Construction of a Fibrobacter succinogenes Genomic Map and Demonstration of Diversity at the Genomic Level

The genomic cleavage map of the type strain Fibrobacter succinogenes S85 was constructed. The restriction enzymes AscI, AvrII, FseI, NotI, and SfiI generated DNA fragments of suitable size distribution that could be resolved by pulsed-field gel electrophoresis (PFGE). An average genome size of 3.6 Mb was obtained by summing the total fragment sizes. The linkages between the 15 AscI fragments of the genome were determined by combining two approaches: isolation of linking clones and cross-hybridization of restriction fragments. The genome of F. succinogenes was found to be represented by the single circular DNA molecule. Southern hybridization with specific probes allowed the eight genetic markers to be located on the restriction map. The genome of this bacterium contains at least three rRNA operons. PFGE of the other three strains of F. succinogenes gave estimated genome sizes close to that of the type strain. However, RFLP patterns of these strains generated by AscI digestion are completely different. Pairwise comparison of the genomic fragment distribution between the type strain and the three isolates showed a similarity level in the region of 14.3% to 31.3%. No fragment common to all of these F. succinogenes strains could be detected by PFGE. A marked degree of genomic heterogeneity among members of this species makes genomic RFLP a highly discriminatory and useful molecular typing tool for population studies. Received: 23 October 1996 / Accepted: 31 December 1996     

Using dense locality sampling resolves the subtle genetic population structure of the dispersive fish species Plecoglossus altivelis

In dispersive species with continuous distributions, genetic differentiation between local populations is often absent or subtle and thus difficult to detect. To incorporate such subtle differentiation into management plans, it may be essential to analyse many samples from many localities using adequate numbers of high‐resolution genetic markers. Here, we evaluated the usefulness of dense locality sampling in resolving genetic population structure in the ayu (Plecoglossus altivelis), a dispersive fish important in Japanese inland fisheries. Genetic variability in, and differentiation between, ayu populations around the Japan–Ryukyu Archipelago were investigated in 4746 individuals collected from 120 localities by genotyping 12 microsatellite markers. These individuals represented the two subspecies of ayu, namely the Ryukyuan subspecies (Plecoglossus altivelis ryukyuensis) and both amphidromous and landlocked forms of the nominotypical subspecies (P. a. altivelis) along the archipelago. We successfully detected an absence of genetic differentiation within the landlocked form and subtle but significant differentiation and clear geographic patterns of genetic variation among populations of the amphidromous form, which had been considered genetically homogeneous. This suggests that dense locality sampling effectively resolves subtle differences in genetic population structure, reducing stochastic deviation in the detection of genetic differentiation and geographic patterns in local populations of this dispersive species. Resampling analyses based on empirical data sets clearly demonstrate the effectiveness of increasing the number of locality samples for stable and reliable estimations of genetic fixation indices. The genetic population structure observed within the amphidromous form provides useful information for identifying management or conservation units in ayu.     

Comparative phylogeography of bagrid catfishes in Taiwan

A phylogeographic analysis of two bagrid catfishes in Taiwan was conducted using sequence data from a portion of the mitochondrial DNA (mtDNA) control region. For Pseudobagrus brevianalis, which is most probably endemic to Taiwan, a total of eight haplotypes were detected in 189 specimens from nine river systems covering its entire distribution range, from northern to central western areas of the island. Obvious genetic differentiation was observed among its populations (average F _ST = 0.753); in particular, the northernmost Tamsui River population was fixed for a single endemic haplotype. Nested clade phylogeographic analysis (NCPA) indicated that the dispersal center of mtDNA was the area around the Touchien River and Holong River, north to the Miaoli Plateau, in northwestern Taiwan, suggesting both northward and southward dispersal in this species. There was no evidence for the validity of P. taiwanensis, the nominal species described from Taiwan, morphologically similar to P. brevianalis. We confirmed that P. adiposalis was distributed discontinuously in three river systems; analysis of 42 specimens from the rivers indicated a total of four haplotypes and population differentiation (average F _ST = 0.876). Fixation into a largely differentiated haplotype in the northernmost Tamsui River population was also found in this species, but different processes for this phylogeographic pattern were implied for the two species. Comparison with P. ussuriensis, a widespread continental species morphologically similar to P. adiposalis, suggested the possibility that P. adiposalis is a group of local populations of P. ussuriensis in Taiwan. Two migration routes vs. random fixation scenarios for the population structure of P. adiposalis are discussed with information on other fishes and the geological history of the island.     

Phylogenetic relationships and natural hybridization in rabbitfishes (Teleostei: Siganidae) inferred from mitochondrial and nuclear DNA analyses

Phylogenetic relationships of rabbitfishes (the family Siganidae), ecologically important components as primary consumers in coral reef communities, were studied using mitochondrial cytochrome b gene and nuclear ITS1 (internal transcribed spacer 1) sequence analyses. The analyses of 19 out of 22 species known in the Western Pacific region revealed that siganids are genetically clustered into three major clades, which are characterized by some morphological and ecological traits. Between closely related species, such as Siganus guttatus-S. lineatus and S. virgatus-S. doliatus, and also between two morphs recognized in S. corallinus, small but discernible genetic differentiation was found, implying that the components of each pair are incipient species. On the other hand, between some species, such as S. fuscescens-S. canaliculatus and S. unimaculatus-S.vulpinus, individuals of the components of each pair were found to construct a genetic mosaic, suggesting that the components are genetic color morphs within a single biological species, respectively. Moreover, evidence from morphological characters, mtDNA, and nuclear DNA gave an inconsistent picture of identity and relationships for several individuals. They were regarded as hybrids or individuals with hybrid origin. Such instances were observed not only between closely related species, such as S. guttatus-S. lineatus, S. virgatus-S. doliatus, and two morphs (incipient species) in S. corallinus, respectively, but also between distantly related ones, such as S. corallinus-S. puellus. In fact, more than half of the species examined (11/20, when treating the two morphs in S. corallinus as independent species) were involved in hybridization. These suggest that hybridization is much more prevalent in marine fishes than previously assumed, and may have some relevance to their diversification.     

Development of the Assessment of Belief Conflict in Relationship-14 (ABCR-14)

Purpose

Nurses and other healthcare workers frequently experience belief conflict, one of the most important, new stress-related problems in both academic and clinical fields.

Methods

In this study, using a sample of 1,683 nursing practitioners, we developed The Assessment of Belief Conflict in Relationship-14 (ABCR-14), a new scale that assesses belief conflict in the healthcare field. Standard psychometric procedures were used to develop and test the scale, including a qualitative framework concept and item-pool development, item reduction, and scale development. We analyzed the psychometric properties of ABCR-14 according to entropy, polyserial correlation coefficient, exploratory factor analysis, confirmatory factor analysis, average variance extracted, Cronbach’s alpha, Pearson product-moment correlation coefficient, and multidimensional item response theory (MIRT).

Results

The results of the analysis supported a three-factor model consisting of 14 items. The validity and reliability of ABCR-14 was suggested by evidence from high construct validity, structural validity, hypothesis testing, internal consistency reliability, and concurrent validity. The result of the MIRT offered strong support for good item response of item slope parameters and difficulty parameters. However, the ABCR-14 Likert scale might need to be explored from the MIRT point of view. Yet, as mentioned above, there is sufficient evidence to support that ABCR-14 has high validity and reliability.

Conclusion

The ABCR-14 demonstrates good psychometric properties for nursing belief conflict. Further studies are recommended to confirm its application in clinical practice.     

Serum Osteopontin Predicts Degree of Hepatic Fibrosis and Serves as a Biomarker in Patients with Hepatitis C Virus Infection

Background & Aims

Osteopontin (OPN) is a matricellular protein that upregulates during pathogenesis of hepatic fibrosis. The present study was aimed to evaluate whether serum OPN could be used as a biomarker to assess the degree of hepatic fibrosis in patients with hepatitis C virus (HCV) infection.

Methods

Needle biopsy was performed on HCV patients and scored as zero fibrosis (F0), mild fibrosis (F1), moderate fibrosis (F2), severe fibrosis (F3) and liver cirrhosis (F4) based on Masson’s trichrome and α-smooth muscle actin (α-SMA) staining. Serum OPN levels were measured using ELISA and correlated with the degree of fibrosis. Furthermore, the OPN values were correlated and evaluated with platelets count, serum hyaluronic acid (HA), and collagen type IV and subjected to receiver operating characteristic (ROC) curve analysis.

Results

Serum OPN levels were remarkably increased from F0 through F4 in a progressive manner and the differences were significant (P < 0.001) between each group. The data were highly correlated with the degree of hepatic fibrosis. The ROC curve analysis depicted that serum OPN is an independent risk factor and an excellent biomarker and a prognostic index in HCV patients.

Conclusions

The results of the present study indicate that serum OPN levels reflect the degree of hepatic fibrosis and could be used as a biomarker to assess the stage of fibrosis in HCV patients which would help to reduce the number of liver biopsies. Furthermore, serum OPN serves as a prognostic index towards the progression of hepatic fibrosis to cirrhosis and hepatocellular carcinoma.     

Cloning, sequencing and complementation analysis of the recA gene from Prevotella ruminicola

Abstract Degenerate PCR primers based on conserved RecA protein regions were used to amplify a portion of recE from Prevotella ruminicola strain 23, which was used as a probe to isolate the full-length recA gene from the P. ruminicola genomic library. The P. ruminicola recA gene encoded a protein of 340 amino acids with a molecular mass of 36.81 kDa. P. ruminicola RecA was highly similar to other RecA proteins and most closely resembled that of Bacteroides fragilis (75% identity). It alleviated the methyl methanesulfonate and mitomycin C sensitivities of Escherichia coli recA mutants, but did not restore the resistance to UV-light irradiation. Mitomycin C treatment of otherwise isogenic E. coli strains showed a higher level of prophage induction in a recA harboring lysogen.