Mikania micrantha extract enhances cutaneous wound healing activity through the activation of FAK/Akt/mTOR cell signaling pathway.

Mikania micrantha (MM) has been traditionally used for various health benefits, including mental health, anti-inflammatory, wound dressing, and healing of sores. However, the molecular mechanisms and dose required for the wound healing activity of MM have yet to be reported. Therefore, a study was conducted to evaluate the wound healing potential of a cold methanolic extract of MM through in vitro and in vivo studies. Human dermal fibroblast adult (HDFa) cells were treated with 0 (control), 75 ng/ml, 125 ng/ml, 250 ng/ml, and 500 ng/ml of MMmethanolic extract (MME) for 24 h. MME at 75 ng/ml has significantly (p˂0.05) promoted HDFa cell proliferation and migration. Further, MME has also been shown to enhance the invasiveness of human umbilical vascular endothelial cells (HUVECs), indicating the neovasculature for wound healing. The tube formation assay demonstrated a significant (p<0.05) increase in the angiogenic effect of the MME starting at a concentration of 75 ng/ml as compared to the control. Treatment of excision wounds in Wistar rats with 5% and 10% MME ointment significantly enhanced wound contraction compared to control animals. Incision wounds in rats treated with 5% and 10% MME showed a significant (p<0.01) increase in tensile strength compared to control. HDFa cells, and granulation tissue collected on day 14 post-wounding, revealed the modulation of the FAK/Akt/mTOR cell signaling pathway during the enhancement of wound healing. The results of gel zymography showed increased activity of MMP-2 and MMP-9 in the HDFa cells after treatment with the extract.  It is concluded that MMEcan potentially accelerate cutaneous wound healing.

Psoroptes mites infestation in a captive Burmese Red Serow (Capricornis sumatraensis subspecies rubidus) of Indo-Burma bio-diversity hotspot.

An 8 years old male Burmese Red Serow (Capricornis sumatraensis sub species rubidus) from Aizawl Zoological Park was presented to the Teaching Veterinary Clinical Complex. An exploratory skin scraping revealed existence of nymphal as well as adult stages of mites of the Psoroptes spp. which were not associated with any overt lesions typical to mite infestation such as pruritus, erythema or scaling of the epidermis. The mites were identified as per their morphology, size and shape. Haemato-biochemical analysis revealed alteration of certain haematological and biochemical parameters. The red blood corpuscles were found to have anucleate cells with mild to absent central pallor. The absolute counts showed neutrophilic leucocytosis with mild monocytosis and lymphocytosis. Eosinophilic count was towards the higher side, indicating that the infestation was mild. The serum calcium, albumin, triglyceride, urea nitrogen (BUN) were found to be lower than normal, whereas serum ALT, AST, LDH, ALP and serum amylase were higher than the established reference indices. Due to lack of haemato-biochemical reference values specific to captive Serow, emphasis was given in this report to establish baseline data for this species.

Theileriasis in crossbred cows and its therapeutic management: first report from Lushai hill district of Mizoram.

To know the prevalence of theileriosis infection, haemato-biochemical alteration and efficacy of Oxytetracycline in combination with Diaminazene aceturate injection against theileriosis in Lushai hill district of Mizoram; fifty five cross bred cattle were screened during July, 2013 to November, 2013. The prevalence of tropical theileriosis was 9.09 % based on blood smear examination. Clinical features of the disease were manifested by high rise of body temperature 104.8 ± 1.2 °F, lymphadenopathy, inability to walk, anorexia, listlessness, moderate anaemia, nasal discharge, edema of eye and corneal opacity. There were marked decline (P < 0.01) in the haemoglobin 5.28 ± 1.81 gm/dl, total erythrocytic count 2.14 ± 0.7 × 10(6)/ul, packed cell volume 18.10 ± 0.8 % and total leukocyte count 6.95 ± 1.05 × 10(3)/ul, respectively. Erythrocyte sedimentation rate was significantly increased (P < 0.01) up to 104.5 ± 12.4 mm/1st hour. The percentage of neutrophils increased 62.75 ± 2.77 and lymphocytes decreased drastically 28.69 ± 1.45 as compared with un-infected cross bred cattle. Clinically infected cattle with Theileria annulata had significantly low levels of total proteins and albumin, (P < 0.05) but AST, total, direct and indirect bilirubin, BUN, creatinine levels and Ck-MB activity were significantly high (P < 0.05). Oxytetracycline in combination with Diaminazene aceturate was found effective against T. annulata and could control fever and temperature returns to normalcy by 7th day post treatment along with some haemato-biochemical profiles. In brief, the study revealed that tropical theileriosis was prevalent in Lushai hill district of Mizoram especially among cross breed cattle and had effect on hepatic, renal and cardiac function which can be effectively controlled by Oxytetracycline and Diaminazene aceturate injection.

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.

First Report on Vancomycin-Resistant Staphylococcus aureus in Bovine and Caprine Milk.

The present investigation was carried out to study the vancomycin resistance pattern of Staphylococcus aureus isolates (n = 274) obtained from 352 milk samples of bovine (269) and caprine (63) clinical and subclinical mastitis from different districts of West Bengal, India. Of them, seven isolates (vancomycin-resistant S. aureus [VRSA] 1-7) exhibited resistance to vancomycin. Minimum inhibitory concentration of vancomycin (MICvan) for VRSA2 and VRSA3 was ≥16 µg/ml; thus categorized as VRSA. For rest of the isolates, MICvan was 8 µg/ml and they were grouped as vancomycin intermediate S. aureus (VISA). Even though all the isolates were resistant to cefoxitin and oxacillin and possessed mecA gene, none of them carried vancomycin resistance gene. Furthermore, all the seven isolates were subjected to Staphylococcal cassette chromosome mec (SCCmec) typing, Staphylococcal protein A (spa) typing, and enterobacterial repetitive intergenic consensus polymerase chain reaction. All the isolates except VRSA3 and VRSA4 from Kolkata district exhibited diverse genetic lineage, irrespective of their host and antibiotic resistance pattern. These two isolates showed clonal similarity (MRSA-SCCmec-V-spa t267) with methicillin-resistant S. aureus (MRSA) strains previously reported in human and animal infection. Isolation of VRSA and VISA could probably be due to intensive use of vancomycin in healthcare premises, which might have led to the development of glycopeptide-resistant strains and thereafter, further disseminated in the environment, including livestock farms. Detection of VRSA in milk is a serious concern as it may further cause health problems in the consumers. This is the first ever report of VRSA in food animals, even though the pathogen is otherwise prevalent in humans.

Co-infection of methicillin-resistant Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus and extended spectrum ß-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 ß-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.

Genotyping of Cryptosporidium parvum isolates in bovine population in Kolkata and characterization of new bovine genotypes.

Cryptosporidium infection may have adverse effect in health and production potential of cattle herd. The exact profile of Cryptosporidium infection in bovine population of India in general, particularly from Kolkata is scarce. We here report systematic investigation of clinical and genetic profiling of promiscuous Cryptosporidium infection in selected representative cattle farms from Kolkata as well as some surrounding local areas. The current study was conducted in the period of October to September, 2000-2001 with 149 diarrhoeic and non-diarrhoeic cattle of different age groups from two Government cattle farms, Harringhata Cattle Unit and Kalyani State Livestock Farm and animals raised by local farmers. Among these 149 samples, diarrhoea was recorded in 79 cases (53%) and non-diarrhoeic in 70 (46.9%). Out of 149 faecal samples screened microscopically, 32.9% from diarrhoeic faecal samples and 7.1% from healthy faecal samples revealed the presence of oocysts. Cryptosporidium genus was confirmed by DNA typing with nested PCR. The PCR-RFLP analysis was carried out for genotype identification. In course of PCR-RFLP, unique band patterns were obtained in two of our samples. The unusual RFLP products were characterized by DNA sequencing and homology analysis with other reported variants. This is the first report of identification and characterization of such a variant from the area of present investigation. Further study will be required to understand the phylogenetic origin and functional significance in virulence and morbidity of this genotype.

Therapeutic management of copper deficiency in buffalo heifers: impact on immune function.

To evaluate the magnitude of copper deficiency in Northern India and to examine the various haematobiochemicals, enzymes, vitamins and immune functions affected by copper deficiency, and to identify the parameters which can be of diagnostic importance in copper deficiency, a survey was conducted in 12 districts of Northern India. Significant deficiency of copper was observed in soil, fodder and serum samples of buffalo heifers. Fifty hypocuperaemic buffalo heifers were selected from these areas and were randomly divided into two groups, A and B. The heifers in group A were provided with mineral mixture containing copper sulphate and in group B without copper sulphate. Significant (p<0.01) improvement in serum ceruloplasmin level was observed within 30 days of treatment, while significant (p<0.01) improvement in monoamine oxidase and liver cytochrome oxidase was observed at the 60th day of treatment in group A animals. Significant improvement was observed in T(3) and T(4), in the animals of group A within 60 days of treatment. The values of vitamin A and E showed significant (p<0.01) improvement within 30 days of treatment. The phagocytic activity of neutrophils against Candida albicans significantly (p<0.01) improved in group A within 60 days of treatment. Similarly, significant improvement in superoxide dismutase activity in red blood cells was observed at the 30th day, and in total leukocytes and whole blood at the 60th day in group A animals. Significant improvement in liver copper level was observed at the 30th day of treatment, while in group B the liver copper was significantly (p<0.01) depleted at the 60th day of experimentation. Additional copper supplementation improved growth performance significantly in group A.