Antimicrobial resistance of <Emphasis Type="Italic">Staphylococcus</Emphasis> from otorrhea in chronic suppurative otitis media and comparison with results of all isolated <Emphasis Type="Italic">Staphylococci</Emphasis>

Staphylococcus is one of the most important pathogenic bacteria in chronic suppurative otitis media (CSOM). The prevalence of pathogenic bacteria in patients with CSOM has not been compared with the prevalence rates in patients from other fields of medicine. We investigated the pathogenic bacteria in CSOM throughout Korea and annual isolation rates of methicillin-resistant Staphylococcus aureus (MRSA) over 6 years. Routine culture results and susceptibility data of CSOM isolated from 2000 to 2005 were collected from six general hospitals in Korea, along with the results of all clinically isolated Staphylococci from one tertiary care teaching hospital. Of the 1,162 bacteria identified in 1,360 CSOM patients, 628 (54.0%) were Staphylococci in CSOM. Of the latter, 288 (45.9%) were MRSA, which accounted for 24.8% of identified bacteria. Of the 5,988 clinically isolated Staphylococci from one tertiary care hospital, 3,712 (61.9%) were MRSA. All MRSA isolated from CSOM patients were sensitive to vancomycin and teicoplanin, and 88.2% were sensitive to sulfamethoxazole/trimethoprim. In contrast, these strains showed little or no sensitivity to oxacillin, clindamycin, penicillin, and erythromycin. Annual MRSA isolation rates showed no tendencies to increase or decrease. MRSA was the most frequently identified Staphylococcus in patients with otorrhea. The isolation rate of MRSA has not changed over 6 years. Continuous and periodic surveillance of MRSA is necessary to reduce the spread of antibiotic-resistant pathogens and to guide appropriate antibacterial therapy.     

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.     

<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.     

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     

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.     

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.     

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.     

Characteristics of <Emphasis Type="Italic">Streptococcus bovis</Emphasis> endocarditis and its differences with <Emphasis Type="Italic">Streptococcus viridans</Emphasis> endocarditis

The purpose of this study was to evaluate the characteristics of infective endocarditis (IE) caused by S. bovis and compare them to those caused by streptococci of the viridans group (SVG). A prospective study was undertaken considering 55 consecutive cases of IE due to S. bovis and 41 to SVG over 18 years. The study was divided into two periods (1988–1996 and 1997–2005). S. bovis caused 24% of the IE in our centre and constituted the main aetiology for this disease, showing an increase of 358% during the second period studied. Biotype I was responsible for 94.5% of cases and there was a high degree of association with colon tumours (53%). Over the period of the study, 107 patients admitted to our hospital had bacteraemia caused by S. bovis and 310 patients had bacteraemia caused by SVG. In the first group, 55 (51%) were endocarditis cases, but only 41 (13%) of the patients with SVG bacteraemia had endocarditis (p < 0.0001). The distinguishing features of endocarditis caused by S. bovis in comparison with those caused by SGV were: a greater increase in cases during the 2nd period studied (from 12 to 43 vs. from 19 to 22, p < 0.01), a higher percentage of males (93% vs. 71%, p < 0.004), patients significantly older (median age 66 vs. 58.5, p < 0.004), less predisposing cardiopathy (42% vs. 76%, p < 0.0009), more bivalvular involvement (42% vs. 22%, p < 0.04), more spondylitis (9% vs. 0%, p < 0.04), a higher association with colonic tumours (53% vs. 5%, p < 0.0001), and a higher percentage of antibiotic resistance: erythromycin 66% vs. 19%, p < 0.0001; clindamycin 67% vs. 11%, p < 0.0001; cotrimoxazole 77% vs. 30.5%, p < 0.0001, respectively. IE due to S. bovis is an emergent disease in our environment, presenting different characteristics to those produced by SVG.     

Complicating infectious foci in patients with <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> or <Emphasis Type="Italic">Streptococcus</Emphasis> species bacteraemia

Complicating infectious foci resulting from haematogenous or local spread of microorganisms are observed frequently in patients with Staphylococcus aureus bacteraemia (SAB) or Streptococcus species bacteraemia (SSB). The aim of this study was to compare the epidemiology of complicating infectious foci during SAB and SSB in a university hospital in The Netherlands. The charts of all adult patients diagnosed with SAB or SSB (except for Streptococcus pneumoniae bacteraemia) from July 2002 until December 2004 were reviewed retrospectively. Overall, 180 immunocompetent patients were identified, 127 with SAB and 53 with SSB. The percentage of patients with complicating infectious foci (39% of SAB patients, 25% of SSB patients) did not differ significantly between the groups. Endocarditis and cerebral involvement, however, were significantly more common in the SSB group. Of all complicating infectious foci, 32% lacked guiding signs or symptoms and 10% were detected only at autopsy. Factors associated with the development of complicating infectious foci were a delay in treatment for more than 48 h after the onset of symptoms, community acquisition, persistently positive blood cultures, congenital heart disease, and the presence of foreign bodies or prosthetic valves. Infection-related mortality was 18% in SAB patients and 11% in SSB patients and was significantly higher in patients with complicating infectious foci (29 vs. 9%). In conclusion, complicating infectious foci develop in approximately one-third of all patients with SAB and SSB. An active approach that entails searching for the complicating infectious foci is warranted in these patients, because only two-thirds of complicated infectious foci have guiding symptoms or signs, and infection-related mortality is significantly increased in patients with complicating infectious foci compared to patients without these infections. Authors M.L.H. Cuijpers and F.J. Vos contributed equally to this work.     

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.     

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.     

Flucloxacillin treatment of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> meningitis

In this paper, we aimed to evaluate the efficacy of flucloxacillin treatment of meningitis caused by methicillin-sensitive Staphylococcus aureus. We identified 33 patients with meningitis due to S. aureus; eight had community-acquired meningitis and 25 had neurosurgical meningitis. Six of the eight patients with community-acquired meningitis were cured. Eighteen of the 22 patients treated with flucloxacillin were cured without relapse (86%, 95%CI 65–97%) and their cerebrospinal fluid (CSF) cultures were sterile after a median of 3 days of treatment. The cure rate for 12 patients who also received an additional antibiotic at the outset of treatment was 75% (95%CI 43–95%). This was not different to the cure rate for the ten patients who received flucloxacillin alone 90% (95%CI 56–100%). We conclude that flucloxacillin is an effective treatment for meningitis caused by S. aureus.     

Catalog of 86 single-nucleotide polymorphisms (SNPs) in three uridine diphosphate glycosyltransferase genes: <Emphasis Type="Italic">UGT2A1</Emphasis>, <Emphasis Type="Italic">UGT2B15</Emphasis>, and <Emphasis Type="Italic">UGT8</Emphasis>

We report here three high-density maps of variations found among 48 Japanese individuals in three uridine diphosphate glycosyltransferase (UGT) genes, UGT2A1, UGT2B15, and UGT8. A total of 86 single-nucleotide polymorphisms (SNPs) were identified through systematic screening of genomic regions containing these genes: 8 in 5′ flanking regions, 7 in coding regions, 67 in introns, 3 in 3′ untranslated regions, and 1 in a 3′ flanking region. We also discovered 14 variations of other types. Of the 86 SNPs, 63 (73%) were considered to be novel on the basis of comparison of our data with the Database of SNPs (dbSNP) of the National Center for Biotechnology Information. Among the seven SNPs identified in exonic sequences, five were non-synonymous changes that would result in amino-acid substitutions. The collection of SNPs derived from this study will serve as an additional resource for studies of complex genetic diseases and responsiveness to drug therapy. Received: June 12, 2002 / Accepted: June 13, 2002     

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.     

A frameshift mutation of the chloroplast <Emphasis Type="Italic">mat</Emphasis>K coding region is associated with chlorophyll deficiency in the <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> virescent mutant <Emphasis Type="Italic">Wogon</Emphasis>-Sugi

Wogon-Sugi has been reported as a cytoplasmically inherited virescent mutant selected from a horticultural variety of Cryptomeria japonica. Although previous studies of plastid structure and inheritance indicated that at least some mutations are encoded by the chloroplast genome, the causative gene responsible for the primary chlorophyll deficiency in Wogon-Sugi, has not been identified. In this study, we identified this gene by genomic sequencing of chloroplast DNA and genetic analysis. Chloroplast DNA sequencing of 16 wild-type and 16 Wogon-Sugi plants showed a 19-bp insertional sequence in the matK coding region in the Wogon-Sugi. This insertion disrupted the matK reading frame. Although an indel mutation in the ycf1 and ycf2 coding region was detected in Wogon-Sugi, sequence variations similar to that of Wogon-Sugi were also detected in several wild-type lines, and they maintained the reading frame. Genetic analysis of the 19 bp insertional mutation in the matK coding region showed that it was found only in the chlorophyll-deficient sector of 125 full-sibling seedlings. Therefore, the 19-bp insertion in the matK coding region is the most likely candidate at present for a mutation underlying the Wogon-Sugi phenotype. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Infection of stromal and hemopoietic precursor cells with lentivirus vector <Emphasis Type="Italic">in vivo</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis>

We developed a method for gene transfer into mesenchymal stromal cells. Lentivirus vector containing green fluorescent protein gene for labeling stromal and hemopoietic precursor cells was obtained using two plasmid sets from different sources. The vector was injected into the femur of mice in vivo and added into culture medium for in vitro infection of the stromal sublayer of long-term bone marrow culture. From 25 to 80% hemopoietic stem cells forming colonies in the spleen were infected with lentivirus vector in vivo and in vitro. Fibroblast colony-forming cells from the femoral bones of mice injected with the lentivirus vector carried no marker gene. The marker gene was detected in differentiated descendants from mesenchymal stem cells (bone cavity cells from the focus of ectopic hemopoiesis formed after implantation of the femoral bone marrow cylinder infected with lentivirus vector under the renal capsule of syngeneic recipient). In in vitro experiments, the marker gene was detected in sublayers of long-term bone marrow cultures infected after preliminary 28-week culturing, when hemopoiesis was completely exhausted. The efficiency of infection of stromal precursor cells depended on the source of lentivirus. The possibility of transfering the target gene into hemopoietic precursor cells in vivo is demonstrated. Stromal precursor cells can incorporate the provirus in vivo and in vitro, but conditions and infection system for effective infection should be thoroughly selected. __________ Translated from Kletochnye Tehnologii v Biologii i Meditsine, No. 1, pp. 25–28, January, 2007     

Comparison of <Emphasis Type="Italic">SYBR Green I</Emphasis> and <Emphasis Type="Italic">TaqMan</Emphasis> real-time PCR formats for the analysis of <Emphasis Type="Italic">her2</Emphasis> gene dose in human breast tumors

We compared two technologies of real-time PCR (with the use of fluorescent SYBR Green I dye and specific TaqMan probe) for quantification of the dose of her2 gene in breast tumors. The maximum increase in the gene dose in TaqMan and SYBR Green I analyses was 10-and 5-fold, respectively. In was found that TaqMan and SYBR Green I technologies allow detection of the matrix in amounts corresponding to 1–100 and 2.5–40.0 ng genomic DNA, respectively. Tenfold increase in the gene dose leads to incorrect evaluation of multiplication ratio in the SYBR Green I analysis. These results suggest that TaqMan technology is more preferable for correct evaluation of her2 gene dose. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 2, pp. 201–205, February, 2008