Synthesis of potent and selective inhibitors of Candida albicans N-myristoyltransferase based on the benzothiazole structure

Two parallel synthetic methods using solid-supported reagents were established to examine the rapid optimization of weak hit compound 1. Several compounds showed high potency in the low nanomolar range against N-myristoyltransferase. The structure-activity relationship (SAR) and antifungal activities of a series of novel 2-aminobenzothiazole N-myristoyltransferase inhibitors are presented.     

Electrolyzed hydrogen-saturated water for drinking use elicits an antioxidative effect: a feeding test with rats

A new type of electrolyzed hydrogen-saturated (EHS) water was produced using a water-electrolyzing device equipped with a special cation exchanger. Use of the EHS water for drinking in a feeding test with rats elicited an antioxidative effect. After intraperitoneal injection of 2,2-azobis-amidinopropane dihydrochloride, urinary secretion of 8-hydroxydeoxyguanosine and hepatic formation of peroxidized lipid were significantly lessened in rats which had received the EHS water for one week. These results suggest the possibility that this drinking water shows an effect in reduction of oxidative stress in the body.     

POPK-1/Sad-1 kinase is required for the proper translocation of maternal mRNAs and putative germ plasm at the posterior pole of the ascidian embryo

Maternal mRNAs localized to specific regions in eggs play important roles in the establishment of embryonic axes and germ layers in various species. Type I postplasmic/PEM mRNAs, which are localized to the posterior-vegetal cortex (PVC) of fertilized ascidian eggs, such as the muscle determinant macho-1 mRNA, play key roles in embryonic development. In the present study, we analyzed the function of the postplasmic/PEM RNA Hr-POPK-1, which encodes a kinase of Halocynthia roretzi. When the function of POPK-1 was suppressed by morpholino antisense oligonucleotides, the resulting malformed larvae did not form muscle or mesenchyme, as in macho-1-deficient embryos. Epistatic analysis indicated that POPK-1 acts upstream of macho-1. When POPK-1 was knocked down, localization of every Type I postplasmic/PEM mRNA examined, including macho-1, was perturbed, showing diffuse early distribution and eventual concentration into a smaller area. This is the probable reason for the macho-1 dysfunction. The postplasmic/PEM mRNAs such as macho-1 and Hr-PEM1 are co-localized with the cortical endoplasmic reticulum (cER) and move with it after fertilization. Eventually they become highly concentrated into a subcellular structure, the centrosome-attracting body (CAB), at the posterior pole of the cleaving embryos. The suppression of POPK-1 function reduced the size of the domain of concentrated cER at the posterior pole, indicating that POPK-1 is involved in the movement of postplasmic/PEM RNAs via relocalization of cER. The CAB also shrank. These results suggest that Hr-POPK-1 plays roles in concentration and positioning of the cER, as well as in the concentration of CAB materials, such as putative germ plasm, in the posterior blastomeres.     

Wsh3/Tea4 is a novel cell-end factor essential for bipolar distribution of Tea1 and protects cell polarity under environmental stress in S. pombe

BACKGROUND: The fission yeast Schizosaccharomyces pombe has a cylindrical cell shape, for which growth is strictly limited to both ends, and serves as an excellent model system for genetic analysis of cell-polarity determination. Previous studies identified a cell-end marker protein, Tea1, that is transported by cytoplasmic microtubules to cell tips and recruits other cell-end factors, including the Dyrk-family Pom1 kinase. The deltatea1 mutant cells cannot grow in a bipolar fashion and show T-shaped morphology after heat shock. RESULTS: We identified Wsh3/Tea4 as a novel protein that interacts with Win1 MAP kinase kinase kinase (MAPKKK) of the stress-activated MAP kinase cascade. Wsh3 forms a complex with Tea1 and is transported to cell tips by growing microtubules. The deltawsh3 mutant shows monopolar growth with abnormal Tea1 aggregate at the non-growing cell end; this abnormal aggregate fails to recruit Pom1 kinase. Consistent with the observed interaction between Win1 and Wsh3, cells lacking Wsh3 or Tea1 show more severe cell-polarity defects under osmolarity and heat-stress stimuli that are known to activate the stress MAPK cascade. Furthermore, mutants of the stress MAPK also exhibit cell-polarity defects when exposed to the same stress. CONCLUSIONS: Wsh3/Tea4 is an essential component of the Tea1 cell-end complex. In addition to its role in bipolar growth during the normal cell cycle, the Wsh3-Tea1 complex, together with the stress-signaling MAPK cascade, contributes to cell-polarity maintenance under stress conditions.     

Induction of callus from a metal hypertolerant fern, Athyrium yokoscense, and evaluation of its cadmium tolerance and accumulation capacity

The callus of a metal hypertolerant fern, Athyrium yokoscense, was induced from the spores generated on a small sectioned frond in vitro. The callus grew vigorously with the periodical medium change, especially in a liquid culture. When the callus and regenerated tissues were exposed to Cd, every tissue tolerated at least 1 mM Cd for >1 month. These tissues accumulated high levels of Cd (maximum 3.3 mg g^–1 dry weight in roots) in accordance with the Cd concentration of the medium, and the Cd concentrations of all parts, except roots, were at a similar level. The data suggest that the Cd tolerance of this fern is basically independent of the plant parts and the developmental stages, although the accumulation ability is higher in roots than in the other plant parts.     

Tongue pressure against hard palate during swallowing in post-stroke patients

Objectives: The tongue plays an important role in swallowing by contacting the palate. The aim of the present study was to investigate the characteristics of tongue pressure production during swallowing in post‐stroke patients using a newly developed sensor sheet. Materials and methods: Ten post‐stroke inpatients with hemiplegia and five healthy volunteers participated in this study. Magnitude of tongue pressure during a dry swallow was measured using a newly developed sensor sheet comprising five sensors applied directly to the palate or to the palatal surface of a maxillary denture using denture adhesive. Swallowing ability was evaluated by measuring the time taken to swallow 30 ml of water. The magnitude of tongue pressure was compared between the post‐stroke patients and healthy subjects as well as between each measuring point in both groups. The relationship between tongue pressure and swallowing ability and that between tongue pressure and state of occlusal support were also examined. Results: The magnitude of tongue pressure in the post‐stroke patients was smaller than that of the healthy subjects at the measuring points along the median line (Welch test, p < 0.05), larger in the non‐paralysed side than in the paralysed side (two‐way anova , p < 0.05), and was influenced by swallowing ability and occlusal support (Welch test, p < 0.05). Conclusions: Measurement of the magnitude of tongue pressure shows promise as a simple, non‐invasive and quantitative method by which tongue activity in post‐stroke patients, in whom swallowing ability is a concern, could be evaluated.     

Studies on the amount of aluminum and calcium in urine following aluminum administration with and without amino acids

The evidence implicating aluminum as a neurotoxin is mounting. Research with animals and humans has linked aluminum with neuro-cognitive dysfunction and, in some cases, death. Although the relationship between aluminum intake and Alzheimer's disease is still unclear, some experts have recently issued a strong warning that human exposure to aluminum should be limited. The results indicate that the amount of aluminum decreased in the urine in mice that were administered glycine or glutamic acid together with aluminum ion. In the mice in which the amounts of aluminum decreased in the urine, the amount of calcium conversely increased.     

Blockade of neutrophil elastase attenuates severe liver injury in hepatitis B transgenic mice

Serine proteinases produced by polymorphonuclear neutrophils play important roles in neutrophil-mediated tissue injury at inflammatory sites. Although neutrophil recruitment to the liver has been shown to be involved in the exacerbation of liver inflammation, the function of neutrophil elastase (NE) in liver injury remains unclear. Here, we found that administration of an NE inhibitor (NEI) reduced serum alanine aminotransferase (sALT) activity and inflammatory cell infiltration into the liver from 8 to 24 h after injection of antigen-specific cytotoxic T lymphocytes (CTLs) into hepatitis B virus transgenic mice. Furthermore, the NEI treatment reduced the expressions of inflammatory cytokines and chemokines in the liver and tumor necrosis factor alpha production by macrophages. In addition, the NEI treatment suppressed the mRNA expressions of CC chemokine ligand 3 (CCL-3), CCL-4, and macrophage inflammatory protein 2 (MIP-2) in neutrophils in the liver at 8 h after the CTL injection. In support of these results, we confirmed that administration of anti-CCL-3, anti-CCL-4, and anti-MIP-2 monoclonal antibodies suppressed sALT activity and leukocyte migration into the liver. In conclusion, the present results suggest that NE contributes to the early step of the inflammatory cascade in acute viral hepatitis and that NEIs may have potential as therapeutic drugs against acute severe viral hepatitis.     

Human herpesvirus 6 open reading frame U14 protein and cellular p53 interact with each other and are contained in the virion

A mass spectroscopic analysis of proteins from human herpesvirus 6 (HHV-6)-infected cells showed that the HHV-6 U14 protein coimmunoprecipitated with the tumor suppressor p53. The binding of U14 to p53 was verified by coimmunoprecipitation experiments in both Molt-3 cells infected with HHV-6 and 293 cells cotransfected with U14 and p53 expression vectors. Indirect immunofluorescence assays (IFAs) showed that by 18 h postinfection (hpi) U14 localized to the dot-like structures observed in both the nucleus and cytoplasm where p53 was partly accumulated. Despite Northern blotting evidence that U14 follows late kinetics, the U14 protein was detected immediately after infection (at 3 hpi) by IFA. In addition, by Western blotting, U14 was detected at 0 hpi or in the presence of cycloheximide which completely abolished the expression of IE1 protein. In addition to U14, p53 was detected at 0 hpi although it was not detected in mock-infected cells. Furthermore, both U14 and p53 were clearly detected in the viral particles by Western blotting and immunoelectron microscopy, supporting the idea that U14 and p53 are incorporated into virions. Our study provides the first evidence of the incorporation of cellular p53 into viral particles and suggests that p53 may play an important role in viral infection.     

In vivo cleavage of alpha2,6-sialyltransferase by Alzheimer beta-secretase

beta-Site amyloid precursor protein-cleaving enzyme 1 (BACE1) is a membrane-bound aspartic protease that cleaves amyloid precursor protein to produce a neurotoxic peptide, Abeta, and is implicated in triggering the pathogenesis of Alzheimer disease. We previously reported that BACE1 cleaved rat beta-galactoside alpha2,6-sialyltransferase (ST6Gal I) that was overexpressed in COS cells and that the NH(2) terminus of ST6Gal I secreted from the cells (E41 form) was Glu(41). Here we report that BACE1 gene knock-out mice have one third as much plasma ST6Gal I as control mice, indicating that BACE1 is a major protease which is responsible for cleaving ST6Gal I in vivo. We also found that BACE1-transgenic mice have increased level of ST6Gal I in plasma. Secretion of ST6Gal I from the liver into the plasma is known to be up-regulated during the acute-phase response. To investigate the role of BACE1 in ST6Gal I secretion in vivo, we analyzed the levels of BACE1 mRNA in the liver, as well as the plasma levels of ST6Gal I, in a hepatopathological model, i.e. Long-Evans Cinnamon (LEC) rats. This rat is a mutant that spontaneously accumulates copper in the liver and incurs hepatic damage. LEC rats exhibited simultaneous increases in BACE1 mRNA in the liver and in the E41 form of the ST6Gal I protein, the BACE1 product, in plasma as early as 6 weeks of age, again suggesting that BACE1 cleaves ST6Gal I in vivo and controls the secretion of the E41 form.