Genetic diversity among VRE isolates from Swedish broilers with the coincidental finding of transferrable decreased susceptibility to narasin.

AIMS: In this study, the molecular diversity among clones of vancomycin resistant Enterococcus faecium with vanA gene (VRE) is investigated. The aims were to better understand why one clone is predominant in Swedish broiler production and to better assess the potential for zoonotic gene transfer from the different clones. METHODS AND RESULTS: Twenty-six isolates were separated into 11 clones. Vancomycin resistance was transferrable from the predominant and five minority clones. Decreased susceptibility to narasin was co-transferred with vancomycin resistance in four clones, including the predominant. The plasmid addiction system axe-txe was not detected, and the ω-ε-ζ system was detected in one of the minority clones but was not co-transferred with vancomycin resistance. CONCLUSIONS: The results do not explain why one clone is predominant among VRE in Swedish broiler production but confirms the potential for zoonotic spread of vancomycin resistance genes. The near absence of investigated plasmid addiction systems indicates that they do not play an important role in the epidemiology of VRE in Swedish broiler production. The finding that decreased susceptibility to narasin can be co-transferred with the vanA gene indicates that the use of narasin might play a role in the persistence of vancomycin resistance in enterococci colonizing Swedish broilers. SIGNIFICANCE AND IMPACT OF THE STUDY: This is, to our knowledge, the first report of transferrable decreased susceptibility to narasin.

High-intensity resistance training in people with multiple sclerosis experiencing fatigue: A randomised controlled trial.

BACKGROUND: Exercise studies including only fatigued persons with multiple sclerosis (PwMS) with fatigue as primary endpoint are lacking. OBJECTIVE: To evaluate the effects of high-intensity resistance training (HIRT) on self-reported fatigue in fatigued PwMS in a single center randomised controlled trial. METHODS: We recruited 71 PwMS scoring ≥ 53 on the Fatigue Scale for Motor and Cognitive Functions (FSMC), who were randomised 1:1 to either twice (group A) or once (group B) weekly supervised HIRT for twelve weeks. A non-randomised FSMC score-matched group (n=69) served as non-intervention control. RESULTS: Between HIRT-group differences were non-significant for primary and most secondary endpoints. Mean difference in FSMC score (95% confidence intervals) was -10.9 (-14.8; -6.9) in group A and -9.8 (-13.2; -6.3) in group B. Corresponding values for combined HIRT groups vs non-intervention control were -10.3 (-12.9; -7.7) and 1.5 (-0.6;3.6), respectively, p<0.001. Secondary endpoints also improved in both HIRT groups, though only Hospital Anxiety and Depression Scale anxiety and MS Impact Scale-29 psychological subscales significantly favoured the twice a week HIRT (group A). As an exploratory endpoint, changes in plasma inflammatory protein markers were associated with reduced FSMC scores in the pooled material. CONCLUSION: The finding that HIRT in fatigued PwMS leads to clinically relevant reductions in self-reported fatigue, associated with changes in plasma inflammatory protein levels, provide evidence for recommending HIRT for fatigued PwMS.

Single PCR and nested PCR with a mimic molecule for detection of Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease in ruminants. The current methods for detection of M. avium subsp. paratuberculosis are slow and insensitive. We report the use of a polymerase chain reaction (PCR) based on IS900 to confirm growth of M. avium subsp. paratuberculosis in primary bacterial cultures from bovine tissue and fecal samples. The use of PCR on single colonies reduced the time for analysis by 2 months compared with conventional methods. We also report the development of a nested PCR based on IS900 and the development of a positive internal control molecule, a so-called mimic. The system was tested with spiked tissue samples, and the sensitivity was estimated to 10 CFU per sample. Seventeen tissue samples, previously found M. avium subsp. paratuberculosis positive by microbiological culture, were analyzed by nested PCR and the efficiency of the PCR was checked by co-amplification of the mimic. Absence of the mimic amplicon indicated inhibition of the amplification. Ten of the samples were positive and five were negative, as judged from the presence or absence of the IS900 PCR product. Two negative samples could not be judged because of inhibition revealed by mimic molecules. It was concluded that the nested PCR, together with the mimic, could be a useful tool in screening tissue materials.

The genetics of neurotic-reactive depression: a reanalysis of Shapiro's (1970) twin study using diagnostic criteria.

The present study explored the role of genetic factors in the development of neurotic depression. Case studies of 16 monozygotic (MZ) and 14 same-sex dizygotic (DZ) twins from Robert Shapiro's 1970 study of non-endogenous depression were rediagnosed by two raters blind to the zygosity and identity of each twin. Diagnoses were made using Research Diagnostic Criteria (RDC) and George Winokur's 1985 criteria for neurotic-reactive depression. When neurotic depression was operationally defined using Winokur's criteria plus RDC major or definite minor depression, the concordance rate for MZ twins was significantly greater than that for DZ twins. Our results contrast with Shapiro's negative findings, probably due to our use of formal diagnostic criteria and Shapiro's requirement that cotwins be hospitalized to be considered concordant. The present results suggest that genetic factors play a role in the etiology of at least some forms of neurotic depression.

Detection of Mycobacterium avium subsp. paratuberculosis in tissue samples by single, fluorescent and nested PCR based on the IS900 gene.

The aim of this study was to determine if fluorescent PCR could be used instead of nested PCR, for the detection of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) in clinical specimens, to improve the sensitivity without increasing the risk for cross-contamination. We investigated and compared the sensitivity of single PCR, fluorescent PCR and nested PCR for the detection of IS900, an insertion sequence specific for M. paratuberculosis. A previously described extraction method for clinical specimens, based on xylene, was evaluated regarding its suitability for routine diagnostic work. The sensitivity of each PCR system was assessed by analysing a serial dilution of M. paratuberculosis DNA. To improve the reliability of the PCR and to facilitate the interpretation of the PCR results, a positive internal control molecule ("mimic") was developed and used for single and fluorescent PCR. In nested PCR, an existing mimic was used. The efficiency of recovering DNA of M. paratuberculosis from clinical specimens by the extraction method and detection of the organism by PCR was studied by analysing spiked ileum mucosa specimens. The final evaluation was performed on seventeen ileum mucosa specimens, previously found positive for M. paratuberculosis by bacterial culture. Twelve of the samples were positive by fluorescent PCR and nested PCR, and 10 samples were positive by single PCR. The use of mimics showed inhibition in specimens harbouring few M. paratuberculosis organisms, illustrating the effect of inhibitory substances in combination with small amounts of M. paratuberculosis DNA. We conclude that the extraction method was not adequate to recover small amounts of M. paratuberculosis and that inhibitory substances were still present in the processed specimens, but that the method is useful for identifying positive samples. Fluorescent PCR was a suitable alternative to both single PCR and nested PCR for the detection of M. paratuberculosis.

Characterization of plasmid-mediated AmpC-producing E. coli from Swedish broilers and association with human clinical isolates.

A selection of plasmid-mediated AmpC-producing Escherichia coli isolates carrying blaCMY-2 from Swedish broilers were characterized to establish their relatedness to and a possible overlap with human clinical E. coli isolates. The results showed diversity among the E. coli isolated from broilers, indicating that the spread in the population was not due to one strain. However, only one type of plasmid belonging to replicon type incK was identified. Furthermore, there were no indications of spread of blaCMY-2 E. coli isolates from broilers to human clinical settings, although Swedish broilers may be a source of blaCMY-2 and/or the plasmid carrying blaCMY-2 .

Sensitive detection of Mycobacterium avium subsp. paratuberculosis in bovine semen by real-time PCR.

AIMS: To develop a fast and sensitive protocol for detection of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine semen and to make a critical evaluation of the analytical sensitivity. METHODS AND RESULTS: Processed semen was spiked with known amounts of MAP. Semen from different bulls as well as semen of different dilutions was tested. The samples were treated with lysing agents and beadbeating and the DNA was extracted with phenol and chloroform. Real-time PCR with a fluorescent probe targeting the insertion element IS900 detected as few as 10 organisms per sample of 100 mul semen. PCR-inhibition was monitored by inclusion of an internal control. Pre-treatment with immunomagnetic separation was also evaluated, but was not shown to improve the overall sensitivity. CONCLUSIONS: Real-time PCR is a sensitive method for detection of MAP in bovine semen. Lysis by mechanical disruption followed by phenol and chloroform extraction efficiently isolated DNA and removed PCR-inhibitors. SIGNIFICANCE AND IMPACT OF THE STUDY: The high sensitivity of the applied method allows reliable testing of bovine semen used for artificial insemination to prevent the spread of Johne's disease, caused by MAP.

Postoperative nausea and vomiting in paediatric ambulatory surgery: sevoflurane versus spinal anaesthesia with propofol sedation.

BACKGROUND: Descriptive data report a very low rate of postoperative nausea and vomiting (PONV) following spinal anaesthesia in children. In an attempt to corroborate this observation, we designed a prospective randomized study to compare spinal anaesthesia with intravenous propofol sedation (SA) (n=21) to inhalational sevoflurane anaesthesia (IA) (n=19) with regard to PONV and postoperative analgesia in children (aged 3-12 years) undergoing ambulatory inguinal surgery. RESULTS: No difference was found concerning the number of patients experiencing PONV in each group (SA 1/21 versus IA 5/19; P=0.085). However, spinal anaesthesia was associated with a reduced number of PONV episodes (1/21) compared with inhalation anaesthesia (8/19) (P=0.014) and the need for supplemental postoperative analgesia with ketoralac was significantly lower in the SA group (3/21) compared to the IA group (14/19) (P < 0.001). Despite these benefits of spinal anaesthesia compared with inhalational anaesthesia, spinal anaesthesia did not decrease the time to discharge from the ambulatory surgery unit [SA 161 (SD 51) min, IA 164 (SD 41) min; P=NS] and the overall PONV experience was rated as "no problem" by all patients, except one, regardless of anaesthetic protocol used. CONCLUSIONS: Despite the reduced number of emetic episodes and the better immediate postoperative analgesia associated with spinal anaesthesia, no difference could be identified between the two different anaesthetic protocols regarding time to discharge or overall patient satisfaction. Thus, despite minor advantages associated with spinal anaesthesia with propofol sedation, both anaesthetic regimen appear equally suitable for use in the paediatric outpatient setting.