Concurrent quantitation of the A and D genotypes of hepatitis B virus

Hepatitis B virus (HBV) infection is a global health problem associated with severe liver disorders. Viral load and HBV genotype affect the clinical outcome, guide antiviral therapy and provide long term prognosis for HBV infected patients. Various types of detection and quantitation assays are currently in use with a different effectiveness. The aim of this study was to develop a method that would provide simultaneous identification and quantitation of genotypes A and D in a single-tube reaction. Sera from infected patients were analyzed by a TaqMan based real time PCR. Optimized reagents were used for HBV DNA quantitation while the genotypes A and D were quantified separately by our design of the assay. Multiplex real time PCR was achieved and was specific for HBV genotypes A and D within a single-tube reaction. Simulation of mixed virus populations was identified reproducibly in vitro. Quantitation of these individual genotypes was exceptionally reliable, so much so that the sum of individual genotypes was equal to the total viral load determined in a separate reaction. Therefore, a straightforward, conceptually simple and reliable approach to issues involving HBV genotypes A and D is submitted. Identity and exact titer of these genotypes in the Caucasian population can now be determined easily. (C) 2009 Elsevier B.V. All rights reserved

Immunohistomorphometric Characteristics Of Pituitary GH Cells In Infant And Peripubertal Female Rats After Treatment With Estradiol Or Human Chorionic Gonadotropin

The effects of estradiol-dipropionate (EDP) or human chorionic gonadotropin (hCG) on immunohistomorphometric characteristics of pituitary GH cells in infant and peripubertal female rats were investigated. The first group of females received five injections of EDP (0.25 mg/kg b.w.) during the neonatal period of life, and was further divided into two subgroups which were sacrificed at the infantile period (17th day) or at the peripubertal period (38th day). The second group received two doses of hCG (50 IU/kg b.w.) on the 15th and 16th day of life in the first subgroup, and on the 36th and 37th days of life in the second subgroup, while they were sacrificed 24 h after the last treatment, respectively. The control females were injected with an equivalent volume of the vehicle and sacrificed according to the appropriate schedules as the hormone treated rats. EDP treatment decreased GH cell volume density in infant and peripubertal females, by 38% and 76% (p<0.05) respectively, in comparison with the controls. The number of GH cells per mm2 in infantile and peripubertal period was decreased in EDP treated animals by 26% and 53% (p<0.05) respectively, compared to the controls. Also, upon EDP treatment in both periods, GH cells were diminished in size and less intensely immunolabelled than in the control groups. The morphometric parameters in animals treated with hCG were insignificantly changed in both analyzed periods, in comparison with the controls. Unlike hCG, EDP manifested clear inhibitory effects on the immunohistomorphometric characteristics of GH cells in examined female rats

IMMUNO-HISTOMORPHOMETRIC AND -FLUORESCENT CHARACTERISTICS OF GH CELLS AFTER TREATMENT WITH GENISTEIN OR DAIDZEIN IN AN ANIMAL MODEL OF ANDROPAUSE

Somatopause, the complex aspect of andropause, is recognizable by reduced growth hormone - GH/insulin-like growth factor 1 axis function in the ageing male. Soy isoflavones (usually genistein and daidzein), which are known for their beneficial effects in the treatment of ageing symptoms, are active in the pituitary, as well. The iromuno-histomorphometric and fluorescent characteristics of pituitary growth hormone secreting cells, in an animal model of andropause, were examined after a treatment with genistein or daidzein. Andropausal Wistar rats were divided into sham operated, orchidectomized and genistein or daidzein treated orchidectomized groups. Genistein or daidzein (30 mg/kg/day) were administered subcutaneously for three weeks, while sham operated and orchidectomized groups received the vehicle alone. Growth hormone secreting cells were identified by the percoxidase-antiperoxidase immuno-histochemical, and inmuno-fluorescent procedure. The main characteristic of growth hormone secreting cells in soy isoflavones treated groups is a weaker immuno-histochemical staining and immuno fluorescent signal compared to sham operated and orchidectomized groups. The growth hormone secreting cell volume in orchidectomized +genistein or +daidzein groups is by 13.8\% and 11.9\% (p<0.05) smaller respectively, in comparison with the orchidectomized group. In orchidectomized +genistein or +daidzein groups, the growth hormone secreting cells relative volume density is by 62.5\% and 61.0\% lower (p<0.05) respectively than for the sham operated group, and decreased by 65.4\% and 64.0\% (p<0.05) respectively, compared to the orchidectomized group. It can be concluded that chronic genistein or daidzein treatment, in an animal model of andropause, attenuates immuno-histomorphometric and fluorescent characteristics of growth hormone secreting cells.Ministry of Education, Science and Technological Development of the Republic of Serbia {[}173009