Design of microarray probes for virus identification and detection of emerging viruses at the genus level

BACKGROUND: Most virus detection methods are geared towards the detection of specific single viruses or just a few known targets, and lack the capability to uncover the novel viruses that cause emerging viral infections. To address this issue, we developed a computational method that identifies the conserved viral sequences at the genus level for all viral genomes available in GenBank, and established a virus probe library. The virus probes are used not only to identify known viruses but also for discerning the genera of emerging or uncharacterized ones. RESULTS: Using the microarray approach, the identity of the virus in a test sample is determined by the signals of both genus and species-specific probes. The genera of emerging and uncharacterized viruses are determined based on hybridization of the viral sequences to the conserved probes for the existing viral genera. A detection and classification procedure to determine the identity of a virus directly from detection signals results in the rapid identification of the virus. CONCLUSION: We have demonstrated the validity and feasibility of the above strategy with a small number of viral samples. The probe design algorithm can be applied to any publicly available viral sequence database. The strategy of using separate genus and species probe sets enables the use of a straightforward virus identity calculation directly based on the hybridization signals. Our virus identification strategy has great potential in the diagnosis of viral infections. The virus genus and specific probe database and the associated summary tables are available a

Phyllanthus urinaria Induces Apoptosis in Human Osteosarcoma 143B Cells via Activation of Fas/FasL- and Mitochondria-Mediated Pathways

Phyllanthus urinaria (P. urinaria), in this study, was used for the treatment of human osteosarcoma cells, which is one of the tough malignancies with few therapeutic modalities. Herein, we demonstrated that P. urinaria inhibited human osteosarcoma 143B cells growth through an apoptotic extrinsic pathway to activate Fas receptor/ligand expression. Both intracellular and mitochondrial reactive oxygen species were increased to lead to alterations of mitochondrial membrane permeability and Bcl-2 family including upregulation of Bid, tBid, and Bax and downregulation of Bcl-2. P. urinaria triggered an intrinsic pathway and amplified the caspase cascade to induce apoptosis of 143B cells. However, upregulation of both intracellular and mitochondrial reactive oxygen species and the sequential membrane potential change were less pronounced in the mitochondrial respiratory-defective 143Bρ0 cells compared with the 143B cells. This study offers the evidence that mitochondria are essential for the anticancer mechanism induced by P. urinaria through both extrinsic and intrinsic pathways

Transmembrane Domain Interactions and Residue Proline 378 Are Essential for Proper Structure, Especially Disulfide Bond Formation, in the Human Vitamin K-Dependent γ-Glutamyl Carboxylase †

We used recombinant techniques to create a two-chain form (residues 1–345 and residues 346–758) of the vitamin K-dependent γ-glutamyl carboxylase, a glycoprotein located in the endoplasmic reticulum containing five transmembrane domains. The two-chain carboxylase had carboxylase and epoxidase activities similar to those of one-chain carboxylase. In addition, it had normal affinity for the propeptide of factor IX. We employed this molecule to investigate formation of the one disulfide bond in carboxylase, the transmembrane structure of carboxylase, and the potential interactions among the carboxylase’s transmembrane domains. Our results indicate that the two peptides of the two-chain carboxylase are joined by a disulfide bond. Proline 378 is important for the structure necessary for disulfide formation. Results with the P378L carboxylase indicate that noncovalent bonds maintain the two-chain structure even when the disulfide bond is disrupted. As we had previously proposed, the fifth transmembrane domain of carboxylase is the last and only transmembrane domain in the C-terminal peptide of the two-chain carboxylase. We show that the noncovalent association between the two chains of carboxylase involves an interaction between the fifth transmembrane domain and the second transmembrane domain. Results of a homology model of transmembrane domains 2 and 5 suggest that not only do these two domains associate but that transmembrane domain 2 may interact with another transmembrane domain. This latter interaction may be mediated at least in part by a motif of glycine residues in the second transmembrane domain

Measuring Intergroup Forgiveness: The Enright Group Forgiveness Inventory

Until recently, researchers operationalized and measured the psychological construct of forgiveness at the individual, rather than the group, level. Social psychologists started applying forgiveness to groups and examining the role intergroup forgiveness may have in conflict resolution and peace efforts. Initial attempts to define and measure forgiveness at the group level either assumed individual and group capacities were the same, or insufficiently described what intergroup forgiveness meant. We developed a new measure of intergroup forgiveness, and a novel group administration process, that operationalized the construct in a philosophically coherent way. Our conceptualization of intergroup forgiveness was rooted in what groups, as opposed to the individuals who compose them, have the capacity to do. We collected data on the psychometric properties of the measure with 595 participants in three different geographic and cultural settings. We assessed the factor structure, internal consistency, and validity of the measure. We also assessed a novel group-based method of administering the measure to better understand the relationship between group based reports and self-reports of intergroup forgiveness. The factor structure of the measure was supported, and the measure had strong internal consistency, as well as convergent and discriminant validity. The group administration process revealed important group dynamics and was not statistically different than a standard self-report administration; this finding has important implications for research and practice

) The Influence of Low-powered Family LED Lighting on Eyes in Mice Experimental Model

Abstract: Ocular tissue damage because of exposure to visible light has been demonstrated by the results of human and animal studies. The short-wavelength visible light between 430 nm to 500 nm (blue light) is especially associated with retina damage. Recently, new powerful sources and relatively inexpensive blue energy of LED (light emitting diodes) family lamps in home illumination are available. The aim of this study is to investigate the effects of illumination source from the low-powered and the conscious spectrum source of LED family lamps on retina tissues. The illumination source of LED family lamps was analyzed from 300 nm to 800 nm using an UVvisible spectrophotometer. In animal experiments, young adult mice were assigned to expose to family LED light for 2h every day ranging 2 to 4 weeks or light environment using LED family lamps for 39 weeks. After LED light treatment, sections of eyes were stained with hematoxylin and examined using histopathology. The data clearly demonstrated irradiation of the white LED is above 400 nm and is not within the ultraviolet light region. However, the analysis of spectrum distribution demonstrated that the family LED lighting exhibited power-peak at 450 nm is within the blue light region. Histological results showed that the photoreceptor layer is significantly reduced in thickness after 4 weeks of LED exposure 2h every day or LED illuminated environment. This study provides important data regarding the efficacy and safety of LED light in family illumination. It is impossible to consider these degenerative changes are related unavoidably part of their mechanism of action or an avoidable toxic effect

A Protective Mechanism against Antibiotic-Induced Ototoxicity: Role of Prestin

Hearing loss or ototoxicity is one of the major side effects associated with the use of the antibiotics, particularly aminoglycosides (AGs), which are the most commonly used antibiotics worldwide. However, the molecular and cellular events involved in the antibiotic-induced ototoxicity remains unclear. In the present study, we test the possibility that prestin, the motor protein specifically expressed in the basolateral membrane of outer hair cells (OHCs) in the cochlea with electromotility responsible for sound amplification, may be involved in the process of AG-induced apoptosis in OHCs. Our results from both mice model and cultured cell line indicate a previously unexpected role of prestin, in mediating antibiotic-induced apoptosis, the effect of which is associated with its anion-transporting capacity. The observed downregulation of prestin mRNA prior to detectable apoptosis in OHCs and hearing loss in the antibiotic-treated mice is interesting, which may serve as a protective mechanism against hearing loss induced by AGs in the early stage