Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates

Enterotoxigenic Escherichia coli (ETEC) produce heat-labile (LT) and/or heat-stable enterotoxins (ST). Despite that, the mechanism of action of both toxins are well known, there is great controversy in the literature concerning the in vitro production and release of LT and, for ST, no major concerns have been discussed. Furthermore, the majority of published papers describe the use of only one or a few ETEC isolates to define the production and release of these toxins, which hinders the detection of ETEC by phenotypic approaches. Thus, the present study was undertaken to obtain a better understanding of ST and LT toxin production and release under laboratory conditions. Accordingly, a collection of 90 LT-, ST-, and ST/LT-producing ETEC isolates was used to determine a protocol for toxin production and release aimed at ETEC detection. for this, we used previously raised anti-LT antibodies and the anti-ST monoclonal and polyclonal antibodies described herein. the presence of bile salts and the use of certain antibiotics improved ETEC toxin production/release. Triton X-100, as chemical treatment, proved to be an alternative method for toxin release. Consequently, a common protocol that can increase the production and release of LT and ST toxins could facilitate and enhance the sensitivity of diagnostic tests for ETEC using the raised and described antibodies in the present work.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Butantan Inst, Bacteriol Lab, BR-05503900 São Paulo, BrazilSão Paulo Trop Med Inst, Seroepidemiol & Immunol Lab, BR-05403000 São Paulo, BrazilFleury Med & Hlth, BR-04344903 São Paulo, BrazilButantan Inst, Immunopathol Lab, BR-05503900 São Paulo, BrazilButantan Inst, Immunochem Lab, BR-05503900 São Paulo, BrazilAdolfo Lutz Inst, Bacteriol Sect, BR-01246000 São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol, BR-04923062 São Paulo, SP, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol, BR-04923062 São Paulo, SP, BrazilWeb of Scienc

Standardization and international multicenter validation of a PulseNet pulsed-field gel electrophoresis protocol for subtyping Shigella flexneri isolates

Fil: Pichel, Mariana. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Brengi, Silvina P. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Cooper, Kara L. F. Centers for Disease Control and Prevention; Estados Unidos.Fil: Ribot, Efrain M. Centers for Disease Control and Prevention; Georgia.Fil: Al-Busaidy, Suleiman. Central Public Health Laboratory; Omán.Fil: Araya, Pamela. Instituto de Salud Pública de Chile; Chile.Fil: Fernández, Jorge. Instituto de Salud Pública de Chile; Chile.Fil: Vaz, Tania Ibelli. Instituto Adolfo Lutz; Brazil.Fil: Kam, Kai Man. Public Health Laboratory Centre; Japón.Fil: Morcos, Myriam. Regional Center at the U.S. Naval Medical Research Unit #3 (NAMRU-3). Global Disease Detection (GDD); Egipto.Fil: Nielsen, Eva M. Statens Serum Institut; Dinamarca.Fil: Nadon, Celine. National Microbiology Laboratory; Canadá.Fil: Pimentel, Guillermo. Regional Center at the U.S. Naval Medical Research Unit #3 (NAMRU-3). Global Disease Detection (GDD); Egipto.Fil: Pérez-Gutiérrez, Enrique. PAHO/WHO. Health Surveillance; Panamá.Fil: Gerner-Smidt, Peter. Centers for Disease Control and Prevention; Georgia.Fil: Binsztein, Norma. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Shigella flexneri is one of the agents most frequently linked to diarrheal illness in developing countries and often causes outbreaks in settings with poor hygiene or sanitary conditions. Travel is one of the means by which S. flexneri can be imported into developed countries, where this pathogen is not commonly seen. A robust and discriminatory subtyping method is needed for the surveillance of S. flexneri locally and regionally, and to aid in the detection and investigation of outbreaks. The PulseNet International network utilizes standardized pulsedfield gel electrophoresis (PFGE) protocols to carry out laboratory-based surveillance of foodborne pathogens in combination with epidemiologic data. A multicenter validation was carried out in nine PulseNet laboratories located in North and South America, Europe, and Asia, and it demonstrated that a new protocol is highly robust and reproducible for subtyping of S. flexneri. This protocol, already approved for PulseNet laboratories, applies NotI and XbaI as primary and secondary restriction enzymes, respectively, under electrophoresis conditions of initial switch time of 5 s to final switch time of 35 s, at 6 volts/cm

Reducing the Clinical and Public Health Burden of Familial Hypercholesterolemia A Global Call to Action

Q1Q1Artículo completoE1-E13IMPORTANCE Familial hypercholesterolemia (FH) is an underdiagnosed and undertreated genetic disorder that leads to premature morbidity and mortality due to atherosclerotic cardiovascular disease. Familial hypercholesterolemia affects 1 in 200 to 250 people around the world of every race and ethnicity. The lack of general awareness of FH among the public and medical community has resulted in only 10% of the FH population being diagnosed and adequately treated. The World Health Organization recognized FH as a public health priority in 1998 during a consultation meeting in Geneva, Switzerland. The World Health Organization report highlighted 11 recommendations to address FH worldwide, from diagnosis and treatment to family screening and education. Research since the 1998 report has increased understanding and awareness of FH, particularly in specialty areas, such as cardiology and lipidology. However, in the past 20 years, there has been little progress in implementing the 11 recommendations to prevent premature atherosclerotic cardiovascular disease in an entire generation of families with FH. OBSERVATIONS In 2018, the Familial Hypercholesterolemia Foundation and the World Heart Federation convened the international FH community to update the 11 recommendations. Two meetings were held: one at the 2018 FH Foundation Global Summit and the other during the 2018 World Congress of Cardiology and Cardiovascular Health. Each meeting served as a platform for the FH community to examine the original recommendations, assess the gaps, and provide commentary on the revised recommendations. The Global Call to Action on Familial Hypercholesterolemia thus represents individuals with FH, advocacy leaders, scientific experts, policy makers, and the original authors of the 1998 World Health Organization report. Attendees from 40 countries brought perspectives on FH from low-, middle-, and high-income regions. Tables listing country-specific government support for FH care, existing country-specific and international FH scientific statements and guidelines, country-specific and international FH registries, and known FH advocacy organizations around the world were created. CONCLUSIONS AND RELEVANCE By adopting the 9 updated public policy recommendations created for this document, covering awareness; advocacy; screening, testing, and diagnosis; treatment; family-based care; registries; research; and cost and value, individual countries have the opportunity to prevent atherosclerotic heart disease in their citizens carrying a gene associated with FH and, likely, all those with severe hypercholesterolemia as well

Serotype diversity as a drawback in the surveillance of Shiga toxin-producing Escherichia coli infections in Brazil

Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Immunol & Parasitol, BR-04023062 São Paulo, BrazilInst Adolfo Lutz Registro, Secao Bacteriol, São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Immunol & Parasitol, BR-04023062 São Paulo, BrazilWeb of Scienc

Vanadyl cationic complexes as catalysts in olefin oxidation

Three new mononuclear oxovanadium(IV) complexes [VO(acac)(R-BIAN)]Cl (BIAN = 1,2-bis{(R-phenyl)imino} acenaphthene, R = H, 1; CH3, 2; Cl, 3) were prepared and characterized. They promoted the catalytic oxidation of olefins such as cyclohexene, cis-cyclooctene, and styrene with both tbhp (tert-butylhydroperoxide) and H2O2, and of enantiopure olefins (S(-)- and R(+)-pinene, and S(-)- and R(+)-limonene) selectively to their epoxides, with tbhp as the oxidant. The TOFs for styrene epoxidation promoted by complex 3 with H2O2 (290 mol mol(-1)V h(-1)) and for cis-cyclooctene epoxidation by 2 with tbhp (248 mol mol(V)(-1) h(-1)) are particularly good. Conversions reached 90% for several systems with tbhp, and were lower with H2O2. A preference for the internal C=C bond, rather than the terminal one, was found for limonene. Kinetic data indicate an associative process as the first step of the reaction and complex [VO(acac)(H-BIAN)](+) (1(+)) was isolated in an FTICR cell after adding tbhp to 1. EPR studies provide evidence for the presence of a V(IV) species in solution, until at least 48 hours after the addition of tbhp and cis-cyclooctene, and cyclic voltammetry studies revealed an oxidation potential above 1 V for complex 1. DFT calculations suggest that a [VO(H-BIAN)(MeOO)](+) complex is the likely active V(IV) species in the catalytic cycle from which two competitive mechanisms for the reaction proceed, an outer sphere path with an external attack of the olefin at the coordinated peroxide, and an inner sphere mechanism starting with a complex with the olefin coordinated to vanadium

Early Hyperglycemia in Very Preterm Infants Is Associated with Reduced White Matter Volume and Worse Cognitive and Motor Outcomes at 2.5 Years

Introduction: Hyperglycemia in very preterm infants is associated with increased morbidity and mortality. We aimed to investigate potential associations between early hyperglycemia, neonatal cerebral magnetic resonance imaging (MRI), and neurodevelopment at 2.5 years. Methods: The study population included 69 infants with gestational age (GA) 22.3-31.9 weeks (n = 29 with GA <28 weeks), born 2011-2014. Plasma glucose concentrations during the first week were checked according to clinical routines. Hyperglycemia was defined as glucose concentrations above 8.3 mmol/L (150 mg/dL) and above 10 mmol/L (180 mg/dL), respectively, categorized as the highest glucose days 0-2, number of days above 8.3 and 10 mmol/L, and prolonged (yes/no) 2 days or more above 8.3 and 10 mmol/L. The MRI analysis included morphological assessment, regional brain volumes, and assessment of apparent diffusion coefficient (ADC). Neurodevelopmental impairment (NDI) developed in 13 of 67 infants with available outcomes, of which 57 were assessed with the Bayley-III. Univariate and multiple linear and logistic regressions were performed with adjustments for GA, birth weight z-scores, and illness severity expressed as days on mechanical ventilation. Results: Hyperglycemia above 8.3 mmol/L and 10 mmol/L was present in 47.8% and 31.9% of the infants. Hyperglycemia correlated independently with lower white matter volume, but not with other regional brain volumes, and was also associated with lower ADC values in white matter. Hyperglycemia also correlated with lower Bayley-III cognitive and motor scores in infants with GA <28 weeks, but there was no significant effect on NDI. Conclusion: Early hyperglycemia is associated with white matter injury and poorer neurodevelopment in very preterm infants

Prevalence of thyroid nodules in 40 years-old or old women

Nódulos de tireóide são uma condição clínica comum e em seu diagnóstico diferencial inclui-se o câncer de tireóide. Como o câncer de tireóide é raro, é importante conhecer a prevalência de nódulos de tireóide em grupos fortemente predispostos para dimensionar seu impacto e adequar estratégias diagnósticas. OBJETIVOS. O objetivo deste estudo foi verificar a prevalência de nódulos de tireóide diagnosticados por ecografia em mulheres na maturidade. MÉTODOS. Entre agosto de 1996 e dezembro de 1997 foram avaliadas, de forma consecutiva, pacientes do sexo feminino com idade de 40 anos ou mais que realizavam ecografia no Serviço de Radiologia do HCPA. Foram excluídas as mulheres encaminhadas para exame de tireóide. As 207 pacientes examinadas tiveram média de idade de 54,7 anos (mediana: 53 anos com mínimo de 40 e máximo de 86 anos). Cento e vinte e três (59,4%) apresentaram tireóide normal e 82 (39,6%) tiveram alguma alteração na glândula. CONCLUSÕES. Alterações nodulares foram encontradas em 73 (35,3%) mulheres, e 35 (16,9%) portavam nódulos tireoidianos com 1 cm ou mais. As mulheres sem nódulos de tireóide eram mais jovens (53,1 ± 10,7 anos) que as que apresentavam nódulos tireoidianos (58,2 ± 10,6 anos), p=0,001 e houve aumento do número de quaisquer nódulos (p=0.001) ou de nódulos maior que 1 cm (p=0.007) com o aumento da idade. Assim sendo, nódulos de tireóide são muito comuns em mulheres com 40 anos ou mais, a sua prevalência aumenta com a idade nesse grupo etário, e eles são potencialmente palpáveis (maiores que 1 cm) em 1 em cada 6 mulheres nesta faixa etária. É importante que tenhamos esses dados em mente quando indicamos avaliação ecográfica da tireóide em mulheres com 40 anos ou mais.OBJECTIVES. Thyroid nodules constitute a very common clinical problem and the differential diagnosis includes thyroid cancer. As thyroid cancer is rare, it is important to know the prevalence of thyroid nodules in cohorts strongly predisposed to this problem to be able to measure its impact on the health care system, and to devise appropriate diagnostic strategies. The aim of this study was to determine the prevalence of thyroid nodules in women 40 year-old or older by ecography. METHODS. Between August 1996 and December 1997, 207 women, 40 year-old or older, were consecutively evaluated by thyroid ecography in the Radiology Division of the Hospital de Clínicas de Porto Alegre. Patients who came to the Division to be submitted to thyroid ecography were excluded. The mean age of patients studied was 54.7 years (median: 53, range: 40-86 years). One hundred and twenty three patients (54.9%) had a normal thyroid and eighty-two had an abnormal gland (39.6%). Thyroid nodules were detected in seventythree women (35.3%) and these were larger than 1 cm in 35 women (16.9%). Women without thyroid nodules were younger (53.1 ±10.7 years) than women with thyroid nodules (58.2 ±10.6 years) p=0.001; and the prevalence of any thyroid nodule (p=0.001) or nodules larger than 1 cm (p=0.007) increased with age. CONCLUSIONS. Thyroid nodules are very common in women 40 year-old or older, their prevalence increases with age in this cohort and these nodules are potentially palpable (larger than one cm) in one of six women in this age group It is important to have these data in mind when ordering thyroid ecographic studies in women 40 year-old or older

Local plant species delimitation in a highly diverse Amazonian forest: Do we all see the same species?

Question: How reliable is the process of delimiting plant species by morphotyping sterile specimens from a highly diverse Amazonian forest plot? Location: Biological Dynamics of Forest Fragments Project (BDFFP), Central Amazon, Manaus, Brazil. Methods: A taxonomic exercise was conducted during a Center for Tropical Forest Science (CTFS) Taxonomy Workshop held in Manaus in April 2011, using specimens collected in a 25-ha forest plot. The plant species from this plot had been previously delimited by morphotyping of ca. 80 000 sterile specimens, a process that resulted in the recognition of 115 cases (accounting for 38% of all trees) in which species delimitation was problematic. For the workshop, we selected a subsample of specimens for eight of these difficult cases (taxonomic groups/complexes) and asked 14 participants with different levels of botanical training to independently sort these specimens into morphospecies. We then compared the classifications made by all participants and explored correlations between botanical training and plant classification. Results: The classification of specimens into morphospecies was highly variable among participants, except for one taxonomic group/complex, for which the median pair-wise similarity was 95%. For the other seven taxonomic groups/complexes, median pair-wise similarity values ranged from 52% to 67%. Training did not increase the similarity in the definition of morphospecies except for two taxonomic groups/complexes, for which there was higher congruence between the classifications made by participants with a high level of botanical training than in comparisons that included less-experienced participants. The total number of morphospecies defined by participants was highly variable for all taxonomic groups/complexes, with the total number varying from 12 to 46 (a 383% difference). Conclusions: Local plant species delimitation by morphotyping sterile specimens is prone to large uncertainties, and botanical training may not reduce them. We argue that uncertainty in species delimitation should be explicitly considered in plant biodiversity inventories as diversity estimates may be strongly affected by such uncertainties. We recommend that species delimitation and identification be treated as separate processes and that difficulties be explicitly recorded, so as to permit error estimates and the refinement of taxonomic data. Poor taxonomic knowledge, lack of tools and high diversity limit our ability to recognize plant species in Amazon forest plot inventories. While the identification of species is a well-known problem, the delimitation of local species is considered unproblematic. We show that local species delimitation is also prone to large uncertainties, and that botanical experience may not reduce them. © 2012 International Association for Vegetation Science

Molecular epidemiology of a nosocomial outbreak due to Enterobacter cloacae and Enterobacter agglomerans in Campinas, São Paulo, Brazil Epidemiologia molecular de um surto de bacteriemia por Enterobacter cloacae e Enterobacter agglomerans ocorrido na região de Campinas, São Paulo, Brasil

A total of 73 isolates (57 Enterobacter cloacae and 16 Enterobacter agglomerans), recovered during an outbreak of bacteremia in the Campinas area, São Paulo, Brazil, were studied. Of these isolates, 61 were from parenteral nutrition solutions, 9 from blood cultures, 2 from a sealed bottle of parenteral nutrition solution, and one was of unknown origin. Of the 57 E. cloacae isolates, 54 were biotype 26, two were biotype 66 and one was non-typable. Of 39 E. cloacae isolates submitted to ribotyping, 87.2% showed the same banding pattern after cleavage with EcoRI and BamHI. No important differences were observed in the antimicrobial susceptibility patterns among E. cloacae isolates exhibiting the same biotype, serotype and ribotype. All E. agglomerans isolates, irrespective of their origin, showed same patterns when cleaved with EcoRI and BamHI. The results of this investigation suggest an intrinsic contamination of parenteral nutrition solutions and incriminate these products as a vehicle of infection in this outbreak.<br>Foram estudadas um total de 73 cepas (57 de E. cloacae e 16 E. agglomerans), isoladas durante um surto de bacteriemia ocorrido na região de Campinas, S. Paulo. Entre estas cepas, 61 foram isoladas de solução de nutrição parenteral, 9 de sangue, 2 de bolsa fechada de solução de nutrição parenteral e uma era de origem desconhecida. Entre as 57 cepas de E. cloacae, a maioria das cepas foram do biotipo 26/sorotipo O3 (39 cepas) e do biotipo 26/OR (13). Entre as 39 cepas de E. cloacae ribotipadas, 87,2% apresentaram o mesmo padrão de bandas com EcoRI e BamHI. Cepas de E. cloacae pertencentes ao mesmo biotipo, sorotipo e ribotipo não apresentaram diferenças significativas em relação ao padrão de sensibilidade aos agentes antimicrobianos. Todas as cepas de E. agglomerans, independente da origem, pertenciam ao mesmo ribotipo após a clivagem com EcoRI e BamHI. Os resultados obtidos sugerem uma contaminação intrínseca das soluções de nutrição parenteral, incriminando-as como o veículo de transmissão dos agentes etiológicos do surto