Evaluation of a side population of canine lymphoma cells using Hoechst 33342 dye

Cancer stem cell (CSC) research has increased exponentially to gain further insight into the mechanisms underlying both carcinogenesis and chemotherapy resistance. The present study was performed to explore the potential value of a side population (SP) assay for identifying and characterizing putative CSCs among canine lymphoma cells. Canine lymphoma cells from cell lines and clinical samples were subjected to the SP assay consisting of Hoechst 33342 staining and subsequent flow cytometric analysis. The SP assay revealed various amounts of a SP fraction among the canine lymphoma cells. The percentages of SP were not affected by inhibitors of membrane transporters, verapamil hydrochloride, or fumitremorgin C. Most of the canine lymphoma cells expressed high levels of Bmi-1 and membrane transporter proteins such as ABCG2 and phosphorylated (p)-glycoprotein. This investigation lays the groundwork for further studies of the biological behaviors and molecular characteristics of CSCs in cases of canine lymphoma.

Comparative effect of candesartan cilexetil and atenolol on glucose metabolism in essential hypertensive patients

To compare the changes on Glucose metabolism in essential hypertensive patients. Randomized, open-labeled, prospective comparative study. The study was carried out in the department of pharmacology and therapeutics, Basic Medical Sciences Institute [BMSI], Jinnah Post Graduate Medical Centre [JPMC], Karachi, from January 2007 to December 2008. In this study [80] newly diagnosed essential hypertensive patients were enrolled and divided into two groups [DR1] and [DR2] respectively. DRI patients received Candesartan 16 mg once a day and DR2 patients received Atenolol 50mg once a day for 3 months duration. At the end of study period the parameters examined were fasting and random blood glucose levels. The results have been expressed as mean +/- SEM [Standard error of Mean]. The mean fasting blood glucose level was decreased [from 90.22+ 1.34 mg/dl to 89.20 +/- 1.70 mg/dl; for DR1] versus DR2 in which mean fasting blood glucose level was increased [from 91.82 +/- 1.34 mg/dl to 97.91 +/- 1.22 mg/dl; p<0.001]. A reduction of 1.13% in case of DR1 and an increase of 6.63% were observed in case of DR2 group. The mean random blood glucose level was decreased for DRI group [from 151.12 +/- 0.99 mg/dl to 149.36 +/- 0.80 mg/dl] versus DR2 in which mean random blood glucose level was increased from [155.47 to +/- 0.96 mg /dl to 168.83 +/- 0.81 mg/dl; p<0.001]. A reduction of 1.16% was observed in case of DR1 group while and increase of 8.59% was observed in case of DR2 group, respectively. Candesartan cilexetil is a newer and safer alternative for the treatment of diabetic hypertensive patients in comparison to Atenolol treatment

Comparative study of PM2.5 - and PM10 - induced oxidative stress in rat lung epithelial cells

Accurate estimation of the exposure-response relationship between ambient urban particulate matters (PM) and public health is important for regulatory perspective of ambient urban particulate matters (PM). Ambient PM contains various transition metals and organic compounds. PM10 (aerodynamic diameter less than 10 microgram) is known to induce diverse diseases such as chronic cough, bronchitis, chest illness, etc. However, recent evaluation of PM2.5 (aerodynamic diameter less than 2.5 microgram) against health outcomes has suggested that the fine particles may be more closely associated with adverse respiratory health effects than particles of larger size. This study was performed to evaluate PM2.5-induced oxidative stress in rat lung epithelial cell in order to provide basic data for the risk assessment of PM2.5. PM2.5 showed higher cytotoxicity than PM10. Also, PM 2.5 induced more malondialdehyde (MDA) formation than PM10. In Hoechst 33258 dye staining and DNA fragmentation assay, apopotic changes were clearly detected in PM2.5 treated cells in compared to PM10. Expression of catalase mRNA was increased by PM2.5 rather than PM10. PM2.5 induced higher Mth1 mRNA than PM10. In pBR322 DNA treated with PM2.5, production of single strand breakage of DNA was higher than that of PM10. In Western blot analysis, PM2.5 induced more Nrf-2 protein, associated with diverse transcriptional and anti-oxidative stress enzymes, compared to PM10. Our data suggest that PM2.5 rather than PM10 may be responsible for PM-induced toxicity. Additional efforts are needed to establish the environmental standard of PM2.5.

Optimization of tumour radiotherapy: Part V--Radiosensitization by 2-deoxy-D-glucose and DNA ligand Hoechst-33342 in a murine tumour.

Radiosensitizing effects of combination of a minor groove DNA ligand, Hoechst-33342, with the glucose analogue and inhibitor of glycolysis, 2-deoxy-D-glucose (2-DG) have been investigated in Ehrlich ascites tumour (EAT) bearing mice following focal irradiation of the tumour with 60Co gamma-rays. Treatment-induced tumour growth delay and tumour free animal survival were evaluated as parameters of radiation response. Focal irradiation of the tumour with a single fraction of 10 Gy induced a moderate delay in tumour growth but did not lead to complete regression in any of the tumours. Intravenous administration of H-342 1 hr before irradiation enhanced radiation-induced growth delay in a dose dependent manner. Complete regression of the tumour was observed only at a dose of 10 mg/kg body wt, leading to a cure (tumour free survival for more than 100 days) rate of 55%. Administration of 2-DG (2 g/kg body wt; i.v.), immediately before irradiation significantly enhanced radiation-induced growth delay and resulted in a cure rate of 45%. In combination with this dose of 2-DG (2 g/kg body wt), H-342 at a lower dose (5 mg/kg body wt) significantly enhanced the cure rate to 66%. H-342 or 2-DG given alone or in combination at the doses investigated here did not show any significant effects on the unirradiated tumour.