Determination of the positions of aluminum atoms introduced into SSZ-35 and the catalytic properties of the generated Brønsted acid sites.

The positions of aluminum (Al) atoms in SSZ-35 together with the characteristics of the generated protons were investigated by 27Al multiple quantum magic-angle spinning (MQ-MAS), 29Si MAS, and 1H MAS NMR data analyses accompanied by a variable temperature 1H MAS NMR analysis. The origin of the acidic -OH groups (Brønsted acid sites) generated by introducing Al atoms into the T sites was investigated and the T sites introduced into the Al atoms were revealed. To further determine the catalytic properties of the acidic protons generated in SSZ-35, the influence of the concentration of the Al atoms on the catalytic activity and selectivity during the transformation of toluene was examined.

Drug uptake pathways of multidrug transporter AcrB studied by molecular simulations and site-directed mutagenesis experiments.

Multidrug resistance has been a critical issue in current chemotherapy. In Escherichia coli , a major efflux pump responsible for the multidrug resistance contains a transporter AcrB. Crystallographic studies and mutational assays of AcrB provided much of structural and overall functional insights, which led to the functionally rotating mechanism. However, the drug uptake pathways are somewhat controversial because at least two possible pathways, the vestibule and the cleft paths, were suggested. Here, combining molecular simulations and site-directed mutagenesis experiments, we addressed the uptake mechanism finding that the drug uptake pathways can be significantly different depending on the properties of drugs. First, in the computational free energy analysis of drug movements along AcrB tunnels, we found a ligand-dependent drug uptake mechanism. With the same molecular sizes, drugs that are both strongly hydrophobic and lipophilic were preferentially taken in via the vestibule path, while other drugs favored the cleft path. Second, direct simulations realized totally about 3500 events of drug uptake by AcrB for a broad range of drug property. These simulations confirmed the ligand-dependent drug uptake and further suggested that a smaller drug favors the vestibule path, while a larger one is taken in via the cleft path. Moreover, the direct simulations identified an alternative uptake path which is not visible in the crystal structure. Third, site-directed mutagenesis of AcrB in E. coli verified that mutations of residues located along the newly identified path significantly reduced the efflux efficiency, supporting its relevance in in vivo function.

Successful treatment with infliximab for inflammatory colitis in a patient with X-linked anhidrotic ectodermal dysplasia with immunodeficiency.

X-linked anhidrotic ectodermal dysplasia with immunodeficiency (X-EDA-ID) is caused by hypomorphic mutations in the gene encoding nuclear factor-κB essential modulator protein (NEMO). Patients are susceptibile to diverse pathogens due to insufficient cytokine and frequently show severe chronic colitis. An 11-year-old boy with X-EDA-ID was hospitalized with autoimmune symptoms and severe chronic colitis which had been refractory to immunosuppressive drugs. Since tumor necrosis factor (TNF) α is responsible for the pathogenesis of NEMO colitis according to intestinal NEMO and additional TNFR1 knockout mice studies, and high levels of TNFα-producing mononuclear cells were detected in the patient due to the unexpected gene reversion mosaicism of NEMO, an anti-TNFα monoclonal antibody was administered to ameliorate his abdominal symptoms. Repeated administrations improved his colonoscopic findings as well as his dry skin along with a reduction of TNFα-expressing T cells. These findings suggest TNF blockade therapy is of value for refractory NEMO colitis with gene reversion.

Effects of vitamin E on the osteoblast differentiation.

Vitamin E is thought to affect bone formation and bone remodeling. In this study, we investigated the effects of vitamin E (alpha-tocopherol and delta-tocopherol) on the osteoblasts isolated from rat calvariae. At 4 and 7 days (Day 4 and 7) after induction of osteoblastic differentiation, treatment of alpha-tocopherol (100 and 200 microM) and delta-tocopherol (2 and 20 microM) for 3 days significantly decreased alkaline phophatase activity of the cultured osteoblasts. At Day 14, however, no significant change was detected in ALP activity and expression of bone sialoprotein mRNA in the osteoblasts treated with alpha-tocopherol or delta-tocopherol for 3 days. Expression of osteocalcin mRNA was decreased by treatment of alpha-tocopherol (100 and 200 microM) and delta-tocopherol (2 and 20 microM) at Day 4 and 7. At Day 14, expression of osteocalcin mRNA was decreased only with treatment of 200 microM alpha-tocopherol. In addition, the noncalcified nodules were decreased by treatment of alpha-tocopherol (200 microM) and delta-tocopherol (20 microM) at Day 7. However, treatment of alpha-tocopherol and delta-tocopherol showed no significant change of formation of calcified nodules at Day 14. These results indicate that vitamin E inhibits differentiation of osteoblasts especially from early stage to osteoid-producing stage.

Clinicopathological characteristics of primary gastric T-cell lymphoma.

AIMS: To investigate the clinicopathological characteristics of 20 primary gastric T-cell lymphoma (GTCL) cases without human T-lymphotropic virus type I infection in Japan, a non-endemic area for coeliac disease. METHODS AND RESULTS: Fifteen cases had no history of persistent diarrhoea or severe hypoproteinaemia. Histologically, 13 cases (65%) consisted of large cell lymphoma and seven (35%) were of medium-sized cells. Intraepithelial lymphoma cell invasion was found in three cases (15%). Two of 10 surgical cases (20%) showed intramucosal tumour cell spreading with enteropathy-like features. Helicobacter pylori CagA gene was detected in three of 10 cases (30%). The lymphoma cells of all 20 cases were positive for CD3 and/or TCRbetaF1 and negative for CD56. CD4- and CD8- lymphoma was found in 11 cases (55%), CD4+ lymphoma in seven (35%) and CD8+ lymphoma in two (10%). CD30+, CD5+ and CD25+ lymphomas were detected in nine (45%), 10 (50%) and 11 (55%) cases, respectively. Five-year survival of the 16 available cases was 54%. Early clinical stage and medium-sized cell lymphoma were significantly (P < 0.05) better prognostic factors. CONCLUSIONS: Patients with GTCL exhibit distinct clinicopathological findings and prognoses from those with enteropathy-associated T-cell lymphomas. GTCL may be mainly derived from lamina propria and parafollicular T cells.

cDNA cloning and mRNA expression of bovine GPR39.

GPR39 is an orphan G protein-coupled receptor that is thought to be involved in gastrointestinal and metabolic function. In this study, we cloned bovine GPR39 cDNA that encoded 462 amino acids showing high sequence homology to other mammalian GPR39 proteins. Real-time PCR showed expression of GPR39 mRNA in the liver, kidney, abomasums, small intestine, colon, rectum and uterus, with the highest level in the abomasums. Significant promoter activity was observed within the -2.3 kb 5'-upstream region of bovine GPR39 gene with human colon carcinoma-derived CACO-2 cells. These findings suggest that GPR39 may have important roles in gastrointestinal and metabolic functions in bovines as in other mammals.

Effects of overfeeding on the fatty acid profile and stearoyl-CoA desaturase-1 indices in the liver and subcutaneous adipose tissue in cats.

To evaluate the effect of overfeeding on fatty acid distribution and metabolism, especially stearoyl-CoA desaturase-1 (SCD-1) indices, 8 cats in the experimental and control groups (4 per group) were evaluated in this study. The experiments involved feeding the experimental group cats twice their daily energy requirement with a commercial diet for 4 weeks. The control group was fed the estimated daily energy requirement with the same diet. Body weight, feline body mass index, body condition score, several zoometry measurements, and plasma metabolites/hepatic injury markers were measured in all the cats before and after the experiment. In addition, the fatty acid profiles in the liver and subcutaneous adipose tissue were measured after the experiment. After 4 weeks of overfeeding, the experimental group demonstrated significant increases in hepatic C18:1, plasma triglyceride, and nonesterified fatty acid (NEFA) concentrations and in alanine aminotransferase activity. Furthermore, hepatic SCD-1 indices were positively correlated with body weight, feline body mass index, body condition score, and plasma NEFA concentration, although subcutaneous adipose tissue did not demonstrate any increase in SCD-1 indices in this study. The increase in hepatic SCD-1 indices might be enhanced by the inflow of plasma NEFA into the liver, and NEFA toxicity might stimulate C18:1 synthesis by SCD-1.

Profile of the ORION (Osaka emergency information Research Intelligent Operation Network system) between 2015 and 2016 in Osaka, Japan: a population-based registry of emergency patients with both ambulance and in-hospital records.

AIM: To describe the registry design of the Osaka Emergency Information Research Intelligent Operation Network system (ORION) and its profile of hospital information, patient and emergency medical service characteristics, and in-hospital outcomes among all patients transported to critical care centers and emergency hospitals in Osaka Prefecture, Japan. METHODS: The Osaka Prefecture Government has developed and introduced an information system for emergency patients (the ORION system) that uses a smartphone application (app) for hospital selection by on-scene emergency medical service personnel and has been accumulating all ambulance records. Since January 2015, medical institutions have obtained information on the diagnosis and outcome of patients transported to medical institutions, and the ORION system merged these data with ambulance records including smartphone app data. RESULTS: From January 2015 to December 2016, 753,301 eligible patients were registered. The mean age was 58.7 years, and 51.5% of patients were male. After hospital arrival, 39.7% were hospitalized, 58.2% were discharged from hospital, 1.1% changed hospital, and 1.0% died. The most common diagnoses were injury, poisoning, and certain other consequences of external causes. Among the hospitalized patients, 29.2% were continuously hospitalized, 59.0% discharged, 5.2% changed hospital, and 5.8% were dead at 21 days after hospitalization. The most common confirmed diagnosis was diseases of the circulatory system. CONCLUSION: Using the ORION system developed and operated by Osaka Prefecture since January 2015, we described the epidemiological data of all emergency patients transported to emergency hospitals. Analysis using the ORION database in the future could lead to improvements in the emergency transport system and patient outcomes.

Effects of butyrate supplementation in antibiotic-free milk replacer and starter on growth performance in suckling calves.

The aim of this study was to evaluate butyrate supplementation of antibiotic-free milk replacer and starter on growth performance in male Holstein calves. Twenty-nine calves were divided into two groups. Group C (n = 13) was fed antibiotic-free milk replacer without supplementation, and Group B (n = 16) was fed antibiotic-free milk replacer supplemented with butyrate (1.6 % DM of Gustor BP70® ). Starter in Group B contained 0.3 % DM of Gustor BP70® . The intake of milk replacer was lower in group B than in C (p = 0.07 for the treatment x week interaction). Body weight (BW) and heart girth (HG) in group B was higher than in C during the experimental period (p = 0.07 and 0.01 for the treatment × week interaction, respectively). The duration of the weaning period in group B was shorter than in group C (p = 0.02). ß-hydroxybutyrate (BHBA) was higher in group B than in C (p = 0.04). Insulin like growth factor-1 (IGF-1) concentrations tended to be higher in group B than in C (p = 0.07 for treatment × week interaction). Our results show that butyrate supplementation in antibiotic-free milk replacer and starter exerted positive effects on growth performance in suckling calves.

Review of hemophagocytic lymphohistiocytosis (HLH) in children with focus on Japanese experiences.

Hemophagocytic lymphohistiocytosis (HLH) is characterized by fever and hepatosplenomegaly associated with pancytopenia, hypertriglyceridemia and hypofibrinogenemia. Increased levels of cytokines and impaired natural killer activity are biological markers of HLH. HLH can be classified into two distinct forms, including primary HLH, also referred to as familial hemophagocytic lymphohistiocytosis (FHL), and secondary HLH. Although FHL is an autosomal recessive disorder typically occurring in infancy, it is important to clarify that the disease may also occur in older patients. It is now considered that FHL is a disorder of T-cell function; moreover, clonal proliferation of T lymphocytes is observed in a few FHL patients, and cytotoxicity of these T lymphocytes for target cells is usually impaired. In 1999, perforin gene (PRF1) mutation was identified as a cause of 20-30% of FHL (FHL2) cases. In Japan, two specific mutations of PRF1 were also detected. Furthermore, in 2003, MUNC13-4 mutations were identified in some non-FHL2 patients (FHL3). Identification of other genes responsible for remaining cases is a major concern. Hematopoietic stem cell transplantation (HSCT) has been established as the only accepted curative therapy for FHL. Thus, appropriate diagnosis and prompt treatment with HSCT are necessary for FHL patients. Genetic analysis for PRF1 and MUNC13-4 and functional assay of cytotoxic T lymphocytes are recommended to be performed in each patient. In those patients displaying impaired cytotoxic function but lacking genetic defects, samples should be employed for identification of unknown genes. In the near future, an entire pathogenesis should be clarified in order to establish appropriate therapies including immunotherapy, HSCT and gene therapy.

Comparison of enzyme activities in plasma and leukocytes in dairy and beef cattle.

Concentrations of plasma glucose, immunoreactive insulin (IRI) and free fatty acid (FFA) and activities of enzymes related to energy metabolism and lactate dehydrogenase (LDH) isoenzyme pattern in plasma and leukocytes were investigated in lactating Holstein cows (dairy cattle) and fattening Japanese Black Wagyu x Holstein steers (beef cattle). IRI concentrations and LDH and malate dehydrogenase (MDH) activities in the plasma of beef cattle were significantly higher than those in dairy cattle. The cytosolic ratio of MDH/LDH activity in the leukocytes of beef cattle was significantly higher than that of dairy cattle. These findings might be associated with the different energy metabolism between dairy and beef cattle.

The effects of feeding a low level of distiller's dried grains with solubles (DDGS) to yearling Holstein steers.

We examined the effects of feeding a low percentage of distiller's dried grains with solubles (DDGS) to yearling Holstein beef steers during the late stage of the fattening period on growth performance and carcass quality. Fifteen animals were fed a total mixed ration (TMR) feed plus a DDGS supplement equal to 10% of the total feed weight for 3 months (DDGS group). The other 15 animals were fed TMR with no DDGS (Control group). The vitamin E (VE) levels in the blood from the DDGS group showed a tendency to be higher in the DDGS group (P=0.056). Blood urea nitrogen levels in the DDGS group were also higher (P<0.05). The Beef Color Standard (BCS) number for the DDGS group was also higher than that for the Control group (P<0.05). Forty-eight hours after cutting, the yellowness (b*) value for Longissimus muscle in the DDGS group was significantly higher (P<0.05) and the redness (a*) value in the DDGS group tended to be higher than that in the Control group (P=0.05). The study results show a potential for adding value to yearling beef by improving the ability to maintain meat color with an increase in VE content.

Effects of feeding dried distillers grains with solubles (DDGS) on meat quality at the late stage of the fattening period of Holstein steers.

Feeding dried distillers grains with solubles (DDGS) during the late stage of the fattening period of Holstein steers was studied in regard to the influence on meat quality. Sixteen Holstein steers approximately 18 months old were used in this study. Eight animals were fed commercial concentrated feed for the entire fattening period. The other eight animals were fed 15% DDGS in the concentrated feed for 3 months before slaughtering. The moisture, ether extract and crude protein from both groups was approximately the same. The thiobarbituric acid reactive substance (TBA) value of storage for 7 days at 5°C from the animals fed DDGS showed a tendency to be lower (P = 0.059). The change in the TBA value during storage was also lower for the animals not fed DDGS (P < 0.05). There were no differences in the subcutaneous fat color between the two groups. The a*(measure of redness) and b*(measure of yellowness) of the M. longissimus from the animals fed DDGS showed a tendency to be lower (P = 0.051, 0.070). The fatty acid composition of the M. longissimus, subcutaneous and perirenal fat were not widely influenced by the feeding of DDGS. It is suggested that feeding 15% DDGS during the late stage of the fattening period for Holstein steers reduced the oxidation of the beef.

Relationship between adipose maturity and fatty acid composition in various adipose tissues of Japanese Black, Holstein and Crossbred (F1) steers.

The amount of monounsaturated fatty acid (MUFA) is intimately related to adipose softness, melting point (MP) and flavor in beef. Stearoyl-CoA desaturase (SCD) is a main gene involved in MUFA synthesis. Mature adipose tends to be highly saturated, whereas immature or maturing adipose is highly unsaturated when chronologically based, so the degree of non-saturation can be an index of adipose maturity. In this study, three different adipose tissues (coelomic (CL), perirenal (PR), and subcutaneous (SC)) from three beef breeds with differing slaughter ages (Japanese Black (29.5 months), Holstein (20.1 month), and F1 crossbreed (25.6 months)) were examined to: (i) determine adipose maturity level as indexed by MUFA %; and (ii) determine SCD and other lipogenic gene messenger RNA (mRNA) expression levels in relation to unsaturated fatty acid content. Fatty acid composition was significantly different between adipose tissues (P < 0.05). MUFA amount was high in the following order: SC > CL > PR. This pattern corresponded to SCD mRNA expression profile showing higher expression in SC than CL and PR. However, Japanese black cattle are an exception with CL adipose containing similar UFA % as SC adipose, yet having the lowest SCD mRNA expression level among all adipose tissues tested. Therefore, SCD mRNA expression and MUFA % appear to be directly related; however, differences in SCD mRNA expression among three adipose tissues may reflect differences in the fat development characteristics affected by chronological age of the cattle breeds.

Productivity and energy partition of late lactation dairy cows during heat exposure.

Three late-lactation Holstein cows were used to determine the effects of environmental temperature on performance and energy partitioning. Each cow was housed in a respiratory chamber for 30 consecutive days and exposed to three different conditions of environmental temperature: (i) 20 degrees C and 20 degrees C (20 degrees C), (ii) 25 degrees C and 20 degrees C (25 degrees C), (iii) 30 degrees C and 25 degrees C (30 degrees C) during the day and night, respectively. The temperature was switched in an interval of 10 days. Humidity in the chamber was maintained at 55-65% through the entire experimental period. The daily mean as well as morning and evening rectal temperatures of Holstein cows increased linearly (P < 0.05) as chamber temperature increased. There was a significant linear reduction in dry matter (DM) intake (P < 0.05) and an increase in DM digestibility (P < 0.05). The response in milk yield, however, was not affected by heat stress. There were no significant differences among treatments for intake energy, heat production, net energy for lactation and net energy for gain. This results of this study disagreed with the assumption that late lactation cows gave priority to increasing body tissue at the expense of milk production under thermal stress.

Cladribine treatment in two-hour intravenous infusion for previously-treated low grade B-cell lymphoma: a pilot study.

Cladribine is approved to be used in 24-hour continuous infusion for the treatment of low-grade lymphoma by the Ministry of Health, Labor and Welfare in Japan. Pharmacokinetic studies showed that the antitumor activity of cladribine by 2-hour infusion should be comparable to that given by continuous infusion. The safety and anti-tumor activity of short infusion of cladribine was shown in hairy cell leukemia, chronic lymphocytic leukemia and low-grade non-Hodgkin's lymphoma in Europe. We therefore underwent a pilot study to confirm the safety and efficacy of cladribine given by 2-hour infusion for Japanese patients with relapsed or refractory indolent B-cell lymphoma. Cladribine at a dose of 0.09 mg/kg was administered in 2-hour intravenous infusion for 5 consecutive days. The treatment was repeated at a 28-day interval for at least 2 cycles, and its efficacy and toxicity were investigated. Fourteen patients were entered into this study. Eight patients (57%) responded to cladribine, including 2 (14%) complete response (CR) and 6 (43%) partial response (PR). The median duration of response was 20+ and 21+ months for CR, and 12 months ranging from 3 to 34 months for PR, respectively. Grade 3 or 4 neutropenia and lymphocytopenia occurred in 43% and 71% of patients, respectively, but there was no febrile neutropenia or opportunistic infection associated with cladribine treatment. No other adverse events greater than grade 3 were encountered. The tumor response and degree of toxicity were comparable with those observed in cladribine treatment given by continuous infusion at a same dose. Cladribine can be administered in 2-hour infusion in an outpatient clinic and is therefore quite convenient for patients.

Adsorption of bisphenol A by lactic acid bacteria, Lactococcus, strains.

Ten strains of the genus Lactococcus were examined for their ability to remove bisphenol A [2, 2-bis(4-hydroxyphenyl)propane; BPA], which is known as an endocrine disrupter. Nine strains of the lactococci tested could remove BPA from media during growth, although the removal ratio was below 9%. When BPA was incubated with lyophilized cells of lactococci for 1 h, the concentration of BPA in the media was decreased by 9-62%. Especially, the highest removal ratio of BPA was observed for Lactococcus lactis subsp. lactis 712. The lactococci could adsorb BPA but not degrade it, because the lactococci maintained the ability to remove BPA from the medium after autoclaving. When the lyophilized cells of L. lactis subsp. lactis 712 were also incubated with six analogues of BPA, they effectively adsorbed hydrophobic compounds such as 2, 2'-diphenylpropane and bisphenol A dimethylether. The BPA-adsorbing ability of lactococci could be due to the hydrophobic binding effect. The removal ratio of BPA by L. lactis subsp. lactis 712 was increased after treatment with sodium dodecyl sulfate and decreased after digestion with trypsin. These results suggest that the hydrophobic proteins on cell surface may be involved in the BPA-adsorbing ability of lactococci.

Contribution of anaphylatoxins to allergic inflammation in human lungs.

The contribution of complement activation to allergic asthma remains controversial. In order to elucidate the role played by the complement split products, anaphylatoxins C3a and C5a, we evaluated their effects on production of cysteinyl-leukotrienes (cysLTs) by human lung fragments following an anaphylactic reaction. The lung tissues obtained from two patients with lung cancer showed C5aR-, C5L2R-, and C3aR-mRNA expression. When the chopped lung fragments passively sensitized with human IgE were incubated with anti-human IgE antibody, a significant amount of cysLTs was generated in comparison with the control (without anti-IgE antibody). The co-addition of human C5a at doses of 0.1 to 10 ng/ml to the anti-IgE antibody potentiated cysLT production. The response was bell-shaped in distribution, significant, and peaked at a C5a concentration of 1 ng/ml. The co-addition of human C3a up to 1,000 ng/ml seemed to increase cysLT production, but not to any significant extent. A novel C5a receptor complementary peptide, acetylated peptide A, dose-dependently inhibited cysLT production by the human lung fragments following the anaphylactic reaction in the presence of 1 ng/ml C5a. However, this peptide did not inhibit cysLT production in the presence of 100 ng/ml C3a. It is suggested that the anaphylatoxin C5a potentiates cysLT production in human lung tissues and contributes to allergic inflammation in disorders such as asthma, thus acetylated peptide A may be useful for suppressing allergic inflammation in the lungs.

Allogeneic stem-cell transplantation with reduced conditioning intensity as a novel immunotherapy and antiviral therapy for adult T-cell leukemia/lymphoma.

Sixteen patients with adult T-cell leukemia/lymphoma (ATL) who were all over 50 years of age underwent allogeneic stem cell transplantation with reduced-conditioning intensity (RIST) from HLA-matched sibling donors after a conditioning regimen consisting of fludarabine (180 mg/m2), busulfan (8 mg/kg), and rabbit antithymocyte globulin (5 mg/kg). The observed regimen-related toxicities and nonhematologic toxicities were all found to be acceptable. Disease relapse was the main cause of treatment failure. Three patients who had a relapse subsequently responded to a rapid discontinuation of the immunosuppressive agent and thereafter achieved another remission. After RIST, the human T-cell leukemia virus type 1 (HTLV-1) proviral load became undetectable in 8 patients. RIST is thus considered to be a feasible treatment for ATL. Our data also suggest the presence of a possible graft-versus-ATL effect; an anti-HTLV-1 activity was also found to be associated with this procedure.

Characterization of Epstein-Barr virus (EBV)-positive NK cells isolated from hydroa vacciniforme-like eruptions.

Recently, the involvement of Epstein-Barr virus (EBV) in hydroa vacciniforme (HV)-like eruptions has been suggested. To elucidate the role of EBV in this disease, we isolated EBV-infected cell clones from peripheral blood mononuclear cells (PBMC) and the skin lesions of a patient with HV-like eruptions; cells isolated from PBMC were designated SNK-12, and those from the eruption SNK-11. Both cells expressed CD16, CD56, and HLA-DR and had germline configurations of the T-cell receptor and the immunoglobulin genes, indicating that the cell clones were of NK cell lineage. The analysis of EBV terminal repeats indicated that the cells were monoclonal, had identical clonality, and originated from EBV-positive cells in the PBMC and eruption. Both clones expressed EBNA-1, but not EBNA-2. Although LMP-1 was weakly detected in SNK-11, no LMP-1 was detected in SNK-12. Interestingly, EBV-infected cells required less IL-2 for in vitro growth in the later phase of this disease and this appeared to correlate with the expression of LMP-1, suggesting that the proliferative capacity of the EBV-positive NK cells increased during the time course of the disease, and LMP-1 expression might be responsible for that. This is the first report of the isolation of EBV-infected cells from the skin lesions of HV-like eruptions and strongly suggests that the HV-like eruption in the patient was caused by clonal NK cells with latent EBV infection.