Approaches used to improve epigenetic reprogramming in buffalo cloned embryos

The reproductive cloning in buffalo in India has been started using a simplified somatic cell nuclear transfer technique named handmade cloning. Since the birth of first cloned female buffalo in 2009, a number of buffalo clones have been produced in India by utilizing different types of donor cells such as ear cells, embryonic stem cells, semen somatic cells and urine somatic cells. The use of buffalo cloning on a large scale is restricted due to low pregnancy rates and poor calf survival. Considerable attempts have been made to improve the overall buffalo cloning efficiency, particularly by modifying epigenetic reprogramming of cloned embryos. Previous studies have demonstrated that chemical epigenetic modifiers such as trichostatin A and 5-aza-2'-deoxycytidine, m-carboxycinnamic acid bishydroxamide can be used to treat donor somatic cells and reconstructed fused embryos to correct the epigenetic reprogramming to enhance the overall cloning efficiency in terms of live birth rates.

Radiation therapy outcomes in muscle invasive urinary bladder cancer: A single institution experience.

INTRODUCTION: To audit the survival outcomes and loco‑regional control in muscle invasive urinary bladder cancer patients treated with external beam radiation therapy (RT). MATERIALS AND METHODS: From November 2008 through December 2011, 50 consecutively diagnosed muscle invasive urinary bladder carcinoma (T2‑4a N0‑2, M0) patients were included in this retrospective study. All these patients received external beam RT to a median dose of 60 Gy (range 30‑66 Gy), and were not suitable for radical surgery due to patients’ preference or medical comorbidities. A stepwise procedure using proportional hazard regression was used to identify prognostic factors with respect to survival. RESULTS: Completion trans‑urethral resection of bladder tumor was done in 38 (76%) patients of the cohort and 47 (94%) had transitional cell carcinoma on histopathology. Clinical stage T2 was diagnosed in 40 (80%) patients. The median follow‑up for the entire cohort was 14 ± 8.9 months (range 1‑36 months). In conclusion, 24 patients (48%) were free of disease, 5 patients (10%) had residual disease, and 13 patients (26%) had died of disease. Two‑year and 3 year overall survival of intact bladder for the entire cohort was 58% and 43.6%, respectively. Cox regression modeling strongly suggested clinical stage (P = 0.01) and RT dose (P = 0.001) as being predictors for overall survival. CONCLUSION: RT shows reliable outcomes and excellent compliance in this advanced disease. Prescribing a higher RT dose could potentially correlate to better intact bladder control rates while maintaining good quality of life in selected patients.

Antiviral activity of crude extracts of Eugenia jambolana Lam. against highly pathogenic avian influenza (H5N1) virus.

Crude extracts of leaves and bark of E. jambolana were tested for antiviral activity against highly pathogenic avian influenza virus (H5N1) by CPE reduction assay in three different layouts to elucidate virucidal, post-exposure and pre-exposure antiviral activity of the extracts. The cold and hot aqueous extracts of bark and hot aqueous extract of leaves of E. jambolana showed significant virucidal activity (100% inhibition) which was further confirmed in virus yield reduction assay (~98 to 99% reduction) and by egg based in ovo assay. The selective index (CC50/EC50) of hot aqueous extract (248) and cold aqueous extract (43.5) of bark of E. jambolana showed their antiviral potential against H5N1 virus. The significant virucidal activity of leaves and bark of E. jambolana merits further investigation as it may provide alternative antiviral agent for managing avian influenza infections in poultry farms and potential avian-human transmission.

Short-term zinc deficiency in diet induces increased oxidative stress in testes and epididymis of rats.

In order to determine the effects of Zinc deficient diet on oxidative stress in testis and epididymis, various parameters viz: total proteins, lipid peroxidation, hydroperoxides, antioxidant capacity and enzymatic activities are evaluated in rats fed on zinc deficient diet for 2, 4 and 6 weeks. Total proteins, water and lipid solouble antioxidant capacity decreased while lipid peroxidation (TBARS) and hydroperoxides concentration increased in testes, caput and cauda epididymis except in 2ZD (testes) where hydroperoxides revealed a significant decrease. GSH decreased in testes and caput and cauda epididymis. GPx and gamma-GT activities increased in testes and caput and cauda epididymis of zinc deficient rats. Further, GST increased in testes but exhibited decreases after 2 and 4 weeks and an increase after 6 weeks in caput and cauda epididymis of zinc deficient rats. GR activities decreased in testes but it increased in caput and cauda epididymis of zinc deficient rats. Thus, zinc deprivation results in increased sensitivity to oxidative stress. All these may have been as a consequence of increased ROS generation and/or decreased zinc dependent antioxidant processes.

Mentha piperita (Linn) leaf extract provides protection against radiation induced alterations in intestinal mucosa of Swiss albino mice.

Intestinal protection in mice against radiation injury by M. piperita (1 g/kg body weight/day) was studied from day 1 to day 20 after whole body gamma irradiation (8 Gy). Villus height, goblet cells/villus section, total cells, mitotic cells and dead cells/crypt section in the jejunum are good parameters for the assessment of radiation damage. There was significant decrease in the villus height, number of total cells and mitotic cells/crypt section, whereas goblet cells and dead cells showed significant increase after irradiation. Mentha pretreatment resulted in a significant increase in villus height, total cells and mitotic cells, whereas goblet cells and dead cells showed a significant decrease from respective irradiated controls at each autopsy day. The results suggest that Mentha pretreatment provides protection against radiation induced alterations in intestinal mucosa of Swiss albino mice.

A study of prevalence of health problems in asymptomatic elderly individuals in Delhi.

A survey was conducted in asymptomatic aged individuals (> or = 60 years) in The National Capital Territory of Delhi for the prevalence of major health problems like hypertension, diabetes mellitus and respiratory diseases. A total of 200 individuals (100 males and 100 females) were studied over a period of three months in 1998-99. Hypertension was defined as BP > or = 140/90 mmHg (JNC VI criteria), while diabetes mellitus was diagnosed if fasting whole blood sugar was 120 mg/dl or more (WHO criteria). Diagnosis of other health problems was based on relevant history and physical examination. Prevalence of hypertension in the study group was 32.5 per cent (more in males). Of these 18 per cent and 4.2 per cent had isolated systolic and diastolic hypertension, respectively. Prevalence of diabetes mellitus in the same population was 13.0 per cent. Both diseases were more prevalent in urban population. A high prevalence of respiratory disorders was observed (pulmonary tuberculosis 16 per cent, COPD 10 per cent, asthma 4.5 per cent). Cataract was present in 7.5 per cent while 1.5 per cent had symptoms of urinary tract infection. History of Jaundice was present in 3.5 per cent. Three per cent each had a history suggestive of IHD and TIA, respectively. Proteinuria and glycosuria was seen in 22.2 and 7.6 percent, respectively. A large percentage of the study group (34.4 per cent) had asymptomatic ECG abnormalities.

Characterization of toxin from cheilanthes fern and its effect on lymphocyte proliferation and DNA fragmentation.

Thin layer chromatography of aqueous extract of whole Cheilanthesfarinosa fern indicated the presence of ptaquiloside or ptaquiloside like compound, coinciding Rf values with that of Pterosin B standard. HPLC analysis revealed the presence of 26.3 mg/kg ptaquiloside. In vitro studies of the aqueous extract on lymphocyte culture revealed a correlation between stimulative indices and concentration of aqueous extract. Stimulation in lymphocyte proliferation was in order of bracken > cheilanthes > ConA> ptaquiloside standard. On incubation of lymphocyte with aqueous extract of ferns, no DNA damage was observed in isolated DNA.

Apoptosis in bovine herpesvirus-1 infected bovine peripheral blood mononuclear cells.

Apoptosis is a process whereby cells die in a controlled manner in response to various stimuli like cytotoxins, viral antigens and normal physiological signals during differentiation and development. Virus induced immunosuppression has been reported for various viral diseases including Bovine Herpesvirus-1 (BHV-1). In the present study, BHV-1 was found to cause apoptosis in ConA stimulated bovine peripheral blood mononuclear cells (PBMCs). Apoptotic index quantified by fluorescent dyes revealed a significant (P < 0.001) increase in percent apoptotic cells at 2, 24 and 48 hr post infection as compared to their respective non-infected controls. Apoptosis specific internucleosomal laddering in DNA from BHV-1 infected PBMCs was seen in agarose gel electrophoresis. No DNA fragmentation was observed in control non-infected PBMCs.

Embryonic/fetal mortality after exposure to tritiated water in pregnant Swiss albino mice during different gestation periods.

Pregnant Swiss albino mice were given a priming injection(im) of tritiated water (HTO) at the dose rate of 2.3 and 5 microCi/ml body water (74, 111 or 185 KBq/ml body water) at 0, 6 and 14 day post conception (d.p.c.) and were subsequently maintained on tritiated drinking water ad libitum during preimplantation (0-5 d.p.c.), organogenetic (6-12 d.p.c.) or fetal (14-18 d.p.c.) period, respectively. On day 18 of gestation the females were sacrificed by cervical dislocation to record the implant sites per dam and embryonic/fetal mortality. Significant reduction was observed in average implant sites per dam when the females were exposed to any of the three doses during the preimplantation period due to embryonic resorption before implantation. However, the same was found to be within the normal range when mothers were exposed during the organogenetic or fetal period. Prenatal mortality (embryonic resorption/fetal death) was higher after in utero exposure to different doses during preimplantation period as compared to organogenetic period, but mortality did not occur after exposure to any of the doses during the fetal period. Occurrence of mortality was found to be dose dependent.