Adenosine A1 receptor-mediated presynaptic inhibition of GABAergic transmission in immature rat hippocampal CA1 neurons

In the mechanically dissociated rat hippocampal CA1 neurons with native presynaptic nerve endings, namely "synaptic bouton" preparation, the purinergic modulation of spontaneous GABAergic miniature inhibitory postsynaptic currents (mIPSCs) was investigated using whole-cell recording mode under the voltage-clamp conditions. In immature neurons, adenosine (10 microM) reversibly decreased GABAergic mIPSC frequency without affecting the mean current amplitude. The inhibitory effect of adenosine transmission was completely blocked by 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 100 nM), a selective Alpha(1) receptor antagonist, and was mimicked by N(6)-cyclopentyladenosine (CPA, 1 microM), a selective Alpha(1) receptor agonist. However, CPA had no effect on GABAergic mIPSC frequency in postnatal 30 day neurons. N-ethylmaleimide (10 microM), a guanosine 5'-triphosphate binding protein uncoupler, and Ca(2+)-free external solution removed the CPA-induced inhibition of mIPSC frequency. K(+) channel blockers, 4-aminopyridine (100 microM) and Ba(2+) (1 mM), had no effect on the inhibitory effect of CPA on GABAergic mIPSC frequency. Stimulation of adenylyl cyclase with forskolin (10 microM) prevented the CPA action on GABAergic mIPSC frequency. Rp-cAMPS (100 microM), a selective PKA inhibitor, also blocked the CPA action. It was concluded that the activation of presynaptic Alpha(1) receptors modulates the probability of spontaneous GABA release via cAMP- and protein kinase A dependent pathway. This Alpha(1) receptor-mediated modulation of GABAergic transmission may play an important role in the regulation of excitability of immature hippocampal CA1 neurons.     

The changing epidemiology of hepatitis A in children and the consideration of active immunization in Korea

Currently, Korea is a low endemicity country for HAV, especially in children. However, recent reports of hepatitis A outbreaks show that there has been a shift of disease incidence to adolescents and young adults, with 2 cases of acute liver failure in one reported outbreak. We need to study the immune status for HAV in order to provide information for the establishment of preventive measures and possible consequences of HAV in Korea. A total of 334 infants, children and adolescents less than 20 years of age living in rural areas of Kyonggi Province, Korea were evaluated for anti-HAV immune status in 1996. Five hundred and eighty-four primary school children living in the same area were separately evaluated for the natural seroconversion rate between 1993 and follow-up samples taken in 1996. Anti-HAV IgG antibody was measured by enzyme immunoassay (HAVAB EIA kit, Abbott Laboratories, Chicago, Illinois, USA). In comparison with previous reports of seroprevalence rates, our data confirmed a dramatic drop in seroprevalence rates among children and adolescents under 20 years of age living in rural areas, from over 63.8% two decades ago to 4.6% in 1996. Natural acquisition of HAV antibody in primary school children rarely occurs, registering only 0.5% during three years. Several outbreaks in young adults during 1996-1998 suggested that immunity against HAV in this population is so low that massive outbreaks are unavoidable. Teenagers and young adults, especially soldiers, who are likely to be exposed to contaminated food or water, would also have a greater risk of hepatitis A. Immunizing children with HAV vaccine as a routine schedule should also be considered in Korea in the future, particularly if the disease burden could be estimated and the cost-effectiveness of the vaccine could be proved.     

PCR and direct fluorescent-antibody staining confirm Chlamydia trachomatis antigens in swabs and urine below the detection threshold of Chlamydiazyme enzyme immunoassay.

In order to test the hypothesis that specimens blocking with a neutralizing reagent below the cutoff of the Chlamydiazyme enzyme immunoassay represent infected patients, we used direct fluorescent-antibody staining for elementary bodies (EBs) and PCR to confirm results for cervical swabs collected from 55,963 women and urethral swabs or first-void urine (FVU) samples collected from 5,781 men attending physicians' offices in the Toronto, Canada, area. Within a grey zone arbitrarily selected to represent values up to 40% below the positive threshold of the test run, 134 cervical swabs, 44 urethral swabs, and 39 FVU specimens exhibited a blocking response ( > 50% reduction in signal). Three or more EBs were observed in each of 98 cervical swabs (73.1%), 38 urethral swabs (86.4%), and 21 FVU specimens (53.8%). Of the 36 cervical swabs with fewer than three EBs, 33 were PCR positive; the positive PCR results for male specimens were 6 of 6 urethral swabs and 17 of 18 FVU samples. Application of the blocking test to specimens negative in the Chlamydiazyme enzyme immunoassay but having optical densities within 40% of the cutoff added 14.2% (217 of 1,531 specimens) more positive results to the survey. A total of 213 of 217 samples (98.2%) were reconfirmed as having EBs or DNA.     

Comparison of a polymer conjugate-enhanced enzyme immunoassay to ligase chain reaction for diagnosis of Chlamydia trachomatis in endocervical swabs

Two endocervical swabs from each of 1,123 women were collected into manufacturer-supplied transport tubes and tested for Chlamydia trachomatis by a polymer conjugate-enhanced (PCE) enzyme immunoassay (EIA) (IDEIA PCE Chlamydia; DAKO) and a ligase chain reaction assay (LCx Chlamydia; Abbott). After confirmation by the EIA blocking test, the sensitivity of the IDEIA PCE remained at 91.8% and the specificity increased from 98.2 to 99.8% compared to LCx.     

Confirmatory polymerase chain reaction testing for Chlamydia trachomatis in first-void urine from asymptomatic and symptomatic men.

First-void urine specimens from 683 men (592 without symptoms) were tested for Chlamydia trachomatis by a polymerase chain reaction (PCR) with KL1 and KL2 plasmid primers and by a Chlamydiazyme enzyme immunoassay (EIA). Thirty-seven specimens were confirmed to be positive by using the EIA blocking reagent and a second set of plasmid primers (T1 and T2). By comparing unconfirmed PCR results (KL1 and KL2 primers only) with the blocked Chlamydiazyme EIA results, the sensitivity and specificity of PCR were 100% (37 of 37 specimens) and 99.5% (643 of 646 specimens), respectively. Three additional specimens were negative by EIA but positive by PCR and were confirmed to be positive with primers T1 and T2. Two of the three specimens were from men with symptoms. The confirmatory PCR assay performed equally well in detecting positive specimens from symptomatic (31 of 31) and asymptomatic (9 of 9) men. Comparison of confirmatory testing of first-void urine specimens by PCR and EIA showed that PCR was 100% sensitive (40 of 40 specimens) and that the EIA was 92.5% sensitive (37 of 40 specimens) but that the assays were equally specific (100%).     

Protective effect of Hypericum perforatum Linn (St. John's wort) against hydrogen peroxide-induced apoptosis on human neuroblastoma cells

The medicinal plant Hypericum perforatum Linn, commonly known as St. John's wort, has been used as an antidepressant. To investigate whether St. John's wort possesses a protective effect against hydrogen peroxide (H(2)O(2))-induced cytotoxicity in neuronal cells, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, 4,6-diamidino-2-phenylindole staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay, flow cytometry analysis, DNA fragmentation assay, and caspase-3 enzyme assay were performed on SK-N-MC human neuroblastoma cells. Cells treated with H(2)O(2) exhibited several apoptotic features, while those pre-treated with St. John's wort prior to H(2)O(2) exposure showed a decreased occurrence of apoptotic features. In addition, pre-treatment with St. John's wort inhibited H(2)O(2)-induced increase in caspase-3 enzyme activity. These results suggest that St. John's wort may exert a protective effect against H(2)O(2)-induced apoptosis in human neuroblastoma cells.     

Cooperative transformation of murine fibroblast NIH3T3 cells by hepatitis C virus core protein and hepatitis B virus X protein

Co-infection with hepatitis B virus (HBV) and hepatitis C Virus (HCV) is associated with increased frequency in the development of hepatocellular carcinoma (HCC). Here, we demonstrated that HBV X protein (HBx) and HCV core cooperate to transform mouse fibroblast NIH3T3 cells. They additively stimulated cell growth, especially in the absence of serum growth factors. In addition, co-expression of HBx and HCV core had additive effects on the induction of anchorage-independent cell growth as well as on the secretion of matrix metalloproteases, which may contribute to increased metastatic potential. Furthermore, the cells expressing both viral proteins exhibited higher tumorigenicity, as demonstrated in athymic nude mice.     

Liver profile in COVID-19: a meta-analysis

Journal of Public Health - Few studies have been performed to investigate multiorgan failure occurring with extrapulmonary symptoms like diarrhea, anorexia, and vomiting. There has been no attempt...     

Associations Between Thyroid Hormone Levels and Urinary Concentrations of Bisphenol A,F, and S in 6-Year-old Children in Korea

ObjectivesBisphenol A (BPA) is used in the electrical, mechanical, medical, and food industries. Previous studies have suggested that BPA is an endocrine disruptor. Regulation of BPA has led to increased use of bisphenol F (BPF) and bisphenol S (BPS). However, few studies have investigated the associations of BPF and BPS with thyroid dysfunction in children. Our study investigated the associations of prenatal BPA and early childhood BPA, BPF, and BPS exposure with thyroid function in 6-year-old children.MethodsPrenatal BPA concentrations were measured during the second trimester of pregnancy in an established prospective birth cohort. We measured urinary BPA, BPF, and BPS concentrations and thyroid hormone levels (thyroid-stimulating hormone, total T₃, and free T₄) in 6-year-old children (n=574). We examined the associations between urinary bisphenol concentrations and percentage change of thyroid hormone concentrations using multivariate linear regression. We also compared thyroid hormone levels by dividing the cohort according to BPA, BPF, and BPS concentrations.ResultsThe associations between prenatal BPA and total T₃ levels were statistically significant in all models, except for girls when using a crude model. The associations between urinary BPA and BPS concentrations and levels of all thyroid hormones were not statistically significant. However, we observed that lower free T₄ levels (−1.94%; 95% confidence interval, −3.82 to −0.03) were associated with higher urinary BPF concentrations in girls only.ConclusionsOur findings identified significant associations between prenatal BPA exposure and total T₃ levels in all children and between BPF exposure and free T₄ levels in girls only.     

Effect of Heat-Killed Lactobacillus casei DKGF7 on a Rat Model of Irritable Bowel Syndrome

Non-viable bacteria, referred to as “paraprobiotics,” have attracted attention as potentially safer alternatives to probiotics. The aim of this study was to investigate the efficacy of heat-killed Lactobacillus casei DKGF7 on the symptomatic improvement of irritable bowel syndrome (IBS) in a rat disease model and to elucidate the underlying mechanisms that contribute to the beneficial effects of heat-killed probiotics. Seven male Wistar rats were induced with IBS by restraint stress and administered heat-killed L. casei DKGF7 for four weeks and then compared with seven rats in the control group. Stool consistency measured four weeks after initial treatment was the primary outcome measure. To investigate the mechanism of action of the heat-killed bacteria on IBS, we measured serum corticosterone levels, inflammatory cytokines in colon tissue, and expression of tight junction proteins (TJPs) in the epithelium. The treatment group showed significantly better stool consistency scores than the control group at week 4, as well as at every measured time point (all p values < 0.05). The treatment group showed lower serum corticosterone levels, lower colonic inflammatory cytokine levels, and higher expression of TJPs compared with the control group. Paraprobiotics such as heat-killed L. casei DKGF7 can improve stool consistency in a rat IBS model, which may indicate a potential therapeutic strategy for IBS treatment.