Gunjan Das

Cucurbitacin B inhibits breast cancer metastasis and angiogenesis through VEGF-mediated suppression of FAK/MMP-9 signaling axis

Available breast cancer therapeutic strategies largely target the primary tumor but are ineffective against tumor metastasis and angiogenesis. In our current study, we determined the effect of Cucurbitacin B (CuB), a plant triterpenoid, on the metastatic and angiogenic potential of breast cancer cells. CuB was found to inhibit cellular proliferation and induce apoptosis in breast cancer cells in a time- and dose-dependent manner. Further, CuB-treatment significantly inhibited the migratory and invasive potential of highly metastatic breast cancer MDA-MB-231 and 4T1 cells at sub-IC50 concentrations, where no significant apoptosis was observed. CuB was also found to inhibit migratory, invasive and tube-forming capacities of HUVECs in vitro. In addition, inhibition of pre-existing vasculature in chick embryo chorioallantoic membrane ex vivo further supports the anti-angiogenic effect of CuB. CuB-mediated anti-metastatic and anti-angiogenic effects were associated with the downregulation of VEGF/FAK/MMP-9 signaling, which has been validated by using FAK-inhibitor (FI-14). CuB-treatment resulted in a significant inhibition of VEGF-induced phosphorylation of FAK and MMP-9 expressions similar to the action of FI-14. CuB was also found to decrease the micro-vessel density as evidenced by the decreased expression of CD31, a marker for neovasculature. Further, CuB-treatment inhibited tumor growth, lung metastasis and angiogenesis in a highly metastatic 4T1-syngeneic mouse mammary cancer. Collectively, our findings suggest that CuB inhibited breast cancer metastasis and angiogenesis, at least in part, through the downregulation of VEGF/FAK/MMP-9 signaling.

Co-infection of methicillin-resistant Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus and extended spectrum β-lactamase producing Escherichia coli in bovine mastitis – three cases reported from India

Emergence of antimicrobial resistance among bovine mastitis pathogens is the major cause of frequent therapeutic failure and a cause of concern for veterinary practitioners. This study describes intra-mammary infection of methicillin-resistant Staphylococcus epidermidis (MRSE), methicillin-resistant Staphylococcus aureus (MRSA) and extended spectrum β-lactamase (ESBL) producing Escherichia coli in two Holstein Friesian crossbred cows with subclinical mastitis and one non-descript cow with clinical mastitis in two different districts of West Bengal, India. In total, three MRSE, one MRSA and three ESBL producing E. coli were isolated from these cases. Both the crossbreds were detected with MRSE (HFSE1 and HFSE2) and ESBL producing E. coli (HFEC1 and HFEC2), whereas, simultaneous infection of three pathogens viz. MRSA (NDSA1), MRSE (NDSE1) and ESBL producing E. coli (NDEC1) was found in the non-descript cow. The methicillin-resistant isolates possessed mecA gene and exhibited resistance to various antibiotics such as amikacin, tetracycline and glycopeptides. The ESBL producers were positive for blaCTX-M and blaTEM genes; in addition, HFEC1 and HFEC2 were positive for blaSHV and possessed the genes for class I integron (int1), sulphonamide resistance (sul1), quinolone resistance (qnrS) and other virulence factors (papC, iucD and ESTA1). All the ESBL producers exhibited resistance to a variety of antibiotics tested including third- and fourth-generation cephalosporins and were also intermediately resistant to carbapenems. This is the first ever report on simultaneous occurrence of MRSE, MRSA and ESBL producing E. coli in bovine mastitis indicating a major concern for dairy industry and public health as well.

Anti-Diarrhoeal Activity and Toxicity Trial of Methanolic Fruit-Pulp Extract of Aegle Marmelos (L.) Correa in Sprague-Dawle Rats

To determine the toxicity as well as anti-diarrhoeal effect of the unripe methanolic fruit-pulp extract of Aegle marmelos in SD rats. Qualitative Phytochemical analysis of methanolic un ripe fruit-pulp extract of Aegle marmelos was done as per standard method. Extracts of methanolic unripe fruit-pulp extract of Aegle marmelos were evaluated at doses of 15mg/kg, 30mg/kg, 120mg/kg and 1600mg/kg in SD rats for anti diarrhoeal effect in castor oil induced diarrhoea. For Toxicity effect, 5000 mg/kg of methanolic extract of Aegle marmelos was administered orally to three female rats at first with 0.02% tween 80 vehicles and observed for any abnormal condition up to 14days .Statistical analysis was conducted by ANOVA following post hoc test. Qualitative analyses of different phytochemicals of methanolic extract of Aegle marmelos showed that the presence of Tannin and Terpenoids were highly positive whereas Alkaloid was moderately positive and Flavonoid was slightly positive. The study revealed that the dose of 30mg/kg BW and 1600 mg/kg BW of methanolic extract of A. marmelos showed same treatment response with Loperamide against castor oil induced diarrhoea in rats. Safety study did not show any side effects of high dose of A. marmelos therapy during rat model study. It can be concluded that methanolic extract of Aegle marmelos may be effective in reducing diarrhoea in animals

Therapeutic Management of Chronic Renal Failure in a Dog

A case of chronic renal failure (CRF) was diagnosed in an eight year old Labrador dog on the basis of history, clinical signs, physical examination, hematology, plasma biochemistry, followed by confirmation with the help of ultrasonography and urinalysis. Urinalysis on the day of presentation revealed turbid straw colored urine, decreased specific gravity, trace amount of non-haemolysed blood, alkaline pH, marked proteinuria, high inflammatory cells, leucocyte count (125cells/µl) and epithelial cast. Basing on the creatinine value, the case was diagnosed as terminal stage 4 CRF. The patient was treated with fluid therapy, antibiotic, antacid, antiemetic, calcium supplement, multivitamin, and erythropoietin. Though the prognosis in case of end stage CRF is guarded, nevertheless, it took 30 days to bring back the animal from stage 4 to stage 2 with clinical signs of improvement and it was followed up to 5 months without any further complication.

Management of Incomplete Uterine Inertia in a Labrador Bitch - A Case Report -

A Labrador bitch having fifty four days of pregnancy was presented at college clinics with the history of no sign of labor after birth of one pup. The bitch was found active with dilated cervix. Abdominal palpation and ultrasonographic imaging revealed presence of foetuses in the gravid uterus. The case was further diagnosed as incomplete uterine inertia. In the present case study it was noted that intravenous administration of oxytocin, calcium gluconate and dextrose saline as effective therapeutic agents to expelled the fetuses.

Therapeutic evaluation of homoeopathic drug Crotalus horridus 200C against Ehrlichiosis-infected dogs in Mizoram

Objective: To study, the effect of a homoeopathic medicine Crotalus horridus 200C on ehrlichiosis in dogs in an endemic area of Aizawl district of Mizoram state of India. Materials and Methods: To evaluate the efficacy of Crotalus horridus 200C against ehrlichiosis dogs. 12 positive cases confirmed by polymerase chain reaction (PCR) were divided into two groups comprising six dogs in each group. One group was treated with standard therapy (doxycycline) and other group was treated with Crotalus horridus 200C at 4 pills orally for 20 days. Clinical improvement of affected dogs was recorded after therapy. Important haemato-biochemical parameters before and after therapy such as haemoglobin (Hb), packed cell volume (PCV), total erythrocyte count (TEC), total leukocyte count (TLC), differential leukocyte count (DLC), platelet count, total protein, albumin, globulin, A:G ratio, total bilirubin, serum creatinine, blood urea nitrogen (BUN), and liver-specific enzymes namely alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were assessed following standard protocol. All the parameters were compared with a control healthy group (T3). All experiment dogs were from different age with different breeds and bloods were collected at forenoon only. Results: PCR test yielded 13 dogs positive out of 67 suspected samples screened (19.40%) with an amplification of 387 bp fragment from 16S rRNA gene of E. Canis. Off total positive, only 8 (61.53%) could be detected in peripheral blood smear. Crotalus horridus-treated group of dogs showed clinical recovery from fever and temperature to normalcy by the 14th day posttreatment. Haemato-biochemical profiles of affected dogs such as Hb, PCV, TEC, TLC, DLC, platelet count, total protein, albumin, globulin, A:G ratio, total bilirubin, serum creatinine, BUN, and liver-specific enzymes namely ALT and ALP were turned to normalcy within 21 days of post-treatment. Conclusion: Nested PCR assay had been shown to be sensitive and specific for detection of Ehrlichia canis. Crotalus horridus 200C may be an effective and choice of drug for control of canine ehrlichiosis.