Kinetic analysis of reverse transcriptase activity of bacterial family A DNA polymerases.

Some bacterial thermostable, wild-type or genetically engineered family A DNA polymerases have reverse transcriptase activity. However, difference in reverse transcriptase activities of family A DNA polymerases and retroviral reverse transcriptases (RTs) is unclear. In this study, comparative kinetic analysis was performed for the reverse transcriptase activities of the wild-type enzyme of family A DNA polymerase (M1pol(WT)) from Thermus thermophilus M1 and the variant enzyme of family A DNA polymerase (K4pol(L329A)), in which the mutation of Leu329→Ala is undertaken, from Thermotoga petrophila K4. In the incorporation of dTTP into poly(rA)-p(dT)(45), the reaction rates of K4pol(L329A) and M1pol(WT) exhibited a saturated profile of the Michaelis-Menten kinetics for dTTP concentrations but a substrate inhibition profile for poly(rA)-p(dT)(45) concentrations. In contrast, the reaction rates of Moloney murine leukemia virus (MMLV) RT exhibited saturated profiles for both dTTP and poly(rA)-p(dT)(45) concentrations. This suggests that high concentrations of DNA-primed RNA template decrease the efficiency of cDNA synthesis with bacterial family A DNA polymerases

Association between acetylcholine receptor characteristics in biceps motor endplates and the epidemiological predictors for conversion from ocular to generalized myasthenia gravis

Objective: Epidemiological studies have identified various predictors of conversion from ocular myasthenia gravis (OMG) to secondary generalized myasthenia gravis (SGMG), but none have been confirmed. We investigated the effects of the epidemiological conversion predictors on the destruction of motor endplates in the biceps of patients with OMG and attempted to identify predictors of conversion to SGMG histologically.Methods: Patients with clinically diagnosed OMG who requested immunohistological diagnosis and who underwent muscle biopsy were included in this study. We immunostained the biceps motor endplate and semi-quantitatively measured the density and number of AChRs to determine their association with the epidemiological predictors of conversion from OMG to SGMG.Results: Thirteen patients with OMG were included, of which two patients with positive AChR antibody and concomitant thymoma converted to SGMG. In the classification according to the presence of AChR antibody, the AChR densities tended to be lower in the antibody-positive group than in the negative group (p=0.079), and the AChR numbers were significantly lesser in the AChR antibody-positive group than in the negative group (p=0.019). There were no differences in AChR densities or numbers according to sex, presence of thymic abnormalities, or presence of comorbid autoimmune diseases.Conclusion: In OMG, the AChR numbers in motor endplates of the biceps were significantly lesser in the AChR antibodypositive group than in the negative group. Since the muscle strength tends to decrease as the number of AChRs decreases, AChR antibody positivity may be a predictor of OMG to SGMG conversion, but further studies are needed to confirm

Stability Testing of Drug Products Approved by the Japanese Government in 2015

2015年に日本で承認された新医療用製剤に対し、安定性試験の現状を調査した。我々は、2015年に安定性試験の記述がある81の新医療用製剤を特定した。長期保存試験としては、5製剤が30±2℃/65±5％相対湿度の条件下で、55製剤が25±2℃/60±5％相対湿度の条件下で、21製剤が5±3℃の条件下で試験が実施されていた。また、加速性試験では、60製剤が40±2℃/75±5％相対湿度の条件下で実施されていた。17製剤は25±2℃/60±5％相対湿度の条件下で実施された。3製剤は温度及び試験期間がマスキングされていて、不明であった。1製剤は半減期が短いため、加速試験は実施されなかった｡これらの結果から、2015年に日本で承認された新医療用製剤は、1つを除いて ICH ガイドラインに従って、適正に承認されていることがわかった

HLA-DRB1 and DQB1 alleles in Japanese type 1 autoimmune hepatitis: The predisposing role of the DR4/DR8 heterozygous genotype

ObjectiveAutoimmune hepatitis (AIH) is a chronic progressive liver disease. AIH is composed predominantly of type 1 in Japanese populations. The genetic and environmental factors are associated with the pathogenesis of AIH. HLA-DRB1*03:01 and *04:01 are associated with type 1 AIH in European and *04:05 in Japanese populations. Here, we conducted an HLA association study in order to find HLA alleles or haplotypes predisposing or protective for Japanese AIH.MethodsHLA-DRB1 and DQB1 genotyping of 360 type 1 AIH patients and 1026 healthy controls was performed.ResultsThe predisposing association of DRB1*04:01 (P = 0.0006, corrected P [Pc] = 0.0193, odds ratio [OR] 2.97, 95% confidence interval [CI] 1.62–5.43), DRB1*04:05 (P = 1.89×10−21, Pc = 5.86×10−20, OR 3.41, 95% CI 2.65–4.38), and DQB1*04:01 (P = 4.66×10−18, Pc = 6.99×10−17, OR 3.89, 95% CI 2.84–5.33) and the protective association of DRB1*13:02 (P = 0.0003, Pc = 0.0080, OR 0.48, 95% CI 0.32–0.72) with Japanese type 1 AIH were observed. An association of the DR4/DR8 heterozygous genotype with Japanese AIH was identified for the first time (P = 3.12×10−9, OR 3.52, 95% CI 2.34–5.29). Susceptible diplotypes were DRB1*04:05-DQB1*04:01/DRB1*08:02-DQB1*03:02 (P = 0.0004, OR 24.77, 95% CI 1.45–424.31) and DRB1*04:05-DQB1*04:01/DRB1*08:03-DQB1*06:01 (P = 1.18×10−6, OR 10.64, 95% CI 3.19–35.46). Serum levels of Immunoglobulin G and Immunoglobulin M, International Autoimmune Hepatitis Group score, positive rate of anti-smooth muscle antibodies, and the rate of definite AIH were higher in AIH patients with DRB1*04:05 than without.ConclusionsThe important roles of specific combinations of DRB1 and DQB1 alleles or haplotypes in the pathogenesis of type 1 AIH were suggested. The association of DR4/DR8 heterozygous genotype suggested the pathologic importance of trans-complementing DQα-β heterodimer molecules encoded by DQA1 allele of one haplotype and the DQB1 allele of the other haplotype, as it was proposed in the HLA association studies of Type 1 diabetes

Development of D-to-D-to-P telemedicine at a remote island hospital using smart glasses

Background: Medical resources on remote islands are limited, which makes it difficult for patients to receive specialized medical care.Purpose: This study aimed to develop and evaluate a method to perform doctor-to-doctor-to-patient (D-to-D-to-P) telemedicine.Methods: The-D-to-D-to-P telemedicine was implemented to provide specialized medical support from a neurologist at Nagasaki University Hospital to a rural physician wearing camera-equipped smart glasses at Goto Chuoh Hospital on a remote island, which was called a virtual neurological outpatient (VNO). For the first six months, the rural physician independently saw patients with Parkinson’s disease (PD), and then for the next six months, VNO was implemented. Comparisons were made before and after the implementation of the VNO. Next, by adding a 4 K overhead camera, in-person examinations of a single outpatient were compared between the rural physician with VNO and another neurologist unrelated to the VNO.Results: The clinical efficacy of VNO was not superior to no VNO, but had a learning effect on rural physicians and was satisfactory for patients. By adding a 4 K overhead camera to the VNO, the accuracy of the in-person examination by the rural physician was shown to be equivalent to that of an in-person neurologist.Conclusion: VNO using smart glasses could be applied for D-to-D-to-P telemedicine in neurology. However, to promote telemedicine on remote islands, it will be necessary to improve the system to make it more accessible to rural physicians

Perfusion abnormality in neuronal intranuclear inclusion disease with stroke-like episode: A case report

Neuronal intranuclear inclusion disease (NIID) is a slowly progressive neurodegenerative disease. Some patients with NIID occasionally present with acute symptoms. However, its mechanism remains unclear. We report a patient with NIID who presented with a stroke-like episode. Arterial spin labeling magnetic resonance imaging revealed hypoperfusion in the focal cerebral region at the onset while no apparent arterial occlusion was observed. The abnormal perfusion area was normalized 6 days after admission. Therefore, the perfusion abnormality was likely the main cause of acute neurologic deficits in NIID. NIID should be considered in the differential diagnosis of stroke mimics

Being Praised for Prosocial Behaviors Longitudinally Reduces Depressive Symptoms in Early Adolescents: A Population-Based Cohort Study

BackgroundDepression is highly prevalent and causes a heavy burden in adolescent life. Being praised for prosocial behavior might be a preventive factor because both being praised and prosocial behavior are protective against depression. Here, we investigated the longitudinal relationship between being praised for prosocial behavior and depressive symptoms in adolescents.MethodsIn Tokyo Teen Cohort study (TTC), an ongoing prospective population-based cohort study, we collected 3,171 adolescents' data on self-reported experiences of being praised for prosocial behavior, depressive symptoms, and caregiver-evaluated prosocial behavior. Ten-year-old children were asked to freely describe answers to the question “What are you praised for?”. Only children who clearly answered that they were praised for their prosocial behavior were designated the “prosocial praise group.” The degree of depression at ages 10 and 12 was measured with the Short Mood and Feelings Questionnaire (SMFQ), a self-report questionnaire about depression. Objective prosocial behavior of the 10 year-old children was assessed by the Strength and Difficulty Questionnaire (SDQ). Multiple linear regression analysis was performed using the SMFQ score at age 12 as the objective variable and being praised for prosocial behavior as the main explanatory variable, and the SMFQ score at age 10 and the objective prosocial behavior at age 10 were included as confounders.ResultsDepressive symptoms (SMFQ scores) in the “prosocial praise group” were significantly lower than those in the other group both at age 10 (4.3 ± 4.4 vs. 4.9 ± 4.6, p < 0.001) and at age 12 (3.4 ± 4.2 vs. 4.0 ± 4.6, p < 0.01). In the single regression analysis, the children who reported being praised for prosocial behavior at age 10 had significantly lower depressive symptoms at age 12 (partial regression variable: −0.57, 95% confidence interval (CI) [−0.96, −0.17]). This association remained significant after adjusting for confounders, including baseline depressive symptoms (partial regression variable: −0.44, 95% CI [−0.80, −0.08]). Prosocial behavior alone was not associated with depressive symptoms.ConclusionsBeing praised for prosocial behavior rather than objective prosocial behavior at 10 years of age predicted lower depressive symptoms 2 years later. Praise for adolescents' prosocial behavior can be encouraged to prevent depression