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This study aimed to investigate the relationship between rumen microbiota and physiological characteristics in Japanese Black cattle during the fattening period, which are known for their distinctive physiological characteristics due to prominent intramuscular fat deposition. The experimental period was categorized into three stages: early (12 to 14 months of age; T1), middle (15 to 22 months; T2), and late fattening phases (23 to 30 months; T3). We analyzed the rumen microbiota of 21 Japanese Black cattle in each phase using 16S rRNA gene amplicon sequencing. In addition, we examined the correlation between the rumen microbiota and various physiological parameters, such as blood metabolites, hormones, and rumen fermentation characteristics. Our results showed that rumen microbial diversity was greater in T1, and the inter-group distribution of rumen microbiota in T1 was differentiated from T2 and T3. At the genus level, Flexilinea, Streptococcus, Butyrivibrio, Selenomonasand Kandleriawere the most abundant in T1. Bifidobacterium, Shuttleworthiaand Sharpeawere the most abundant in T2. Acetitomaculum, Mycoplasma, Atopobiumand Howardellawere the most abundant in T3. In correlation analysis, Monoglobusand Olsenellanegatively correlated with IGF-1 and positively correlated with cortisol in T1. Firmicutes/Bacteroidotaratio positively correlated with daily gain, blood total cholesterol, and lactic acid dehydrogenase in T3. Planctomycetotacorrelated with volatile fatty acid components during the entire period, while Verrucomicrobiotapositively correlated with carcass traits in T1. Additionally, functional genetic profiles indicated that rumen microbiota related to carbohydrate metabolism and nucleotide biosynthesis was significantly enriched in T1 and T3, respectively. The composition of rumen microbial in Japanese Black cattle varied depending on fattening stages, and this variation is likely attributable to changes in feed composition. Furthermore, specific microorganisms were significantly associated with physiological parameters. Our findings could help in the development of strategies to enhance gut health and productivity by manipulating the rumen microbiota through the changes in diet compositions considering the fattening stages of Japanese Black cattle.

Several algorithms for localizing accessory pathways (APs) are based on the delta wave morphology, R/S ratio, and QRS polarity. However, they are somewhat complicated, and an accurate determination of the delta wave morphology is occasionally difficult. The aims of this study were to develop a simple algorithm for localizing APs using only the R/S ratio, and to test the accuracy of the algorithm prospectively.

Several algorithms for localizing accessory pathways (APs) are based on the delta wave morphology, R/S ratio, and QRS polarity. However, they are somewhat complicated, and an accurate determination of the delta wave morphology is occasionally difficult. The aims of this study were to develop a simple algorithm for localizing APs using only the R/S ratio, and to test the accuracy of the algorithm prospectively. We studied 142 patients with a single anterogradely conducting AP on a 12-lead ECG. R/S ratios were analyzed in leads V1, V2, and aVF (R/S-V1, R/S-V2, and R/S-aVF). AP locations were divided into five regions based on fluoroscopic anatomy. A new algorithm was developed by correlating R/S-V1, R/S-V2, and R/S-aVF with successful ablation sites in 88 initial consecutive patients. All 55 patients with left free wall APs showed R/S-V1 =0.5, and 47 (98%) of 48 patients with left anterior or lateral APs showed R/S-aVF =1. In contrast, all seven patients with left posterolateral or posterior APs showed R/S-aVF <1. All nine patients with right-and-left midseptal or posteroseptal APs showed R/S-V1 <0.5 and R/S-V2 =0.5. Of 12 patients with right anterior, lateral or anteroseptal APs, 10 (83%) showed R/S-V1 <0.5, R/S-V2 <0.5 and R/S-aVF =1. Finally, nine (75%) of 12 patients with right posterolateral or posterior APs showed R/S-V1 <0.5, R/S-V2 <0.5, and R/S-aVF <1. Then this algorithm was tested prospectively in 54 patients. Overall, the sensitivity was 94%, and the specificity was 98%. This ECG algorithm provides a simple and accurate way to identify the AP localization.

Previously, we reported that Co-Ce-O composite films were successfully deposited from the reaction solution by a newly developed chemical method called the "metal-oxide co-electro/electroless deposition method". In the present study, we first constructed potential-pH diagrams of the Co-Ce-H2O system and Co-Ce-glycine-H2O system using a set of thermodynamic data at 298K and 333K to obtain the fundamental thermodynamic consideration of the electro/electroless deposition conditions of Co-Ce-O films. As a result, we revealed that the addition of glycine to the Co-Ce-O electro/electroless deposition mixture solution is thermodynamically suitable for the fabrication of Co-Ce-O films.

ABSTRACTLoop-mediated isothermal amplification (LAMP) is a novel method that rapidly amplifies target DNA with high specificity under isothermal conditions. It has been applied as a diagnostic tool for several infectious diseases, including viral, bacterial, and parasitic diseases. In the present study, we developed a LAMP method for the molecular diagnosis of Plasmodium knowlesiinfection (PkLAMP) and evaluated its sensitivity, specificity, and clinical applicability. We designed three sets of PkLAMP primers for the species-specific ß-tubulin gene. The primer sets for PkLAMP specifically amplified the autologous DNA extracts of P. knowlesi, and the sensitivity of the test was 100-fold that of single-PCR assay. These results indicate that our PkLAMP method can be used to efficiently distinguish between P. knowlesiand other malaria parasites. To evaluate the feasibility of using in vivomaterials, comparisons of PkLAMP and the conventional nested PCR (nPCR) method and microscopic examination were made with blood samples from two experimentally infected monkeys. These studies showed that P. knowlesiinfection can be identified much earlier with PkLAMP than with nPCR and microscopy. Moreover, the detection performance of PkLAMP using whole blood as the template was identical to that of PkLAMP when genomic DNA extracts were used. These results suggest that the PkLAMP method is a promising tool for molecular diagnosis of P. knowlesiinfection in areas of endemicity.

Abstract: Synthesis of photoluminescent Si nanoparticles has been successfully prepared using an intense pulsed ion-beam evaporation (IBE) technique in vacuum. Si nanoparticles are produced by the IBE method without any post-annealings. Photoluminescence (PL) mainly in blue range with a peak of 455 nm and a shoulder near 510 nm is observed in as-deposited Si nanoparticles at room temperature. The blue light emission is relatively stable with no noticeable change, as the samples have already stored in air more than 4 months. The observed PL does not fit the quantum confinement model, since a majority of particle size is around ~20 nm, estimated by SEM and XRD measurements. Moreover, hydrofluoric acid (HF) corrosion tests on the Si nanoparticles also indicate a correlation between the presence of the surface oxide layers and the PL. Oxide-related luminescence is likely the source of this blue light emission.

Abstract Introduction: Pulmonary vein (PV) isolation (PVI) has been recently proposed as an effective technique to cure atrial fibrillation (AF).

Plasminogen activator inhibitor-1 (PAI-1) plays a critical role in tissue fibrosis by inactivating matrix metalloproteinases, which might effect on the progression of left ventricular dysfunction. However, little has been known about the expression of PAI-1 during cardiac remodeling. We used a mouse model of myocardial infarction (MI) by coronary ligation, in which the progression of left ventricular remodeling was confirmed by echocardiography. Histological examination showed that interstitial and perivascular fibrosis progressed in the post-MI (PMI) heart at 4 weeks after the procedure. We observed the dramatic induction of cardiac PAI-1 mRNA and PAI-1 antigen in plasma in the PMI mice, as compared with the sham-operated (sham) mice. In situhybridization analysis demonstrated that strong signals for PAI-1 mRNA were localized to cardiomyocytes in the boarder of infarct area and around fibrous lesions, and to perivascular mononuclear cells, which seemed to be mast cells, only in hearts of the PMI mice. Importantly, less development of cardiac fibrosis after MI was observed in mice deficient in PAI-1 as compared to wild-type mice. The mRNA expression of cytokines, transforming growth factor-β, and tumor necrosis factor-α, was also increased in hearts of the PMI mice, but not in the sham mice. These observations suggest that cardiomyocytes and mast cells contribute to the increased PAI-1 expression, resulting in the development of interstitial and perivascular fibrosis in the PMI heart, and that the regional induction of cytokines may be involved in this process.

Ligand-receptor internalization has been traditionally regarded as part of the cellular desensitization system. Low-density lipoprotein receptor-related protein (LRP) is a large endocytosis receptor with a diverse array of ligands. We recently showed that LRP binds heparin-binding growth factor midkine. Here we demonstrate that LRP mediates nuclear targeting by midkine and that the nuclear targeting is biologically important. Exogenous midkine reached the nucleus, where intact midkine was detected, within 20 min. Midkine was not internalized in LRP-deficient cells, whereas transfection of an LRP expression vector restored midkine internalization and subsequent nuclear translocation. Internalized midkine in the cytoplasm bound to nucleolin, a nucleocytoplasmic shuttle protein. The midkine-binding sites were mapped to acidic stretches in the N-terminal domain of nucleolin. When the nuclear localization signal located next to the acidic stretches was deleted, we found that the mutant nucleolin not only accumulated in the cytoplasm but also suppressed the nuclear translocation of midkine. By using cells that overexpressed the mutant nucleolin, we further demonstrated that the nuclear targeting was necessary for the full activity of midkine in the promotion of cell survival. This study therefore reveals a novel role of LRP in intracellular signaling by its ligand and the importance of nucleolin in this process.

ABSTRACTLigand-receptor internalization has been traditionally regarded as part of the cellular desensitization system. Low-density lipoprotein receptor-related protein (LRP) is a large endocytosis receptor with a diverse array of ligands. We recently showed that LRP binds heparin-binding growth factor midkine. Here we demonstrate that LRP mediates nuclear targeting by midkine and that the nuclear targeting is biologically important. Exogenous midkine reached the nucleus, where intact midkine was detected, within 20 min. Midkine was not internalized in LRP-deficient cells, whereas transfection of an LRP expression vector restored midkine internalization and subsequent nuclear translocation. Internalized midkine in the cytoplasm bound to nucleolin, a nucleocytoplasmic shuttle protein. The midkine-binding sites were mapped to acidic stretches in the N-terminal domain of nucleolin. When the nuclear localization signal located next to the acidic stretches was deleted, we found that the mutant nucleolin not only accumulated in the cytoplasm but also suppressed the nuclear translocation of midkine. By using cells that overexpressed the mutant nucleolin, we further demonstrated that the nuclear targeting was necessary for the full activity of midkine in the promotion of cell survival. This study therefore reveals a novel role of LRP in intracellular signaling by its ligand and the importance of nucleolin in this process.

This article aims to clarify the clinical significance of changes in electrocardiographic (ECG) R-wave voltage on chest leads from 1 to 4 weeks in patients with acute anterior myocardial infarction (MI) in combination with echocardiographic findings and dual scintigraphic findings. Seventy-one patients with acute anterior MI who underwent emergency revascularization were subjected to ECG and echocardiography, at both 1 and 4 weeks, and to thallium-201 (TI) and iodine-123-[beta ]-methyl-p-iodophenyl pentadecanoic acid (BMIPP) single-photon emission computed tomography (SPECT) about 1 week after the onset of MI. The total sum of ECG R-wave voltage on each chest lead was calculated. The mean defect ratio on TI and that on BMIPP derived from circumferential profile curve analysis were calculated. The percentage defect-discordant ratio of both SPECT images [(%) discordance on TI/BMIPP] was obtained. The percentage increase ratio of ECG R-wave voltage on chest leads [(%) increase of R wave] and the increase of left ventricular ejection fraction ([Delta ]EF) from 1 to 4 weeks were obtained. There were significant correlations between the (%) increase of R wave and the [Delta ]EF as well as between the (%) increase of R wave and the (%) discordance on TI/BMIPP (r = .63, P[lt ] .001; r = .74, P[lt ] .001). The reversibility of ECG R-wave voltage was related to cardiac functional improvement in addition to the discordance on the 2 images. Monitoring of changes in ECG R-wave voltage on chest leads is useful to detect the presence of myocardial viability and to evaluate functional evolution in patients with acute anterior MI.

Gamete activation factor (GAF) induces exflagellation of Plasmodium microgametes. We found GAF in the salivary glands of female mosquitoes, Anopheles stephensi. The exflagellation was induced in a concentration-dependent manner in the supernatant of salivary gland's crude homogenate. The exflagellation-inducing activity in the salivary gland was higher than that in the midgut and the head. GAF in the salivary glands was found to be heat stable and low molecular weight (<3000 molecular weight). Analysis of the supernatant by capillary electrophoresis and UV absorbance profile showed that the salivary glands contained xanthurenic acid, which was previously identified as GAF in the head of A. stephensi. The exflagellation-inducing activity in the salivary gland declined immediately after a blood meal, implying that GAF was in the saliva, and was delivered into the midgut together with the blood and induced exflagellation in the midgut.

Background. Few studies have focused on sudden death among apparently healthy workers, and the risk factors have not been fully discussed.Methods. A nested case-control study was conducted among 164,017 male employees receiving annual medical checkups in Japan. Most recent medical checkup data of 242 sudden death victims (mean age, 48.0 years) were compared with corresponding data of 505 age-, workplace-, and job-type-matched male controls. Odds ratios (ORs) and their 95% confidence intervals (CIs) for each variable were calculated by logistic regression.Results. OR (95% CI) significantly increased with advancing blood pressure, reaching 6.6 (3.4–13.1) for systolic blood pressure ≥ 160 mm Hg relative to that <120 mm Hg. Hypo-HDL-cholesterolemia, hyperuricemia, increased aminotransferases, and abnormal urinary findings were associated with the risk in a dose-dependent manner. The presence of arrhythmias and ST -T abnormalities as well as abnormal Q waves on electrocardiograms yielded a 3.5 to 4.8 times greater risk of sudden death. As for lifestyles, heavy smoking was a positive, and light drinking was a negative risk factor. Multivariate analysis revealed that hypertension, proteinuria, glucosuria, arrhythmias, ST-T abnormalities, and light drinking were independent predictors for sudden death.Conclusion. These findings suggest that periodic medical checkups can help to predict and prevent employee sudden death.

A recent long-term multicenter trial has shown that pimobendan is more effective when administered in low doses. However, no data are available concerning the effect of a low dose of pimobendan on the systolic and diastolic pressure-volume relations in patients with heart failure. Therefore we examined the effects of a single low dose of oral pimobendan, a calcium sensitizer, on systolic and diastolic hemodynamics in patients with cardiomyopathy and congestive heart failure. We measured the left ventricular (LV) pressure-volume relations using a conductance catheter with a micromanometer tip in 10 patients with chronic congestive heart failure resulting from idiopathic cardiomyopathy before and 45 and 90 min after administration of a single oral dose of 2.5 mg of pimobendan. End-systolic elastance was used as an index of LV contractility and was measured during transient occlusion of the inferior vena cava. End-systolic elastance increased significantly by 25% at 45 min (p < 0.05) and by 55% at 90 min (p < 0.01) without an increase in myocardial oxygen consumption. The inotropic effect was accompanied by improved ventriculoarterial coupling. This effect was attenuated in patients with severely impaired myocardial contractility. LV relaxation, assessed by the time constant of isovolumic pressure decay (T1/2), was significantly shortened at 90 min (from 47.7 ± 1.9 to 41.2 ± 1.7 ms; p < 0.01), although it remained unchanged at 45 min. The diastolic pressure-volume relation showed a leftward and downward shift in all patients. These results indicate that low-dose oral pimobendan had favorable short-term inotropic and lusitropic effects in patients with congestive heart failure caused by idiopathic dilated cardiomyopathy, and may thus be a useful alternative to traditional agents. Further study in a large-scale trial is merited.

Ultrafine γ-alumina (γ-Al2O3) powders have been successfully produced by laser ablation (aluminum target, Nd:YAG laser, 1.06 μm, 7 ns, filter-target distance 6 cm, oxygen flow rate 1 l/min, pressure 200 Torr, fluence ∼16 J/cm2). The structural properties of the Al2O3powders have been studied by X-ray diffraction, transmission electron microscopy and Brunauer, Emmet, Teller analysis. The behavior of luminous plume in background oxygen has been investigated by streak and fast photography, and emission spectroscopy. Stoichiometric γ-Al2O3powder has been obtained at an oxygen pressure of 200 Torr, with a diameter of 11.8 nm and a specific surface area of 160 m2/g. At lower oxygen pressures the average diameter decreased to 6.1 nm, and the surface particle area increased to over 300 m2/g, but the powder composition was altered by the presence of some unreacted aluminum. The measurements on plume expansion revealed initial plume velocities of 0.04–0.18 cm/μs, slowing down with pressure and distance. Emission spectroscopy indicates the presence of both neutral and ionized species, the most prominent lines being Al(I) 394.4 and 396.1 nm, Al(I) 358.7 nm, Al(III) 360.1 and 361.2 nm and Al(II) 466.7 nm; in time, only Al(I) 394.4 and 396.1 nm lines persisted for more than 2 μs. Much weaker lines could be observed for O(II) ions. γ-Al2O3(0.9 g) powders have been obtained for 1 kWh (laser output energy), with a powder collection efficiency of ∼75%.

Juvenile hormone esterase (JHE) from hemolymph of the silkworm moth Bombyx moriwas characterized for substrate specificity and inhibitor sensitivity. B. moriJHE hydrolyzed the juvenile hormone surrogate substrate methyl n-heptylthioacetothioate (HEPTAT) more efficiently than p-nitrophenyl acetate and 1-naphthyl acetate substrates widely used to assay total carboxylesterase activity. B. moriJHE was sensitive to 3-octylthio-1,1,1-trifluoro-2-propanone (OTFP), which was developed as a selective inhibitor for lepidopteran JHE, and relatively insensitive to diisopropyl fluorophosphate (DFP), an inhibitor of serine esterases but not of all JHEs. Affinity purification with a trifluoromethyl ketone ligand was more efficient for purification of B. moriJHE than DEAE ion exchange chromatography.

Vitamin K-dependent carboxylase catalyzes the posttranslational conversion of glutamic acid to γ-carboxyglutamic acid in vitamin K-dependent proteins. The clustered charged-to-alanine scanning mutagenesis of bovine carboxylase has identified five distinct candidate regions (I. Sugiuraet al., J. Biol. Chem.271, 17837–17844, 1996) with significant loss-of-function phenotype. To further specify the residues essential for the structure and function of the enzyme, Lys-217, Lys-218, Arg-359, His-360, Lys-361, Arg-513, and Lys-515 were analyzed by substituting to alanine individually. All the mutants except for K217A were expressed in Chinese hamster ovary cells. The carboxylase activities of R359A, H360A, and R513A decreased in parallel with the vitamin K epoxidase activities. Both carboxylations by R359A and H360A were stimulated saturatively at 1 μM factor IX propeptide (proFIX18) concentration, but that by R513A was not at a concentration up to 128 μM. K218A completely lost the enzyme activities but it cross-linked to the propeptide, suggesting that Lys-218 is critical for enzyme activity without affecting propeptide binding. We conclude that Lys-218, Arg-359, and His-360 are involved in the catalytic event, and Arg-513 participates in propeptide binding.

Chemical properties of the ice-nucleating active agents (INAs) distributed in the muscle of diapausing larvae of the rice stem borer, Chilo suppressalis Walker were studied. Sensitivity to protease indicated that the INAs were proteinaceous. The INAs were stable at at least 50°C for 5 min, and active over a broad pH range (pH 2.0 to 8.0) and after delipidation treatments. These results suggest that the chemical characteristics of the INAs of the muscle in the diapausing larvae are similar to those of the exogenous agents in the nondiapausing ones in Fusarium sp. After differential centrifugation of muscle homogenate of the diapausing larvae, the crystallization temperature of the 105,000g pellet was the highest of all fractions separated, implying that INAs are present in microsome membrane. Copyright 1995, 1999 Academic Press

Objectives. We investigated the usefulness of QRST values obtained from 12-lead electrocardiograms (ECGs) for identification of repolarization abnormalities before and after radiofrequency ablation in patients with Wolff-Parkinson-White syndrome.

To evaluate the diagnostic value of the postexercise systolic blood pressure (SBP) response for detecting and evaluating the presence of coronary artery disease (CAD), treadmill testing was conducted in 130 subjects with normal blood pressure and 51 patients with hypertension, each of whom underwent selective coronary angiography. A total of 48 subjects with normal blood pressure and 27 patients with hypertension had no significant narrowing of the coronary artery (control subjects), whereas 82 subjects with normal blood pressure and 24 patients with hypertension had significant narrowing (patients with CAD). The postexercise SBP response was defined on the basis of the SBP ratio (i.e., the SBP at a minutes of recovery divided by that at peak exercise). An SBP ratio that exceeded 0.90 (cutoff point for discriminating control subjects from patients with CAD) was considered to be an abnormal SBP response. In the subjects with normal blood pressure, the abnormal SBP response identified CAD as accurately as did ST-segment depression. In the patients with hypertension, the diagnostic accuracy was increased significantly by combining the abnormal SBP response and ST-segment depression (p< 0.01). The SBP ratio increased with the number of diseased coronary arteries. Ten of the 14 patients with a narrowing of the left main coronary artery had an SBP ratio higher than 1.00. The postexercise SBP response may be useful for detecting CAD in patients with and without hypertension and for evaluating the severity of CAD.

Co-Ce-O films were synthesized by a newly developed chemical method called the metal-oxide co-electroless deposition method. The Co content in the Co-Ce-O deposited films was subject to change from 16 to 100 at.%, depending on the concentration of the CoSO4 in the reaction solution. About 10 at.% boron, including in the Co-Ce-O films, therefore meant Co existed as an amorphous phase. Co-Ce-O film was synthesized with saturation magnetization of 0.57 T and minimum coercivity of 2.3 kA/m using this method. This result is the first step for obtaining metal-oxide nanogranular films with good magnetic properties at high frequency by chemical method.

We made a diagnosis of glucose-6-phosphate dehydrogenase (G6PD) deficiency with a new mutation of 848A→G (exon 8) in a 16-year-old male patient presenting with severe hemolysis. He was administered a diclofenac sodium suppository (50 mg) at the time of first visit to our hospital because of pyrexia. In the acute phase, pyrexia, severe general fatigue, lumbar back pain, hemoglobinuria, and jaundice developed. Laboratory blood examinations showed hemolysis, and remarkable increases in serum ferritin and cytosol leucine aminopeptidase levels. Serum interleukin-6 and interferon- levels were also increased. No liver injury was found. He had neonatal jaundice persisting over 3 weeks. He did not have a history of chronic hemolysis or hyperbilirubinemia. Increases in serum ferritin or cytosol leucine aminopeptidase levels in G6PD-deficient patients were not reported earlier. In this case, it is presumed that infection and administration of anti-inflammatory agents induce the hemolytic episode and that hypercytokinemia deteriorates the disease condition.

It is important to evaluate the haemodynamics of cerebral arteriovenous malformations (AVMs) in order to predict and avoid complications following surgical intervention. This study measured the transit time of each component of AVMs by high-speed digital subtraction angiography (DSA) to understand the haemodynamics and to evaluate which parameter is an appropriate indicator for the monitoring. Eleven cases of cerebral AVMs were studied. Haemodynamics were evaluated by analysis of tracer transit using DSA. Tracer transit parameters including transit time and peak time were calculated by application of gamma fitting to time density curve, and measured in the following compartments; feeding artery, nidus and draining vein. Significant correlations of tracer transit parameters were present among compartments. Although there was no significant correlation of nidus volume and tracer transit parameters, the peak time was shorter in haemorrhage cases than in non-haemorrhage cases. Measurements of tracer transit parameters by high-speed DSA may be a useful tool for characterising and monitoring the haemodynamics of AVMs following surgical intervention.