Simultaneous real-time PCR detection of <Emphasis Type="Italic">Bacillus anthracis</Emphasis>, <Emphasis Type="Italic">Francisella tularensis</Emphasis> and <Emphasis Type="Italic">Yersinia pestis</Emphasis>

This report describes the development of in-house real-time PCR assays using minor groove binding probes for simultaneous detection of the Bacillus anthracis pag and cap genes, the Francisella tularensis 23 KDa gene, as well as the Yersinia pestis pla gene. The sensitivities of these assays were at least 1 fg, except for the assay targeting the Bacillus anthracis cap gene, which showed a sensitivity of 10 fg when total DNA was used as a template in a serial dilution. The clinical value of the Bacillus anthracis- and Francisella tularensis-specific assays was demonstrated by successful amplification of DNA from cases of cow anthrax and hare tularemia, respectively. No cross-reactivity between these species-specific assays or with 39 other bacterial species was noted. These assays may provide a rapid tool for the simultaneous detection and identification of the three category A bacterial species listed as biological threats by the Centers for Disease Control and Prevention.     

SCC<Emphasis Type="Italic">mec</Emphasis> and <Emphasis Type="Italic">spa</Emphasis> types of methicillin-resistant <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> strains in Israel. Detection of SCC<Emphasis Type="Italic">mec</Emphasis> type V

Molecular characterization of methicillin-resistant Staphylococcus aureus isolates from three hospitals in Israel was the aim of the study presented here. We identified 11 distinct genetic clones by pulsed-field gel electrophoresis. Molecular typing identified four different SCCmec types—I, II, IV, and V−and nine spa types. Spa type t002 was the most common. An erratum to this article can be found at     

<Emphasis Type="Italic">Trichomonas vaginalis</Emphasis> harboring <Emphasis Type="Italic">Mycoplasma hominis</Emphasis> increases cytopathogenicity in vitro

The parasite Trichomonas vaginalis causes one of the most common non-viral sexually transmitted infections in humans. Mycoplasmas are frequently found with trichomonads but the consequences of this association are not yet known. In the present study, the effects of T. vaginalis harboring M. hominis on human vaginal epithelial cells and on MDCK cells are described. The results were analyzed by light, scanning and transmission electron microscopy, as well as using cell viability assays. There was an increase in the cytopathic effects on the epithelial cells infected with T. vaginalis associated with M. hominis compared to T. vaginalis alone. The epithelial cells exhibited an increase in the intercellular spaces, a lesser viability, and increased destruction provoked by the infected T. vaginalis. In addition, the trichomonads presented a higher amoeboid transformation rate and an intense phagocytic activity, characteristics of higher virulence behavior.     

Heterogeneity of the <Emphasis Type="Italic">ospA</Emphasis> gene structure from isolates of <Emphasis Type="Italic">Borrelia garinii</Emphasis> and <Emphasis Type="Italic">Borrelia afzelii</Emphasis> from Western Siberia and Mongolia

In this work, identification and analyses of 48 full-length sequences of the ospA gene isolates of Borrelia garinii and Borrelia afzelii from Western Siberia and Mongolia has been made. It was shown that B. garinii isolates was of its high genetic heterogeneity of the ospA gene. Four genetic groups of the ospA gene from the Ixodes persulcatus tick collected in of Western Siberia and Mongolia were defined. The basic differences in the genetic variants of the ospA gene considered are seen in regions which code for antibody determinants of thhe OspA protein.     

Community-acquired <Emphasis Type="Italic">Acinetobacter</Emphasis> infections

Acinetobacter infections have been attracting increasing attention during recent years because they have become common in hospitalized patients, especially in the intensive care unit (ICU) setting. However, the available literature suggests that the pathogen has another fearful potential; it can cause community-acquired infections. We searched PubMed and the reference lists of the initially identified articles and identified six case series regarding a total of 80 patients with community-acquired Acinetobacter baumannii infections; from these, 51 had pneumonia and 29 had bacteremia. Of these 80 patients, 45 (56%) died of the infection. In addition, we identified 26 case reports regarding 43 patients with community-acquired Acinetobacter infections; from these, 38 had pneumonia, two had meningitis, one had soft-tissue infection, one had ocular infection, and one had native valve endocarditis. Comorbidity was commonly present in patients reported in the case series as well as the case reports, mainly, chronic obstructive pulmonary disease, renal disease, and diabetes mellitus; heavy smoking and excess alcohol consumption were also common. Most of the studies originated from China, Taiwan, and tropical Australia. We also identified 12 retrospective or prospective studies (seven from the Far East, two from Oceania, one from N. Guinea, one from Palestine, and one from USA/Canada) that reported the frequency of community-acquired Acinetobacter infections; the range of isolation of Acinetobacter from patients with community-acquired pneumonia in these studies was 1.3%-25.9%. In conclusion, most community-acquired Acinetobacter infections have been reported from countries with tropical or subtropical climate, and mainly affect patients with some form of comorbidity or are associated with heavy smoking and excess alcohol consumption.     

Antioxidative and <Emphasis Type="Italic">in vitro</Emphasis> antiproliferative activity of <Emphasis Type="Italic">Arctium lappa</Emphasis> root extracts

Background  

Arctium lappa, known as burdock, is widely used in popular medicine for hypertension, gout, hepatitis and other inflammatory disorders. Pharmacological studies indicated that burdock roots have hepatoprotective, anti-inflammatory, free radical scavenging and antiproliferative activities. The aim of this study was to evaluate total phenolic content, radical scavenging activity by DPPH and in vitro antiproliferative activity of different A. lappa root extracts.     

Effects of <Emphasis Type="Italic">Sophora japonica</Emphasis> flowers (<Emphasis Type="Italic">Huaihua</Emphasis>) on cerebral infarction

The dried flowers and buds of Sophora japonica are used as a medicinal herb in China, Japan and Korea to treat bleeding hemorrhoids and hematemesis. This article presents an overview of the effects of Sophora japonica on cerebral infarction based on literature searched from Medline, PubMed, Cochrane Library and the China National Knowledge Infrastructure (CNKI). Sophora japonica contains both anti-hemorrhagic and anti-hemostatic substances. Sophora japonica reduces cerebral infarction partly as a result of its anti-oxidative and anti-inflammatory activities. Previous studies found that Sophora japonica reduced the size of cerebral infarction and neurological deficits and reduced microglial activation, interleukin-1β release and number of apoptotic cells in ischemia-reperfusion injured Sprague-Dawley rats. Further study is required to determine the relationship between Sophora japonica-mediated reduction in cerebral infarction size and the effects of Sophora japonica on platelet aggregation and cardiovascular function.     

Histopathologic and electron microscopic characterization of cutaneous leishmaniasis in <Emphasis Type="Italic">Tatera indica</Emphasis> and <Emphasis Type="Italic">Gerbillus</Emphasis> spp. infected with <Emphasis Type="Italic">Leishmania major</Emphasis>

Cutaneous leishmaniasis (CL) is a zoonotic disease transmitted between rodents and canines, mainly by Phlebotomus sand flies and man. In southern Iran, the incidence of this protozoan disease has doubled over the last decade. The present study deals with histopathological features of CL in Tatera indica and Gerbillus spp. that participate in the epidemiology of CL in southern Iran. Thirty-two trapped rodents were evaluated for any Leishmania infection using enzyme electrophoresis and polymerase chain reaction and were concomitantly studied for any histopathological changes. Histopathological studies showed that bone marrow was the tissue of choice for light and electron microscopic study of Leishmania, demonstrating the macrophages infected with the amastigote form of the parasite. This is the first report of the histopathological detection of L. major in naturally infected T. indica and Gerbillus spp in the Larestan region.     

Differentiating the growing nosocomial infectious threats <Emphasis Type="Italic">Ralstonia pickettii</Emphasis> and <Emphasis Type="Italic">Ralstonia insidiosa</Emphasis>

Differentiation of the growing nosocomial infectious threats, Ralstonia pickettii and Ralstonia insidiosa, based on nitrate reduction, desferrioxamine susceptibility, arabinose, N-acetyl-glucosamine and phenylacetate assimilation is described. These tests can be used for preliminary identification of Ralstonia pickettii and Ralstonia insidiosa resulting in more accurate identification of these species.     

Structural and functional analysis of <Emphasis Type="Italic">araC</Emphasis> gene in <Emphasis Type="Italic">Yersinia pestis</Emphasis> of various origins

Structural and functional analyses of the araC gene, which is involved in regulation of expression of diagnostically important characteristics, such as arabinose fermentation in the plague microbe strain, were carried out. It was established that the reason for the lack of arabinose fermentation in the altaica and hissarica subspecies and strains of talassica groups is the presence of the insertion of a single nucleotide in the araC gene at position 773 bp. The strains of the basic, caucasia, and ulegeica subspecies do not contain this mutation in the araC gene, which correlates to their ability to ferment arabinose.     

Characterization of the siderophore transport protein AsbJ in <Emphasis Type="Italic">Aeromonas salmonicida</Emphasis> subsp. <Emphasis Type="Italic">salmonicida</Emphasis>

Iron is an important nutrient for all living systems. Bacteria have developed multiple mechanisms to obtain iron from the host. Aeromonas salmonicida subsp. salmonicida produces catechol-type siderophore under iron starvation conditions. This study attempted to characterize and ascertain the role of a siderophore ABC transporter protein AsbJ in siderophore transporter system in this bacterial pathogen. Prediction of protein domains was carried out, and deleted asbJ gene mutant strain was constructed. Results showed that this protein contains special domain related to permease and ATP-binding protein, and it is essential for the siderophore transport process in this bacterial fish pathogen.     

Genetic diversity of porcine <Emphasis Type="Italic">Norovirus</Emphasis> and <Emphasis Type="Italic">Sapovirus</Emphasis>: Canada, 2005–2007

Noroviruses and sapoviruses are members of the family Caliciviridae and emerging enteric pathogens of humans and animals. Since their discovery and characterization in swine, relatively few strains have been described in detail. In order to investigate their genetic diversity, a total of 266 fecal samples collected in the province of Quebec, Canada, between 2005 and 2007 were screened for the presence of caliciviruses by RT-PCR using broadly reactive primers. Genetically heterogeneous caliciviruses were detected on the majority of farms. Typical noroviruses related to known swine genotypes were present on 20% of the farms. Sapoviruses were detected on 75% of the farms and were the most heterogeneous group. Further characterization of selected strains in their 3′ end parts was carried out for their classification and unveiled possibly new clusters of sapoviruses. No human-like noroviruses or sapoviruses were detected in the present study. GenBank accession numbers of all sequences described in this study are indicated in the figure legends.     

Chromosome pairing in allotetraploid hybrids of <Emphasis Type="Italic">Festuca pratensis</Emphasis> × <Emphasis Type="Italic">Lolium perenne</Emphasis> revealed by genomic <Emphasis Type="Italic">in situ</Emphasis> hybridization (GISH)

Genomic in situ hybridization (GISH) was used to make a detailed study of chromosome pairing at metaphase I (MI) of meiosis in six F(1) hybrid plants of the allotetraploid Festuca pratensis x Lolium perenne (2n = 4x = 28; genomic constitution FpFpLpLp). The mean chromosome configurations for all hybrids analysed were 1.13 univalents + 11.51 bivalents + 0.32 trivalents + 0.72 quadrivalents, and the mean chiasma frequency was 21.96 per cell. GISH showed that pairing was predominantly intragenomic, with mean numbers of L. perenne (Lp/Lp) and F. pratensis (Fp/Fp) bivalents being virtually equal at 5.41 and 5.48 per cell, respectively. Intergenomic pairing between Lolium and Festuca chromosomes was observed in 33.3% of Lp/Fp bivalents (0.62 per cell), in 79.7% of trivalents - Lp/Lp/Fp and Lp/Fp/Fp (0.25 per cell), and in 98.4% of quadrivalents - Lp/Lp/Fp/Fp and Lp/Lp/Lp/Fp (0.71 per cell). About 4.0% of the total chromosome complement analysed remained as univalents, an average 0.68 Lp and 0.45 Fp univalents per cell. It is evident that in these hybrids there is opportunity for recombination to take place between the two component genomes, albeit at a low level, and this is discussed in the context of compromising the stability of Festulolium hybrid cultivars and accounting for the drift in the balance of the genomes over generations. We speculate that genotypic differences between hybrids could permit selection for pairing control, and that preferences for homologous versus homoeologous centromeres in their spindle attachments and movement to the poles at anaphase I could form the basis of a mechanism underlying genome drift.     

Use of cross-species <Emphasis Type="Italic">in-situ</Emphasis> hybridization (ZOO-FISH) to assess chromosome abnormalities in day-6 <Emphasis Type="Italic">in-vivo</Emphasis>- or <Emphasis Type="Italic">in-vitro</Emphasis>-produced sheep embryos

Causes of chromosomal differences such as mosaicism between embryos developed in vivo and in vitro may be resolved using animal models to compare embryos generated in vivo with those generated by different production systems. The aims of this study were: (1) to test a ZOO-FISH approach (using bovine painting probes) to detect abnormal chromosome make-up in the sheep embryo model, and (2) to examine the extent of chromosome deviation in sheep embryos derived in vivo and in vitro. Cytogenetic analysis was performed on day 6 in-vivo and in-vitro derived sheep embryos using commercially available bovine chromosome painting probes for sex chromosomes X–Y and autosomes 1–29. A total of 8631 interphase and metaphase nuclei were analyzed from 49 in-vitro-derived and 51 in-vivo-derived embryos. The extent of deviation from normal ovine chromosome make-up was higher (p < 0.05) in in-vitro-produced embryos relative to in-vivo-derived embryos (65.3% vs. 19.6% respectively) mainly due to diploid–polyploid mosaicism. Polyploid cells ranged from 3n to 8n with tetraploids most predominant among non-diploid cells. The proportions of polyploid cells per mixoploid embryo in in-vitro-produced embryos ranged from 1.4% to 30.3%, in contrast to less than 10% among the in-vivo-derived embryos. It was concluded that in-vitro-derived embryos are vulnerable to ploidy change compared to their in-vivo counterparts. The application of ZOO-FISH to domestic animal embryos is an effective approach to study the chromosome complement of species for which DNA probes are unavailable.