A role for nickel-iron cofactors in biological carbon monoxide and carbon dioxide utilization

Ni-Fe containing enzymes are involved in the biological utilization of carbon monoxide, carbon dioxide, and hydrogen. Interest in these enzymes has increased in recent years due to hydrogen fuel initiatives and concerns over development of new methods for CO2 sequestration. One Ni-Fe enzyme called carbon monoxide dehydrogenase (CODH) is a key player in the global carbon cycle and carries out the interconversion of the environmental pollutant CO and the greenhouse gas CO2. The Ni-Fe center responsible for this important chemistry, the C-cluster, has been the source of much controversy, but several recent structural studies have helped to direct the field toward a unifying mechanism. Here we summarize the current state of understanding of this fascinating metallocluster.     

The 3' UTR variants in the GRP78 are not associated with overall survival in resectable hepatocellular carcinoma

Background

Elevated glucose-regulated protein 78 (GRP78) levels in tissues have been known to be related with poor prognosis in hepatocellular carcinoma (HCC) patients. Though the variants in the 3′ untranslated region (UTR) of GRP78 gene were not associated with HCC risk, we wonder whether these polymorphisms affect survival of HCC patients.

Methodology/Principal Findings

Blood samples of HCC patients were maintained in our specimen bank between 1996 to 2003. DNA from 576 unrelated and resectable patients with HCC was typed for rs16927997 (T>C), rs1140763 (T>C) and rs12009 (T>C) by TaqMan assays. The Kaplan-Meier method and log-rank test were used to estimate overall survival. Linkage disequilibrium (LD) analysis identified a total of 3 haplotypes and 6 diplotypes in this region. The distribution of haplotype was not related to the clinical characteristics. Univariate analysis showed that the allele, genotype, haplotype and diplotype did not effect the survival. None of the clinical features show a significant association (P _correced>0.05) with overall patient outcome in multiple comparisons.

Conclusions/Significance

There is no noteworthy influence of 3′ UTR variants in the GRP78 on prognosis of resectable HCC in the Chinese population.     

Identification of reference genes for normalization of gene expression in thoroughbred and Jeju native horse(Jeju pony) tissues

Quantitative analysis of horse gene expression profiles under diverse experimental conditions is limited by the lack of reliable reference genes for normalization of mRNA levels. Therefore, in this study, the expression of potential reference genes was compared between thoroughbred and Jeju native horse (Jeju pony). We compared the expression of nine genes by quantitative real-time RT-PCR in fourteen tissues between the two horse breeds and analyzed their stability using the geNorm and NormFinder programs. The data obtained in this study suggest that the UBB gene could serve as a reference gene in gene expression analysis of thoroughbred and Jeju native horses.     

Identifying Hendra virus diversity in pteropid bats

Hendra virus (HeV) causes a zoonotic disease with high mortality that is transmitted to humans from bats of the genus Pteropus (flying foxes) via an intermediary equine host. Factors promoting spillover from bats to horses are uncertain at this time, but plausibly encompass host and/or agent and/or environmental factors. There is a lack of HeV sequence information derived from the natural bat host, as previously sequences have only been obtained from horses or humans following spillover events. In order to obtain an insight into possible variants of HeV circulating in flying foxes, collection of urine was undertaken in multiple flying fox roosts in Queensland, Australia. HeV was found to be geographically widespread in flying foxes with a number of HeV variants circulating at the one time at multiple locations, while at times the same variant was found circulating at disparate locations. Sequence diversity within variants allowed differentiation on the basis of nucleotide changes, and hypervariable regions in the genome were identified that could be used to differentiate circulating variants. Further, during the study, HeV was isolated from the urine of flying foxes on four occasions from three different locations. The data indicates that spillover events do not correlate with particular HeV isolates, suggesting that host and/or environmental factors are the primary determinants of bat-horse spillover. Thus future spillover events are likely to occur, and there is an on-going need for effective risk management strategies for both human and animal health.     

Testing Equality between Two Diagnostic Procedures in Paired‐Sample Ordinal Data

When a new diagnostic procedure is developed, it is important to assess whether the diagnostic accuracy of the new procedure is different from that of the standard procedure. For paired‐sample ordinal data, this paper develops two test statistics for testing equality of the diagnostic accuracy between two procedures without assuming any parametric models. One is derived on the basis of the probability of correctly identifying the case for a randomly selected pair of a case and a non‐case over all possible cutoff points, and the other is derived on the basis of the sensitivity and specificity directly. To illustrate the practical use of the proposed test procedures, this paper includes an example regarding the use of digitized and plain films for screening breast cancer. This paper also applies Monte Carlo simulation to evaluate the finite sample performance of the two statistics developed here and notes that they can perform well in a variety of situations. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Efficient and seamless DNA recombineering using a thymidylate synthase A selection system in Escherichia coli

λ-Red system-based recombinogenic engineering is a powerful new method to engineer DNA without the need for restriction enzymes or ligases. Here, we report the use of a single selectable marker to enhance the usefulness of this approach. The strategy is to utilize the thymidylate synthase A (thyA) gene, which encodes an enzyme involved in the synthesis of thymidine 5′-triphosphate, for both positive and negative selection. With this approach, we successfully created point mutations in plasmid and bacterial artificial chromosome (BAC) DNA containing the mouse Col10a1 gene. The results showed that the thyA selection system is highly efficient and accurate, giving an average of >90% selection efficiency. This selection system produces DNA that is free from permanent integration of unwanted sequences, thus allowing unlimited rounds of modifications if required.     

Tetrahydroisoquinoline 1-carboxamides as growth hormone secretagogues

Several novel series of tetrahydroisoquinoline 1-carboxamides were prepared and shown to be potent growth hormone (GH) secretagogues. Among them, carbamate 12a-E2 displays excellent in vivo activity by increasing plasma GH 10-fold in an anesthetized IV rat model.