TB-related technical enquiries received in Japan, 2017-2019.

SETTING: Japan, an intermediate TB burden country. OBJECTIVE: To review TB-related technical enquiries received at the Research Institute of Tuberculosis, Japan, from January 2017 to December 2019. DESIGN: This was a cohort study. RESULTS: A total of 2,197 enquiries were analysed. On average, 61.0 enquiries/month (range: 42-81) were received. The enquiry rates were highest for the Yamanashi (4.65/100,000 population) and Ishikawa (4.55) Prefectures, and lowest in the Yamagata (0.46) and Tochigi (0.56) Prefectures. The main organisations the enquirers belonged to were local governments (n = 1,585, 72.1%) and healthcare facilities (n = 307, 14.0%). The enquirers were medical doctors (n = 391, 17.8%), nurses (n = 1,207, 54.9%), other healthcare professionals (n = 57, 2.6%), the general public (n = 168, 7.6%) and others/unknown (n = 374, 17.0%). The most frequent enquiries were about TB diagnosis and treatment (n = 501, 22.8%), including laboratory diagnosis (n = 88, 4.0%), TB treatment in general (n = 93, 4.2%) and management of comorbidities (n = 86, 3.9%), followed by contact investigations (n = 385, 17.5%) and TB in foreigners (n = 344, 15.7%). CONCLUSION: As the most frequent enquiries were about diagnosis and treatment of TB, the health ministry of Japan should maintain a few specialised TB institutions with TB physicians to provide technical assistance.

LIEU: Le Japon, un pays à charge intermédiaire en matière de TB. OBJECTIF: Examiner les demandes de renseignements techniques liées à la TB reçues au Research Institute of Tuberculosis, au Japon, de janvier 2017 à décembre 2019. METHODE: Il s'agissait d'une étude de cohorte. RÉSULTATS: Au total, 2 197 demandes ont été analysées. En moyenne, 61,0 demandes de renseignements/mois (fourchette : 42­81) ont été reçues. Les taux de demande étaient les plus élevés dans les préfectures de Yamanashi (4,65/100 000 habitants) et d'Ishikawa (4,55), et les plus faibles dans les préfectures de Yamagata (0,46) et de Tochigi (0,56). Les principales organisations auxquelles appartiennent les enquêteurs sont les administrations locales (n = 1 585 ; 72,1%) et les établissements de santé (n = 307; 14,0%). Les enquêteurs étaient des médecins (n = 391 ; 17,8%), des infirmières (n = 1 207 ; 54,9%), d'autres professionnels de la santé (n = 57 ; 2,6%), le grand public (n = 168 ; 7,6%) et autres/inconnus (n = 374 ; 17,0%). Les demandes les plus fréquentes concernaient le diagnostic et le traitement de la TB (n = 501 ; 22,8%), y compris le diagnostic en laboratoire (n = 88 ; 4,0%), le traitement de la TB en général (n = 93 ; 4,2%) et la prise en charge des comorbidités (n = 86 ; 3,9%), suivis par les enquêtes sur les contacts (n = 385 ; 17,5%) et la TB chez les étrangers (n = 344 ; 15,7%). CONCLUSION: Comme les demandes de renseignements les plus fréquentes concernaient le diagnostic et le traitement de la TB, le ministère de la santé du Japon devrait maintenir quelques institutions spécialisées dans la TB avec des médecins spécialistes de la TB pour fournir une assistance technique.

Optical high-power nonlinearity comparison between the National Institute of Standards and Technology and the National Metrology Institute of Japan at 1480 nm.

We compare the results of measurements of the nonlinearity of high-power optical fiber powermeters (OFPMs) by two national metrology institutes (NMIs): the National Institute of Standards and Technology (NIST-USA) and the National Metrology Institute of Japan/National Institute of Advanced Industrial Science and Technology (NMIJ/AIST-Japan) at a wavelength of 1480 nm. The nonlinearity and range discontinuity of a commercial OFPM were measured from 1 mW to 500 mW by use of a superposition method (both laboratories) and from 1 mW to 250 mW by use of a comparison method (NMIJ only). Measurement results showed largest differences of less than 1.6 parts in 10(3), which is within the combined expanded (k = 2) uncertainty for both laboratories.

Association between the Human Development Index and Millennium Development Goals 6 Indicators in Sub-Saharan Africa from 2000 to 2014: Implications for the New Sustainable Development Goals.

It is important to assess whether regional progress toward achieving the millennium development goals (MDGs) has contributed to human development and whether this has had an effect on the triple burden of disease in the continent. This analysis investigates the association between the human development index (HDI) and co-occurrence of HIV/AIDS, tuberculosis (TB), and malaria as measured by MDG 6 indicators in 35 selected sub-Saharan African countries from 2000 to 2014. The analysis used secondary data from the United Nations Development Programme data repository for HDI and disease data from WHO Global Health observatory data repository. Generalized Linear Regression Models were used to analyze relationships between HDI and MDG 6 indicators. HDI was observed to improve from 2001 to 2014, and this varied across the selected sub-regions. There was a significant positive relationship between HDI and HIV prevalence in East Africa (ß = 0.048 [95% CI: 0.040-0.056], p < 0.001) and Southern Africa (ß = 0.032 [95% CI: 0.002-0.062], p = 0.034). A significant positive relationship was observed with TB incidence (ß = 0.009 [95% CI: 0.003-0.015], p = 0.002) and a significant negative relationship was observed with malaria incidence (ß = -0.020 (95% CI: -0.029 to -0.010, p < 0.001) in East Africa. Observed improvements in HDI from the year 2000 to 2014 did not translate into commensurate progress in MDG 6 goals.

A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo.

We have developed a method for visualizing phosphorylation of proteins in living cells using a novel fluorescent indicator composed of two green fluorescent protein (GFP) variants joined by the kinase-inducible domain (KID) of the transcription factor cyclic adenosine monophosphate (cAMP)-responsive element binding protein (CREB). Phosphorylation of KID by the cAMP-dependent protein kinase A (PKA) decreased the fluorescence resonance energy transfer (FRET) among the flanking GFPs. By transfecting COS-7 cells with an expression vector encoding this indicator protein (termed ART for cAMP-responsive tracer), we were able to visualize activation dynamics of PKA in living cells.

Genotype frequency of plasminogen activator inhibitor-1 (PAI-1) 4G/5G polymorphism in healthy Japanese males and its relation to PAI-1 levels.

Plasminogen activator inhibitor-1 (PAI-1) plays an inhibitory role in the fibrinolytic enzyme system and is associated with thrombotic diseases. The gene for PAI-1 has an insertion/deletion polymorphism at the promoter region, the 4G/5G polymorphism, which is related to differences in transcription activity in vitro. Association of the 4G/5G polymorphism with plasma PAI-1 levels, however, has not been uniformly reported. We evaluated the relationship between the 4G/5G polymorphism and plasma PAI-1 antigen levels in 104 Japanese males not taking lipid-lowering drugs and without non-insulin-dependent (type 2) diabetes mellitus or coronary artery disease. The genotype frequency was 37.5%, 50.0%, and 12.5% for 4G/4G, 4G/5G, and 5G/5G genotypes, respectively, which differs from that reported for healthy Caucasian males (P < 0.01), with the 4G allele more frequently found in the Japanese population. No association was found between the 4G/5G polymorphism and plasma PAI-1 antigen levels. Multiple regression analysis revealed a significant (P < 0.01) contribution of triglyceride (TG) levels to variations in plasma PAI-1 antigen levels. The correlation between TG levels and plasma PAI-1 antigen levels was not 4G/5G genotype-specific. These findings suggest that PAI-1 4G/5G polymorphism is not associated with plasma PAI-1 antigen levels among healthy Japanese males and that TG levels correlate to plasma PAI-1 antigen levels in all PAI-1 4G/5G genotypes.

Low prevalence of activated protein C resistance and coagulation factor V Arg506 to Gln mutation among Japanese patients with various forms of thrombosis, and normal individuals.

Resistance to activated protein C (APC), recently reported to be the most prevalent inherited cause of thrombosis among Caucasians, is associated with a single point mutation in the coagulation factor V gene. We investigated the prevalence of APC resistance and the factor V gene mutation (R506Q) in 34 consecutive Japanese patients with venous thrombosis or pulmonary thromboembolism and 63 control subjects. Three of the 33 patients examined (9%) had an APC ratio below the 5th percentile of control values (2.27), but all were above 2.0. The factor V mutation (R506Q) was not detected in the 29 patients studied, including the 3 patients whose APC ratios were below 2.27, or in 53 controls. In a tissue factor-based factor V assay to detect APC resistance recently described by Le et al. (Blood 1995;85:1704-1711), all patients studied were found to be normal including the three with a low APC ratio. We conclude that APC resistance and factor V gene mutation are less prevalent in Japan than in several European countries.

Beamline for calibration of transfer standard light sources in the UV and VUV regions.

A beamline which serves for calibrating transfer standard light sources (deuterium lamps, excimer lamps, Xe lamps etc.) in the UV and VUV regions is being constructed. The synchrotron radiation from the electron storage ring TERAS (750 MeV) is used as a primary standard of spectral radiant intensity. In order to use synchrotron radiation as a primary standard, the electron beam and synchrotron radiation beam parameters need to be evaluated. Uncertainties of synchrotron radiation flux evaluated by measurements of the magnetic flux density, the position of the electron orbital plane, the electron beam size and the distance from the synchrotron radiation tangent point to the detector system are expected to be about 0.003, 0.01, 0.05 and 0.1%, respectively.

Cloning of human PRP4 reveals interaction with Clk1.

Prp4 is a protein kinase of Schizosaccharomyces pombe identified through its role in pre-mRNA splicing, and belongs to a kinase family including mammalian serine/arginine-rich protein-specific kinases and Clks, whose substrates are serine/arginine-rich proteins. We cloned human PRP4 (hPRP4) full-length cDNA and the antiserum raised against a partial peptide of hPRP4 recognized 170-kDa polypeptide in HeLa S3 cell extracts. Northern blot analysis revealed that hPRP4 mRNA was ubiquitously expressed in multiple tissues. The extended NH(2)-terminal region of hPRP4 contains an arginine/serine-rich domain and putative nuclear localization signals. hPRP4 phosphorylated and interacted with SF2/ASF, one of the essential splicing factors. Indirect immunofluorescence analysis revealed that endogenous hPRP4 was distributed in a nuclear speckled pattern and colocalized with SF2/ASF in HeLa S3 cells. Furthermore, hPRP4 interacted directly with Clk1 on its COOH terminus, and the arginine/serine-rich domain of hPRP4 was phosphorylated by Clk1 in vitro. Overexpression of Clk1 caused redistribution of hPRP4, from the speckled to the diffuse pattern in nucleoplasm, whereas inactive mutant of Clk1 caused no change of hPRP4 localization. These findings suggest that the NH(2)-terminal region of hPRP4 may play regulatory roles under an unidentified signal transduction pathway through Clk1.

Identification of a novel kinesin-related protein, KRMP1, as a target for mitotic peptidyl-prolyl isomerase Pin1.

Mitosis utilizes a number of kinesin-related proteins (KRPs). Here we report the identification of a novel KRP termed KRMP1, which has a deduced 1780-amino acid sequence composed of ternary domains. The amino-terminal head domain is most similar to the kinesin motor domain of the MKLP-1 subfamily and has an intrinsic ATPase activity that is diminished by substituting the consensus Lys-168 with Arg. The central stalk domain is predicted to form a long alpha-helical coiled-coil, and can interact with each other in vivo. An in vivo labeling experiment revealed that KRMP1 is phosphorylated, and we also found that the region within the tail domain containing Thr-1604 as the cdc2 kinase phosphorylation site differs from the bimC box conserved in the bimC subfamily of KRPs. Immunofluorescence analysis showed that endogenous KRMP1 was localized predominantly to the cytoplasm during interphase and dispersed throughout the cell during mitosis. Consistent with this finding, overexpressed KRMP1 was detected in a complicated nuclear or cytoplasmic pattern reflecting multiple nuclear localization/export signals. Furthermore, KRMP1 interacted with the mitotic peptidyl-prolyl isomerase Pin1 in vivo, and an in vitro interaction was detected between the tail domain of KRMP1 and the WW domain of Pin1. Overexpression of KRMP1 caused COS-7 cells to arrest at G(2)-M, and co-expression of Pin1 reversed this effect, indicating their physiological interaction. Together, our results suggest that KRMP1 is a mitotic target regulated by Pin1 and vice versa.

Coronary artery disease and polymorphisms in a receptor mediating shear stress-dependent platelet activation.

BACKGROUND: Platelets play pivotal roles in coronary thrombosis, and antiplatelet therapies are widely used for coronary artery disease (CAD). However, the effects of genetic variation in platelets on CAD are poorly understood. We have assessed the association between CAD and polymorphisms in a platelet receptor for von Willebrand factor, the glycoprotein (GP) Ib/IX complex, which mediates shear stress-dependent platelet activation. METHODS AND RESULTS: Genotypes of the alpha-chain of the receptor (GP Ib alpha, 145Thr/Met) were determined in 91 patients with myocardial infarction (MI) or angina pectoris whose lesions were confirmed by coronary angiography as well as in 105 individuals from the general population with no history of angina or other heart diseases and normal resting ECGs. There was no homozygote for Met/Met in either the control or patient groups. The prevalence of the Thr/Met genotype (T/M) in all patients was not significantly different from that in the control group. However, the frequency of T/M was significantly higher in patients aged < or = 60 years (31.8%) than in control subjects aged < or = 60 years (16.0%; P<.05, odds ratio=2.5). An association was also demonstrated between CAD and the other polymorphism of GP Ib alpha, a variable number of tandem repeats of a 13-amino acid sequence, which is known to be linked to the 145Thr/Met polymorphism. There was an association between the frequency of the T/M genotype and the angiographic severity of CAD: 11.1% for Gensini score < 40 versus 50.0% for Gensini score > or = 40 (P=.0015). There was no difference in the distribution of GP Ib alpha genotypes between patients with MI and those with angina pectoris. CONCLUSIONS: This study suggests that the presence of the Met allele in GP Ib alpha is a risk factor for the prevalence and severity of CAD in individuals aged < or = 60 years. The results need to be confirmed in a large-scale study of incident case subjects and matching control subjects.

Genotype distribution of estrogen receptor polymorphisms in men and postmenopausal women from healthy and coronary populations and its relation to serum lipid levels.

The cardiovascular protective effects of estrogen are known to be mediated by its beneficial effects on lipid metabolism and its direct actions on the vessel wall. The latter can be mediated by a specific receptor for estrogen present on smooth muscle cells and endothelial cells. The gene for the receptor (the classic estrogen receptor [ER]) has three known polymorphisms, Pvu II, Xba I, and B-variant polymorphisms, which are reportedly associated with receptor expression and altered receptor function and with some disorders including breast cancer, hypertension, and spontaneous abortion. However, the significance of genetic variations of the ER in vascular diseases has not been reported. We have examined the association between coronary artery disease (CAD) and the three polymorphisms in ER. Genotypes (P1/P2, X1/X2, and B-wild type/B-variant type) were determined in 87 men and postmenopausal women with myocardial infarction or angina pectoris whose lesions were confirmed by coronary angiography, as well as from 94 control individuals from the general population with no coronary heart disease and normal resting ECG. For B-variant polymorphism, all individuals examined had B-wild type, which contrasts with the reported allele frequency for B-variant type (0.1) in the white population. Genotype distributions and allele frequencies of Pvu II or Xba I polymorphisms were not significantly different between control subjects and patients (P > .05 for Pvu II or Xba I genotypes; P > .05 for Pvu II or Xba I allele frequencies). When the allele frequencies were analyzed separately by sex, there was still no statistically significant difference for both polymorphisms (P > .05 for men; P > .05 for women). No association was found between the polymorphisms and the angiographic severity of CAD. Total cholesterol, triglyceride, or HDL-cholesterol levels were not significantly different among ER genotypes. These findings suggest that the three polymorphisms in ER are not associated with the prevalence and severity of CAD and that the polymorphisms are unrelated to the serum lipid levels in control subjects and patients.

A family with hereditary factor X deficiency with a point mutation Gla32 to Gln in the Gla domain (factor X Tokyo).

We report a new family with hereditary factor X deficiency. The propositus had a markedly prolonged prothrombin time, a mild prolongation of activated partial thromboplastin time and a clotting time activated by Russell's viper venom. Factor X activity in plasma was 3 u/dl (normal range 56-138 u/dl). Factor X antigen level was 61 u/dl. Molecular analysis revealed a homozygous mutation, Glu (GAG) to Gln (CAG) at residue 32 which normally undergoes gamma-carboxylation within the gamma-carboxyglutamic acid rich domain. The genotypes of family members completely correlated with their factor X activities. It is suggested that the Glu32 to Gln mutation is the molecular basis for the abnormal factor X in this family.

A 192Arg variant of the human paraoxonase (HUMPONA) gene polymorphism is associated with an increased risk for coronary artery disease in the Japanese.

Recent reports have suggested that polymorphisms in the human paraoxonase (HUMPONA) gene may be a genetic risk factor for coronary artery disease (CAD) in white populations. However, this association has not yet been confirmed in other ethnic populations. We studied 75 Japanese patients with CAD, whose coronary lesions were confirmed by angiography, and 115 Japanese control subjects with no history of CAD and a normal resting electrocardiogram. The assays for genotyping the two polymorphisms in the HUMPONA gene (192Arg/Gln and 55Leu/Met) were based on changes in restriction enzyme digestion patterns. For codon 192, the frequencies of the Arg-coding allele (B allele) in both patients and control subjects were much higher than those from published results of whites (.26 to .31), and the difference between patients (.74) and control subjects (.59) was statistically significant (P = .002). The patient group had a higher proportion of Arg/Arg (B/B) homozygotes (52.0% vs 32.2%, P = .006). For codon 55, the frequencies of the Leu-coding allele in control subjects and patients were much higher (.91 and .93, respectively) than those published results for whites, but there was no difference between Japanese control subjects and Japanese patients. When subjects with the 55Leu/Leu genotype only were analyzed, 192Arg/Arg homozygotes were still significantly more frequent in the patients than in the control subjects (55.4% vs 37.2%, P = .024), and the frequency of the 192Arg allele was also higher in patients than control subjects (P = .013). Logistic regression analysis including conventional coronary risk factors revealed that 192Arg is an independent risk factor for CAD. Thus, in the Japanese, the association of CAD with the 192Arg variant of HUMPONA (B-type enzyme) is similar to that reported for whites, although the allele frequencies for 192Arg and 55Leu are much higher in the former than the latter population.

Proteolytic enzymes at various stages of oncogenic transformation of rat fibroblasts. I. Aspartyl and cysteine proteinases.

Aspartyl and cysteine proteinases at distinct stages of carcinogenesis were analyzed in rat embryo fibroblasts, sequentially immortalized and transformed by 2 different genes: the early region of simian adenovirus SA7 and c-Ha-ras oncogene. The dynamics of expression and distribution of proteinases throughout the transformation process were examined. It was shown that in immortalized and transformed cells the activities of the aspartyl and cysteine proteinases were expressed to a variable degree and that the expression was dependent on cell-propagation time in vitro. The increase in activity both of cathepsin-D-like aspartyl proteinase and of cathepsin-L- and -B-like cysteine proteinases in cell lysates was correlated with the stages of fibroblast transformation (immortalization and tumorigenic transformation). In all cell types the major part of cysteine proteinases was localized inside the cell, while the cathepsin-D-like proteinase was apparently predominant among secreted proteinases. The cathepsin-L-like proteinase accounts for the major part of the cysteine-proteinase activity as measured by Z-Phe-Arg-MCA hydrolysis. We suggest that considerable portions of the cathepsin-D- and -L-like proteinases in all cell lines studied are secreted as a complex with inhibitor(s) and that inhibitor expression plays an important role in regulating the activity of cathepsin-D-like proteinase at different stages of transformation. Cathepsin-L-like proteinase is probably secreted in the precursor form.