A Method for Systematic Assessment of Intrinsically Disordered Protein Regions by NMR

Intrinsically disordered proteins (IDPs) that lack stable conformations and are highly flexible have attracted the attention of biologists. Therefore, the development of a systematic method to identify polypeptide regions that are unstructured in solution is important. We have designed an “indirect/reflected” detection system for evaluating the physicochemical properties of IDPs using nuclear magnetic resonance (NMR). This approach employs a “chimeric membrane protein”-based method using the thermostable membrane protein PH0471. This protein contains two domains, a transmembrane helical region and a C-terminal OB (oligonucleotide/oligosaccharide binding)-fold domain (named NfeDC domain), connected by a flexible linker. NMR signals of the OB-fold domain of detergent-solubilized PH0471 are observed because of the flexibility of the linker region. In this study, the linker region was substituted with target IDPs. Fifty-three candidates were selected using the prediction tool POODLE and 35 expression vectors were constructed. Subsequently, we obtained ¹⁵N-labeled chimeric PH0471 proteins with 25 IDPs as linkers. The NMR spectra allowed us to classify IDPs into three categories: flexible, moderately flexible, and inflexible. The inflexible IDPs contain membrane-associating or aggregation-prone sequences. This is the first attempt to use an indirect/reflected NMR method to evaluate IDPs and can verify the predictions derived from our computational tools.     

Positive-temperature-coefficient effect of electrical resistivity below melting point of poly(vinylidene fluoride) (PVDF) in Ni particle-dispersed PVDF composites

This paper discusses the positive-temperature-coefficient effects of resistivity in Ni particle-dispersed poly(vinylidene fluoride) (PVDF) composites based on experiment results from SEM, DSC, and pressure-volume-temperature (PVT) measurements. The melting points of composites with Ni content of 20, 30, 40, and 50vol.% were equal to that of pure PVDF. The PTC effects in composites with Ni content of 40 and 50vol.% occurred at temperatures near the melting point of the PVDF matrix, whereas those in composites with Ni content of 20 and 30vol.% occurred at temperatures below the melting point of the PVDF matrix. We found that the PTC effect occurs even without melting of the matrix polymer. Moreover, we determined that a slight increase in specific volume at temperatures below the melting point of the matrix polymer acts fully as a driving force for forming a gap between fillers. This suggestion was backed up by theoretical analyses using percolation theory and a thermal-fluctuation-induced tunneling model.     

Preparation and Properties of Carbon/Zeolite Composites with Corrugated Structure

Composites of carbon/zeolite with corrugated structures were prepared by carbonization, steam activation and/or hydrothermal treatment of corrugated paper. No zeolite formation resulted from conventional hydrothermal treatment of the carbonized and activated samples in NaOH solution but zeolite Na P1 was formed by addition of silica to the solution. By contrast, zeolites Na P1 and Na A were formed by the in-situ crystallization method (hydrothermal treatment of solid samples impregnated with NaOH solution). With higher impregnating NaOH concentrations, longer reaction times and higher reaction temperatures, the products changed to sodalite- and cancrinite-type compounds. Hydrothermal treatment was effective in increasing the specific surface area of the products by the formation of zeolite from amorphous calcium aluminosilicate, and also increased the mechanical strength by gluing together the carbon fibers in the samples. The resulting samples showed enhanced adsorption for polar molecules such as ammonia, water vapor and methanol due to the formation of composites of activated carbon with hydrophilic zeolites.     

Effect of Lipase Activity and Specificity on the DAG Content of Olive Oil from the Shodoshima-Produced Olive Fruits

Olive oils have a higher relative diacylglycerol (DAG) content than other plant oils. The lipase in olive fruits is involved in DAG production and is directly related to the acidity of the olive oil. However, the lipase activity and positional selectivity have not been clarified. To investigate the properties of olive fruit lipase, olive fruits of the Mission variety harvested during mid-December of 2005 on Shodoshima Island (Japan) were stored at 20, 30 or 40 °C for 4 weeks. Changes in the acidity and acylglycerol content of the oils extracted from the stored fruits were analyzed. The acidity and DAG content of the olive oils increased due to triacylglycerol (TAG) hydrolysis during storage. sn-1,2-DAGs preferentially increased during the early stages of storage, indicating that the olive fruit lipase is enantioselective for the sn-3 position, while non-enzymatic isomerization of sn-1,2-DAGs was observed throughout the entire duration of storage. Kinetic analysis revealed that the enantioselectivity of olive fruit lipase for the sn-3 position was approximately four times greater than for the sn-1 position. The lipase was gradually inactivated at temperatures of 30 °C or higher, and the ratios of the rate constant for inactivation to TAG hydrolysis at the sn-3 position was 0.2, 13, and 23 at 20, 30, and 40 °C, respectively.     

Potato and soy peptides alter caecal fermentation and reduce serum non‐HDL cholesterol in rats fed cholesterol

We examined the effect of potato peptides (PPC) in rats fed a cholesterol‐enriched diet, in comparison with two cholesterol‐enriched diets containing soy peptides (SPC) or casein (CNC), and a cholesterol‐free diet containing casein (CN) for 4 weeks. The serum non‐high‐density lipoprotein (HDL) cholesterol level was lower in the PPC‐ (–18.39%) and SPC‐fed (–32.76%) groups (p <0.05) than in the CNC‐fed group at the end of the feeding period. The low‐density lipoprotein receptor mRNA level in the PPC‐fed group, and cholesterol 7α‐hydroxylase and scavenger receptor class B type 1 mRNA levels in the SPC‐fed group, were higher (p <0.05) than in the CN‐ and CNC‐fed groups. Faecal neutral sterol and caecal short‐chain fatty acid concentrations in the PPC‐ and SPC‐fed groups were higher (p <0.05) than in the CN‐ and CNC‐fed groups. The faecal total acidic sterol concentration was higher in the SPC‐fed group than in the CN‐ and PPC‐fed groups. Caecal anaerobe and Bifidobacterium populations were higher (p <0.05) in the PPC‐ and SPC‐fed groups than in the CN‐fed group. This study suggests that potato peptides, as soy peptides, alter caecal fermentation and steroid absorption and reduce the serum non‐HDL cholesterol level in rats fed cholesterol.     

Preparation of ultrafine polyurethane fiber web by laser‐electrospinning combined with air blowing

Thermoplastic polyurethane fiber webs were prepared using a laser‐heated electrospinning process combined with air blowing. The effect of spinning conditions such as air flow rate and air temperature on fiber diameter and molecular weight was investigated. Although the average fiber diameter decreased with increased air flow rate at each air temperature, the diameter increased when the air flow rate was >15 NL min^?1. In addition, the fiber was comparatively thicker with an increase in the air temperature. The variation in the fiber diameter tends to increase with the air flow rate, and a reduction in the molecular weight of the fiber by thermal degradation was suppressed. The thinnest and most uniform fiber with a diameter of 0.9 µm and a diameter coefficient variation of 15% was obtained at an air temperature of 25°C under an air flow rate of 15 NL min^?1. This fiber also had a minimum of decreased molecular weight. POLYM. ENG. SCI., 54:2605–2609, 2014. © 2013 Society of Plastics Engineers     

Enhanced Development of Azoxymethane-Induced Colonic Preneoplastic Lesions in Hypertensive Rats

Metabolic syndrome is associated with an increased risk of colorectal cancer. This study investigated the impact of hypertension, a component of metabolic syndrome, on azoxymethane (AOM)-induced colorectal carcinogenesis using SHRSP/Izm (SHRSP) non-diabetic/hypertensive rats and SHRSP.Z-Lepr^fa/IzmDmcr (SHRSP-ZF) diabetic/hypertensive rats. Male 6-week-old SHRSP, SHRSP-ZF, and control non-diabetic/normotensive Wister Kyoto/Izm (WKY) rats were given 2 weekly intraperitoneal injections of AOM (20 mg/kg body weight). Two weeks after the last injection of AOM, the SHRSP and SHRSP-ZF rats became hypertensive compared to the control WKY rats. Serum levels of angiotensin-II, the active product of the renin-angiotensin system, were elevated in both SHRSP and SHRSP-ZF rats, but only the SHRSP-ZF rats developed insulin resistance, dyslipidemia, and hyperleptinemia and exhibited an increase in adipose tissue. The development of AOM-induced colonic preneoplastic lesions and aberrant crypts foci, was significantly accelerated in both SHRSP and SHRSP-ZF hypertensive rats, compared to WKY normotensive rats. Furthermore, induction of oxidative stress and exacerbation of inflammation were observed in the colonic mucosa and systemically in SHRSP and SHRSP-ZF rats. Our findings suggest that hypertension plays a role in the early stage of colorectal carcinogenesis by inducing oxidative stress and chronic inflammation, which might be associated with activation of the renin-angiotensin system.     

Numerical computation of two-loop box diagrams with masses

A new approach is presented to evaluate multi-loop integrals, which appear in the calculation of cross-sections in high-energy physics. It relies on a fully numerical method and is applicable to a wide class of integrals with various mass configurations. As an example, the computation of two-loop planar and non-planar box diagrams is shown. The results are confirmed by comparisons with other techniques, including the reduction method, and by a consistency check using the dispersion relation.