Molecular epidemiology of Coxsackievirus A16 strains isolated from children in Yamagata,Japan between 1988 and 2011

To clarify the longitudinal molecular epidemiology of coxsackievirus A16, phylogenetic analysis based on the VP1 region of 220 isolates in Yamagata, Japan was performed. The resultant phylogenetic tree indicates that the Yamagata isolates and reference strains can be readily genotyped into three genogroups, and 0, 12 and 208 isolates belonged to the first, second, and third genogroups, respectively. The first genogroup includes only the prototype strain, the second strains that had disappeared by the end of the 20th century and the third comprises those that have been circulating since then in local communities, such as Yamagata.     

The Relationship of the Feces Protein Particles to Rice Protein Bodies

Feces protein particles (FPP), spherical or oval in shape, 1.0~3.5 μ in diameter, which were observed in the fresh feces of the Japanese, in an average density of 5 × 10⁹/g, resembled the rice protein bodies (RPB) in electron-microscopical fine structure. The FPP decreased in the feces of the man who had been given a rice-free diet consisting of steamed sweet potato; when boiled rice had been fed, the FPP increased, and reached the level of the original density within 5 days. The FPP were concluded to be derived from an indigestible fraction of the RPB which were supplied to the Japanese with the daily intake of rice.     

Sucrose-induced Autolysis and Development of Protoplast-like Cells of Clostridium saccharoperbutylacetonicum

A growing culture of Clostridium saccharoperbutylacetonicum partially lost its turbidity in the presence of 0.3 to 0.6 m sucrose without any extraneous supplements for cell wall degradation. The maximum effect was shown at 0.35 m of sucrose and the culture lost 40 to 50% of initial turbidity. The rate of lysis depended on the age of culture. The most rapid lysis occurred in the organisms of early exponential growing cultures, but no lysis was observed on those of late exponential and stationary phase cultures. The optimal pH was 5.5 to 6.0, and the optimal temperature 30 to 35°C. The sucrose-induced lysis was inhibited by bivalent cations (such as Ca²⁺, Mg²⁺), heavy metal cations (such as Cu²⁺, Pb²⁺), enzymic inhibitors (such as PCMB) and fixative agents (such as formalin, glutaraldehyde), while organisms whose growth had been inhibited by antibiotics (such as chloramphenicol, tetracycline) were also resistant to sucrose-induced lysis. The sucrose-induced lysis was accompanied by striking morphological conversion from original rod cells (3.0~6.0}0.4~0.6 μ) to spherical cells (1.0~ 1.2 μ diameter). The sucrose-induced lysis was also observed on the relative strains of C. saccharoperbutylacetonicum and C. sporogenes, but not observed on many other species of Clostridium and aerobic bacteria tested. It was suggested that sucrose-induced lysis was a kind of bacterial autolysis which was induced by sucrose treatment. The bacterial spheres developed during the lysis may be the protoplasts.     

Bacteriophages of Clostridium saccharoperbutylacetonicun

Some characteristics of the twelve phages were given. Phages were obtained from the twelve abnormal broths in the industrial production of acetone and butanol by use of Cl. saccharoperbutylacetonicum, producing a high proportion of butanol. These new phages were designated as HM and numbered consecutively in the order of appearance. The HM-phages were highly specific for the strains of this bacterium. Each phage could be distinguished from the others by its differential host specificity against various phage-resistant mutants of this bacterium. The HM-phages were not temperate ones under our experimental conditions. They were divided into two groups on the basis of their stability in the salts solution. While one group was slightly less stable in 0.85 percent saline or 0.067 m phosphate buffer, the other was strikingly unstable. The addition of magnesium ion was effective for increase in stability of both groups.

The double-layer method similar to that described by Adams, with modifications to incubate the plates anaerobically, was applied to assay the phages of butanol-producing bacteria, Cl. saccharoperbutylacetonicum, after the studies on some factors influencing the plaque formation. Factors influencing the number and size of plaque were as follows; agar concentration and amount of overlayer medium, pH of media, age and number of host cells, temperature and period of incubation, and so on. Plaques formed by this method were medial size and easily counted. Assay of viable cells of this anaerobic bacterium was also possible by this method with slight modifications.     

Intrinsic Temperature Sensitivity of Influenza C Virus Hemagglutinin-Esterase-Fusion Protein

Influenza C virus replicates more efficiently at 33°C than at 37°C. To determine whether hemagglutinin-esterase-fusion protein (HEF), a surface glycoprotein of influenza C virus, is a restricting factor for this temperature sensitivity, we analyzed the biological and biochemical properties of HEF at 33°C and 37°C. We found that HEF exhibits intrinsic temperature sensitivities for surface expression and fusion activity.     

CD4+ CD25+ regulatory T cells impair HIV-1-specific CD4 T cell responses by upregulating interleukin-10 production in monocytes

T cell dysfunction in the presence of ongoing antigen exposure is a cardinal feature of chronic viral infections with persistent high viremia, including HIV-1. Although interleukin-10 (IL-10) has been implicated as an important mediator of this T cell dysfunction, the regulation of IL-10 production in chronic HIV-1 infection remains poorly understood. We demonstrated that IL-10 is elevated in the plasma of individuals with chronic HIV-1 infection and that blockade of IL-10 signaling results in a restoration of HIV-1-specific CD4 T cell proliferation, gamma interferon (IFN-γ) secretion, and, to a lesser extent, IL-2 production. Whereas IL-10 blockade leads to restoration of IFN-γ secretion by HIV-1-specific CD4 T cells in all categories of subjects investigated, significant enhancement of IL-2 production and improved proliferation of CD4 T helper cells are restricted to viremic individuals. In peripheral blood mononuclear cells (PBMCs), this IL-10 is produced primarily by CD14(+) monocytes, but its production is tightly controlled by regulatory T cells (Tregs), which produce little IL-10 directly. When Tregs are depleted from PBMCs of viremic individuals, the effect of the IL-10 signaling blockade is abolished and IL-10 production by monocytes decreases, while the production of proinflammatory cytokines, such as tumor necrosis factor alpha (TNF-α), increases. The regulation of IL-10 by Tregs appears to be mediated primarily by contact or paracrine-dependent mechanisms which involve IL-27. This work describes a novel mechanism by which regulatory T cells control IL-10 production and contribute to dysfunctional HIV-1-specific CD4 T cell help in chronic HIV-1 infection and provides a unique mechanistic insight into the role of regulatory T cells in immune exhaustion.     

Effects of N-[N-(3, 5-difluorophenacetyl-l-alanyl)]-S-phenylglycine t-butyl ester (DAPT) on cell proliferation and apoptosis in Ishikawa endometrial cancer cells

Endometrial cancer is one of the most common gynecological malignancies in Japan, where the disease shows an increasing morbidity. However, surgical therapy remains the treatment of choice for endometrial cancers that tend to be insensitive to radiation therapy and chemotherapy. Therefore, novel therapeutic strategies are required. The Notch signaling pathway regulates embryogenesis and cellular development, but deregulated Notch signaling may contribute to tumorigenesis in several cancers. Moreover, γ-secretase inhibitors have been shown to be potent inhibitors of the Notch signaling pathway; they suppress cellular proliferation and induce apoptosis in several cancer cells. In the present study, we investigated the effect of N-[N-(3, 5-difluorophenacetyl-L-alanyl)]-S-phenylglycine t-butyl ester (DAPT, γ-secretase inhibitor) on the cell proliferation and apoptosis in Ishikawa endometrial cancer cells. Real-time PCR detected mRNA derived from NOTCH1 and HES1, which are target genes of the Notch signaling pathway, in Ishikawa endometrial cancer cells. After blocking Notch signaling, cellular proliferation decreased, accompanied by increased expression of p21 mRNA and decreased expression of the cyclin A protein. Furthermore, blockade of Notch signaling induced apoptosis. These results suggest that the Notch signaling pathway may be involved in cell proliferation through cell cycle regulation and apoptosis in Ishikawa endometrial cancer cells. Inhibition of the Notch signaling pathway by γ-secretase inhibitors is expected to be a potential target of novel therapeutic strategies for endometrial cancer.     

Cosmetic benefits of astaxanthin on humans subjects

Two human clinical studies were performed. One was an open-label non-controlled study involving 30 healthy female subjects for 8 weeks. Significant improvements were observed by combining 6 mg per day oral supplementation and 2 ml (78.9 μM solution) per day topical application of astaxanthin. Astaxanthin derived from the microalgae, Haematococcus pluvialis showed improvements in skin wrinkle (crow's feet at week-8), age spot size (cheek at week-8), elasticity (crow's feet at week-8), skin texture (cheek at week-4), moisture content of corneocyte layer (cheek in 10 dry skin subjects at week-8) and corneocyte condition (cheek at week-8). It may suggest that astaxanthin derived from H. pluvialis can improve skin condition in all layers such as corneocyte layer, epidermis, basal layer and dermis by combining oral supplementation and topical treatment. Another was a randomized double-blind placebo controlled study involving 36 healthy male subjects for 6 weeks. Crow's feet wrinkle and elasticity; and transepidermal water loss (TEWL) were improved after 6 mg of astaxanthin (the same as former study) daily supplementation. Moisture content and sebum oil level at the cheek zone showed strong tendencies for improvement. These results suggest that astaxanthin derived from Haematococcus pluvialis may improve the skin condition in not only in women but also in men.     

The location of the arginine-specific carboxypeptidase in the membrane of Mycoplasma salivarium and its physiological functions

A non-penetrating probe, 2,4,6-trinitrobenzenesulfonate, inhibited the activity of the carboxypeptidase purified from the cell membranes of Mycoplasma salivarium and the same enzymatic activity of intact Mycoplasma cells as well. Growth of the organism in medium containing benzoylglycyl-L-arginine resulted in a higher pH and higher turbidity than growth in the same medium without this supplement. It was concluded that the enzyme existed in the outer surface of the membrane of the cells and probably functioned to supply the organism with arginine as an energy source.