Loss of protein kinase C inhibition in the beta-T594M variant of the amiloride-sensitive Na+ channel

We previously reported the presence of a novel variant (beta-T594M) of the amiloride-sensitive Na+ channel (ASSC) in which the threonine residue at position 594 in the beta-subunit has been replaced by a methionine residue. Electrophysiological studies of the ASSC on Epstein-Barr virus (EBV)-transformed lymphocytes carrying this variant showed that the 8-(4-chlorophenylthio) adenosine 3':5'-cyclic monophosphate (8cpt-cAMP)-induced responses were enhanced when compared to wild-type EBV-transformed lymphocytes. Furthermore, in wild-type EBV-transformed cells, the 8cpt-cAMP-induced response was totally blocked by the phorbol ester, phorbol 12-myristate 13-acetate (PMA). This inhibitory effect of PMA was blocked by a protein kinase C inhibitor, chelerythrine. We now have identified individuals who are homozygous for this variant, and showed that PMA had no effect on the 8cpt-cAMP-induced responses in the EBV-transformed lymphocytes from such individuals. Cells heterozygous for this variant showed mixed responses to PMA, with the majority of cells partially inhibited by PMA. Our results demonstrate that an alteration in a single amino acid residue in the beta-subunit of the ASSC can lead to a total loss of inhibition to PMA, and establish the beta-subunit as having an important role in conferring a regulatory effect on the ASSC of lymphocytes.     

Ubiquitin-mediated regulation of 3-hydroxy-3-methylglutaryl-CoA reductase

Regulation of the sterol-synthesizing mevalonate pathway occurs in part through feedback-regulated endoplasmic reticulum degradation of 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-R). In yeast, the Hmg2p isozyme of HMG-R is regulated in this manner. We have tested the involvement of ubiquitination in the regulated degradation of Hmg2p, by using both genetic and direct biochemical approaches. Hmg2p degradation required the UBC7 gene, and Hmg2p protein was directly ubiquitinated. Hmg2p ubiquitination was dependent on UBC7 and was specific for the degraded yeast Hmg2p isozyme. Furthermore, Hmg2p ubiquitination was regulated by the mevalonate pathway in a manner consistent with regulation of Hmg2p stability. Thus, regulated ubiquitination appeared to be the mechanism by which Hmg2p stability is controlled in yeast. Finally, our data indicated that the feedback signal controlling Hmg2p ubiquitination and degradation was derived from farnesyl diphosphate, and thus implied conservation of an HMG-R degradation signal between yeast and mammals.     

Parameter optimization of ferro-sonication pre-treatment process for degradation of bisphenol A and biodegradation from wastewater sludge using response surface model

In this study, the application of response surface model in predicting and optimizing the ferro-sonication pre-treatment for degradation of bisphenol A (BPA), an endocrine disrupter compound from wastewater sludge (WWS) was investigated. The ferro-sonication pre-treatment process was carried out according to central composite design (CCD) with four independent variables such as wastewater sludge solids concentration, pH, ultrasonication time and FeSO(4) concentration. The effect of ferro-sonication pre-treatment was assessed in terms of increase in sludge solids (suspended solids (SS) and volatile solids (VS)) and organic matter (chemical oxygen demand (COD) and soluble organic carbon (SOC)) solubilization and simultaneous BPA degradation from WWS. It was observed that among all the variables studied, ultrasonication time had more significantly affected the efficiency of the ferro-sonication pre-treatment process followed by FeSO(4) and solids concentration. Through this optimization process, it was found that maximum BPA degradation of 88% could be obtained with 163 min ultrasonication time, 2.71 mg/L FeSO(4) concentration, pH 2.81 with 22 g/L SS. Further, the effect of ferro-sonication pre-treatment on biodegradation of WWS was also studied. It was observed that ultrasonication time had significant effect and the higher biodegradation (32.48%) was observed at 180 min ultrasonication time.     

Heparin-induced thrombocytopenia: pathophysiology

We have used computer modeling of insulin 3-D structure and experimental data about action of site point mutation on insulin activity to design functionally important domain with signaling activity and synthesized peptide than might be sufficient for the binding to insulin receptor. The designed and synthesized peptide consist of ten residues and may be obtained in two forms: oxidized and reduced (with or without disulfide bond). The synthesized decapeptide peptide represents functionally important site for binding to the insulin receptor. Amino acid residues at position 1-8 correlate with B-chain of insulin at position (B19-B26). Residues at position 9.10 correlate with A-chain at position A-10-A21. This peptide was tested with cell culture L-929 (glucose uptake) in comparison with bioactive commercial peptide (R-G-FF) and insulin. It was shown that synthesized peptide exhibit biological activity at molar concentration 0.01-1 mkM. Our results successfully demonstrate the synthetic insulin fragment have insulin-like biological activity.     

Self-sustained pH oscillations in immobilized proteolytic enzyme systems

The efficacy of doramectin, a novel avermectin, was assessed against both naturally-acquired and experimentally-induced infections of gastrointestinal roundworms, lungworms, kidneyworms, lice and mites in studies conducted across North America and Europe. Twenty-two studies evaluated efficacy against fourth larval and adult stages of the following nematode species: Hyostrongylus rubidus, Ascaris suum, Strongyloides ransomi, Oesophagostomum dentatum, Oesophagostomum quadrispinulatum, Trichuris suis, Metastrongylus spp. and Stephanurus dentatus. Efficacy was evaluated against the louse Haematopinus suis in six studies and against the mite Sarcoptes scabiei in four studies. A common study design was employed for each study type. In all studies, animals were allotted at random to a doramectin-treated or a saline-treated group. The doramectin-treated group received the drug at 300 micrograms kg-1 by intramuscular injection while the saline-treated group received saline by the same route. In the nematode studies, worm burdens were determined for each animal at slaughter 14-16 days after treatment. Efficacy against each nematode species/stage was assessed on the basis of percentage reduction in geometric mean worm burden in doramectin-treated animals compared with saline-treated controls. In louse and mite studies, counts were made immediately before treatment and then at weekly intervals for four weeks. Efficacy was based on a comparison of the level of infestation on the day of treatment with that on the last day of test. Data from individual studies were combined to derive a single estimate of efficacy against each of the parasite species represented in the study program. Efficacy of doramectin was 98% or greater against all nematode species except T. suis for which the efficacy was 87% and 79% against adult and fourth larval stage, respectively. Efficacy was 100% against both Haematopinus suis and Sarcoptes scabiei.     

Factors affecting young children's mental development and their significance in intervention

Factors affecting their developmental quotients (DQ) in 48 normal young children, who were involved in regular development assessments at child health care outpatient visits and whose parents received child-rearing guidance at Chaoyangmen Subdistrict of Dongcheng District, Beijing, and 43 controls aged 36 months were analyzed. Factors, such as their parents' receiving child-rearing guidance, paternal education, maternal education, paternal occupation, type of nursery schools, home environment quality score (HEQS) at 24 and 36 months of age, and mothers' knowledge of child-rearing were associated with children's DQ at their age of 36 months, with statistical significance at a level of 0.05 in univariate analysis. With multiple regression analysis, HEQS at the age of 36 months, family size at the age of 24 months, paternal education, children's sex, and mothers' age had influences on DQ at age of 36 months with statistical significance, and the degree of correlation of HEQS with DQ at 36 months was much higher than that of other factors, and it suggested that HEQS was an important link in child developmental intervention.     

School-based health care for urban minority junior high school students

OBJECTIVE: To describe the use of school-based health clinics by urban minority junior high school students. DESIGN: Review of demographic and utilization data collected by service providers during clinic visits. SETTINGS AND PARTICIPANTS: Health clinics in four junior high schools that enrolled predominantly Hispanic students who were residing in an economically disadvantaged, medically underserved New York (NY) school district. RESULTS: Of 5757 students who were enrolled in the schools, 5296 (92%) obtained parental consent to use the clinics, and 3723 (65%) used the clinics during the 1991-1992 academic year. Clinic users were 11 to 15 years old, 50% male and 50% female, 81% Hispanic and 14% black, and 29% sixth graders, 33% seventh graders, and 38% eighth graders. Clinic users made 16,340 clinic visits during the 1991-1992 academic year. Presenting complaints were mental health problems (32%), illness (14%), injury (12%), physical examination (5%), immunization (3%), follow-up (21%), and other (13%). Referral sources were clinic outreach (48%), self (44%), and school personnel (8%). Disposition of visits was on-site treatment (92%), referral to an affiliated hospital (5%), and referral elsewhere (3%). Compared with a nationwide group of high school-based clinics that served predominantly black adolescents, these clinics provided more mental health care (31% vs 21%), similar illness/injury care (32% vs 30%), and less preventive (10% vs 24%) and reproductive/contraceptive (7% vs 12%) care. CONCLUSIONS: Junior high school-based clinics can provide a wide range of primary and preventive health care services for large numbers of medically underserved youths. The provision of mental health services may fill a critical need among inner-city adolescents. Clinic outreach may be necessary to maximize utilization, especially among high-risk students.     

Suprachiasmatic nucleus organization

OBJECTIVE: To assess the effect of a combined exercise and motivation program on the compliance and level of disability of patients with chronic and recurrent low back pain. DESIGN: A double-blind prospective randomized controlled trial. SETTING: Physical therapy outpatient department, tertiary care. PATIENTS: Ninety-three low back pain patients were randomly assigned to either a standard exercise program (n = 49) or a combined exercise and motivation program (n = 44). INTERVENTIONS: Patients were prescribed 10 physical therapy sessions and were advised to continue exercising after treatment termination. The motivation program consisted of five compliance-enhancing interventions. Follow-up assessments were performed at 3 1/2 weeks, 4 months, and 12 months. MAIN OUTCOME MEASURES: Disability (low back outcome score), pain intensity, physical impairment (modified Waddell score, fingertip-to-floor distance, abdominal muscle strength), working ability, motivation, and compliance. RESULTS: The patients in the motivation group were significantly more likely to attend their exercise therapy appointments (p = .0005). Four and 12 months after study entry there was a significant difference in favor of the motivation group with regard to the disability score (p = .004) and pain intensity (p < or = .026). At 4 months, there was a significant advantage for the motivation group in the fingertip-to-floor distance (p = .01) and in abdominal muscle strength (p = .018). No significant differences were found in motivation scores, self-reported compliance with long-term exercise, and modified Waddell score. In terms of working ability, there was a trend favoring the combined exercise and motivation program. CONCLUSION: The combined exercise and motivation program increased the rate of attendance at scheduled physical therapy sessions, ie, short-term compliance, and reduced disability and pain levels by the 12-month follow-up. However, there was no difference between the motivation and control groups with regard to long-term exercise compliance.     

Thrombocytopenia following liver transplantation is associated with platelet consumption and thrombin generation

PSA-based screening substantially increases the prostate cancer detection rate and the percentage of organ-confined tumors. It appears that there is some benefit from screening for prostate cancer because of the increased amount of potentially curable disease discovered and the fact that 96% of the pathologically staged tumors detected have histologic features associated with aggressive cancer. Additional evidence that nearly all tumors detected on the basis of initial PSA screening are apt to be clinically significant may be derived from the information that PSA-based screening decreases the incidence of incidental A1 grade III and A2 tumors but does not increase the detection of clinically insignificant A1 grade I and II tumors. At this time, PSA represents the most effective and valuable tool to detect early prostate cancer; therefore, PSA should be used to improve early diagnosis of prostate cancer. Some advances have been made with the introduction of age-specific reference ranges and the ability to measure free to total PSA ratios. The data presented support the clinical usefulness of age-specific reference ranges for serum PSA. Calculation of the free to total PSA ratio is valuable in deciding which screening volunteers require further evaluation, increases the specificity of PSA screening, and as demonstrated may be useful in deciding which patients with isolated PIN should undergo repeat biopsies. Based on these facts, PSA truly can be described as the most important and useful marker for adenocarcinoma of the prostate. Based on these encouraging results and the obligingness of the social insurances, we will be able to continue PSA screening for early detection of prostate cancer for all concerned Tyrolean men in the future.