Epimorphin is involved in differentiation of rat hepatic stem-like cells through cell-cell contact

Epimorphin, a mesenchymal cell surface-associated molecule, is detected on hepatic stellate cells (HSCs) in the liver. Here, we show the involvement of epimorphin in differentiation of rat hepatic stem-like cells (HSLCs) through contact with HSCs. HSLCs, isolated from adult rats, cultured in stellate cell-conditioned medium had no phenotypic and morphological changes, whereas HSLCs co-cultured with HSCs expressed albumin, transferrin, and tyrosine aminotransferase. An anti-epimorphin antibody inhibited hepatocytic differentiation of HSLCs in co-culture. Furthermore, epimorphin induced mRNA expression of albumin, transferrin, tyrosine aminotransferase, and gamma-glutamyl transpeptidase with decrease of c-kit and musashi-1. Morphologically, HSLCs piled up when co-cultured with HSCs, which was dramatically inhibited by an anti-epimorphin antibody. HSLCs contact with epimorphin started piling up, changed their shape from flat to cuboidal, and subsequently developed bile-canaliculi-like structures. In conclusion, epimorphin is a factor that induces differentiation of hepatic stem-like cells through epithelial-mesenchymal cell contact.     

Small molecule pharmacological chaperones: From thermodynamic stabilization to pharmaceutical drugs

A great deal of attention has been paid to so-called amyloid diseases, in which the proteins responsible for the cell death and resultant diseases undergo conformational changes and aggregate in vivo, although whether aggregate formation is the cause or the result of the cell death is controversial. Recently, an increasing attention is given to protein folding diseases tightly associated with mutations. These mutations result in temperature-dependent misfolding and hence inactivation of the proteins, leading to loss of function, at physiological temperature; at low so-called permissive temperatures, the mutant proteins correctly fold and acquire functional structure. Alternatively, activation can be induced by use of osmolytes, which restores the folding of the mutant proteins and hence are called chemical chaperones. The osmolytes are compatible with macromolecular function and do stabilize the native protein structure. However, chemical chaperones require high concentrations for effective folding of mutant proteins and hence are too toxic in in-vivo applications. This limitation can be overcome by pharmacological chaperones, whose functions are similar to the chemical chaperones, but occur at much lower concentrations, i.e., physiologically acceptable concentrations. Although the research and clinical importance of pharmacological chaperones has been emphasized, the initial and central concept of osmolytes is largely ignored. Here we attempt to bridge the concept of osmolytes to applications of pharmacological chaperones.     

Geospatial analysis of the associations between environmental contamination with livestock feces and children with chronic fascioliasis in the Anta province of Cusco,Peru

Fasciola hepatica is a neglected parasitic infection with significant human health and livestock industry impact. The Andean Altiplano harbors an estimated 50% of the Fasciola’s world infection burden. There is scarce data regarding the spatial associations between different Fasciola hosts. In this project, we aimed to determine the geospatial relationships between Fasciola eggs passed in feces of different livestock species and the risk of infection among each household as a unit. We used data from a cross-sectional study evaluating children and livestock feces for Fasciola infection around households in three districts of Anta province, in the Cusco region of Peru. Each sample was geographically tagged and evaluated for fascioliasis using microscopy methods. A total of 2070 households were included, the median age was 9.1 years (6.7–11.8), 49.5% were female, and 7.2% of the households had at least one infected child. A total of 2420 livestock feces samples were evaluated. The infection rate in livestock samples was 30.9%. The highest infection rate was found in sheep with 40.8%, followed by cattle (33.8%), and swine (26.4%). The median distance between a household with an infected child to a positive animal sample was 44.6 meters (IQR 14.7–112.8) and the distance between a household with no infected children to a positive animal sample was 62.2 meters (IQR 18.3–158.6) (p = 0.025). The multivariable logistic regression adjusted by presence of poor sanitation, unsafe water consumption, altitude, and presence of multiple infected children per household demonstrated an association between household infection and any cattle feces at a 50 meters radius (Uninfected: OR 1.42 (95%CI 1.07–1.89), p = 0.017. Infected: OR 1.89 (95%CI 1.31–2.73), p = 0.001), positive cattle feces at a 100 meters radius (OR 1.35 (95% CI 1.08–1.69), p = 0.008), and negative cattle feces at a 200 meters radius (OR 1.08 (95% CI 1.01–1.15), p = 0.022). We identified potential hot and cold spots for fascioliasis in the Anta province. An association between environmental contamination with feces from different livestock species and infected children in rural households was found in our study. Local health authorities may apply this strategy to estimate the risk of infection in human populations and apply targeted interventions.     

Regional neural activation defines a gateway for autoreactive T cells to cross the blood-brain barrier

Although it is believed that neural activation can affect immune responses, very little is known about the neuroimmune interactions involved, especially the regulators of immune traffic across the blood-brain barrier which occurs in neuroimmune diseases such as multiple sclerosis (MS). Using a mouse model of MS, experimental autoimmune encephalomyelitis, we show that autoreactive T?cells access the central nervous system via the fifth lumbar spinal cord. This location is defined by IL-6 amplifier-dependent upregulation of the chemokine CCL20 in associated dorsal blood vessels, which in turn depends on gravity-induced activation of sensory neurons by the soleus muscle in the leg. Impairing soleus muscle contraction by tail suspension is sufficient to reduce localized chemokine expression and block entry of pathogenic T?cells at the fifth lumbar cord, suggesting that regional neuroimmune interactions may offer therapeutic targets for a variety of neurological diseases.     

Photophysiological responses of phytobenthic communities to the strong light and UV in Antarctic shallow lakes

Light environment, community structure, pigments, and photophysiological properties of mat-forming phytobenthos were studied in four shallow Antarctic lakes in 2007 at maximum water depths of 1.7–2.5 m. All lakes were oligotrophic, and water transparencies were high, enabling 45–60% of photosynthetically active radiation (PAR, 400–700 nm) and 20–40% of ultraviolet radiation (300–400 nm) to reach the lake beds. Phytobenthic mats were dominated by cyanobacteria and green algae. Little PAR_L (500–700 nm) penetrated through the firm mat in the shallowest lake, while in the other lakes more (>20%) PAR_L got through the mats to the subsurface mat layers. Photochemical activities indicated almost no photoinhibition but low photosynthetic efficiency in all mat surface layers. Non-photochemical quenching was rarely detected, suggesting excess energy dissipation may not be efficient in the UV-rich environment. There was a positive correlation between photo-protective substances and incident radiation in the mats, and an inverse correlation between such substances and photochemical efficiency, suggesting that the phytobenthos survive by changing a light-protection/utilization balance. The communities under strong UV-B and PAR had firm mat textures and were characterized by high UV/photo-protective substance ratios that make them less transparent. Maximum relative electron transportation rates (rETR_max) and photochemical efficiencies, however, were low, possibly because the protective substances prevent efficient light usage. In contrast, communities under mild light were characterized by lower substance ratios and softer textures, while rETR_max values and photochemical efficiencies were greater. The phytobenthic mat surface seems to act as a filter for strong and harmful light, typically penetrating through the clear water of Antarctic lakes, and produces a milder light environment for the subsurface mat organisms.     

Enhanced Selection of High Affinity DNA-Reactive B Cells Following Cyclophosphamide Treatment in Mice

A major goal for the treatment of patients with systemic lupus erythematosus with cytotoxic therapies is the induction of long-term remission. There is, however, a paucity of information concerning the effects of these therapies on the reconstituting B cell repertoire. Since there is recent evidence suggesting that B cell lymphopenia might attenuate negative selection of autoreactive B cells, we elected to investigate the effects of cyclophosphamide on the selection of the re-emerging B cell repertoire in wild type mice and transgenic mice that express the H chain of an anti-DNA antibody. The reconstituting B cell repertoire in wild type mice contained an increased frequency of DNA-reactive B cells; in heavy chain transgenic mice, the reconstituting repertoire was characterized by an increased frequency of mature, high affinity DNA-reactive B cells and the mice expressed increased levels of serum anti-DNA antibodies. This coincided with a significant increase in serum levels of BAFF. Treatment of transgene-expressing mice with a BAFF blocking agent or with DNase to reduce exposure to autoantigen limited the expansion of high affinity DNA-reactive B cells during B cell reconstitution. These studies suggest that during B cell reconstitution, not only is negative selection of high affinity DNA-reactive B cells impaired by increased BAFF, but also that B cells escaping negative selection are positively selected by autoantigen. There are significant implications for therapy.     

Clarification of pathway-specific inhibition by Fourier transform ion cyclotron resonance/mass spectrometry-based metabolic phenotyping studies

We have developed a metabolic profiling scheme based on direct-infusion Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). The scheme consists of: (1) reproducible data collection under optimized FT-ICR/MS analytical conditions; (2) automatic mass-error correction and multivariate analyses for metabolome characterization using a newly developed metabolomics tool (DMASS software); (3) identification of marker metabolite candidates by searching a species-metabolite relationship database, KNApSAcK; and (4) structural analyses by an MS/MS method. The scheme was applied to metabolic phenotyping of Arabidopsis (Arabidopsis thaliana) seedlings treated with different herbicidal chemical classes for pathway-specific inhibitions. Arabidopsis extracts were directly infused into an electrospray ionization source on an FT-ICR/MS system. Acquired metabolomics data were comprised of mass-to-charge ratio values with ion intensity information subjected to principal component analysis, and metabolic phenotypes from the herbicide treatments were clearly differentiated from those of the herbicide-free treatment. From each herbicide treatment, candidate metabolites representing such metabolic phenotypes were found through the KNApSAcK database search. The database search and MS/MS analyses suggested dose-dependent accumulation patterns of specific metabolites including several flavonoid glycosides. The metabolic phenotyping scheme on the basis of FT-ICR/MS coupled with the DMASS program is discussed as a general tool for high throughput metabolic phenotyping studies.     

Intramolecular acyl migration and enzymatic hydrolysis of a novel monoacylated ascorbic acid derivative, 6-O-dodecanoyl-2-O-α-d-glucopyranosyl-l-ascorbic acid

A stable ascorbic acid derivative, 2-O-α-d-glucopyranosyl-l-ascorbic acid (AA-2G), exhibits vitamin C activity in vitro and in vivo after enzymatic hydrolysis to ascorbic acid. AA-2G has been approved by the Japanese Government as a quasi-drug principal ingredient in skin care and as a food additive. In order to achieve efficient action as an ascorbic acid source, a pro-vitamin C agent, on a variety of cells or tissues, we have synthesized a series of monoacyl AA-2G derivatives. Our previous studies indicate that a series of the derivatives is a readily available source of AA activity in vitro and in vivo, and suggested that intramolecular acyl migration of the derivatives might have occurred in a neutral aqueous solution. In this study, intramolecular acyl migration and enzymatic hydrolysis of a monoacyl AA-2G derivative, 6-O-dodecanoyl-2-O-α-d-glucopyranosyl-l-ascorbic acid (6-sDode-AA-2G), were investigated. 6-sDode-AA-2G underwent an intramolecular acyl migration to yield ca. 10% of an isomer in neutral aqueous solutions, and the acyl-migrated isomer was isolated and characterized as 5-O-dodecanoyl-2-O-α-d-glucopyranosyl-l-ascorbic acid (5-sDode-AA-2G). In some tissue homogenates from guinea pigs as well as in neutral aqueous solutions, 6-sDode-AA-2G underwent partial acyl migration to give 5-sDode-AA-2G. 6-sDode-AA-2G and the resulting 5-sDode-AA-2G were predominantly hydrolyzed with esterase to AA-2G and then with α-glucosidase to ascorbic acid in the tissue homogenates. The results will provide a further basis for its use as an ingredient in skin care, as an effective pharmacological agent and as a promising food additive.