Identification of fish species by 5S rRNA gene amplification

The 5S ribosomal RNA is a very suitable target for easy, rapid and inexpensive fish species identification due to its structure, consisting of a conserved region followed by a species-specific noncoding region called ‘nontranscribed spacer’. We have exploited this species-specificity in length and sequence to discriminate among fish species which can be subjected to substitution in the fish markets. After sequencing and alignment of the corresponding portions of the 5S rRNAs of different fish species, we have designed the primer pairs necessary for PCR amplification on the DNA traits which most diverged and a primer pair on conserved regions. Our results have shown the feasibility, simplicity and reliability of the proposed approach for the detection of mislabelling or fraudulent substitution of fish species.     

Report from the Maryland Epidemiology Schizophrenia Linkage Study: no evidence for linkage between schizophrenia and a number of candidate and other genomic regions using a complex dominant model

Our collaborative group has undertaken a linkage study of schizophrenia, using a systematic sample of patients admitted to Maryland hospitals. An initial sample of 39 families, each having two or more affecteds, was available for genotyping candidate genes, candidate regions, and highly polymorphic markers randomly distributed throughout the genome. We used a single complex dominant model (with a disease gene frequency of 0.005 and age-dependent penetrance for affected phenotype: for under 35, penetrance = .45; for 35 and older, penetrance = .85). We report here 130 markers, which met the exclusion criteria of LOD score < -2.00 at theta > 0.01 in at least 10 informative families, and no evidence for heterogeneity. We also report here markers that were tested as candidates for linkage to the schizophrenic phenotype. They were selected based on the following criteria: a) proximity to reported chromosomal rearrangements (both 5q and 11q), b) suggestions of linkage from other families (5q), or c) presence of a candidate gene (5q, 11q, 3q: Dopamine receptors 1, 2, and 3, respectively). We also tested for mutations of codon 717 in exon 17 of the amyloid precursor protein (APP) gene and were unable to detect the C to T substitution in our schizophrenic group.     

Autonomous growth in serum-free medium and production of hepatocellular carcinomas by differentiated hepatocyte lines that overexpress transforming growth factor alpha 1

Transforming growth factor alpha (TGF-alpha) is a polypeptide closely associated with hepatocyte proliferation in vivo and in vitro. In order to investigate the mechanisms by which TGF-alpha contributes to hepatocyte replication and transformation, we isolated hepatocytes from mice bearing a human TGF-alpha transgene and examined their growth properties and gene expression in defined, serum-free culture. The transgenic hepatocytes continued to overexpress human TGF-alpha mRNA and peptide, and were able to proliferate without exogenous growth factors in primary culture, in contrast to nontransgenic mouse hepatocytes. In short-term culture the transgenic hepatocytes underwent 1 wave of DNA replication at 72-96 h in culture before senescing, similar to nontransgenic hepatocytes supplemented with epidermal growth factor. Constitutive expression of TGF-alpha rendered the transgenic hepatocytes unresponsive to further growth stimulation by exogenous TGF-alpha, as well as other mitogens such as epidermal growth factor and hepatocyte growth factor. However, it did not alter their sensitivity to growth inhibition by TGF beta 1, 2 and 3. The addition of nicotinamide to the culture medium enabled both transgenic and epidermal growth factor-supplemented normal hepatocytes to replicate repeatedly and survive for > or = 2 months in primary culture while maintaining differentiated traits. From these long-term primary cultures of transgenic and nontransgenic hepatocytes, we established immortalized cell lines (designated TAMH and NMH lines, respectively). Both lines continued to express differentiated adult hepatocytic markers such as albumin, alpha-1-antitrypsin, transferrin, and connexin 26 and 32 mRNAs, but also expressed mRNAs for the oncofetal markers alpha-fetoprotein and insulin-like growth factor II. Unlike the near-diploid NMH hepatocyte line, the transgenic TAMH hepatocyte line was quasi-tetraploid, strongly expressed human TGF-alpha mRNA, and was highly tumorigenic in nude mice. Well-differentiated hepatocellular carcinomas developed in nude mice given injections of the TAMH line, and these appeared similar to the primary liver tumors seen in TGF-alpha transgenic mice with regard to histology and strong expression of mouse and human TGF-alpha, insulin-like growth factor II, and alpha-fetoprotein mRNAs. Our data show that TGF-alpha overexpression causes autonomous hepatocyte proliferation and contributes to neoplasia but that additional cellular alterations must occur for carcinogenesis. Inappropriate expression of insulin-like growth factor II may constitute one of these steps. The TGF-alpha transgenic mouse hepatocyte line TAMH appears to undergo transformation in a similar manner to that of hepatocytes overexpressing TGF-alpha in vivo, and should serve as an ideal system in which to study hepatocarcinogenesis.     

A digital filter-based approach to the remote condition monitoring of railway turnouts

Railway operations in Europe have changed dramatically since the early 1990s, partly as a result of new European Union Directives. Performance targets have become more and more exacting, due to reductions in state support for railways and the need to increasing traffic. More intensive operations also place greater demands on the hardware of the railway. This is true for both rolling stock and infrastructure subsystems and components, particularly so in the case of the latter where the time available for maintenance is being reduced. The authors of this paper focus on the railway infrastructure, and more specifically on points. These are critical elements whose reliability is key to the operation of the whole system. Using intelligent monitoring systems, it is possible to predict problems and enable quick recovery before component failures disrupt operations. The authors have studied the application of remote condition monitoring to point mechanisms and their operation, and have identified algorithms which may be used to identify incipient failures. In this paper, the authors propose a Kalman filter for the linear discrete data filtering problem encountered when using current sensor data in a point condition monitoring system. The reason for applying Kalman filtering in this study was to increase the reliability of the model presented to the rule-based decision mechanism.     

Thermophiles as Potential Source of Novel Endotoxin Antagonists: the Full Structure and Bioactivity of theLipo‐oligosaccharide from Thermomonas hydrothermalis

Thermomonas hydrothermalis is a Gram‐negative thermophilic bacterium that is able to live at 50 °C. This ability is attributed to chemical modifications, involving those to bacterial cell‐wall components, such as proteins and (glyco)lipids. As the main component of the outer membrane of Gram‐negative bacteria, lipopolysaccharides (LPSs) are exposed to the environment, thus they can undergo structural chemical changes to allow thermophilic bacteria to live at their optimal growth temperature. Furthermore, as one of the major target of the eukaryotic innate immune system, LPS elicits host immune response in a structure‐dependent mode; thus the uncommon chemical features of thermophilic bacterial LPSs might exert a different biological action on the innate immune system—an antagonistic effect, as shown in studies of LPS structure–activity relationship in the ongoing research into antagonist LPS candidates. Here, we report the complete structural and biological activity analysis of the lipo‐oligosaccharide isolated from Thermomonas hydrothermalis, achieved by a multidisciplinary approach (chemical analysis, NMR, MALDI MS and cellular immunology). We demonstrate a tricky and interesting structure combined with a very interesting effect on human innate immunity.     

The Fight against the Influenza A Virus H1N1: Synthesis,Molecular Modeling,and Biological Evaluation of Benzofurazan Derivatives as Viral RNA Polymerase Inhibitors

The influenza RNA polymerase complex, which consists of the three subunits PA, PB1, and PB2, is a promising target for the development of new antiviral drugs. A large library of benzofurazan compounds was synthesized and assayed against influenza virus A/WSN/33 (H1N1). Most of the new derivatives were found to act by inhibiting the viral RNA polymerase complex through disruption of the complex formed between subunits PA and PB1. Docking studies were also performed to elucidate the binding mode of benzofurazans within the PB1 binding site in PA and to identify amino acids involved in their mechanism of action. The predicted binding pose is fully consistent with the biological data and lays the foundation for the rational development of more effective PA–PB1 inhibitors.     

{\alpha^{\prime}}_{\text{H}} -Dicalcium silicate bone cement doped with tricalcium phosphate: characterization,bioactivity and biocompatibility

The influence of phosphorus doping on the properties of $ \alpha^{\prime}_{\text{H}} $ -dicalcium silicate (C₂S) bone cement was analyzed, in addition to bioactivity and biocompatibility. All the cements were composed of a solid solution of TCP in C₂S ( $ \alpha^{\prime}_{\text{H}} $ -C₂S_ss) as the only phase present. The compressive strength ranged from 3.8–16.3 MPa. Final setting times ranged from 10 to 50 min and were lower for cements with lower L/P content. Calcium silicate hydrate was the principal phase formed during the hydration process of the cements. The cement exhibited a moderate degradation and could induce carbonated hydroxyapatite formation on its surface and into the pores. The cell attachment test showed that the $ \alpha^{\prime}_{\text{H}} $ -Ca₂SiO₄ solid solution supported human adipose stem cells adhesion and spreading, and the cells established close contacts with the cement after 24 h of culture. The novel $ \alpha^{\prime}_{\text{H}} $ -C₂S_ss cements might be suitable for potential applications in the biomedical field, preferentially as materials for bone/dental repair.     

Performance evaluation of wavelet-based distributed video coding schemes

In this paper we propose and compare different distributed video coding (DVC) schemes based on the use of the wavelet transform, which naturally allows for spatial and other forms of scalability. In particular, we propose a hybrid encoder which utilizes channel codes, and evaluate its performance in the absence of a feedback channel. The proposed scheme uses statistical models for the estimation of the required bitrate at the encoder. We also propose a scheme that is based on a modulo reduction procedure and does not use channel codes at the receiver/transmitter. These schemes are compared with more conventional coders that do not or only partially exploit the distributed coding paradigm. Experimental results show that the considered schemes have good performance when compared with similar asymmetric video compression schemes, and that DVC can be an interesting option in appropriate scenarios.     

An Investigation of the Electrical Degradation of GaN High-Electron-Mobility Transistors by Numerical Simulations of DC Characteristics and Scattering Parameters

The effects of direct-current (DC) stress on GaN high-electron-mobility transistors (HEMTs) are investigated by means of numerical simulations, by which the creation of an acceptor trap in the AlGaN barrier layer was correlated to the observed experimental degradation. An increase in the trap concentration induces a worsening of the saturated current I _DSS, transconductance g _m, and output conductance g _O. An increase in the length of the trapping region induces a degradation of I _DSS and g _m, but can reduce g _O. Analysis of scattering parameters in the saturation region shows that the cutoff frequency f _T matches the trend of g _m.