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Background Vitamin K (VK) is a co-factor of the γ-glutamyl carboxylase that catalyzes the conversion of glutamate residues to γ-carboxyglutamate in VK-dependent proteins. The carboxylation reaction imparts the essential calcium-binding residues for the biological function of several proteins involved in the process of coagulation and bone metabolism. VK deficiency is frequently encountered in newborns and can lead to fatal hemorrhagic complications. This review describes and discusses the clinical application of VK deficiency testing. Methods References and data were researched in PubMed and reviewed. Results In adults, VK deficiency is associated with uncontrolled bleeding, liver dysfunction, osteoporosis, and coronary diseases. An improved understanding of the role of VK deficiency in health and illness can be achieved by setting a gold-standard in the inter-laboratory estimations of VK. However, conventional methods used to measure the VK deficiency based upon the coagulation time lack sensitivity and specificity. Recently, the alterations in proteins induced by VK absence or antagonism (PIVKA) have proven to be suitable biomarkers for detecting VK deficiency. The measurement of PIVKA-II exhibits an enhanced sensitivity and specificity in comparison to other methods conventionally used for the assessment of VK deficiency in newborns and adults. Conclusions PIVKA-II could potentially be employed as an effective biomarker in the diagnosis of VK deficiency.

Population-based screening for CGG-repeat expansions in the fragile X mental retardation 1 (FMR1) gene that cause fragile X syndrome can now be performed more cost-effectively and simply by combining direct triplet-primed PCR (dTP-PCR) with melting curve analysis (MCA). We have now performed a detailed technical validation to define the operational parameters for achieving robust and reliable performance of the FMR1 dTP-PCR MCA assay. We compared the assay's performance on 2 real-time PCR platforms and determined its analytic sensitivity and specificity. We also assessed the assay's performance on DNA isolated from different sources, the effect of differences in CGG-repeat length and AGG-interruption pattern on melt peak temperature (Tm), and the effect of common substances found in DNA solutions on Tms. The assay performed well in distinguishing normal from expansion-carrying samples. The assay had detection sensitivity down to 1 ng and an analytical specificity beyond 150 ng. In addition to peripheral blood DNA, analysis could also be performed on DNA from saliva, buccal swabs, and dried blood spots. Salt increased Tms, glycogen contamination had minimal effect, whereas AGG interruptions lowered Tms. The FMR1 dTP-PCR MCA screening assay is highly sensitive and specific, performs well using DNA from different sources, and is robust and reproducible when reagent concentrations are maintained across all tested samples.

NF-κB controlled transcriptional regulation plays a central role in inflammatory and immune responses. Currently understanding about NF-κB activation mechanism emphasizes IκB-tethered complex inactivation in the cytoplasm. In the case of NF-κB activation, IκB phosphorylation leads to its degradation followed by NF-κB relocating to the nuclear and transactivation of NF-κB targeted genes. Pre-translational mechanism mediated NF-κB activation remains poorly understood. In this study, we studied NF-κB pre-translational regulation by performed a series of data-base mining using experimental data-based NCBI Unigene EST profile database, Gene Expression Omnibus (GEO) database, Transcription Element Search System (TESS) database, AceView database, Epigenomics databases, and TargetScan software. We reported the following findings: 1) NF-κB family/signaling genes are differentially expressed in 20 human and 19 mouse tissues; 2) Heart and vessels are the inflammation privilege tissues and less easy to be inflamed because lacking of NF-κB signaling key molecular expression; 3) NF-κB activation is readily induced by cardiovascular disease risk factors oxidized-LDL and pro-inflammatory cytokines in endothelial cells; 4) Transcription factors C/EBPs and NF-κB have higher binding site frequencies in the promoters of Pro-inflammatory cytokine-induced NF-κB genes; 5) Most NF-κB signaling genes have multiple alternative promoters and alternatively spliced isoforms; 6) NF-κB family genes have DNA methylation characteristics and may be subjected to epigenetic regulation; 7) 27 out of 38 NF-κB signaling gene can be regulated by microRNAs, with the binding qualities to mRNA targets being equivalent to what have been approved experimentally. Our findings provide important insight in the mechanism of NF-κB activation, which may contribute to cardiovascular disease, inflammatory diseases and immunological disorders.

NF-κB-controlled transcriptional regulation plays a central role in inflammatory and immune responses. Currently, understanding about NF-κB activation mechanism emphasizes IκB-tethered complex inactivation in the cytoplasm. In the case of NF-κB activation, IκB phosphorylation leads to its degradation, followed by NF-κB relocation to the nucleus and trans-activation of NF-κB-targeted genes. Pretranslational mechanism mediated NF-κB activation remains poorly understood. In this study, we investigated NF-κB pretranslational regulation by performing a series of database mining analyses and using six large national experimental databases (National Center of Biotechnology Information UniGene expressed sequence tag profile database, Gene Expression Omnibus database, Transcription Element Search System database, AceView database, and Epigenomics database) and TargetScan software. We reported the following findings: 1) NF-κB-signaling genes are differentially expressed in human and mouse tissues; 2) heart and vessels are the inflammation-privileged tissues and less easy to be inflamed because lacking in key NF-κB-signaling molecular expression; 3) NF-κB-signaling genes are induced by cardiovascular disease risk factors oxidized phospholipids and proinflammatory cytokines in endothelial cells; 4) transcription factors CCAAT/enhancer-binding proteins and NF-κB have higher binding site frequencies in the promoters of proinflammatory cytokine-induced NF-κB genes; 5) most NF-κB-signaling genes have multiple alternative promoters and alternatively spliced isoforms; 6) NF-κB family genes can be regulated by DNA methylation; and 7) 27 of 38 NF-κB-signaling genes can be regulated by microRNAs. Our findings provide important insight into the mechanism of NF-κB activation, which may contribute to cardiovascular disease, inflammatory diseases, and immunological disorders.

Herpes keratitis is one of the most severe pathologies associated with the herpes simplex virus-type 1 (HSV-1). Herpes keratitis is currently the leading cause of both cornea-derived and infection-associated blindness in the developed world. Typical presentation of herpes keratitis includes infection of the corneal epithelium and sometimes the deeper corneal stroma and endothelium, leading to such permanent corneal pathologies as scarring, thinning, and opacity 1. Corneal HSV-1 infection is traditionally studied in two types of experimental models. The in vitro model, in which cultured monolayers of corneal epithelial cells are infected in a Petri dish, offers simplicity, high level of replicability, fast experiments, and relatively low costs. On the other hand, the in vivo model, in which animals such as rabbits or mice are inoculated directly in the cornea, offers a highly sophisticated physiological system, but has higher costs, longer experiments, necessary animal care, and a greater degree of variability. In this video article, we provide a detailed demonstration of a new ex vivo model of corneal epithelial HSV-1 infection, which combines the strengths of both the in vitro and the in vivo models. The ex vivo model utilizes intact corneas organotypically maintained in culture and infected with HSV-1. The use of the ex vivo model allows for highly physiologically-based conclusions, yet it is rather inexpensive and requires time commitment comparable to that of the in vitro model.

This report is a review of the development testing of mission-critical software for the U.S. Army Prophet Engineering and Manufacturing Development (EMD) System and the Prophet Block I System. Prophet is a Division-Level ground based electronic surveillance system, which provides protection in a direct support role to the maneuver brigade; either stationary, or while on the move. The system monitors and exploits signals of interest and determines the area of signal origin by providing direction finding and line-of-bearing in the frequency range of 20 Megahertz to 2000 Megahertz. Prophet is operated in either the dismounted or mounted mode. In the dismounted mode Prophet is man-packed with a portable antenna. In the mounted mode Prophet is installed in an equipment enclosure carried on a High Mobility Multi-Purpose Wheeled Vehicle with the antennas attached to a retractable mast.