron microscopy, which showed that the size of the bacterium increased with increasing concentrations of oxacillin. These changes in cell size were dependent on the concentration of oxacillin and occurred only after addition of non-lethal concentrations. In the presence of lethal concentrations (≥35 μg/ml) that completely inhibited cell growth, the cell sizes were smaller than those of wild-type cells and irregular in shape. This stepwise-adapted methicillin (oxacillin) resistant S. aureus (MRSA) mutant showed a greater acid tolerance response (ATR) to lactic and citric acids than the parent susceptible strain. These data indicates that methicillin resistance alters the morphology and ATR in stepwise-adapted MRSA mutant cells.     

Molecular diversity of internal transcribed spacer among the monoconidial isolates of Magnaporthe oryzae isolated from rice in Southern Karnataka,India

Blast disease of rice plant is caused by Magnaporthe oryzae (anamorph Pyricularia oryzae). This disease is recognized to be one of the most serious diseases of rice crop around the world. A total of 72 monoconidial isolates of M. oryzae obtained from blast disease samples collected around Southern Karnataka were characterized using internal transcribed spacers of the ribosomal DNA sequences. These were analyzed by comparing with already deposited sequences in GenBank database. It helped in diagnosing the invasive pathogen in all locations. Variability of rDNA sequences was found to be highly polymorphic with 0.068962 nucleotide diversity showing 6 distinct clades. 33 haplotype groups were identified with haplotype diversity of 0.8881 and Tajima's neutrality test with a D value of ?1.96827 with P?<?0.05 showing the presence of variations among the sequences of pathogen isolates. The Tajima’s D value of less than one indicates the presence of a high number of rare alleles. Our study indicates that the pathogen might have undergone recent selection pressure because of the exposure to a large number of cultivars resulting in the evolution of rare alleles. This shows the importance of characterizing internal transcribed spacer (ITS) to know pathogen diversity and its fitness which has potential to contribute to the field of breeding for blast disease resistance.     

Building a Hybrid Physical-Statistical Classifier for Predicting the Effect of Variants Related to Protein-Drug Interactions

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