Challenges in the identification of Chryseobacterium indologenes and Elizabethkingia meningoseptica in cases of nosocomial infections and patients with cystic fibrosis.

Rare nonfermenting Gram-negative bacilli, such as Chryseobacterium indologenes and Elizabethkingia meningoseptica, have clinical importance in nosocomial infections and cystic fibrosis (CF), and their identification is a challenge to microbiology laboratories. Thus, the objective of this study was to verify the performance of phenotypic and mass spectrometry (matrix-assisted desorption ionization-time of flight mass spectrometry, MALDI-TOF MS) methods to identify C. indologenes and E. meningoseptica. In this context, the results obtained with phenotypic methods-namely manual biochemical and automated VITEK 2 (bioMérieux, Marcy l'Etoile, France) and Phoenix tests (Becton Dickinson (BD), San Diego, CA, USA)-and by MALDI-TOF MS-namely MALDI-TOF VITEK MS (MALDI-MS; bioMérieux) and MALDI-TOF BioTyper (MALDI-BD; BD)-of 22 isolates (blood cultures of patients with nosocomial infection (n = 15) and from patients with CF (n = 7)), initially identified as C. indologenes and E. meningoseptica, were compared. As result, using the manual phenotypic method, it was possible to identify the species level in 18/22; no identification was found in 4/22. There was a low agreement level between manual and VITEK 2 automated phenotypic methods when considering the genus level. The greatest agreement for genus-level identification occurred in MALDI-TOF MS equipment (15/22). When comparing all methods to identify the 22 isolates, there was agreement of 4/22 at the genus level and of 4/22 at the species level. In conclusion, there is low agreement level among identification methods of C. indologenes and E. meningoseptica. Although MALDI-TOF MS equipment shows a higher agreement level among them, results present low levels of confidence.

Evaluation of PCR in the diagnosis of canine leishmaniasis in two different epidemiological regions: Campinas (SP) and Teresina (PI), Brazil.

SUMMARY Using the polymerase chain reaction (PCR) test for diagnosis of canine leishmaniasis has greater sensitivity and specificity than culture and visualization of the parasite. This study compares PCR for the diagnosis of the genus and species of Leishmania with serological techniques used for the control of canine visceral leishmaniasis (CVL) in Brazil, considering two regions. We analysed peripheral blood samples collected from 195 dogs in the Campinas (SP) and Teresina (PI) regions. ELISA was performed as a serological method and PCR was performed using specific primers for the genus Leishmania spp. and the species Leishmania chagasi. In Campinas, a greater sensitivity of PCR (88.24%) (P = 0.0455) compared to Teresina (14.71%) (P < 0.0001) was observed, and an agreement was observed for Cohen's kappa index (0.9096). Both PCR and ELISA showed discordance for sensitivity (Campinas 100%, Teresina 21.74%), specificity (Campinas 30.77%, Teresina 100%), positive predictive value (Campinas 68.97%, Teresina 100%), negative predictive value (Campinas 100%, Teresina 37.94%) and Cohen's kappa index (0.1238). This study confirms the importance of PCR in analysis of the canine reservoir, and as an effective method for the detection of active and recent infection.

UriSed as a screening tool for presumptive diagnosis of urinary tract infection.

BACKGROUND: Although a quantitative urine culture is essential for the final diagnosis of urinary tract infection, it is time-consuming and an expensive procedure. Effective screening tests would be a promising alternative to provide immediate results for the clinician and eliminate unnecessary culturing for most of the negative samples. The aim of this study was to evaluate the performance of an automated sediment analyzer (UriSed) as screening tool for presumptive diagnosis of urinary tract infection. METHODS: We studied 1379 fresh midstream clean-catch urine samples from children to elderly. All samples were submitted to automated sediment analysis (UriSed) and quantitative urine culture (CLED medium agar). RESULTS: The sediment analyzer detected leukocyturia and/or significant bacteriuria with sensitivity of 97%, specificity of 59%, positive predictive value of 27%, negative predictive value of 99%, and accuracy of 64% at cutoff values of bacteria count ≥12.6 elements/hpf and WBC ≥6 cells/hpf. These data suggest a potential 52% reduction of unnecessary urine cultures. CONCLUSION: The UriSed seems to be an efficient tool for screening UTI with high sensitivity and low rate of false-negative results.

Laryngoscope blades and handles as sources of cross-infection: an integrative review.

The lack of standardization of efficient procedures to clean and disinfect laryngoscope blades and handles, which may be important sources of infection during their clinical use, has been reported previously, revealing contamination with blood, body fluids and micro-organisms. This paper aimed to evaluate the evidence available in the literature regarding the risk of laryngoscope blades and handles as a source of patient contamination. An integrative review of the literature was performed using databases such as Medline, LILACS, SciELO, Cochrane Library, BDENF and PubMed, and keywords in Medical Subject Headings (MeSH). The sample comprised 20 articles published between 1994 and 2012. The studies demonstrated risk of cross-infection and no consensus in current guidelines regarding cleaning and disinfection of this equipment. It was concluded that there are important gaps to be filled and urgent investigations required in order to facilitate standardization of efficient procedures to clean and disinfect laryngoscope blades and handles, and in turn to reduce the potential risk to which the patient and/or health team is exposed.

Changes in vancomycin-resistant Enterococcus faecium causing outbreaks in Brazil.

Enterococci have been implicated in severe human infections as a consequence of associated determinants of virulence and antimicrobial resistance. The majority of vancomycin-resistant Enterococcus faecium (VRE(fm)) connected to outbreaks worldwide pertains to the clonal complex 17 (CC17). In Brazil, the majority of VRE(fm) involved in outbreaks reported so far are not related to CC17. VRE(fm) strains responsible for an outbreak and sporadic cases in hospitals located in the city of Campinas, Brazil, were compared to other VRE(fm) strains in the country. Twenty-two out of 23 E. faecium were vancomycin-resistant and harboured the vanA gene. One vancomycin-susceptible E. faecium (VSE(fm)) strain was included in this study because it was isolated from a patient who one week later harboured a VRE(fm). All strains, except VSE, showed the same alteration in the VanA element characterised by deletion of the left extremity of the transposon and insertion of IS1251 between the vanS and vanH genes. Genes codifying virulence factors such as collagen-adhesin protein, enterococcal surface protein and hyaluronidase were detected in the VRE(fm) and VSE(fm) studied. Both pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) revealed that VRE(fm) and VSE(fm) strains have a clonal relationship. New sequence types (STs) were identified by MLST as ST447, ST448, ST478 and ST412 but all belonged to the CC17. The present study revealed that VRE(fm) outbreaks in Brazil were caused by strains that did not share a common evolutionary history, and that VRE(fm) strains belonging to CC17 could be predominant in Brazil as in other countries.

Evaluation of antimicrobial effectiveness of C-8 xylitol monoester as an alternative preservative for cosmetic products.

Xylitol is a natural sugar derived from plants, fruits and vegetables, whose antimicrobial properties are described in the literature. This study aimed to evaluate the antimicrobial effectiveness of C-8 xylitol monoester, for its use as a preservative in cosmetic formulations. The minimum inhibitory concentration (MIC) was determined by the broth macrodilution method, and the antimicrobial effectiveness of C-8 xylitol monoester was determined by using challenge test method. The results obtained in the determination of minimum inhibitory concentration are between 1.0% and 1.25% for Staphylococcus aureus, Escherichia coli and Candida albicans and between 1.0% and 1.5% for Pseudomonas aeruginosa and Aspergillus niger. The amount of 1% of C-8 xylitol monoester was added to the lotion used in the challenge test, observing a rapid decline in the number of CFU g(-1) in stages of evaluation after contamination of the product by all bacteria. The same occurs in relation to C. albicans, which shows a 90% reduction in the number of CFU g(-1). Regarding A. niger, similar reduction is observed when pH value of the lotion is adjusted from 5.5 to 7.0. The results indicate that under the tests conditions, C-8 xylitol monoester has antimicrobial activity and could be considered as an alternative preservative for cosmetic formulations.

Controlling a vancomycin-resistant enterococci outbreak in a Brazilian teaching hospital.

The purpose of this article was to describe a 2.5-year interventional program designed to control the dissemination after a large hospital outbreak of vancomycin-resistant enterococci (VRE) in a tertiary-care university hospital. A VRE working group was designated to work specifically on controlling VRE intrahospital dissemination after the detection of the first VRE infection at in our hospital in June 2007. The intervention consisted in the interruption of new admissions during a period of 15 days and closure of the index case unit, microbiological surveillance of rectal swabs for VRE, cohorting patients and staff, immediate application of contact precautions, and continuous education. From July 2007 to December 2009, 8,692 rectal swabs were cultured for VRE and 321 (3.7%) were positive. An expressive reduction of the detection of new positive rectal swabs cultures was seen during the year 2009 (1.5%) when compared to 2008 (4.2%) and 2007 (7.2%) (p < 0.005). The annual ratio of VRE per 1,000 admissions reduced from 20.3 in 2007 to 10.07 and 3.82 in 2008 and 2009, respectively (p < 0.001). The continuous microbiologic surveillance for VRE and strict and prompt contact precautions for VRE patients were the fundamental aids in the control of VRE.

Cytotoxic activity of clinical Stenotrophomonas maltophilia.

AIMS: To determine the potential virulence factors produced by culture supernatants of clinical isolates of Stenotrophomonas maltophilia. METHODS AND RESULTS: Culture supernatants of clinical isolates of S. maltophilia were assayed for haemolytic, enzymatic (lipase, protease and phospholipase) and cytotoxic activity. Cytotoxic activity was assayed in Vero (African green monkey), HeLa (human cervix) and HEp-2 (human larynx epidermoid carcinoma) cells. Microscopic analyses revealed intensive rounding, loss of intercellular junctions and membrane alterations (blebbing) followed by death of HEp-2 cells. In Vero and HeLa cells, the cytotoxic effects were characterized by vigorous endocytosis and cell aggregation. The viability of cultured mammalian cells was determined with neutral red and demonstrated that the sensitivity among the cells was different. This activity was inactivated by heating at 56 degrees C for 15 min and protease inhibitors did not inhibit cytotoxic activity. The clinical S. maltophilia presented a cell-free haemolytic activity similar to the 'hot-cold' haemolysins. CONCLUSIONS: S. maltophilia culture supernatants caused vigorous endocytosis and cell aggregation in HeLa and Vero cells, produced haemolytic and enzymatic activities. SIGNIFICANCE AND IMPACT OF THE STUDY: This work revealed the presence of putative virulence factors that could be associated with human infections involving Stenotrophomonas maltophilia strains.

Characterization of the Brazilian endemic clone of methicillin-resistant Staphylococcus aureus (MRSA) from hospitals throughout Brazil.

The objective of this study was to characterize patterns of the Brazilian endemic clone of methicillin-resistant Staphylococcus aureus (MRSA) from hospitals throughout Brazil. We studied 83 MRSA strains isolated from patients hospitalized in 27 public and private hospitals in 19 cities located in 14 Brazilian states from September, 1995, to June, 1997. The MRSA strains were typed using antibiograms, bacteriophage typing and pulsed field gel electrophoresis (PFGE). The analysis of genomic DNA by PFGE showed that 65 isolates presented the same PFGE pattern. This pattern was present in all of the hospitals studied indicating the presence of an endemic MRSA clone widely disseminated throughout Brazilian hospitals (BEC). All isolates belonging to the BEC proved to be resistant to ciprofloxacin, erythromycin, lincomycin, trimethoprim-sulphamethoxazole, and tetracycline. Variable susceptibility to these drugs was found only in isolates belonging to clones other than the BEC. The results show that, among MRSA, the BEC is common in Brazil. The best method for mapping changes in the frequency of this clone among MRSA is pulsed field gel electrophoresis. Use of molecular mapping is an important tool for monitoring the spread of potentially dangerous microbes.

Isolation in Brazil of nosocomial Staphylococcus aureus with reduced susceptibility to vancomycin.

OBJECTIVE: To evaluate the possible presence of vancomycin-resistant Staphylococcus aureus (VRSA) in a Brazilian hospital. DESIGN: Epidemiological and laboratory investigation of nosocomial VRSA. METHODS: 140 methicillin-resistant S aureus strains isolated between November 1998 and October 1999 were screened for susceptibility to vancomycin. The screening was carried out by using brain-heart infusion agar (BHIA) supplemented with 4, 6, and 8 microg/mL of vancomycin. The minimum inhibitory concentration (MIC) determination was carried out as standardized by the National Committee for Clinical Laboratory Standards using the broth macrodilution, agar-plate dilution, and E-test methods. PATIENTS: Hospitalized patients exposed to vancomycin. RESULTS: 5 of the 140 isolates had a vancomycin MIC of 8 microg/mL by broth macrodilution, agar plate dilution, and E-test methods. Four VRSA strains were isolated from patients in a burn unit who had been treated with vancomycin for more than 30 days, and one from an orthopedic unit patient who had received vancomycin treatment for 7 days. Pulsed-field gel electrophoresis characterized four of the VRSA strains as belonging to the Brazilian endemic clone. All five strains were negative for vanA, vanB, and vanC genes by polymerase chain reaction. Transmission electron microscopy of the five strains revealed significantly thickened cell walls. One patient died due to infection caused by the VRSA strain. CONCLUSIONS: This is the first report of isolation of VRSA in Brazil and the first report of isolation of multiple VRSA strains from one facility over a relatively short period of time. This alerts us to the possibility that VRSA may be capable of nosocomial transfer if adequate hospital infection control measures are not taken.

Resident research education in physical medicine and rehabilitation: a practical approach.

The Accreditation Council for Graduate Medical Education includes training in research as a required component of physical medicine and rehabilitation residency programs. Unfortunately, there is a lack of practical information on how to meet this requirement. In this paper, information is provided for individuals involved in resident education on how to teach residents about research.

Acquired limb deficiencies. 3. Prosthetic components, prescriptions, and indications.

UNLABELLED: This self-directed learning module highlights indications for prosthetic components and prescription formulation for adults with acquired limb deficiency. It is part of the chapter on acquired limb deficiencies in the Self-Directed Physiatric Education Program for practitioners and trainees in physical medicine and rehabilitation. Advantages and disadvantages of specific components of upper and lower limb prostheses are discussed, and a sample prescription sheet for upper limb devices is included. Recent innovations in terminal devices for upper limb prostheses are reviewed. Special considerations for the adult with acquired multilimb deficiency are also examined. OVERALL ARTICLE OBJECTIVE: To describe indications for prosthetic components and prescription formulation for adults with acquired limb deficiency.

Acquired limb deficiencies. 4. Troubleshooting.

UNLABELLED: This self-directed learning module offers practical analyses of and solutions for common clinical problems of amputees. It is part of the chapter on acquired limb deficiencies in the Self-Directed Physiatric Education Program for practitioners and trainees in physical medicine and rehabilitation. The information presented here has been designed to be useful also to other interested professionals, including prosthetists, physical therapists, occupational therapists, and nurses. Topics covered include the management of typical obstacles encountered in upper limb amputees, and the diagnosis and treatment of phantom and residual limb pain. Diagnostic and treatment approaches to skin breakdown in the transtibial amputee and to knee instability in the transfemoral amputee are also presented. OVERALL ARTICLE OBJECTIVE: To analyze common clinical problems of amputees.

Sports and recreation for persons with limb deficiency.

UNLABELLED: Opportunities for persons with limb deficiency to participate in sport and recreational activities have increased dramatically over the past 20 years. Various factors have contributed to this phenomenon, including an increased public interest in sports and fitness as well as improvements in disability awareness. An even more essential element has been a consumer-driven demand for advances in prosthetic technology and design. Whether the activity is a music performance, a friendly round of golf, or a high-level track-and-field competition, the benefits of participation in sports and recreation are numerous both at the individual and at the societal level. This article provides an overview of the development and scope of sport and recreational opportunities available to persons with limb deficiency. In addition, specific prosthetic considerations for several common sport and recreational activities are presented in a case-discussion format. OVERALL ARTICLE OBJECTIVE: To review the development and scope of sport and recreational opportunities available to persons with limb deficiency.

Functional MRI evidence of cortical reorganization in upper-limb stroke hemiplegia treated with constraint-induced movement therapy.

OBJECTIVE: The purpose of this pilot study was to test constraint-induced movement therapy for chronic upper-limb stroke hemiparesis and to investigate the neural correlates of recovery with functional magnetic resonance imaging (MRI) in two subjects. Both subjects had been discharged from traditional therapy because no further improvement was anticipated. DESIGN: Constraint-induced movement therapy consisted of 6 hr of daily upper-limb training for 2 wk; a restrictive mitt was worn on the nonparetic limb during waking hours. Functional MRI was performed on a 1.5-T MRI with echo-planar imaging; at the same time, the subjects attempted sequential finger-tapping. RESULTS: Compared with baseline, performance time improved an average of 24% immediately after training and also continued to improve up to 33% 3 mo after training. Lift, grip strength, and Motor Activity Log scores likewise improved. Initially, on functional MRI, subject 1 activated scattered regions in the ipsilateral posterior parietal and occipital cortices. Subject 2 showed almost no areas of significant activation. After training, subject 1 showed activity bordering the lesion, bilateral activation in the association motor cortices, and ipsilateral activation in the primary motor cortex. Subject 2 showed activation near the lesion site. CONCLUSION: Constraint-induced movement therapy produced significant functional improvement and resulted in plasticity as demonstrated by functional MRI.

Using the Internet to enable access to medical conferences.

OBJECTIVE: The goal of expanding access to individuals with disabilities to scientific and medical conferences is supported by both the Americans with Disabilities Act and the National Institutes of Health. DESIGN: Live-streaming video broadcast over the internet is widely available, although it has been used only in a limited fashion by the medical community. A consumer-oriented medical and rehabilitation conference concerning the rare disabling disease, fibrodysplasia ossificans progressiva, was broadcast via the world wide web. The address of a web page was announced before the conference to three computer users' groups that were considered likely to have an interest in the conference. The web page presented a live-streaming video broadcast of the conference. A phone line was installed, thereby allowing viewers to ask questions of the presenters during the question and answer periods. RESULTS: Sixteen users logged in 83 times to view the conference over a 2-day period. Five (23%) of 22 members of a fibrodysplasia ossificans progressiva internet users' group tuned in from distant places as The Netherlands and Israel. CONCLUSIONS: The internet is a viable tool to expand access to and increase the participation of individuals with disabilities in scientific and medical conferences. This technology should be used routinely in conferences of interest.

Clinical relevance and virulence factors of pigmented Serratia marcescens.

Pigmented Serratia marcescens isolated in a Brazilian hospital were studied with respect to frequency of isolation, serotyping, antibiotic resistance and virulence factors. The serotype most frequent was O6:K14 (53%) and all isolates were resistant to ampicillin, cephalothin and tetracycline. The majority of the isolates (92%) were resistant to the action of human serum and all produced cytotoxins on Vero, CHO, HEp-2 and HeLa cells. These isolates were virulent for mice (LD(50)=10(7) bacteria ml(-1)) and showed virulence factors, but were isolated with low frequency (3. 4%) and caused infection in only 31% of cases. Analysis of serotyping, phage typing and chromosomal DNA revealed at least 13 unrelated strains among pigmented S. marcescens. In conclusion, this work describes a low frequency of isolation of pigmented S. marcescens from clinical specimens, indicating that non-pigmented strains are clinically more significant.

Comparison of three methods for recording tibial H reflex: a clinical note.

Recording the tibial H reflex from the calf is routine in clinical electromyography. Two widely used methods are Hugon's and Braddom and Johnson's. Both methods produce an H wave with an initial positive deflection. Another method, the "half and half" method, generates an H wave with an initial negative deflection. To compare these methods, H-reflex responses were recorded in the right calf of 20 normal adults with each method using identical amplifier and stimulator settings. Stimulus current was increased by increments of 0.5-1.0 mA until the H reflex and motor (M) wave responses reached maximum amplitudes (Hmax, Mmax). H wave to M wave (H/M) amplitude ratios were calculated. The half-and-half method produced greatest Hmax, Mmax, and H/M ratio. Wave form configuration for the half-and-half method was biphasic with an initial negative deflection; it was triphasic with an initial positive deflection with the other methods. The shapes of M and H waves were similar in two methods but dissimilar in one. Because of the greater amplitude and the initial negative deflection, the H wave may be easier to detect with the half-and-half method and may present a latency that is more easily determined. For these reasons, the half-and-half method is recommended for measuring amplitude and latency in clinical settings.

Nosocomial infections caused by multiresistant Pseudomonas aeruginosa.

A case-control study was done to evaluate factors associated with nosocomial infections by multiresistant Pseudomonas aeruginosa (MRPA). Results showed that MRPA was associated with the use of immunosuppressive and antimicrobial drugs. Five typing methods indicated that the MRPA infections were due to multiple strains rather than a single strain.

Conductive hearing loss in individuals with fibrodysplasia ossificans progressiva.

Fibrodysplasia ossificans progressiva (FOP) is a very rare genetic disorder that is characterized by progressive heterotopic ossification of soft tissues and congenital malformation of the great toes. Although previous case studies have reported hearing loss in individuals with FOP, there have been no large-scale studies regarding the nature or cause of the hearing loss. Here, we report the findings of a two-part study. In Part I, we report the findings of a postal survey regarding hearing loss that was sent to 102 individuals with FOP. In Part II, we report the findings of on-site hearing evaluations of eight individuals with FOP. The findings of both studies indicate that individuals with FOP are at risk for hearing loss and that the type of loss is predominantly conductive in nature, similar to that seen in individuals who have otosclerosis.