Nitroprusside increases intracellular Zn2+ concentration without affecting cellular thiol content : A model experiment using rat thymocytes and FluoZin-3

Nitric oxide (NO) is cytotoxic under some conditions although it has physiological roles. It is recently proposed that the cytotoxicity of NO is resulted from its interaction with glutathione and zinc. Since we have revealed that a decrease in cellular content of non-protein thiols, presumably glutathione, induces intracellular Zn2+ release, there is a possibility that the cytotoxicity of nitroprusside, a donor of NO, is resulted from the interaction of NO with cellular thiols, leading to an increase in intracellular Zn2+ concentration. To test the possibility, the effects of nitroprusside on cell lethality, intracellular thiol content, and intracellular Zn2+ concentration were examined in rat thymocytes by using a flow cytometer with propidium iodide and FluoZin-3. Nitroprusside at concentrations of 0.3 mM or more (up to 10 mM) significantly augmented FluoZin-3 fluorescence, indicating an increase in intracellular Zn2+ concentration. It was also the case under external Zn2+-free condition, suggesting nitroprusside-induced release of intracellular Zn2+. However, nitroprusside at 10 mM did not affect cell lethality and cellular thiol content. Thus, it can be concluded that nitroprusside-induced increase in intracellular Zn2+ concentration is not related to its cytotoxicity

Tetracaine decreases intracellular Zn2+ concentration by inhibiting Zn2+ influx in rat thymocytes

In this study to examine the cytotoxic property of tetracaine, we cytometrically examined the effect of tetracaine on intracellular Zn2+ concentration by the use of FluoZin-3, a fluorescent indicator of intracellular Zn2+. Lidocaine was used as a reference drug. The incubation of rat thymocytes with tetracaine decreased the intensity of FluoZin-3 fluorescence while that with lidocaine increased the intensity. The incubation with 10 μM DTPA, a chelator for extracellular Zn2+, attenuated the tetracaine-induced decrease in fluorescence intensity. The application of ZnCl2 augmented FluoZin-3 fluorescence. The augmentation by ZnCl2 was a temperature-sensitive. Tetracaine attenuated the ZnCl2-induced augmentation of FluoZin-3 fluorescence. Taken together, the results suggest that tetracaine attenuates membrane Zn2+ influx, resulting in a decrease in intracellular Zn2+ concentration in rat thymocytes. Although the cells in this study are not targets for actions of local anesthetics, the result may give one clue to explain the difference between the cytotoxicity of local anesthetics since the action of tetracaine on FluoZin-3 fluorescence was opposite to that of lidocaine

The Late Phase/Early Phase Ratio of Pancreatic CT Values as a Novel Predictor of Pancreatic Fistula after Distal Pancreatectomy

Post-operative pancreatic fistula (POPF) remains the most common complication after distal pancreatectomy (DP). In this retrospective study, we reviewed the data from patients who underwent DP between 2008 and 2019 in our institute to determine whether the late phase/early phase ratio (L/E ratio) by preoperative computed tomography (CT) scan in the pancreas could predict POPF occurrence after DP. We examined the relationship between preoperative or intraoperative factors and the occurrence of POPF after DP using statistical methods in 23 males and 21 females with a mean age of 73. The mean L/E ratio was significantly lower in the POPF group than the non-POPF group (p=0.035). The L/E ratio had moderate diagnostic accuracy, with a calculated optimal cutoff value of 0.77. In univariate analysis, a significant association was noted between POPF and stump thickness ≥ 16.9, body mass index ≥ 27.5, and L/E ratio ≤ 0.77. In the multivariate analysis, the L/E ratio (odds ratio, 5.96; p=0.036) was an independent risk factor for POPF. Our findings suggest that the pancreatic L/E ratio may predict the occurrence of POPF after DP. This measure may be useful in preoperative risk stratification, patient counseling, and perioperative patient management, improving clinical outcomes after DP

Phosphorylation of the RSRSP stretch is critical for splicing regulation by RNA-Binding Motif Protein 20 (RBM20) through nuclear localization

RBM20 is a major regulator of heart-specific alternative pre-mRNA splicing of TTN encoding a giant sarcomeric protein titin. Mutation in RBM20 is linked to autosomal-dominant familial dilated cardiomyopathy (DCM), yet most of the RBM20 missense mutations in familial and sporadic cases were mapped to an RSRSP stretch in an arginine/serine-rich region of which function remains unknown. In the present study, we identified an R634W missense mutation within the stretch and a G1031X nonsense mutation in cohorts of DCM patients. We demonstrate that the two serine residues in the RSRSP stretch are constitutively phosphorylated and mutations in the stretch disturb nuclear localization of RBM20. Rbm20 S637A knock-in mouse mimicking an S635A mutation reported in a familial case showed a remarkable effect on titin isoform expression like in a patient carrying the mutation. These results revealed the function of the RSRSP stretch as a critical part of a nuclear localization signal and offer the Rbm20 S637A mouse as a good model for in vivo study

ケニア キョウワコク ノ ショトウ ガッコウ ニオケル ジュギョウ ケンキュウ : ゲンジョウ ト カダイ

Japan International Cooperation Agency (JICA) implemented a project titled Strengthening Mathematics and Science Education Project (SMASE:2009-2013) to improve the quality of teaching by introducing lesson study in clusters as well as in schools. A total of 80 plus Teacher Advisory Centre (TAC) tutors and District Quality Assurance and Standard Officers (DQASO) were trained between 2010 and 2013 in Naruto University of Education to enhance their skills to observe lessons and make constructive comments for lesson improvement. As a follow-up，the authors visited a few INSET sites and confirmed that the lesson study is an effective tool for improving teaching methods in classroom， but further capacity building on subject content is crucial to make an effective change in lesson presentation

Adult patients with Ph+ ALL benefit from conditioning regimen of medium‐dose VP16 plus CY/TBI

The medium-dose etoposide (VP16) added on cyclophosphamide (CY)/total body irradiation (TBI) is one of the intensified myeloablative conditioning regimens used in allogenic hematopoietic stem cell transplantation (allo-HSCT) for acute lymphoblastic leukemia (ALL). However, the patient subgroups who can actually benefit from VP16/CY/TBI compared to CY/TBI have not been precisely defined. Therefore, we conducted a multi-center retrospective study using the Japanese nationwide registry database to elucidate the efficacy of VP16/CY/TBI on post-transplant prognosis. Biological and clinical distinct subtypes (i.e., Philadelphia chromosome-positive (Ph+) and -negative (Ph−) ALL) were evaluated separately, which included 820 Ph+ and 1463 patients with Ph− ALL, respectively. Compared with the CY/TBI group, the VP16/CY/TBI group showed superior progression-free survival (PFS) in patients with Ph+ ALL (65% vs. 57% at 3 years after HSCT; adjusted hazard ratio (HR), 0.73; 95% confidence interval (CI), 0.55–0.98; p = 0.03), along with significantly reduced incidence of relapse (adjusted HR, 0.58; 95% CI, 0.37–0.90; p = 0.02) without the increase of non-relapse mortality (NRM). By contrast, in patients with Ph− ALL, VP16/CY/TBI did not improve PFS nor incidence of relapse; addition of VP16 reduced relapse (HR, 0.65; p = 0.06) in patients with Ph− ALL transplanted at CR1, while improved PFS was not observed (HR, 0.90; p = 0.52) due to increased NRM. This study demonstrated that VP16/CY/TBI is a more effective and well-tolerated regimen in comparison with CY/TBI in patients with myeloablative allo-HSCT for adult Ph+ ALL. Our findings can provide a novel algorithm for conditioning regimen selection in patients with adult ALL

Mutant analyses reveal different functions of fgfr1 in medaka and zebrafish despite conserved ligand–receptor relationships

AbstractMedaka (Oryzias latipes) is a small freshwater teleost that provides an excellent developmental genetic model complementary to zebrafish. Our recent mutagenesis screening using medaka identified headfish (hdf) which is characterized by the absence of trunk and tail structures with nearly normal head including the midbrain–hindbrain boundary (MHB). Positional-candidate cloning revealed that the hdf mutation causes a functionally null form of Fgfr1. The fgfr1hdf is thus the first fgf receptor mutant in fish. Although FGF signaling has been implicated in mesoderm induction, mesoderm is induced normally in the fgfr1hdf mutant, but subsequently, mutant embryos fail to maintain the mesoderm, leading to defects in mesoderm derivatives, especially in trunk and tail. Furthermore, we found that morpholino knockdown of medaka fgf8 resulted in a phenotype identical to the fgfr1hdf mutant, suggesting that like its mouse counterpart, Fgf8 is a major ligand for Fgfr1 in medaka early embryogenesis. Intriguingly, Fgf8 and Fgfr1 in zebrafish are also suggested to form a major ligand–receptor pair, but their function is much diverged, as the zebrafish fgfr1 morphant and zebrafish fgf8 mutant acerebellar (ace) only fail to develop the MHB, but develop nearly unaffected trunk and tail. These results provide evidence that teleost fish have evolved divergent functions of Fgf8–Fgfr1 while maintaining the ligand–receptor relationships. Comparative analysis using different fish is thus invaluable for shedding light on evolutionary diversification of gene function

A New Serum Biomarker Set to Detect Mild Cognitive Impairment and Alzheimer’s Disease by Peptidome Technology

Background: Because dementia is an emerging problem in the world, biochemical markers of cerebrospinal fluid (CSF) and radio-isotopic analyses are helpful for diagnosing Alzheimer’s disease (AD). Although blood sample is more feasible and plausible than CSF or radiological biomarkers for screening potential AD, measurements of serum amyloid- β (Aβ), plasma tau, and serum antibodies for Aβ1 - 42 are not yet well established. Objective: We aimed to identify a new serum biomarker to detect mild cognitive impairment (MCI) and AD in comparison to cognitively healthy control by a new peptidome technology. Methods: With only 1.5μl of serum, we examined a new target plate “BLOTCHIP®” plus a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) to discriminate control (n = 100), MCI (n = 60), and AD (n = 99). In some subjects, cognitive Mini-Mental State Examination (MMSE) were compared to positron emission tomography (PET) with Pittsburgh compound B (PiB) and the serum probability of dementia (SPD). The mother proteins of candidate serum peptides were examined in autopsied AD brains. Results: Apart from Aβ or tau, the present study discovered a new diagnostic 4-peptides-set biomarker for discriminating control, MCI, and AD with 87% of sensitivity and 65% of specificity between control and AD (***p  Conclusion: The present serum biomarker set provides a new, rapid, non-invasive, highly quantitative and low-cost clinical application for dementia screening, and also suggests an alternative pathomechanism of AD for neuroinflammation and neurovascular unit damage

Sense of coherence as a predictor of onset of depression among Japanese workers: a cohort study

<p>Abstract</p> <p>Background</p> <p>The ability to predict future onset of depression is required for primary prevention of depression. Many cross-sectional studies have reported a correlation between sense of coherence (SOC) and the presence of depressive symptoms. However, it is unclear whether SOC can predict future onset of depression. Therefore, whether measures to prevent onset of depression are needed in for persons with low SOC is uncertain. Thus, the aim of this cohort study was to determine whether SOC could predict onset of depression and to assess the need for measures to prevent onset of depression for persons with low SOC.</p> <p>Methods</p> <p>A total of 1854 Japanese workers aged 20-70 years in 2005 who completed a sense of coherence (SOC) questionnaire were followed-up until August 2007 using their sick-pay records with medical certificates. Depression was defined as a description of "depression" or "depressive" as a reason for sick leave on the medical certificates. The day of incidence of depression was defined as the first day of the sick leave. Risk ratios of SOC for onset of depression were calculated using a multivariate Cox proportional hazards model.</p> <p>Results</p> <p>Of the 1854 participants, 14 developed depression during a mean of 1.8 years of follow-up. After adjustment for gender and age, the risk ratio of high SOC compared with low SOC for sick leave from depression was 0.18 (95% confidence interval [CI], 0.04 to 0.79). The area under the receiver operating characteristic curve of SOC was 0.70 (95% CI, 0.58 to 0.82).</p> <p>Conclusions</p> <p>The SOC may be able to predict onset of depression in Japanese workers. Measures to prevent onset of depression for persons with low SOC might be required in Japanese workplaces. Thus, SOC could be useful for identifying persons at high risk for future depression.</p