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Objective: Hepatic steatosis, the early stage of nonalcoholic fatty liver disease (NAFLD), currently lacks targeted pharmacological treatments. G protein-coupled receptors (GPCRs) in hepatocytes differentially regulate lipid metabolism depending on their coupling profile of G protein subtypes. Unlike G s , G i , and G q signaling, the role of G 12 signaling in hepatic steatosis remains elusive. The objective of this study was to investigate the effect of G 12 signaling on hepatic steatosis and obesity and its mechanisms., Methods: We generated mice expressing a G 12 -coupled designer GPCR in a liver-specific manner. We performed phenotypic analysis in the mice under the condition of fasting (acute hepatic steatosis model) or high-fat diet feeding (chronic hepatic steatosis model)., Results: In acute and chronic hepatic steatosis models, chemogenetic activation of hepatic G 12 signaling suppressed the progression of hepatic steatosis. The treatment led to an increased triglyceride secretion with little effect on mitochondrial respiratory activity, fatty acid oxidation, de novo lipogenesis, and fatty acid uptake. Furthermore, in a high-fat-diet-induced obesity model, activation of the G 12 -coupled designer GPCR exerted anti-obesity effects with increased whole-body energy expenditure and fat oxidation. Anti-FGF21 antibody treatment showed that the anti-obesity effects of the hepatic G 12 D activation relied in part on the hepatokine FGF21., Conclusions: Our findings indicate that the activation of G 12 signaling in the liver has the potential to prevent hepatic steatosis and obesity. This discovery provides a strong rationale for the development of drugs targeting G 12 -coupled GPCRs expressed in the liver., Competing Interests: Declaration of competing interest None., (Copyright © 2024. Published by Elsevier B.V.)

Animal trypanosomosis, caused by Trypanozoon trypanosomes (Trypanosoma evansi and T. equiperdum), and Trypanosoma vivax, is endemic to South American countries and has a negative impact on the livestock industry. However, the risk factors for trypanosomosis in Paraguay remain unknown. This study aimed to determine the risk factors for equine trypanosomosis in Paraguay based on a PCR-based molecular survey and individual horse sampling data. In this study, 739 blood samples were collected from horses in 16 departments of Paraguay between August 2019 and November 2020. To elucidate the risk factors for trypanosome infection, the relationship between trypanosome infection status detected by PCR and the location, sex, age, breed of horses, and season of sample collection was analyzed. There were no significant differences in trypanosome prevalence in horses between the eastern and western regions, ages, or breeds of horses in Paraguay. Sex and season were identified as risk factors for trypanosome infection in horses in Paraguay in the current study. These results suggest that the rainy-summer season, when vectors increase in number and their blood-sucking activity, could be the most important risk factor for trypanosome infection in Paraguay horses. Preventive measures and treatments should be developed to address these factors., (Copyright © 2022 Elsevier B.V. All rights reserved.)

We report here a liquid chromatography/electrospray ionization-tandem mass spectrometry assay for the quantification of three isoforms of antimicrobial peptide (AMP), chrysophsin-1, -2 and -3, in the red sea bream, Pagrus (Chrysophrys) major. Chrysophsin-1 was mainly distributed in the pyloric caeca and gills, followed by intestine and stomach. Chrysophsin-2 was detected in the gills and stomach, but chrysophsin-3 was only in the gills. The present procedure is valuable as a general method for simultaneous determination of the level of multiple AMP isoforms in fish tissues, and the data give important information for understanding the significance of each AMP isoform in host defense., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Although fractional flow reserve (FFR) measurements during coronary angiography are performed in routine clinical practice, few studies have evaluated FFR measurements in dialysis patients., Methods: We retrospectively studied 42 hemodialysis patients with suspected or known coronary artery disease (CAD) who underwent stress myocardial perfusion imaging and coronary angiography with FFR measurements for 61 coronary lesions. The cut-off value for FFR to detect myocardial ischemia was determined by receiver operating characteristic (ROC) curve analysis., Results: There were 61 coronary vessels measured by FFR. The FFR range was 0.34-0.93 with a mean of 0.74±0.13. The ROC curve analysis revealed that the best cut-off value of FFR for detecting myocardial ischemia was 0.76 (p<0.0001), with 70% sensitivity, 86% specificity, and 76% accuracy for myocardial ischemia. Compared with patients who had positive myocardial ischemia and an FFR≤0.76, those who had negative myocardial ischemia despite an FFR≤0.76 had less left ventricular (LV) mass index, whereas patients who had positive myocardial ischemia despite an FFR>0.76 had greater LV mass indexor serum calcium-phosphorus product., Conclusions: The cut-off value of FFR for myocardial ischemia in chronic hemodialysis patients is similar to that in other CAD patients. However, caution is necessary when FFR measurements are applied to dialysis patients with significantly increased LV mass index or serum calcium-phosphorus product., (Copyright © 2017 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.)

β-Lactam antibiotics including penicillin G, nafcillin, cefazolin, cefotaxime, and oxacilline in water were rapidly removed and degraded by using didodecyldimethylammonium bromide (DDAB)-montmorillonite (MT) organoclay. Removal of antibiotics increased with increasing the amount of organoclay added and the amount of DDAB sorbed on MT. Extents of organoclay sorption of antibiotics were represented by the binding constants to DDAB molecules and correlated to the aqueous-octanol distribution coefficients. The degradation rate of β-lactam antibiotics was found to significantly increase by the organoclay sorption. Even under the mild conditions (25°C and pH 7), penicillin G (m/z=335) nearly completely (>98%) degraded into penicilloic acid (m/z=353) missing β-lactam ring within 2h. The first-order reaction rate of the primary degradation increased with increasing in temperature. The activation energy estimated from the Arrhenius plot was 49kJmol(-1) and lower than the value (83.5kJmol(-1)) in water, strongly suggesting catalytic activity of DDAB-MT organoclay. The applicability to wastewater treatment was demonstrated by using secondary effluents of municipal sewage treatment plants and synthesized hospital wastewaters., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The detailed epidemiology of meningococcal diseases in Japan has yet to be determined and, moreover, the healthy carriage rate is also unknown. In this study, to obtain insight into the carriage rate of Neisseria meningitidis in healthy individuals in Japan, we developed a new method to detect the N. meningitidis-specific ctrB gene, one of the genes encoding enzymes for capsule synthesis, by Loop-Mediated Isothermal Amplification (LAMP) and examined the meningococcal carriage rate by using self-collected oral throat wash specimens from 836 students at a university. Examination by LAMP showed that 7 out of 836 samples were positive for N. meningitidis DNA, and the results were also verified by the nested PCR method for the meningococcus specific ggt gene. The N. meningitidis carriage rate in healthy individuals was estimated to be 0.84%. Moreover, we further confirmed by the nested-PCR-based serogroup typing method that 5 of the positive samples belonged to serogroup Y, 1 belonged to group B and 1 was unidentifiable. Considering the epidemiology for meningococcal diseases in Japan, the carriage rate and the serogroup profile seem to be consistent with low incidence of meningococcal diseases and serogroup distribution of clinical meningococcal isolates in Japan, respectively., (Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Background: In Japan, intravenous digoxin was previously recommended as a standard drug for acute control of the heart rate in atrial fibrillation (AF)/atrial flutter (AFL) patients with chronic heart failure (CHF). Treatment alternatives for such cases were limited and new drugs for this purpose are needed. In November 2013, landiolol hydrochloride (Onoact(®) 50 for Injection, Ono Pharamaceutical, Osaka, Japan) was approved with the indication for "tachyarrhythmia (AF/AFL) in patients with cardiac dysfunction." However, clinical experience with this condition is still insufficient. Therefore, it is important to conduct a surveillance of landiolol under actual clinical settings. In addition, collecting data on the mid- and long-term outcomes in patients treated with landiolol which have not been collected in clinical trials are indispensable., Methods: This prospective survey will involve patients treated with landiolol for the treatment of tachyarrhythmia (AF/AFL) in cardiac dysfunction at Japanese medical facilities from June 2014 to May 2017. The planned number of patients for analysis is approximately 500. The evaluations will be made not only to identify the adverse events and clinical effectiveness of the drug, but also to characterize the mid- and long-term outcomes of patients receiving and switching to oral-β-blockers after the discontinuation of landiolol., Results: This study was started in June 2014 (registration period 2 years, survey period 3 years) and will end in May 2017., Conclusions: This survey will clarify both the characteristics and mid- and long-term outcome of using landiolol to treat AF/AFL patients with cardiac dysfunction in clinical practice. Moreover, this survey will simultaneously provide important data that will reveal the possible gap between clinical trials and clinical practice in these patients., (Copyright © 2014 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.)

Objective: We have performed laparoscopic adrenalectomy including retroperitoneoscopic adrenalectomy via a single large port (RASLP) and conventional laparoscopic adrenalectomy (CLA) for adrenal tumors since 1992, and report our experience to date., Methods: The study population consisted of 134 patients who underwent laparoscopic adrenalectomy from 1992 to 2012. Fifty-eight patients (18 aldosterone-producing adenomas, 13 adenomas with Cushing's syndrome, 1 adenoma with preclinical Cushing's syndrome, and 26 nonfunctioning tumors) were treated using RASLP, and 76 patients (33 aldosterone-producing adenomas, 17 adenomas with Cushing's syndrome, 6 adenomas with preclinical Cushing's syndrome, 17 pheochromocytomas, and 3 nonfunctioning tumors) were treated using CLA. Complications were graded according to the modified Clavien system., Results: The majority of RASLPs were performed during the 1990s, whereas all patients underwent CLA after 2000. The mean operation times (166 vs. 205 minutes, p < 0.01) and intraoperative estimated blood loss (85 vs. 247 mL, p < 0.01) were significantly lower in the CLA group. Conversion to open surgery was required in three patients (5%) in the RASLP group and five patients (7%) in the CLA group (p = 0.73). Postoperative complications were grade 1 in three patients and grades 4 and 5 in one patient each in the RASLP group, whereas grade 2 in one patient was observed in the CLA group (p = 0.085)., Conclusion: Although this study included biases such as different eras and indications, CLA resulted in decreased operative times, blood loss, and postoperative complications compared with RASLP. CLA has so far become our preferred procedure for patients with adrenal tumor in our experience., (Copyright © 2014. Published by Elsevier Taiwan.)

It is essential to know the detailed structure of the thin filament to understand the regulation mechanism of striated muscle contraction. Fluorescence resonance energy transfer (FRET) was used to construct an atomic model of the actin-tropomyosin (Tm)-troponin (Tn) core domain complex. We generated single-cysteine mutants in the 167-195 region of Tm and in TnC, TnI, and the β-TnT 25-kDa fragment, and each was attached with an energy donor probe. An energy acceptor probe was located at actin Gln41, actin Cys374, or the actin nucleotide-binding site. From these donor-acceptor pairs, FRET efficiencies were determined with and without Ca(2+). Using the atomic coordinates for F-actin, Tm, and the Tn core domain, we searched all possible arrangements for Tm or the Tn core domain on F-actin to calculate the FRET efficiency for each donor-acceptor pair in each arrangement. By minimizing the squared sum of deviations for the calculated FRET efficiencies from the observed FRET efficiencies, we determined the location of Tm segment 167-195 and the Tn core domain on F-actin with and without Ca(2+). The bulk of the Tn core domain is located near actin subdomains 3 and 4. The central helix of TnC is nearly perpendicular to the F-actin axis, directing the N-terminal domain of TnC toward the actin outer domain. The C-terminal region in the I-T arm forms a four-helix-bundle structure with the Tm 175-185 region. After Ca(2+) release, the Tn core domain moves toward the actin outer domain and closer to the center of the F-actin axis., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Tissue infiltration of activated lymphocytes is a hallmark of transplant rejection and organ-specific autoimmune diseases. Migration and activation of lymphocytes depend on DOCK2, an atypical Rac activator predominantly expressed in hematopoietic cells. Although DOCK2 does not contain Dbl homology domain typically found in guanine nucleotide exchange factors, DOCK2 mediates the GTP-GDP exchange reaction for Rac through its DHR-2 domain. Here, we have identified 4-[3'-(2″-chlorophenyl)-2'-propen-1'-ylidene]-1-phenyl-3,5-pyrazolidinedione (CPYPP) as a small-molecule inhibitor of DOCK2. CPYPP bound to DOCK2 DHR-2 domain in a reversible manner and inhibited its catalytic activity in vitro. When lymphocytes were treated with CPYPP, both chemokine receptor- and antigen receptor-mediated Rac activation were blocked, resulting in marked reduction of chemotactic response and T cell activation. These results provide a rational of and a chemical scaffold for development of the DOCK2-targeting immunosuppressant., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Fluorescence resonance energy transfer (FRET) was used to construct an atomic model of the actin-tropomyosin (Tm) complex on a reconstituted thin filament. We generated five single-cysteine mutants in the 146-174 region of rabbit skeletal muscle α-Tm. An energy donor probe was attached to a single-cysteine Tm residue, while an energy acceptor probe was located in actin Gln41, actin Cys374, or the actin nucleotide binding site. From these donor-acceptor pairs, FRET efficiencies were determined with and without Ca(2+). Using the atomic coordinates for F-actin and Tm, we searched all possible arrangements for Tm segment 146-174 on F-actin to calculate the FRET efficiency for each donor-acceptor pair in each arrangement. By minimizing the squared sum of deviations for the calculated FRET efficiencies from the observed FRET efficiencies, we determined the location of the Tm segment on the F-actin filament. Furthermore, we generated a set of five single-cysteine mutants in each of the four Tm regions 41-69, 83-111, 216-244, and 252-279. Using the same procedures, we determined each segment's location on the F-actin filament. In the best-fit model, Tm runs along actin residues 217-236, which were reported to compose the Tm binding site. Electrostatic, hydrogen-bonding, and hydrophobic interactions are involved in actin and Tm binding. The C-terminal region of Tm was observed to contact actin more closely than did the N-terminal region. Tm contacts more residues on actin without Ca(2+) than with it. Ca(2+)-induced changes on the actin-Tm contact surface strongly affect the F-actin structure, which is important for muscle regulation., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The lack of the Hes1 gene leads to the failure of cranial neurulation due to the premature onset of neural differentiation. Hes1 homozygous null mutant mice displayed a neural tube closure defect, and exencephaly was induced at the mid/hindbrain boundary. In the mutant mesencephalon, the roof plate was not formed and therefore the ventricular zone showing cell proliferation was displaced to the brain surface. Furthermore, the telencephalon and ventral diencephalon were defective. Despite the severe defects of neurogenesis in null mutants, the mesencephalic dopaminergic (mesDA) neurons were specified at the midline of the ventral mesencephalon in close proximity to two important signal centers - floor plate and mid/hindbrain boundary (i.e., the isthmic organizer). Using mesDA neuronal markers, tyrosine hydroxylase (TH) and Pitx3, the development of mesDA neurons was studied in Hes1 null mice and compared with that in the wild type. At early stages, between embryonic day (E) 11.5 and E12.5, mesDA neurons were more numerous in null mutants than in the wild type. From E13.5 onward, however, the cell number and fiber density of mesDA neurons were decreased in the mutants. Their distribution pattern was also different from that of the wild type. In particular, mesDA neurons grew dorsally and invaded the rostral hindbrain. 5-HT neurons were also ectopically located in the mutant midbrain. Thus, the loss of Hes1 resulted in disturbances in the inductive and repulsive activities of the isthmic organizer. It is proposed that Hes1 plays a role in regulating the location and density of mesDA neurons., (Copyright © 2011 Elsevier Inc. All rights reserved.)

A chitosan-conjugated thermo-responsive polymer containing 15% chitosan, PNIPAAm-15CS, was used for the removal of different phenols in water. The polymer was synthesized by a 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide-mediated condensation of poly(N-isopropylacrylamide-co-acrylic acid) and chitosan in the aqueous solution (pH 6). At 30 °C, phenol, 4-methylphenol, 4-methoxyphenol, and 4-chlorophenol were converted to dark brown oxidized compounds by the tyrosinase-induced enzymatic reaction and subsequently bound to the amino moiety of PNIPAAm-15CS. In the presence of 1.0 g L(-1) PNIPAAm-15 CS and 50 k UL(-1) tyrosinase, phenols (20 mg L(-1)) decreased to undetectable levels (<0.01 mg L(-1)) within 2h. By the vigorous mixing of the solution at 40 °C, the polymer deposited and became a small coagulate that can be easily taken up from water. Accompanying the polymer deposition, the oxidized compounds were completely (>98%) removed. The proposed method was successfully applied to the removal of phenols from wastewaters., (Copyright © 2010 Elsevier B.V. All rights reserved.)

We developed a simple and easy method for solution mixing based on the heat-induced regulation of capillary action in thermo-responsive polymer-coated microchannels. The channels having two T-junctions were fabricated on a glass plate by a sand-blast technique and then coated with a poly(N-isopropylacrylamide) film. The polymer-coating was performed by the modification with allyltrimethoxysilane and the subsequent radical polymerization of N-isopropylacrylamide and N,N'-methylenebisacrylamide. When the channel was warmed by a Peltier device, a capillarity-based solution flow completely stopped because of the water-repellency of channel surfaces. On the other hand, the cooling of the channels allowed the restart of the solution flow through hydrophilic channels. Solution mixing downstream a T-junction was readily conducted by a Peltier device that had placed at the junction. The technique was applied to the fluorometric analysis of polyamines in saliva. The saliva sample was mixed with nickel(II) chloride solution at the first junction to mask amino acids and then mixed with o-phthalaldehyde solution at the second junction to form the fluorometric derivatives of polyamines. Blue fluorescence was observed downstream the second junction. Linear correlation was obtained between the emission intensity and the spermine concentration in the range of 20-100 microM. No mechanical pump or valve was required for the fluid manipulation.

A simple and rapid method for the highly sensitive determination of polycyclic aromatic hydrocarbons (PAHs) in water was developed. Benzo[a]pyrene, benzo[k]fluoranthene, perylene, and pyrene in water were concentrated into sodium dodecyl sulfate (SDS)-alumina admicelles. The collection was performed by adding SDS and alumina particles into the sample solution at pH 2. After gentle mixing, the resulting suspension was passed through a membrane filter to collect the SDS admicelles containing highly concentrated PAHs. The filter was placed on a slide glass and then covered admicellar layer with a fused silica glass plate before setting in a fluorescence spectrometer. Benzo[a]pyrene, benzo[k]fluoranthene, perylene, and pyrene were selectively determined by the synchronous fluorescence scan (SFS) analysis with keeping wavelength intervals between excitation and emission to 98, 35, 29, and 45 nm, respectively. Because of the minimum spectral overlapping, 1-40 ng l(-1) of benzo[a]pyrene, benzo[k]fluoranthene, and perylene as well as 10-150 ng l(-1) of pyrene were selectively determined with eliminating the interferences of other 12 PAHs. The detection limits were 0.3 ng l(-1) for benzo[a]pyrene, benzo[k]fluoranthene, and perylene, and 1 ng l(-1) for pyrene. They were 2-3 orders of magnitude lower than the detection limits in normal aqueous micellar solutions. The application to water analysis was studied.

Apoptosis in Xenopus egg extracts is carried out by maternally stockpiled materials, but the contributions of endogenous apoptosis regulators are still poorly characterized. Here we examined the physiological role of Xenopus Bid (xBid), a pro-apoptotic BH3-only member of Bcl-2 family proteins. We found that endogenous xBid was a physiological accelerator of apoptosis in egg extracts. Interestingly, xBid was mono-/diubiquitylated but not degraded by proteasome in egg extracts, and we identified three ubiquitylated Lys residues in the N-terminal propeptide region. Comparison with human Bid suggested that mono-/diubiquitylation is a specific feature of xBid.

Spontaneous baroreflex function can be altered by acute changes in central blood volume. Both a reduction in spontaneous baroreflex function at central hypovolemia and augmentation at hypervolemia suggest a dose-effect relationship between central blood volume and spontaneous baroreflex function. However, this relationship has not been quantified over stepwise widespread changes in central blood volume. Twelve individuals underwent central hypovolemia at two levels of lower body negative pressure (LBNP) (-15 mm Hg, LBNP15; -30 mm Hg, LBNP30) and hypervolemia with two discrete infusions of normal saline (NS) (15 ml kg(-1), NS15; total 30 ml kg(-1), NS30). Spontaneous baroreflex function was assessed using transfer function analysis and the sequence method between blood pressure and R-R interval. Both central venous pressure (-0.6-7.9 mm Hg) and left ventricular end-diastolic volume (72.4-133.1 ml) decreased during LBNP and increased after saline infusion. Both spontaneous baroreflex indices of high-frequency transfer function gain (LBNP30, 17.4+/-3.2; LBNP15, 22.3+/-3.8; baseline, 25.6+/-4.1; NS15, 28.5+/-4.2 ms mm Hg(-1), ANOVA P=0.001) and of the sequence slope (LBNP30, 14.4+/-2.2; LBNP15, 17.2+/-2.5; baseline, 20.5+/-2.8; NS15, 24.5+/-3.1 ms mm Hg(-1), ANOVA P=0.001) increased stepwise from hypovolemia of LBNP30 to hypervolemia of NS15. However, these indices were lower at NS30 (high-frequency transfer function gain, 22.0+/-2.2 ms mm Hg(-1), post-hoc P=0.071; sequence slope, 17.7+/-1.7 ms mm Hg(-1), post-hoc P<0.05) than NS15 during hypervolemia. These results indicated that the relationship between central blood volume and spontaneous baroreflex function is apparently bell-shaped, with maximal augmentation at moderate hypervolemia.

Esterified drugs such as imidapril, derapril, and oxybutynin hydrolyzed by carboxylesterase 1 (CES1) are extensively used in clinical practice. The kinetics using the CES1 substrates have not fully clarified, especially concerning species and tissue differences. In the present study, we performed the kinetic analyses in humans and rats in order to clarify these differences. The imidaprilat formation from imidapril exhibited sigmoidal kinetics in human liver microsomes (HLM) and cytosol (HLC) but Michaelis-Menten kinetics in rat liver microsomes and cytosol. The 2-cyclohexyl-2-phenylglycolic acid (CPGA) formation from oxybutynin were not detected in enzyme sources from rats, although HLM showed high activity. The kinetics were clarified to be different among species, tissues, and preparations. In individual HLM and HLC, there was large interindividual variability in imidaprilat (31- and 24-fold) and CPGA formations (15- and 9-fold). Imidaprilat formations exhibited Michaelis-Menten kinetics in HLM and HLC with high activity but sigmoidal kinetics in those with low activity. CPGA formations showed sigmoidal kinetics in high activity HLM but Michaelis-Menten kinetics in HLM with low activity. We revealed that the kinetics were different between individuals. These results could be useful for understanding interindividual variability and for the development of oral prodrugs.

Objectives: Cardiac involvement is an important prognostic factor in patients with sarcoidosis. We evaluated the usefulness of delayed enhancement MRI (DE-MRI) for diagnosing cardiac sarcoidosis by comparing with nuclear imaging and studying the correlation between DE area and left ventricular (LV) function., Methods: Twelve patients (male:female 3:9) diagnosed as having sarcoidosis underwent Gd-MRI, myocardial perfusion SPECT (Tl-201, Tc-99m sestamibi), Ga-67 scintigraphy, and/or F-18 FDG-PET., Results: DE was observed in 5 patients, and was positive in 39 (39%) of 100 LV segments. The corresponding perfusion defects in myocardial perfusion SPECT were undetectable in 14 (36%) segments. DE distributed mainly in mid- to epi-myocardium, and the lack of perfusion defects in myocardial perfusion SPECT was more prominent in less transmural DE segments. Two patients with diffuse DE and 1 case with focal DE exhibited positive cardiac uptake in Ga-67 scintigraphy, and 2 other cases with focal DE showed cardiac uptake in F-18 FDG-PET. In 7 patients without DE, there were no significant findings in nuclear imaging. Both LV end-diastolic and end-systolic volume were positively and LV ejection fraction was negatively correlated with the extent of DE area. Four patients treated with corticosteroid showed improvement in nuclear imaging and slight decreases in DE area but no recovery in LV function., Conclusions: DE-MRI is useful to diagnose the cardiac involvement of sarcoidosis and to evaluate cardiac function. It is likely that the distribution of DE in mid- to epi-myocardium is the characteristic of cardiac sarcoidosis, and the larger DE area may be correlated with poor LV function.

The nuclear factor-kappaB (NF-kappaB) transcription factors play important roles in cancer development by preventing apoptosis and facilitating the tumor cell growth. However, the precise mechanisms by which NF-kappaB is constitutively activated in specific cancer cells remain largely unknown. In our current study, we now report that NF-kappaB-inducing kinase (NIK) is overexpressed at the pretranslational level in adult T-cell leukemia (ATL) and Hodgkin Reed-Sternberg cells (H-RS) that do not express viral regulatory proteins. The overexpression of NIK causes cell transformation in rat fibroblasts, which is abolished by a super-repressor form of IkappaBalpha. Notably, depletion of NIK in ATL cells by RNA interference reduces the DNA-binding activity of NF-kappaB and NF-kappaB-dependent transcriptional activity, and efficiently suppresses tumor growth in NOD/SCID/gammac(null) mice. These results indicate that the deregulated expression of NIK plays a critical role in constitutive NF-kappaB activation in ATL and H-RS cells, and suggest also that NIK is an attractive molecular target for cancer therapy.

Transcription factors known as CCAAT enhancer binding proteins (C/EBPs) are involved in hematopoietic differentiation, including myelopoiesis and granulopoiesis. C/EBPbeta-deficient mice develop normally; however, they exhibit defective macrophage function, resulting in increased susceptibility to infection. Little is known about the role of C/EBPbeta in granulopoiesis; therefore, we examined granulopoiesis in C/EBPbeta-deficient mice. Morphology, the number of peripheral blood and bone marrow cells, and the expression of genes specific for the myeloid lineage were normal in C/EBPbeta-deficient mice. Interestingly, the hematopoietic progenitor cells of C/EBPbeta-deficient mice did not respond normally to granulocyte/macrophage-colony stimulating factor and granulocyte colony stimulating factor. In addition, C/EBPbeta-deficient neutrophils displayed enhanced apoptosis compared with wild-type neutrophils. Our present results indicate that C/EBPbeta helps regulate survival of neutrophils, downstream of the granulocyte colony stimulating factor receptor.

Mycobacterium avium-intracellulare (MAI) is, among acid-fast bacilli, the most common cause of nontuberculous pulmonary diseases, and MAI infections are often treated according to the guidelines of the American Thoracic Society. However, despite the use of multiple drugs, patients sometimes do not recover with the initial round of treatment. Other kinds of nontuberculous mycobacteria are sometimes found in patients' respiratory samples, even during such treatment for MAI pulmonary disease. We experienced three patients with pulmonary disease due to Mycobacterium abscessus (MA) in whom the disease was difficult to treat because of resistance to all the antituberculous agents used. MA infection had occurred in these patients after long-term treatment with multiple drugs for previous MAI pulmonary disease. We considered that the MA infection in these patients appeared to be the result of insufficient efficacy of the drugs used for MAI and insufficiently aggressive use of antituberculous agents. It thus appeared that MA infection was the result of microbial substitution, and that, if this is the case, the guidelines for the treatment of MAI may need to be modified to eliminate microbial substitution.

Chlorophenols including monochlorophenol, dichlorophenol, trichlorophenol, tetrachlorophenol, and pentachlorophenol in water were extracted into dialkylated cationic surfactant-silica gel admicelles. The dialkylated cationic surfactants such as didecyldimethylammonium bromide (DC10) and didodedyldimethylammonium bromide (DC12) sorbed on silica gel surfaces to form admicelles at pH 9. Approximately 200mg of DC10 was quantitatively sorbed on 1g of silica gel. The sorption further increased by further addition of DC10. This is in contrast to the fact that the maximum sorption of mono-alkylated cetyltrimethyammonium chloride (CTAC) was only ca. 100mg. Based on the fluorescent spectra of a molecular probe, N-phenyl-1-naphthylamine, DC10- and DC12-silica gel admicelles were more hydrophobic than CTAC-silica gel admicelles. The extents of the extraction of chlorophenols into DC10-silica gel admicelles were greater than those into CTAC-silica gel admicelles. However, the extractions to DC12-silica gel admicelles were insufficient due to leakage of DC12 vesicles. Consequently, DC10-silica gel admicelles were the most adequate for concentrating chlorophenols in water. An admicelle column was prepared by passing aqueous buffer solution of DC10 through a Bond Elut Jr. silica gel solid-phase extraction cartridge. It was successfully applied to the 500-fold concentration of chlorophenols including hydrophilic mono-substituted chlorophenol in water samples prior to their HPLC analysis.

The main objective of this study was to investigate the biological function of beta amyloid precursor protein (APP), in particular its nerve growth factor-like activity. We hypothesize that the extracellular domain containing the sequence RERMS, amino acids 328-332 of APP(695), represents the active site for this function. Binding assays using peptide fragments of this domain have demonstrated specific and saturable binding to the cell surface with affinity in the low nanomolar range. This induced our quest for an APP-specific receptor. We chose different peptide fragments of the RERMS domain as ligands and displacing agents on affinity columns to purify APP-binding molecules. Amino acid microsequencing yielded partial sequences of serum albumin, actin, two novel proteins of 41 and 63kDa, and human Collapsin Response Mediator Protein-2 (hCRMP-2). Because both APP and hCRMP-2 promote neuronal outgrowth and use a common signaling pathway, APP could be acting through a semaphorin receptor as well.

An extraction method based on the thermo-responsive precipitation of a water-soluble polymer, poly(N-isopropylacrylamide) [PNIPAAm], was applied to the concentration of dimedone (5,5-dimethylcycrohexane-1,3-dion) derivatives for the highly sensitive determination of aldehydes in air. Aliphatic aldehydes including formaldehyde, acetaldehyde, propanal, 1-butanal, 1-heptanal, and 1-hexanal in air were well solubilized into the aqueous solution of dimedone and ammonium acetate by mixing the solution and air sample in a polyvinyl fluoride bag. Fluorescent derivatives of aldehydes that had formed by the Hantzsch reaction with dimedone were concentrated by polymer-mediated extraction. The recoveries of the fluorescent compounds increased with increasing the carbon number of aldehyde and were more than 80% for the derivatives from aldehydes having more than three carbon atoms under the optimal conditions. Microgram per m3 (sub-ppb) levels of the aliphatic aldehydes, propanal, 1-butanal, 1-heptanal, and 1-hexanal, in ambient air were successfully determined by HPLC separation with fluorometric detection. The sampling volume and time required were only 1l and 20 s, respectively.

The solid phase extraction of gold(III), platinum(II), and palladium(II) to surfactant-impregnated polystyrene-divinylbenzene porous resin (XAD-4) was studied. The extracting media could be prepared just by mixing the resin in aqueous surfactant solutions. XAD-4 impregnated with a nonionic surfactant, polyethylene glycol monooleyl ether, was useful for extracting gold(III) from hydrochloric acid. The extractions of platinum(II) and palladium(II) were improved in the use of XAD-4 impregnated with a nitrogen-containing nonionic surfactant, polyethylene glycol stearyl amine. On the other hand, base metals such as copper(II), cobalt(II), nickel(II) and zinc(II), were hardly extracted.

Most traumatic carotid artery aneurysms occur at or close to its bifurcation, and traumatic aneurysm of the intrathoracic carotid arteries are rare. We describe a case of false aneurysm at the origin of the left common carotid artery (LCCA) after blunt trauma. A 53-year-old man suffered a blow from a broken steel plate, which flew from a working concrete crusher over his neck when he looked down the machine. Chest computed tomography revealed aneurysm of the LCCA, and aortic arch arteriography demonstrated a false aneurysm of about 3 x 5 cm at the origin of the LCCA, with loss of arterial continuity and abnormal tortuosity above the aneurysm. An ascending aorta to LCCA bypass graft was placed during the cooling period of cardiopulmonary bypass, and mattress sutures were placed in the normal aorta to close the origin of the LCCA under hypothermic circulatory arrest because of the extreme danger of dissection. The LCCA was transected partially at its origin from the aorta. We speculated that the direct lifting force which caused the carotid artery to move upward might produce a tear at the junction of the LCCA and the aortic arch.

Polyaromatic hydrocarbons (PAHs) in water were concentrated into sodium dodecyl sulfate (SDS)-gamma-alumina and di-2-ethylhexyl sodium sulfosuccinate (Aerosol-OT, AOT)-gamma-alumina admicelles. The comparison of the binding constants (Kad[={adsorbed concentration of the solute (mol/g surfactant)}/{the concentration in the bulk aqueous phase (mol/ml)}] indicated almost the same extraction abilities of the both admicelles. However, better and more reproducible recovery was obtained in the concentration of PAHs into the SDS-gamma-alumina admicelle. PAHs in tobacco smoke that were trapped in water were successfully concentrated into SDS-gamma-alumina admicelle for the HPLC analysis.

Transferrin receptor 2 (TfR2) is a membrane glycoprotein that mediates cellular iron uptake from holotransferrin. Homozygous mutations of this gene cause one form of hereditary hemochromatosis in humans. We recently reported that homozygous TfR2(Y245X) mutant mice, which correspond to the TfR2(Y250X) mutation in humans, showed a phenotype similar to hereditary hemochromatosis. In this study, we further analyzed the phenotype as well as iron-related gene expression in these mice by comparing the TfR2-mutant and wild-type siblings. Northern blot analyses showed that the levels of expression of hepcidin mRNA in the liver were generally lower, whereas those of duodenal DMT1, the main transporter for uptake of dietary iron, were higher in the TfR2-mutant mice as compared to the wild-type siblings. Expression of hepcidin mRNA in the TfR2 mutant mice remained low even after intraperitoneal iron loading. In isolated hepatocytes from both wild-type and TfR2 mutant mice, interleukin-6 and lipopolysaccharide each induced expression of hepcidin mRNA. These results suggest that up-regulation of hepcidin expression by inflammatory stimuli is independent of TfR2 and that TfR2 is upstream of hepcidin in the regulatory pathway of body iron homeostasis.

A novel medium for protein separation, namely affinity admicelle, was prepared by mixing of octadecylsilyl (ODS) silica gels, a polyoxyethylene-type nonionic surfactant (Triton X-100), and a surfactant-conjugated substrate (affinity ligand) in an aqueous solution. The ligand was synthesized by mixing a triazine dye (Cibacron Blue 3GA, CB) and a polyethylene glycol monooleyl ether (C18EO7, C18EO10, or C18EO20) having different length of polyoxyethylene moiety in weakly alkaline solutions. The amount of Triton X-100 sorbed on 1 g of ODS silica was 0.2 mmol. Affinity ligands having highly hydrophobic oleyl group were predominantly sorbed on ODS silica. The losses of Triton X-100 and affinity ligand were within 0.3% and negligible by washing the admicelles were with a 25-fold volume of 1 mM Tris-HCl solution (pH 7.4). The coating ODS silica with Triton X-100 was effective to prevent the irreversible sorption of albumin (bovine, serum). An NADH-dependent enzyme, alcohol dehydrogenase (ADH, yeast), was successfully collected on the admicelles involving CB-conjugated ligands (CB-C18EO20). The maximum collection of ADH to 90 mg/ml of affinity admicelles was 68+/-4%. However, CB-C18EO7 and CB-C18EO10 having shorter polyoxyethylene unit were not available, suggesting the requirement of the spacer moiety in the affinity ligand. The recovery and purification factor based on the ratio of activity (unit)/protein (mg) from Whatman DE52-treated yeast extract was 27% and 12, respectively.

A novel admicelle composing of a dialkylated anionic surfactant, di-2-ethylhexyl sodium sulfosuccinate (Aerosol-OT, AOT) and gamma-alumina was prepared by mixing them in acidic aqueous solution. The amount of the maximum sorption of AOT on 1 g of alumina at pH 2 was ca. 130 mg. By comparing the fluorescence spectra of N-phenyl-1-naphthylamine in different solvents, the solvent property of AOT-gamma-alumina admicelles was corresponding to that of toluene or diethyl ether. Thus, the AOT-gamma-alumina admicelles had greater hydrophobicity than SDS-gamma-alumina admicelles having similar hydrophobicity to 1-octanol or ethyl acetate. Hydrophobic organic compounds, chlorophenols having more than three chloro substituents, octylphenol, nonylphenol, dibutyl phthalate was almost quantitatively (98% or more) collected onto AOT admicelles composing of 1.5 g gamma-alumina and 150 mg AOT. The greater collection yields rather than those in SDS-admicellar system were ascribable to greater hydrophobicity and stability of AOT admicelles. After the 500-fold concentration, traces (nM) of organic contaminants in water samples were successfully detected with an HPLC having a photometric detector.

Through mixing of porous polystyrene particles (Amberlite XAD-4), non-ionic surfactants, and surfactant-conjugated substrates (affinity ligand) in an aqueous solution led to the formation of a novel medium (affinity admicelle) for protein separation. The ligand (CB-Triton) was synthesized by mixing a triazine dye (Cibacron Blue 3GA (CB)) and a polyoxyethylene-type non-ionic surfactant (Triton X-100) in weakly alkaline solutions. Triton X-100 and CB-Triton were competitively sorbed onto XAD-4. Albumin (bovine serum), alcohol dehydrogenase (yeast), and lysozyme (chicken egg) having specific interaction to CB were collected onto the affinity admicelle. On the other hand, the collection of ovalubmin (chicken egg white), having no binding ability to CB, was negligibly small. Lysozyme in 100 microl of chicken egg white, diluted with 900 microl of 10 mM Tris-HCl (pH 7.4), was successfully collected on 18 mg of CB-Triton admicelles and, then, it was eluted with 1 ml of aqueous solution of 100 mM phosphate (pH 7.4). The recovery based on the activity for the lysis of micrococcus and the concentration factor were 60% and 40 (n = 3), respectively.

The primary objective was to determine the effect of supplemental progesterone, administered via an intravaginal device (CIDR), on conception rates to timed-artificial insemination (timed-AI) in postpartum suckled Japanese Black beef cows treated with the Ovsynch protocol. A secondary objective was to compare the effects of treatments on plasma concentrations of progesterone and estradiol. Cows in the control group (Ovsynch, n=38) received a standard Ovsynch protocol (100 microg GnRH analogue on Day 0, 500 microg PGF2alpha analogue on Day 7, and 100 microg GnRH analogue on Day 9), with AI on Day 10, approximately 20 h after the second GnRH treatment. Cows in the treatment group (Ovsynch+CIDR; n=40) received a standard Ovsynch protocol plus a CIDR for 7 days (starting on Day 0). Plasma progesterone concentrations were determined on Days 0, 1, 7, 9, 10, and 17 and plasma estradiol-17beta concentrations were determined on Days 7, 9, 10, and 17. The odds ratio for likelihood of conception was 3.29 times greater (P=0.02) in the Ovsynch+CIDR group compared to Ovsynch group. The conception rate was greater (P=0.03) in the Ovsynch+CIDR group than in the Ovsynch group (72.5% versus 47.7%). Insertion of a CIDR device significantly increased plasma progesterone concentrations only on Days 1 and 7 (P<0.001 and P=0.05, respectively), but had no significant effect on plasma estradiol-17beta concentrations. Including a CIDR with the Ovsynch protocol significantly improved conception rates in postpartum suckled Japanese Black beef cows.

Objective: To investigate the relationship between the efficacy of carbamazepine (CBZ) and the presence of the trigger zone for diagnosis of trigeminal neuralgia (TN). Study design CBZ was administered to 61 patients with suspected TN. All patients underwent intracranial examination by magnetic resonance imaging or computed tomography. The final diagnosis was established by oral and maxillofacial surgeon and neurosurgeon., Results: Of the 61 patients, 50 were finally diagnosed as having TN and 6 as having atypical facial pain. CBZ was effective for pain relief in 45 of the 50 TN patients (90%), and in 5 of the 11 patients (45%) with other diseases (P <.005). However, CBZ also relieved pain in some patients other than TN. Thirty of the 31 patients (97%) with a distinct trigger zone and 20 of the 30 (67%) without a trigger zone were diagnosed as having TN (P<0.005)., Conclusion: The efficacy of CBZ is an auxiliary indicator of TN and the presence of a distinct trigger zone is a strong indicator of TN.

Thermoresponsive polymers, poly(N-isopropylacrylamide) (PNIPAAm), having chelating functionalities were synthesized. PNIPAAm-imidazole (-Im) was precipitated and formed a gum-like aggregate in the neutral pH region at 50 degrees C, while PNIPAAm-carboxylic acid (COOH) and PNIPAAm-iminodiacetic acid (-IDA) remained soluble even at pH 7. An addition of a paired ion, dodecyltrimethylammonium ion, was effective for inducing the precipitation of those polymers. PNIPAAm-Im was useful for collecting copper(II), nickel(II), cobalt(II), and lead(II), but was ineffective for cadmium(II) recovery. In contrast, PNIPAAm-COOH collected cadmium(II), while insufficiently recovered cobalt(II) and nickel(II). PNIPAAm-IDA was the best choice for collecting all metal ions in neutral pH's. After 20-folds concentration, the metal ions in river and seawater were successfully determined by graphite furnace atomic absorption spectrometry (GFAAS).

The etiology of type 2 diabetes (DM) is polygenic. We investigated here genes and polymorphisms that associate with DM in the Japanese population. Single-nucleotide polymorphisms (SNPs) of 398 derived from 120 candidate genes were examined for association with DM in a population-based case-control study. The study group consisted of 148 cases and 227 controls recruited from Funagata, Japan. No evident subpopulation structure was detected for the tested population. The association tests were conducted with standard allele positivity tables (chi(2) tests) between SNP genotype frequency and case-control status. The independent association of the SNPs from serum triglyceride levels and body mass index was examined by multiple logistic regression analysis. A value of P<0.01 was accepted as statistically significant. Six genes (met proto-oncogene, ATP-binding cassette transporter A1, fatty acid binding protein 2, LDL receptor defect C complementing, aldolase B, and sulfonylurea receptor) were shown to be associated with DM.

Chlorophenols in water were sorbed onto sodium dodecylsulfate (SDS)-alumina (gamma-form) admicelles. The extent of sorption increased with increasing amount of SDS and decreasing solution pH. Conditions for good recovery were obtained when 100 mg SDS and 1.5 g alumina was used at pH 2. However, the yield decreased with a further increase in the SDS concentration due to the formation of normal SDS micelles. The extent of sorption also increased with increasing hydrophobicity of the chlorophenol, indicating that hydrophobic interactions predominate for the collection of analytes. When a cartridge column filled with admicelles was used, >90% of tetrachlorophenol and pentachlorophenol in 200 ml of water samples were rapidly recovered. The sorbed analytes were eluted with 1 ml acetonitrile. The accuracy and precision of the present method were demonstrated for the HPLC analysis with ultraviolet (290 nm) detection of microg l(-1) levels of tetrachlorophenol and pentachlorophenol in river water samples.