チュウトウド コウネツセイ サイキン Geobacillus stearothermophilus セイイク オンド ヘンイカブ ニ オケル タンパクシツ ハツゲン カイセキ

Optimum growth of moderately thermophile Geobacillus stearothermophilus (Bst) has been observed at 55ºC, and suppressed by the downshift to lower temperature. In order to understand the relationship between growth temperature and protein expression, we made Bst mutant strains which were adapted to lower temperature (30ºC) compared with wild type strain, and then we examined their protein expression under various growth temperatures. When they were cultured at 55ºC, different expression patterns of six proteins (P1-P6) were observed between wild type and mutant strains, whereas five proteins (A1-A5) were found only in mutant strains cultured at 37ºC. Furthermore, expression of five proteins (DS1-DS5) was increased according to the downshift of growth temperature to 37ºC. Among all 16 proteins, abundant three proteins (P1, DS3, and DS5) were electroblotted to PVDF membrane, followed by analyzed N-terminal amino acid sequences. As a result, it was suggested that P1 must be Surface-layer protein A (sbsA), whereas DS3 and DS5 were unknown proteins

Changes in biotin levels during production of natto, Japanese fermented soybean

The change of biotin level during production of natto (Japanese fermented soybean) was investigated in this study.  The total biotin level was measured by an agar plate bioassay using Lactobacillus plantarum ATCC 8014.  The total biotin level decreased during water soaking, but increased after the fermentation of soybeans using Bacillus subtilis var. natto (B. natto) and reached a maximum level.  The increase of total biotin was not affected by Asp, Arg, and Ile which promoted the growth of L. plantarum in high concentrations.  The peak level of biotin in the fermented soybeans was significantly higher than that of dry soybeans.  The fermented soybeans at the biotin peak level were adequate for food.  In addition, we detected 9 and 4 biotinylated polypeptides in the soybeans and B. natto used in this study, respectively.  We speculated that the increase of biotin level may depend on the increase of the 4 biotinylated polypeptides and free biotin in B. natto

アミトロール テイコウセイ ダイチョウキン カブ ニオケル タンパクシツ ハツゲン カイセキ

It was reported that the E.coli mutant strain exhibited the resistance against amitrole (AT) which had used as a pesticide and inhibits the bacterial growth. In order to elucidate the mechanism on the AT-resistance in E.coli, we prepared the E.coli mutant strain by treating with chemical mutagen, MNNG, and then selected the AT-resistant E. coli strain. The obtained mutant strain was possible for proliferating even in the M9 minimal medium containing 2 mg/ml AT. Therefore, we examined the expressed proteins in the mutant strain, which was cultured under various conditions. As a result, it was suggested the expression of 22 kDa protein (P1) was suppressed as the AT concentration was increased in M9 medium. By the amino acid sequence analysis, it was proved that P1 must be alkyl hydroperoxide reductase C22 protein (ahpC). On the other hand, the expression of P2 protein in mutant strain has increased in the M9 medium containing 2mg/ml AT, and partial amino acid sequence of P2 was consistent with zinT (yodA) protein. From these results, it is likely that the amitrole-resistance in E.coli might be regulated by soxRS regulon, not oxy R, like adaptation for the oxidative stress

D-Tagatose Effectively Reduces the Number of Streptococcus mutans and Oral Bacteria in Healthy Adult Subjects: A Chewing Gum Pilot Study and Randomized Clinical Trial

We examined the effect of D-Tagatose on the growth of oral bacteria including Streptococcus mutans (S. mutans). Saliva collected from 10 healthy volunteers was plated on BHI medium (to culture total oral bacteria) and MBS medium (to culture S. mutans, specifically). Agar plates of BHI or MBS containing xylitol or D-Tagatose were cultured under aerobic or anaerobic conditions. We then counted the number of colonies. In BHI plates containing D-Tagatose, a complete and significant reduction of bacteria occurred under both aerobic and anaerobic conditions. In MSB medium, significant reduction of S. mutans was also observed. We then performed a doubleblind parallel randomized trial with 19 healthy volunteers. They chewed gum containing xylitol, D-Tagatose, or both for 4 weeks, and their saliva was collected weekly and plated on BHI and MSB media. These plates were cultured under anaerobic conditions. Total bacteria and S. mutans were not effectively reduced in either the D-Tagatose or xylitol gum group. However, S. mutans was significantly reduced in volunteers chewing gum containing both D-Tagatose and xylitol. Thus, D-Tagatose inhibited the growth of S. mutans and many types of oral bacteria, indicating that D-Tagatose intake may help prevent dental caries, periodontitis, and many oral diseases

Loco-regional therapy for isolated locoregional lymph node recurrence of breast cancer: Focusing on surgical treatment with combined therapy

Advances have been made in systemic as well as locoregional treatment of primary breast cancer. Evidence, based established therapeutic strategies, for isolated locoregional lymph node recurrence is not yet sufficient. In this series, we focused especially on isolated axillary lymph node recurrence (AR) and supraclavicular lymph node recurrence (SR) in patients receiving systemic and/or radiation therapy combined with surgery. Disease free survival (DFS) in patients with AR ranged from 20 to 36 months. From 69% to 77% of all patients underwent surgical excision. The 5-year overall survival (OS) ranged from 39% to 46%. Positive lymph node metastases of primary cancer, size of the primary tumor, and R0 resection were associated with good outcomes. Longer DFS is associated with good outcomes. Limited SR data showed DFS to range from 25–27%. Median progression free survival (PFS) was 18 months, 5-year OS rates were 24–42%, and 5-year OS were 29–34 months. Combination therapy was an independent factor associated with better PFS as compared to local therapy only. Salvage treatment and grade of the primary tumor significantly were associated with OS on multivariate analysis. Available data, retrospective and not randomized, showed therapy combining systemic treatments and/or radiotherapy with surgery might contribute to good local control, better PFS, and longer OS

A combination of routine laboratory findings and vital signs can predict survival of advanced cancer patients without physician evaluation: a fractional polynomial model

IntroductionThere have been no reports about predicting survival of patients with advanced cancer constructed entirely with objective variables. We aimed to develop a prognostic model based on laboratory findings and vital signs using a fractional polynomial (FP) model.MethodsA multicentre prospective cohort study was conducted at 58 specialist palliative care services in Japan from September 2012 to April 2014. Eligible patients were older than 20 years and had advanced cancer. We developed models for predicting 7-day, 14-day, 30-day, 56-day and 90-day survival by using the FP modelling method.ResultsData from 1039 patients were analysed to develop each prognostic model (Objective Prognostic Index for advanced cancer [OPI-AC]). All models included the heart rate, urea and albumin, while some models included the respiratory rate, creatinine, C-reactive protein, lymphocyte count, neutrophil count, total bilirubin, lactate dehydrogenase and platelet/lymphocyte ratio. The area under the curve was 0.77, 0.81, 0.90, 0.90 and 0.92 for the 7-day, 14-day, 30-day, 56-day and 90-day model, respectively. The accuracy of the OPI-AC predicting 30-day, 56-day and 90-day survival was significantly higher than that of the Palliative Prognostic Score or the Prognosis in Palliative Care Study model, which are based on a combination of symptoms and physician estimation.ConclusionWe developed highly accurate prognostic indexes for predicting the survival of patients with advanced cancer from objective variables alone, which may be useful for end-of-life management. The FP modelling method could be promising for developing other prognostic models in future research

マウス胸腺の脂肪細胞分化過程の速度論による解析

The differential process of adipocytes was investigated in the thymus and spleen of BALB/c mice by the real time PCR method. The transcriptions of PPARγ (PP) ,adiponectin (Ad) and resistin (Re) were considered as indications of differential stages of adipocytes and the transcription ratios, log Ad/PP and log Re/PP , were measured. These logarithmic ratios increased rapidly, at the border between fetal stage and neonatal, from negative value to positive in the thymus and to nearly 0 in the spleen. Subsequently, they were remained virtually constant throughout the experimental period. Kinetics was applied to the differential model of adipocytes in which the quantity of each transcription was assumed to be proportional to the cell number, and the changes of log Ad/PP and log Re/PP with age were interpreted successfullyBALB/cマウスの胸腺および脾臓での脂肪細胞の分化過程を,PPARγ(PP)に対するアディポネクチン(Ad),およびレジスチン(Re)の転写量を指標として,リアルタイムPCR法により検討した。加齢に伴い成熟脂肪細胞が増加してくる胸腺においては,アディポネクチンおよびレジスチンの転写量は,胎生期にはPPARγ の転写量より少なかったが,出生を境に急激に増大し,PPARγ の転写量の数十倍となり,その後ほぼ一定となった。成熟した脂肪細胞が認められない脾臓では,アディポネクチンおよびレジスチンの転写量は,出生を境に増大し,PPARyの転写量とほぼ等しくなった。各遺伝子の転写量は細胞数に比例すると仮定し,脂肪細胞系列の幹細胞を考えることにより,これらの結果を,速度論により説明することができた

Rapid HER2 cytologic fluorescence in situ hybridization for breast cancer using noncontact alternating current electric field mixing

Background: Human epidermal growth factor receptor 2-in situ hybridization (HER2-ISH) is widely approved for diagnostic, prognostic biomarker testing of formalin- fixed paraffin-embedded tissue blocks. However, cytologic ISH analysis has a potential advantage in tumor samples such as pleural effusion and ascites that are difficult to obtain the histological specimens. Our aim was to evaluate the clinical reliability of a novel rapid cytologic HER2 fluorescence ISH protocol (rapid-CytoFISH). Materials and Methods: Using a new device, we applied a high-voltage/frequency, noncontact alternating current electric field to tissue imprints and needle rinses, which mixed the probe within microdroplets as the voltage was switched on and off (AC mixing). Cytologic samples (n = 143) were collected from patients with immunohistochemically identified HER2 breast cancers. The specimens were then tested using standard dual-color ISH using formalin-fixed paraffin-embedded tissue (FFPE-tissue DISH) for HER2-targeted therapies, CytoFISH, and rapid-CytoFISH (completed within 4 h). Results: All 143 collected cytologic specimens (50 imprinted cytology specimens from resected tumors and 93 liquid-based cytology specimens from needle rinses) were suitable for FISH analysis. The HER2/chromosome enumeration probe (CEP) 17 ratios did not significantly differ between FFPE-tissue DISH and either CytoFISH protocol. Based on HER2 scoring criteria, we found 95.1% agreement between FFPEtissue DISH and CytoFISH (Cohen\u27s kappa coefficient = 0.771 and 95% confidence interval (CI): 0.614–0.927). Conclusion: CytoFISH could potentially serve as a clinical tool for prompt determination of HER2 status in breast cancer cytology. Rapid-CytoFISH with AC mixing will enable cancer diagnoses and HER2 status to be determined on the same day a patient comes to a clinic or hospital

Self-Construals, Anger Regulation, and Life Satisfaction in the United States and Japan

Previous studies have reported evidence that indicates differences between Western and East Asian cultures in anger regulation and its psychological consequences. However, many of these studies have focused on a specific anger regulation strategy and its relation with a psychological consequence. Here, we developed an integrated model that can comprehensively examine three different anger regulation strategies (anger suppression, expression, and control), independent and interdependent self-construals as the psychological antecedent, and life satisfaction as the psychological consequence. We estimated the model using large samples of American and Japanese adults to examine the associations between the two self-construals, three anger regulation strategies, and life satisfaction. We compared the difference in the patterns of relationships among the key constructs between the American and Japanese samples. The results confirmed previously suggested cultural differences while also discovering new culturally different paths. The results generally suggest that individual-level self-construals matter more when anger is a culturally condoned emotion (vs. condemned). The implications and limitations of the integrated model are discussed