Production of cloned mice by nuclear transfer of cumulus cells

Over the past several years, mammals have been successfully cloned by either the splitting of an early stage embryo or nuclear transfer of adult somatic cells [NT] into oocytes. Although it has been 15 years since the generation of the first cloned mammals from somatic cells by NT, the success rate for producing live offspring by this technique is low regardless of the cell type and animal species used. However, these techniques have the potential to be important tools for future research in basic biology. In the present study, we described our experiences in producing successfully cloned mouse using NT method and piezo-actuated micromanipulator. B6D2F1 mice, 8-12 weeks old, were superovulated with injections of 5 IU of pregnant mare serum gonadotropin and 5 IU of human chorionic gonadotropin administered 48 hr apart. Enucleation and donor nuclei cumulus cell injection were performed with a piezo-actuated micromanipulator after which activation and trichostatin A treatment were used for reconstructed oocytes. Two-cell stage cloned embryos that developed in the mWM medium were transferred into the oviducts of pseudopregnant NMRI mice. Of 367 oocytes collected, 131 [69%] developed into 2-cell stage embryos. Of these, 5 [1%] live pups were successfully delivered. We used NMRI foster mother to raise the pups by lactation. One adult cloned mouse was mated, after which she delivered and raised normal offspring. For mouse cloning, the present study also successfully tested the capability of somatic cell nuclear transfer SCNT using a piezo unit

Alteration in antioxidant capacity in patients with chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease [COPD] is a major public health problem that needs greater attention. Variability in the susceptibility to develop COPD is related to both genetic and environmental factors. Oxidative stress and inflammation are the major hallmarks of COPD and antioxidant status can be used as a biomarker to assess the risk of chronic diseases. We used the FRAP [ferric reducing ability of plasma] assay as a simple and powerful test for determination of the total antioxidant capacity of plasma of patients and normal subjects. The patients were selected by cross-sectional method. The mean average age +/- SD of normal subjects and patients was 56 +/- 4 and 60 +/- 2 years respectively. The spectrophotometeric method was used for this assay. The means of the FRAP assays in the patients were higher [about twice] than those of normal subjects. The differences were significant [p < 0.01]. The high levels of antioxidant capacity in the patient group indicated that the antioxidant defense system had been activated due to the oxidative stress and hypoxic condition. A though, FRAP assay can probably be used for demarcation of severity and risk of developing COPD, clinical follow-up and further investigation are required for the assessment of this hypothesis