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PURPOSE: Our objective was to elucidate host dependent factors of disease severity in invasive group A Streptococcal disease (iGAS) using transcriptome profiling of iGAS cases of varying degrees of severity at different timepoints. To our knowledge there are no previous transcriptome studies in iGAS patients. METHODS: We recruited iGAS cases from June 2018 to July 2020. Whole blood samples for transcriptome analysis and serum for biomarker analysis were collected at three timepoints representing the acute (A), the convalescent (B) and the post-infection phase (C). Gene expression was compared against clinical traits and disease course. Serum chemokine ligand 5 (CCL5, an inflammatory cytokine) concentration was also measured. RESULTS: Forty-five patients were enrolled. After disqualifying degraded or impure RNAs we had 34, 31 and 21 subjects at timepoints A, B, and C, respectively. Low expression of the CCL5 gene correlated strongly with severity (death or need for intensive care) at timepoint A (AUC = 0.92), supported by low concentrations of CCL5 in sera. CONCLUSIONS: Low gene expression levels and low serum concentration of CCL5 in the early stages of an iGAS infection were associated with a more severe disease course. CCL5 might have potential as a predictor of disease severity. Low expression of genes of cytotoxic immunity, especially CCL5, and corresponding low serum concentrations of CCL5 associated with a severe disease course, i.e. death, or need for intensive care, in early phase of invasive group A Streptococcal disease.
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A case of listeriosis occurred in a hospitalised patient in England in July 2017. Analysis by whole genome sequencing of the Listeria monocytogenes from the patient's blood culture was identified as clonal complex (CC) 121. This culture was indistinguishable to isolates from sandwiches, salads and the maufacturing environment of Company X which supplied these products widely to the National Health Service. Whilst an inpatient, the case was served sandwiches produced by this company on 12 occasions. No other cases infected by this type were detected in the UK between 2016 and 2020. Between 2016 and 2020, more than 3000 samples of food, food ingredients and environmental swabs from this company were tested. Listeria monocytogenes contamination rates declined after July 2017 from 31% to 0.3% for salads and 3% to 0% for sandwiches. A monophyletic group of 127 L. monocytogenes CC121 isolates was recovered during 2016-2019 and was used to estimate the time of the most recent common ancestor as 2014 (95% CI of between 2012 and 2016). These results represent persistent contamination of equipment, food contact surfaces and foods at a food manufacturer by a single L. monocytogenes strain. Colonisation and persistent contamination of food and production environments are risks for public health.
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Microbiología de Alimentos/estadística & datos numéricos , Servicio de Alimentación en Hospital , Listeria monocytogenes/aislamiento & purificación , Listeriosis/etiología , Inglaterra , Manipulación de Alimentos/normas , Enfermedades Transmitidas por los Alimentos/epidemiología , Enfermedades Transmitidas por los Alimentos/etiología , Humanos , Listeria monocytogenes/genética , Listeriosis/epidemiología , Masculino , Persona de Mediana Edad , Secuenciación Completa del GenomaRESUMEN
A gastrointestinal outbreak was reported among 154 diners who attended a Christmas buffet on the 9 and 10 December 2016. A retrospective cohort study was undertaken. Faecal samples, water, ice and an air ventilation device were tested for indicators and routine pathogens. Altogether 26% (24/91) fulfilled the case definition of having typical viral gastrointestinal symptoms. Norovirus genogroup I was detected in faecal samples from three cases. One of these cases tested positive also for sapovirus and had a family member testing positive for both norovirus and sapovirus. A diner who drank water or drinks with ice cubes (risk ratios (RR) 6.5, 95% confidence intervals (CI) 1.5-113.0) or both (RR 8.2, 95% CI 1.7-145.5) had an increased risk in a dose-response manner. Ice cubes from three vending machines had high levels of heterotrophic bacteria. A faulty air ventilation valve in the space where the ice cube machine was located was considered a likely cause of this outbreak. Leaking air ventilation valves may represent a neglected route of transmission in viral gastrointestinal outbreaks.
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In July 2014, an outbreak of gastroenteritis occurred among visitors to lakes in Tampere, Finland. We conducted a retrospective cohort study using an internet-based survey, solicited by public announcement, to identify source of infection and to implement control measures. Of 1453 persons enrolled in the study, 244 met the case definition (attack rate, 17%). In the pooled univariate analysis, risk factors for gastroenteritis included getting water in the mouth while swimming (Risk ratio (RR) 3.32; 95% Confidence interval (CI), 2.36-4.68) and playing on the wet sand at the beach (RR 1.90; 95% CI 1.50-2.41). In a multivariable analysis (logistic regression), the source of the infection was likely at two lakes (lake A Odds ratio (OR) 1.66; 95% CI 1.15-2.39 and lake B, OR 2.35; 95% CI 1.49-3.72). Norovirus (NoV) was found in 19 stool samples. All water samples from implicated beaches had acceptable values of fecal indicator bacteria and were negative for NoV. The likely source of the outbreak was lake-water contaminated with NoV at two popular lakes. Closure of swimming beaches, advice on hygienic precautions and rapid outbreak alerts were efficient in controlling the outbreak. Results suggest a need for new indicators of water quality and development of evidence-based recommendations regarding timing of safe reopen of recreational water venues associated with outbreaks.
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Playas , Infecciones por Caliciviridae/epidemiología , Brotes de Enfermedades/estadística & datos numéricos , Gastroenteritis/epidemiología , Lagos/virología , Natación , Adolescente , Adulto , Anciano , Niño , Preescolar , Estudios de Cohortes , Femenino , Finlandia/epidemiología , Gastroenteritis/virología , Humanos , Lactante , Masculino , Persona de Mediana Edad , Recreación , Estudios Retrospectivos , Factores de Riesgo , Adulto JovenRESUMEN
Respiratory viruses cause seasonal epidemics every year. Several respiratory pathogens are circulating simultaneously and typical symptoms of different respiratory infections are alike, meaning it is challenging to identify and diagnose different respiratory pathogens based on symptoms alone. mariPOC® is an automated, multianalyte antigen test which allows the rapid detection of nine respiratory infection pathogens [influenza A and B viruses, respiratory syncytial virus (RSV), human metapneumovirus, adenovirus, parainfluenza 1-3 viruses and pneumococci] from a single nasopharyngeal swab or aspirate samples, and, in addition, can be linked to laboratory information systems. During the study period from November 2010 to June 2014, a total of 22,485 multianalyte respi tests were performed in the 14 participating laboratories in Finland and, in total, 6897 positive analyte results were recorded. Of the tested samples, 25 % were positive for one respiratory pathogen, with RSV (9.8 %) and influenza A virus (7.2 %) being the most common findings, and 0.65 % of the samples were multivirus-positive. Only small geographical variations in seasonal epidemics occurred. Our results show that the mariPOC® multianalyte respi test allows simultaneous detection of several respiratory pathogens in real time. The results are reliable and give the clinician a picture of the current epidemiological situation, thus minimising guesswork.
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Infecciones del Sistema Respiratorio/epidemiología , Infecciones del Sistema Respiratorio/virología , Antígenos Virales/inmunología , Finlandia/epidemiología , Geografía , Historia del Siglo XXI , Humanos , Inmunoensayo/métodos , Infecciones del Sistema Respiratorio/diagnóstico , Infecciones del Sistema Respiratorio/historia , Sensibilidad y Especificidad , Virosis/diagnóstico , Virosis/epidemiología , Virosis/historia , Virosis/virologíaRESUMEN
In this study, the performances of two matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) systems, MALDI Biotyper (Bruker Daltonics) and VITEK MS (bioMérieux), were evaluated in the identification of viridans group streptococci. Two collections of isolates were tested with both methods. From a panel of type collection strains (n = 54), MALDI Biotyper gave correct species-level identification for 51/54 (94 %) strains and 37/54 (69 %) strains for the VITEK MS in vitro diagnostic (IVD) method. Additionally, a collection of blood cultures isolates which had been characterized earlier with partial sequencing of 16S rRNA (n = 97) was analyzed. MALDI Biotyper classified 89 % and VITEK MS 93 % of these correctly to the group level. Comparison of species-level identification from the blood culture collection was possible for 36 strains. MALDI Biotyper identified 75 % and VITEK MS 97 % of these strains consistently. Among the clinical isolates, MALDI Biotyper misidentified 36 strains as Streptococcus pneumoniae. Nevertheless, our results suggest that the current MALDI-TOF methods are a good alternative for the identification of viridans streptococci and do perform as well as or better than commercial phenotypical methods.
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Técnicas de Laboratorio Clínico/métodos , Técnicas Microbiológicas/métodos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Infecciones Estreptocócicas/diagnóstico , Estreptococos Viridans/clasificación , Estreptococos Viridans/aislamiento & purificación , Humanos , Infecciones Estreptocócicas/microbiología , Estreptococos Viridans/químicaRESUMEN
Mosquito-borne Sindbis virus (SINV) causes rash-arthritis syndrome in Finland. Major outbreaks with approximately 7-year cycles have caused substantial burden of illness. Forest dwelling grouse are suspected to be amplifying hosts, with the infection transmitted to humans by mosquito bites. SINV infection surveillance data for 19842010 were used to create a negative binomial hurdle model, with seasonality, long-term cycles, climatic, ecological and socioeconomic variables. Climatic factors during early summer and amount of snow in April described the occurrence and incidence of SINV infections. Regulated water shore and hatch-year black grouse density described the occurrence, while population working in agriculture, agricultural land(negative) and income (negative) described the incidence of the disease. The prediction for 2009 was 85 cases (95% prediction interval 2-1187), while the actual occurrence was 106. We identified novel and known risk factors. The prevention of SINV infections in regulated water areas by infected mosquito populations should be targeted.
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Infecciones por Alphavirus/epidemiología , Virus Sindbis/aislamiento & purificación , Adulto , Agricultura , Infecciones por Alphavirus/transmisión , Animales , Clima , Culicidae/crecimiento & desarrollo , Ecosistema , Femenino , Finlandia/epidemiología , Humanos , Incidencia , Insectos Vectores , Masculino , Persona de Mediana Edad , Modelos Estadísticos , Exposición Profesional , Factores de Riesgo , Factores SocioeconómicosRESUMEN
A new automated closed tube PCR assay, the GenomEra(™) MRSA/SA Diagnose (Abacus Diagnostica Oy, Finland) was evaluated for rapid confirmation of methicillin-resistant Staphylococcus aureus (MRSA) from cultured screening specimens. The ability of the assay to detect genotypically different MRSA strains was studied with a collection of 304 MRSA isolates covering 68 spa types. The specificity was investigated with a collection of 146 non-MRSA staphylococcus isolates. The usefulness of the assay for clinical purposes was assessed by a sequential combination of MRSA screening culture and confirmation of the colonies with the GenomEra MRSA/SA Diagnose assay. A total of 145 suspected MRSA colonies on chromogenic plates were analyzed this way. All MRSA isolates from the culture collection and from the clinical screening specimens were confirmed as MRSA with the GenomEra MRSA/SA Diagnose assay and none of the non-MRSA staphylococci caused false-positive results, which indicates both sensitivity and specificity of 100%. The combination of GenomEra MRSA/SA Diagnose with preceding culture on selective MRSA agar permitted MRSA confirmation within 24 h. This practice offers a reliable and quick detection of MRSA that is also suitable in areas where several strain types cause epidemics.
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Medios de Cultivo/química , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena de la Polimerasa/métodos , Infecciones Estafilocócicas/microbiología , Agar , Compuestos Cromogénicos/metabolismo , Humanos , Pruebas de Sensibilidad Microbiana/métodos , Sensibilidad y EspecificidadRESUMEN
A method for the rapid detection of methicillin-sensitive and -resistant Staphylococcus aureus (MSSA and MRSA, respectively) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) with a straightforward sample preparation protocol of blood cultures using an automated homogeneous polymerase chain reaction (PCR) assay, the GenomEra™ MRSA/SA (Abacus Diagnostica Oy, Turku, Finland), is presented. In total, 316 BacT/Alert (bioMérieux, Marcy l'Etoile, France) and 433 BACTEC (Becton Dickinson, Sparks, MD, USA) blood culture bottles were analyzed, including 725 positive cultures containing Gram-positive cocci in clusters (n = 419) and other Gram stain forms (n = 361), as well as 24 signal- and growth-negative bottles. Detection sensitivities for MSSA, MRSA, and MRCoNS were 99.4 % (158/159), 100.0 % (9/9), and 99.3 % (132/133), respectively. One false-positive MRSA result was detected from a non-staphylococci-containing bottle, yielding a specificity of 99.8 %. The lowest detectable amount of viable cells in the blood culture sample was 4 × 10(4) CFU/mL. The results were available within one hour after microbial growth detection and the two-step, time-resolved fluorometric (TRF) measurement mode employed by the GenomEra CDX™ instrument showed no interference from blood, charcoal, or culture media. The method described lacks all sample purification steps and allows reliable and simplified pathogen detection also in clinical microbiology laboratory settings without specialized molecular microbiology competence.
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Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Infecciones Estafilocócicas/diagnóstico , Staphylococcus aureus/aislamiento & purificación , Proteínas Bacterianas/análisis , Técnicas de Tipificación Bacteriana/métodos , Supervivencia Celular , Coagulasa , Fluorometría/instrumentación , Fluorometría/métodos , Humanos , Staphylococcus aureus Resistente a Meticilina/enzimología , Proteínas de Unión a las Penicilinas , Reacción en Cadena de la Polimerasa/instrumentación , Juego de Reactivos para Diagnóstico/normas , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Infecciones Estafilocócicas/sangre , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/enzimología , Staphylococcus aureus/genética , Factores de TiempoRESUMEN
In October 2011 in Finland, two persons fell ill with symptoms compatible with botulism after having eaten conserved olives stuffed with almonds. One of these two died. Clostridium botulinum type B and its neurotoxin were detected in the implicated olives by PCR and mouse bioassay, respectively. The olives were traced back to an Italian manufacturer and withdrawn from the market. The public and other European countries were informed through media and Europe-wide notifications.
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Botulismo/diagnóstico , Clostridium botulinum , Alimentos en Conserva/microbiología , Olea/microbiología , Adulto , Anciano , Animales , Botulismo/etiología , Resultado Fatal , Finlandia , Contaminación de Alimentos , Alimentos en Conserva/efectos adversos , Humanos , Internacionalidad , Ratones , Olea/efectos adversosRESUMEN
BACKGROUND: Longer duration from symptom onset is associated with increased risk of perforation in appendicitis. In previous studies, in-hospital delay to surgery has had conflicting effects on perforation rates. Although preoperative antibiotics have been shown to reduce postoperative infections, there are no data showing that administration of antibiotics while waiting for surgery has any benefits. The aims of this study are to evaluate the role of both in-hospital delay to surgery and antibiotic treatment while waiting for surgery on the rate of appendiceal perforation. METHODS: This prospective, open-label, randomized, controlled non-inferiority trial compares the in-hospital delay to surgery of less than 8 hours versus less than 24 hours in adult patients with predicted uncomplicated acute appendicitis. Additionally, participants are randomized either to receive or not to receive antibiotics while waiting for surgery. The primary study endpoint is the rate of perforated appendicitis discovered during appendicectomy. The aim is to randomize 1800 patients, that is estimated to give a power of 90 per cent (χ2) for the non-inferiority margin of 5 percentage points for both layers (urgency and preoperative antibiotic). Secondary endpoints include length of hospital stay, 30-day complications graded using Clavien-Dindo classification, preoperative pain, conversion rate, histopathological diagnosis and Sunshine Appendicitis Grading System classification. DISCUSSION: There are no previous randomized controlled studies for either in-hospital delay or preoperative antibiotic treatment. The trial will yield new level 1 evidence.EU Clinical Trials Register, EudraCT Number: 2019-002348-26; registration number: NCT04378868 (http://www.clinicaltrials.gov).
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Antibacterianos , Apendicitis , Adulto , Antibacterianos/uso terapéutico , Apendicectomía , Apendicitis/tratamiento farmacológico , Apendicitis/cirugía , Estudios de Equivalencia como Asunto , Humanos , Tiempo de Internación , Estudios Prospectivos , Ensayos Clínicos Controlados Aleatorios como AsuntoRESUMEN
BACKGROUND: The usefulness of induced sputum in searching for causative agents of pneumonia in children has not been studied. METHODS: The study involved 101 children, aged 6 months to 15 years, treated for community-acquired pneumonia at Turku University Hospital (Turku, Finland) from January 2006 to April 2007. Nasopharyngeal aspirate samples were first collected through both nostrils. Sputum production was then induced by inhalation of 5.0% hypertonic saline for 5-10 min and a sputum sample was either aspirated or expectorated. The presence and amount of bacteria and viruses in paired nasopharyngeal aspirate and sputum specimens was analysed and compared using semiquantitative bacterial culture and quantitative PCR techniques. RESULTS: A good quality sputum specimen was obtained from 76 children. The possible causative agent was found in 90% of cases. Streptococcus pneumoniae (46%) and rhinovirus (29%) were the most common microbes detected. Newly discovered viruses human bocavirus and human metapneumovirus were detected in 18% and 13% of the children, respectively. One-quarter of all bacterial findings were only detected in sputum, and the amount of bacteria in the remainder of the sputum specimens compared with nasopharyngeal aspirate was higher in 14% and equal in 70%. The amount of rhinovirus in sputum was higher than in nasopharyngeal aspirate in 82%. CONCLUSIONS: Sputum induction provides good quality sputum specimens with high microbiological yield in children with community-acquired pneumonia. Induced sputum analysis can be useful in the microbiological diagnosis of childhood community-acquired pneumonia.
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Infecciones Comunitarias Adquiridas/diagnóstico , Neumonía Bacteriana/diagnóstico , Neumonía Viral/diagnóstico , Esputo/microbiología , Adolescente , Bacterias/aislamiento & purificación , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Nasofaringe/microbiología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Virus/aislamiento & purificaciónRESUMEN
The Vitek 2 system was compared with conventional assimilation, fermentation and morphological methods for its ability to identify yeast isolates from among 151 clinical specimens and 16 known type culture or quality control strains. An unequivocal identification was obtained for 155 (92.8%) isolates, with low discrimination for nine (5.4%) and false identification for three (1.8%) isolates. All isolates of Candida albicans, Candida glabrata and Candida krusei were identified correctly. It was concluded that the Vitek 2 system offers an excellent alternative for the identification of yeasts in a clinical laboratory.
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Candida/aislamiento & purificación , Técnicas de Tipificación Micológica/métodos , Técnicas de Tipificación Micológica/normas , Candida/clasificación , Humanos , Técnicas de Tipificación Micológica/instrumentación , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Ready to use dry-reagent PCR assays for Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas spp. and for broad-range bacteria detection were developed. The assays were based on novel switchable lanthanide probes that provide sensitive target DNA detection with exceptionally high signal-to-background ratio, thus enabling clear discrimination between positive and negative results. For example, sensitivity of three S. aureus and two S. pneumonia bacteria (colony forming units) per PCR assay was measured with fluorescence signal more than 30 times over the background signal level. The rapid and easy-to-use assays are suitable for routine clinical diagnostics without molecular biology expertise and facilities.
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Infecciones Bacterianas/diagnóstico , Técnicas Bacteriológicas/métodos , Elementos de la Serie de los Lantanoides/metabolismo , Mediciones Luminiscentes , Técnicas de Diagnóstico Molecular/métodos , Sondas de Oligonucleótidos/genética , Reacción en Cadena de la Polimerasa/métodos , Escherichia coli/genética , Escherichia coli/aislamiento & purificación , Humanos , Pseudomonas/genética , Pseudomonas/aislamiento & purificación , Sensibilidad y Especificidad , Staphylococcus aureus/genética , Staphylococcus aureus/aislamiento & purificación , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/aislamiento & purificaciónRESUMEN
BACKGROUND: Lovastatin is a cholesterol-lowering drug that can cause myopathy as a rare side effect. Concomitant use of certain drugs (e.g., cyclosporine) increases the risk of skeletal muscle toxicity. Lovastatin is metabolized by CYP3A4. Because itraconazole is a potent inhibitor of CYP3A4, we wanted to study a possible interaction between these drugs. METHODS: In this double-blind, randomized, two-phase crossover study, 12 healthy volunteers received either 200 mg itraconazole or placebo orally once a day for 4 days. On day 4, each subject ingested a single 40 mg dose of lovastatin. Plasma concentrations of lovastatin, lovastatin acid, itraconazole, hydroxyitraconazole, and creatine kinase were measured up to 24 hours. RESULTS: On average, itraconazole increased the peak concentration (Cmax) of lovastatin and the area under the lovastatin concentration-time curve (AUC) more than twentyfold (p < 0.001). The mean Cmax of the active metabolite, lovastatin acid, was increased 13-fold (range, tenfold to 23-fold; p < 0.001) and the AUC(0-24) twentyfold (p < 0.001). In one subject plasma creatine kinase was increased tenfold within 24 hours of lovastatin administration during the itraconazole phase but not during the placebo phase. No increase in creatine kinase was observed in the other subjects. CONCLUSIONS: Itraconazole greatly increases plasma concentrations of lovastatin and lovastatin acid. Inhibition of CYP3A4-mediated metabolism probably explains the increased toxicity of lovastatin caused not only by itraconazole but also by cyclosporine, erythromycin, and other inhibitors of CYP3A4. Their concomitant use with lovastatin and simvastatin should be avoided, or the dose of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors should be reduced accordingly.
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Anticolesterolemiantes/sangre , Antifúngicos/farmacología , Inhibidores Enzimáticos/sangre , Inhibidores de Hidroximetilglutaril-CoA Reductasas , Itraconazol/farmacología , Lovastatina/sangre , Adulto , Estudios Cruzados , Sistema Enzimático del Citocromo P-450/fisiología , Método Doble Ciego , Interacciones Farmacológicas , Femenino , Humanos , Itraconazol/sangre , MasculinoRESUMEN
BACKGROUND: Felodipine, a dihydropyridine calcium antagonist, is extensively metabolized by CYP3A4. Itraconazole strongly interacts with some of the substrates of CYP3A4 (e.g., terfenadine, triazolam and lovastatin); hence it is important to uncover the possible interaction of itraconazole with felodipine. METHODS: A double-blind, randomized, two-phase crossover design was used to investigate the interaction between felodipine and itraconazole. Nine healthy volunteers received either 200 mg itraconazole or placebo orally once a day for 4 days. On day 4, each ingested a single 5 mg oral dose of felodipine. Plasma concentrations of felodipine and itraconazole were determined and systolic and diastolic blood pressures and heart rate were measured up to 32 hours. RESULTS: On average, itraconazole increased the peak plasma concentration (Cmax) of felodipine nearly eightfold (p < 0.001), the areas under the felodipine concentration-time curve [AUC(0-32) and AUC(0-infinity)] about sixfold (p < 0.001), and the elimination half-life twofold (p < 0.05). In seven of the nine subjects, even the Cmax of felodipine was lower without itraconazole than the 32-hour concentrations during the itraconazole phase. The decreases in blood pressure and the increases in heart rate were significantly greater during the itraconazole phase than during the placebo phase. CONCLUSIONS: Itraconazole greatly increases plasma concentrations and effects of oral felodipine. The inhibition of CYP3A4 during the first-pass and elimination phases of felodipine seems to be the mechanism of the observed interaction. The concomitant use of itraconazole and some other azole antifungals with felodipine and other dihydropyridine calcium antagonists should be avoided or their doses should be reduced accordingly.
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Antifúngicos/farmacología , Bloqueadores de los Canales de Calcio/farmacocinética , Felodipino/farmacocinética , Itraconazol/farmacología , Adulto , Antifúngicos/farmacocinética , Área Bajo la Curva , Bloqueadores de los Canales de Calcio/sangre , Estudios Cruzados , Método Doble Ciego , Sinergismo Farmacológico , Felodipino/sangre , Femenino , Humanos , Itraconazol/farmacocinética , Masculino , Valores de ReferenciaRESUMEN
We evaluated PCR for the detection of Bacillus anthracis DNA from simulated clinical specimens relevant for the microbiological diagnosis of anthrax or exposure to B. anthracis spores. In simulated blood specimens, the lowest limit of detection was 400 CFU per mL of blood, which may be sufficient for samples from patients with septic anthrax. Screening nasal swabs by PCR may not be sensitive enough to rule out dangerous exposure to anthrax spores, as a minimum of 2000 spores per sample was required for detectable amplification. As spores survived some standard DNA purification methods, special attention should be paid to laboratory safety when preparing samples possibly containing live spores.
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Carbunco/microbiología , Bacillus anthracis/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Carbunco/sangre , Bacillus anthracis/genética , ADN Bacteriano/química , ADN Bacteriano/genética , Humanos , Mucosa Nasal/microbiología , Sensibilidad y Especificidad , Esporas Bacterianas , Células MadreRESUMEN
H. bizzozeronii CCUG 35045, a new canine gastric Helicobacter spp. was used for experimental infection of four weaned puppies at 7 weeks of age. Controls were four nonchallenged puppies. The puppies originated from two dams which had Helicobacter salomonis infection in biopsy samples taken 3 weeks before the delivery but which had urease, brush cytology and culture-negative biopsy samples taken 7 weeks after antimicrobial treatment (metronidazole, amoxicillin, bismuth subcitrate). Both dams were detected urease- and Helicobacter-positive again three and a half months after therapy. Dam B was shown to be colonised with the similar genotype of H. salomonis for more than 2 years. Unexpectedly, H. salomonis was also cultured from gastric biopsy samples of the nonchallenged puppies three times during 7 months. When H. salomonis isolates of dams and puppies were studied by ribotyping (HaeIII, ClaI or PstI) they were shown to be identical although the HaeIII and PstI REA patterns of dam A differed from the patterns of dam B and nonchallenged group by one fragment. PFGE pattern analysis of NotI digests, however, revealed that the isolates of the puppies were identical with the isolates of dam B, and differed from the isolates of dam A. The isolates of the dams and puppies in the nonchallenged group were metronidazole-resistant. The antimicrobial therapy had merely suppressed, but not eradicated, the infection from dams. These studies suggested that puppies may acquire gastric Helicobacter infection from dams during the lactation period and puppies can infect each other during their early life. PFGE pattern analysis was shown to be a more distinguishing method than ribotyping to study the similarity of the isolates.
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Enfermedades de los Perros/transmisión , Mucosa Gástrica/microbiología , Infecciones por Helicobacter/veterinaria , Helicobacter/clasificación , Gastropatías/veterinaria , Animales , Antibacterianos/farmacología , Biopsia/veterinaria , Modelos Animales de Enfermedad , Enfermedades de los Perros/microbiología , Perros , Electroforesis en Gel de Agar/veterinaria , Electroforesis en Gel de Campo Pulsado/veterinaria , Heces/microbiología , Femenino , Mucosa Gástrica/fisiopatología , Helicobacter/efectos de los fármacos , Helicobacter/genética , Infecciones por Helicobacter/microbiología , Infecciones por Helicobacter/transmisión , Metronidazol/farmacología , Sondas ARN , Mapeo Restrictivo/veterinaria , Gastropatías/microbiologíaRESUMEN
The cell morphology, the number of flagella, the occurrence of periplasmic fibrils and ultrastructural structures of five groups of cultured canine gastric Helicobacter spp. were compared. The study included four strains of Helicobacter felis, four strains of Helicobacter bizzozeronii, one strain of 'Flexispira', six strains of an unnamed spiral organism 2 and one strain of an unnamed spiral organism 3 which were isolated from gastric biopsies. Cultures were studied with negative staining, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Bacterial dimensions were measured from the negative staining samples and values were tested with ANOVA and Bonferroni tests. The organisms studied differed from each other morphologically. H. felis was a slightly spiraled organism with periplasmic fibrils. 'Flexispira' was a thin and straight organism with periplasmic fibrils. H. bizzozeronii was a tightly spiraled organism. Spiral organism 2 was loosely spiraled and thicker than the other organisms. Spiral organism 3 was a short curved rod having a single bipolar flagellum. The other species had multiple flagella. As a conclusion the canine gastric Helicobacter spp. can be differentiated from each other morphologically with an electron microscope. The morphological differences were mainly found in the structures involved in motility. The importance of the differences may lie in their impact on the colonization in a gastric mucous environment.
Asunto(s)
Mucosa Gástrica/microbiología , Helicobacter/clasificación , Animales , Biopsia , Perros , Flagelos/ultraestructura , Helicobacter/ultraestructura , Microscopía Electrónica , Periplasma/ultraestructura , Ácido Fosfotúngstico , Coloración y EtiquetadoRESUMEN
Effects of ethanol (EtOH, 0.65 + 0.35 g.kg-1), diazepam (DZ, 15 and 30 mg), lorazepam (LZ, 2 mg) on divided attention were measured in two placebo-controlled crossover studies with healthy young subjects. The test comprised four parallel computer screens with a ball moving along a circular obstacle course on each screen at different rates. When the ball entered an obstacle on any screen, the subject had to press the respective button. The obstacles varied in numbers and shapes, and randomly changed their location every 10 s. Concomitant aural stimuli were responded to by pushing the foot pedals. The primary visual variables were the absolute and percent numbers of correct responses on each screen. Concentrated attention was measured with a symbol digit substitution (SDST) and digit copying (DDCT) tests, for 3 min each. In Study I, with 12 subjects, the tests (4 min) were made before the treatment (placebo, EtOH, DZ) and 1, 3, and 6 h after intake. EtOH impaired attention on the lateral but not on medial screens, with and without aural stimuli, the "special" obstacles of deviating shape being the most sensitive targets to EtOH effects. DZ 15 mg did not modify divided attention whereas it impaired SDST performance and was subjectively slightly more potent than EtOH on visual analog scales. DZ 30 mg impaired attention on the lateral screens, with and without aural stimuli, but without preference to "special" obstacles. It also reduced responses to aural stimuli, strongly impaired SDST and DDCT, and caused subjective sedation. In Study II, with 9 subjects, the test run without aural stimuli was easier but lasted for 15 min.(ABSTRACT TRUNCATED AT 250 WORDS)