Effects of wood kraft pulp as a partial replacement for roughage on rumen fermentation and productivity in dairy cows.

In this study, we evaluated the partial replacement of roughage with wood kraft pulp (KP) on rumen fermentation and productivity of dairy cows. Eighteen cows were divided into control and KP groups. The KP group started adaptation to KP 3 weeks before calving; after calving, they were fed a total mixed ration for 12 weeks, wherein 18% Timothy hay was replaced with KP. The dry matter intake, body weight, and milk yield and composition were similar in the control and KP groups. The average daily rumen pH was higher with KP feeding, and the average daily ruminal temperature remained lower at 16 days after calving (P < 0.05). The concentration of volatile fatty acids remained unaltered, the molar proportion of acetic acid decreased, and the molar proportion of propionic acid increased, indicating a low acetic acid:propionic acid ratio (P < 0.05). Lipopolysaccharide activity in the rumen fluid was higher in the KP group (P < 0.05); however, the rumen microbiota were unaffected. The digestibility of dry matter and neutral detergent fiber increased 12 weeks after calving, whereas excretion of urinary nitrogen decreased (P < 0.05). Partial replacement of roughage with KP did not suppress rumen fermentation and maintained postpartum productivity.

Long working hours and skipping breakfast concomitant with late evening meals are associated with suboptimal glycemic control among young male Japanese patients with type 2 diabetes.

AIMS/INTRODUCTION: To assess the associations of working conditions, eating habits and glycemic control among young Japanese workers with type 2 diabetes. MATERIALS AND METHODS: This hospital- and clinic-based prospective study included 352 male and 126 female working patients with diabetes aged 20-40 years. Data were obtained from June to July 2012 and June to July 2013. Logistic regression analysis was used to estimate multivariable-adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for suboptimal glycemic control (glycosylated hemoglobin level of ≥7%) obtained from June to July 2013. RESULTS: Multivariable logistic regression analysis showed that disease duration of ≥10 years (OR 2.43, 95% CI 1.02-5.80), glycosylated hemoglobin level of ≥7% in 2012 (OR 8.50, 95% CI 4.90-14.80), skipping breakfast and late evening meals (OR 2.50, 95% CI 1.25-5.00) and working ≥60 h/week (OR 2.92, 95% CI 1.16-7.40) were predictive of suboptimal glycemic control in male workers, whereas a glycosylated hemoglobin level of ≥7% in 2012 (OR 17.96, 95% CI 5.93-54.4), oral hyperglycemic agent therapy (OR 12.49, 95% CI 2.75-56.86) and insulin therapy (OR 11.60, 95% CI 2.35-57.63) were predictive of suboptimal glycemic control in female workers. CONCLUSIONS: Working ≥60 h/week and habitual skipping breakfast concomitant with late evening meals might affect the ability of young male workers with type 2 diabetes to achieve and maintain glycemic control.

Heterologous and High Production of Ergothioneine in Escherichia coli.

Ergothioneine (ERG) is a histidine-derived thiol compound suggested to function as an antioxidant and cytoprotectant in humans. Therefore, experimental trials have been conducted applying ERG from mushrooms in dietary supplements and as a cosmetic additive. However, this method of producing ERG is expensive; therefore, alternative methods for ERG supply are required. Five Mycobacterium smegmatis genes, egtABCDE, have been confirmed to be responsible for ERG biosynthesis. This enabled us to develop practical fermentative ERG production by microorganisms. In this study, we carried out heterologous and high-level production of ERG in Escherichia coli using the egt genes from M. smegmatis. By high production of each of the Egt enzymes and elimination of bottlenecks in the substrate supply, we succeeded in constructing a production system that yielded 24 mg/L (104 µM) secreted ERG.

Socioeconomic status and type 2 diabetes complications among young adult patients in Japan.

OBJECTIVE: To assess the relationship between socioeconomic status (SES) and complications of type 2 diabetes among young adults in Japan. DESIGN: A cross-sectional study. SETTING: Outpatient wards of 96 member hospitals and clinics of the Japan Federation of Democratic Medical Institutions. PARTICIPANTS: A total of 782 outpatients with type 2 diabetes (525 males, 257 females), aged 20-40 years as of March 31, 2012. After excluding 110 participants whose retinopathy diagnosis was in question, 672 participants were analyzed. MEASUREMENTS: We examined the relations between SES (educational level, income, type of public healthcare insurance, and employment status) and diabetes complications (retinopathy and nephropathy) using a multivariate logistic regression analysis. RESULTS: The prevalence of type 2 diabetic retinopathy was 23.2%, while that of nephropathy was 8.9%. The odds of having retinopathy were higher among junior high school graduates (OR 1.91, 95% CI 1.09-3.34), patients receiving public assistance (OR 2.19, 95% CI 1.20-3.95), and patients with irregular (OR 1.72, 95% CI 1.03-2.86) or no employment (OR 2.23, 95% CI 1.36-3.68), compared to those with a higher SES, even after covariate adjustment (e.g., age, gender, body mass index). Similarly, the odds of having nephropathy were higher among patients with middle (OR 3.61, 95% CI 1.69-8.27) or low income levels (OR 2.53, 95% CI 1.11-6.07), even after covariate adjustment. CONCLUSIONS: Low SES was associated with a greater likelihood of type 2 diabetes complications in young adults. These findings suggest the necessity of health policies that mitigate socioeconomic disparity and thereby reduce the prevalence of diabetic complications.

Detainment of Tam3 Transposase at Plasma Membrane by Its BED-Zinc Finger Domain.

Transposable elements (TEs) are considered to be parasites of host genomes because they act as powerful mutagens. If not kept in check, they can cause gene disruption, genome rearrangement, and genomic takeover. Hence, activities of TEs are under the rigid control of hosts. To date, all identified TE regulations have been epigenetic dependent, with the exception of the DNA transposon Tam3. Blocking nuclear translocation of Tam3 transposase (TPase) is consistent with the suppression of Tam3 in Antirrhinum majus In this article, we discovered that epigenetic-independent regulation of Tam3 is mediated by the BED-zinc finger (Znf-BED) domain of Tam3 TPase. The host targets the N terminus of the Znf-BED domain, which contains two highly conserved aromatic amino acids, to detain Tam3 TPase at the plasma membrane and to silence Tam3. Zinc finger proteins perform broader functions in transcriptional regulation through their DNA binding ability. Our data revealed that the posttranslational epigenetic-independent silencing against TEs was a result of the protein binding ability of the Znf-BED domain.

Evaluation of protein-DNA binding affinity by electrospray ionization mass spectrometry.

Binding affinity of complexes between a DNA-binding domain (DBD) of a transcription factor, c-Myb, and several double-stranded DNA (dsDNA) were evaluated by collision-induced dissociation (CID) of the multiply protonated molecules generated by electrospray ionization mass spectrometry (ESI-MS). Complexes of the c-Myb DBD and dsDNA were prepared in solution and analyzed by ESI-MS. Multiply protonated molecules of a high-affinity complex, the c-Myb DBD and dsDNA with a specific sequence, were clearly observed in ESI mass spectrum. Protonated molecules of the complex were quite stable in the gas-phase, and not easily dissociated even if high cone voltage was applied in the first vacuum chamber source when the sample was prepared in 10 mM ammonium acetate. As for the sample prepared in buffer with higher concentration of ammonium acetate, such as 500 mM ammonium acetate, protein-dsDNA complexes could easily be dissociated with an increase in the cone voltage, giving multiply protonated molecules of free c-Myb DBD and some DNA fragments. Systematic CID experiments were carried out on seven complexes between the c-Myb DBD and 22-mer dsDNA with different solution-Kd values in the range of 10(-9) M to 10(-7) M. For each complex dissociation curve as a function of cone voltage was plotted, and the cone voltage where 50% of the complex was dissociated (V(50%)) was calculated. Consequently, positive correlation was obtained between V(50%) and relative binding free energy change (DeltaDeltaG) in complex formation in solution. This suggests that ESI-CID experiments can provide quantitative evaluation of the stability of protein-DNA complexes based on proper calibration.

Selective dissociation of non-covalent bonds in biological molecules by laser spray.

The laser spray developed in our laboratory was applied to the analysis of bovine serum albumin (BSA), double-stranded DNA (dsDNA) and a protein-DNA complex. The tip of a stainless-steel capillary was irradiated with a 10.6 micro m infrared laser by increasing the laser power from 0 W (electrospray) to 1.4 W. The laser beam was focused to about 0.3 mm at the tip of the stainless-steel capillary. When BSA aqueous solution was irradiated by the laser, highly charged monomer ions were newly observed in addition to the multiply charged ions of non-denatured monomer, dimer and trimer moieties. This indicates that BSA suffers from denaturation on irradiation with an infrared laser in solution. A 1.4 W laser power is not sufficient to cause the complete denaturation of BSA under the present experimental conditions. Whereas dsDNA was found to dissociate almost completely to single-stranded DNA constituents on laser irradiation with a power of 1.2 W, no fragmentation of DNA molecules was observed. For a protein-DNA complex, i.e. a complex of c-Myb DNA binding domain and dsDNA, dissociation of the complex to the component moieties was observed. These findings indicate that the laser spray can selectively dissociate non-covalent complexes into subunits without causing dissociation of the covalent bonds of the subunits. The laser spray will be a versatile method for the investigation of the structures and stabilities of biomolecules including non-covalent complexes.