Designing and engineering of DNA-vaccine construction encoding multiple CTL-epitopes of major HIV-1 antigens

A synthetic T cell immunogen (TCI) has been designed as a candidate DNA-based vaccine against Human immunodeficiency virus (HIV)-1 using cytotoxic T lymphocytes (CD8(+) CTL) and T-helper lymphocytes (CD4(+) Th) epitopes retrieved from the Los Alamos HIV Molecular Immunology Database. The protein 392 amino acids in length contains about eighty CTL-epitopes, many of which are overlapping and are totally restricted by ten different HLA class I molecules. To be able to detect CTL responses induced by a DNA vaccine in experimental animals, additional epitopes, restricted by mouse and Macaque rhesus major histocompatibility complex (MHC) class I molecules, were included in the target immunogen. The gene encoding the TCI protein was assembled, cloned into vector plasmids and expressed in a prokaryotic and a eukaryotic system. The presence of HIV-1 protein fragments in the immunogen structure was ascertained by ELISA and immunoblotting using panels of HIV-1-positive sera and monoclonal antibodies to p24. It has been demonstrated that DNA vaccine can induce both specific T cell responses (CTL and blast transformation) and specific antibodies in mice immunized with pcDNA-TCI.     

S100B protein stimulates calcineurin activity

S100B is a calcium binding protein from astrocytes that regulates protein phosphorylation by binding to substrates and protein kinases. S100B might also regulate protein phosphatases and this was investigated for protein phosphatase 2B (calcineurin). The results indicate that S100B (5-10 microM) increased the activity of both purified and cytoskeletal calcineurin in a Ca-dependent manner. This effect was blocked by a specific inhibitor of calcineurin activity, but not by TRTK-12 (an inhibitor of S100B binding to other protein targets). The present results and the known co-localization of S100B and calcineurin in the astrocyte cytoskeleton suggest that S100B may play a role in the phosphorylation state of cytoskeletal proteins.     

The treatment of occupational fluorosis by acupuncture reflexotherapy]

The article contains the electrodiagnostic data on 26 auricular points in 24 fluorosis-affected patients. It was established that the greatest electrical polarity asymmetry was found in points 13, 25, 28, 31, 54, 37, 39 and 40. Palpation of the auricular and corporal points also showed unfavorable conditions in the respective painful point of the organ. Basing on the results of the acupuncture procedures, needle therapies of 80 occupational fluorosis cases were performed. The therapeutic patterns elaborated by the author were also proposed, which proved perfect results at different stages of the disease.     

The interdependence between neurodynamic indices and the state of the hearing in blast trauma]

The authors have proved that in the blast trauma diagnosis of the organ of hearing in slight or middle-gravity cases it was necessary to applicate such important additional methods of researching as pure-tone and speech audiometry, and EEG. It gives the possibility to reveal the affections in primary and secondary cortical fields of temporal, occipital and frontal lobes, prognosticate the possible outcome of hearing pathology, elaborate optimal schemes of treatment, control the efficiency of therapy, and also make expert evidence conclusions. It was proved that the diminished hearing in patients who had suffered from blast injuries have a direct correlative dependency upon pathological deviations in EEG indexes. The severe neurodynamic disorders form an unfavourable background for recovery of hearing.     

Noninvasive determination of the optical properties of adult brain: near-infrared spectroscopy approach

The basic parameters for physiological measurements provided by near-infrared spectroscopy are the local absorption and scattering coefficients. For the adult human head, they have been difficult to measure noninvasively because of the layered structure of the head. The results of measurements of absorption and reduced scattering coefficients through the forehead on 30 adult volunteers using a multidistance frequency domain method are reported. The optode separation distance ranged from 10 to 80 mm and measurements were recorded at 758 and 830 nm. The measured absorption and reduced scattering coefficients of the forehead were used to evaluate the hemoglobin content in the scalp and brain as well as cerebral oxygen saturation. We found that cerebral oxygenation was relatively narrowly distributed within the subject group (the standard deviation was about 3% for scalp and 6% for brain, respectively), whereas hemoglobin concentrations had a relatively broader distribution. We found that as the optode distance increased, the absorption coefficients increased and the scattering coefficients decreased, retrieving the optical values of scalp and brain for shorter and longer optode distances, respectively. We present the transition curves of the absorption and reduced scattering coefficients as functions of the optode distance. In order to verify the values for each layer, a comparison between the experimental data and a prediction based on the two-layer model of the adult head was carried out. The thicknesses of scalp and skull for the two-layer model were obtained by magnetic resonance imaging of a subject's head. The optical parameters obtained from the two-layer model agreed very well with those measured by the multidistance method.     

Inhibition of p38 by vitamin D reduces interleukin-6 production in normal prostate cells via mitogen-activated protein kinase phosphatase 5: implications for prostate cancer prevention by vitamin D

Although numerous studies have implicated vitamin D in preventing prostate cancer, the underlying mechanism(s) remains unclear. Using normal human prostatic epithelial cells, we examined the role of mitogen-activated protein kinase phosphatase 5 (MKP5) in mediating cancer preventive activities of vitamin D. Up-regulation of MKP5 mRNA by 1,25-dihydroxyvitamin-D3 (1,25D) was dependent on the vitamin D receptor. We also identified a putative positive vitamin D response element within the MKP5 promoter that associated with the vitamin D receptor following 1,25D treatment. MKP5 dephosphorylates/inactivates the stress-activated protein kinase p38. Treatment of prostate cells with 1,25D inhibited p38 phosphorylation, and MKP5 small interfering RNA blocked this effect. Activation of p38 and downstream production of interleukin 6 (IL-6) are proinflammatory. Inflammation and IL-6 overexpression have been implicated in the initiation and progression of prostate cancer. 1,25D pretreatment inhibited both UV- and tumor necrosis factor alpha-stimulated IL-6 production in normal cells via p38 inhibition. Consistent with inhibition of p38, 1,25D decreased UV-stimulated IL-6 mRNA stabilization. The ability of 1,25D to up-regulate MKP5 was maintained in primary prostatic adenocarcinoma cells but was absent in metastases-derived prostate cancer cell lines. The inability of 1,25D to regulate MKP5 in the metastasis-derived cancer cells suggests there may be selective pressure to eliminate key tumor suppressor functions of vitamin D during cancer progression. These studies reveal MKP5 as a mediator of p38 inactivation and decreased IL-6 expression by 1,25D in primary prostatic cultures of normal and adenocarcinoma cells, implicating decreased prostatic inflammation as a potential mechanism for prostate cancer prevention by 1,25D.     

Preconception serum DDT and pregnancy loss: a prospective study using a biomarker of pregnancy

Previous studies of pregnancy losses and 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) were limited because they did not include losses prior to clinical detection of pregnancy and because exposures were measured after the pregnancies of interest. The authors examined the association of preconception serum total DDT (sum of DDT isomers and metabolites) concentration and subsequent pregnancy losses in 388 newly married, nonsmoking, female textile workers in China between 1996 and 1998. Upon stopping contraception, subjects provided daily urine specimens and records of vaginal bleeding for up to 1 year or until clinical pregnancy. Daily urinary human chorionic gonadotropin was assayed to detect conception and early pregnancy losses, and pregnancies were followed to detect clinical spontaneous abortions. There were 128 (26%) early pregnancy losses in 500 conceptions and 36 (10%) spontaneous abortions in 372 clinical pregnancies. Subjects were grouped in tertiles by preconception serum total DDT concentration (group 1: 5.5-22.9 ng/g; group 2: 23.0-36.5 ng/g; group 3: 36.6-113.3 ng/g). Compared with group 1, group 2 had adjusted relative odds of early pregnancy losses of 1.23 (95% confidence interval (CI): 0.72, 2.10), and group 3 had adjusted odds of 2.12 (95% CI: 1.26, 3.57). The relative odds of early pregnancy losses associated with a 10-ng/g increase in serum total DDT were 1.17 (95% CI: 1.05, 1.29). The small number of spontaneous abortions following clinical detection of pregnancy were not associated with serum total DDT. In this population, there was a positive, monotonic, exposure-response association between preconception serum total DDT and the risk of subsequent early pregnancy losses.     

Evidence of interaction between polychlorinated biphenyls and phthalates in relation to human sperm motility

Previously, we reported evidence of inverse associations between exposure to some polychlorinated biphenyls (PCBs) and some phthalate monoesters in relation to semen parameters, specifically sperm motility. Because humans are exposed to both phthalates and PCBs and because experimental studies suggest that PCBs may interact with glucuronidative enzymes that are responsible for phthalate metabolism, we explored the potential interaction between phthalates and PCBs in relation to human semen quality. We studied 303 men who were partners in subfertile couples seeking infertility diagnosis from the andrology laboratory at Massachusetts General Hospital. Semen parameters were dichotomized based on World Health Organization reference values, and phthalate and PCB levels were dichotomized at their respective medians. After adjusting for age and abstinence time, for below reference sperm motility there was a greater than additive interaction between monobenzyl phthalate and PCB-153 [relative excess risk due to interaction (RERI) = 1.40; 95% confidence interval (CI), 0.41-3.22], sum of PCBs (RERI = 1.24; 95% CI, 0.15-2.94), and cytochrome P450 (CYP450)-inducing PCBs (RERI = 1.30; 95% CI, 0.21-3.06). For below-reference sperm motility, there was also a greater than additive interaction between monobutyl phthalate (MBP) and PCB-153 (RERI = 1.42; 95% CI, 0.09-3.76) and CYP450-inducing PCBs (RERI = 1.87; 95% CI, 0.56-4.52) and a suggestive interaction between MBP and sum of PCBs (RERI = 1.35; 95% CI, -0.11 to 3.48). In conclusion, because there are important risk assessment and public health implications of interactions between these two ubiquitous classes of compounds, further studies need to be conducted to confirm these results and identify potential mechanisms of interactions.     

The toxicity of SCH 351591, a novel phosphodiesterase-4 inhibitor, in Cynomolgus monkeys

SCH351591, a novel phosphodiesterase-4 inhibitor under investigation as a potential therapeutic for asthma and chronic obstructive pulmonary disease (COPD), was evaluated in a 3-month rising-dose study in Cynomolgus monkeys. Four groups, containing four monkeys/sex, received vehicle control or rising doses up to 12, 24, or 48 mg/kg of SCH351591 daily. Although initial exposure produced clinical signs of emesis, reduced food intake, and reduced body weight, tachyphylaxis to the emesis allowed dose escalation up to 48 mg/kg/day. Two monkeys died and 3 were sacrificed in moribund condition over the course of the study. Early mortality, involving monkeys dosed with 12 or 24 mg/kg, was attributed to sepsis (2 monkeys) or colon inflammation (3 monkeys). Leukocyte function assays on low- and mid-dose group survivors revealed an inhibition of T lymphocyte proliferation for 12 mg/kg group males and 24 mg/kg group monkeys of both sexes. Necropsy findings, unassociated with early mortality, included reduced size and weight of the thymus, depletion of body fat, red discoloration of the gastric mucosa, and perivascular hemorrhage of the stomach and heart. Stomach and heart gross findings were present in the high-dose group only. Histopathologic lesions, in addition to those attributed to concurrent bacterial infection, included thymic atrophy, serous atrophy of fat, myocardial degeneration and acute to chronic inflammation of small to medium-sized arteries in various organs and tissues including the heart, kidneys, stomach, salivary glands, pancreas, esophagus, gallbladder, and mesentery. The findings of this study demonstrate the potential of a PDE4 inhibitor to alter immunologic response as well as to produce arteriopathy in nonhuman primates.