Risk assessment procedures in Argentina for the safe import, quarantine and release of biological control agents against pests.

The decision to import and release biological control agents is a national one that may involve various authorities. It will be overseen by the national plant protection organisation that implements the responsibilities described in the International Plant Protection Convention. This article provides an overview of the decision processes and roles of these authorities in Argentina. Argentina has a long history of coordination with the other Southern Cone nations on plant protection and other technical sanitary and phytosanitary matters associated with trade. This article reports on 25 years of evaluation, import and release of exotic biological control agents. Not a single import with a permit has faced rejection on the basis of phytosanitary requirements. This record highlights the region's commitment to integrated pest management approaches that rely on international shipments of live insects and other beneficial organisms.

La décision d'importer et d'autoriser des lâchers d'agents de lutte biologique est du ressort national, avec la participation de plusieurs autorités compétentes. Elle est supervisée par l'organisation nationale de protection des végétaux, qui est chargée d'exercer les responsabilités prévues au titre de la Convention internationale pour la protection des végétaux. Les auteurs donnent une vue d'ensemble des procédures décisionnelles et des fonctions respectives des autorités compétentes en Argentine. L'Argentine et les autres nations du Cône Sud ont une longue histoire commune de coordination de la protection des végétaux et d'autres questions sanitaires et phytosanitaires en lien avec les échanges internationaux. Les auteurs font le point sur 25 années d'évaluations, d'importations et de lâchers d'agents de lutte biologique exotiques dans le pays. Aucune importation ayant fait l'objet d'une autorisation préalable n'a été rejetée pour des motifs de conformité phytosanitaire. Ce constat souligne l'engagement fort de la région en faveur de méthodes intégrées de gestion des nuisibles reposant sur le transport international d'insectes vivants et d'autres organismes utiles.

La de importar y liberar al medio agentes de control biológico es una decisión de carácter nacional en la que pueden intervenir diversas autoridades y que supervisará la organización nacional de protección fitosanitaria encargada de trasladar a la práctica las responsabilidades enunciadas en la Convención Internacional de Protección Fitosanitaria. Los autores presentan a grandes líneas los procesos decisorios y las funciones de dichas autoridades en la Argentina, país que tiene una larga trayectoria de coordinación con otras naciones del Cono Sur en materia de protección de los vegetales y otras cuestiones técnicas de índole sanitaria y fitosanitaria ligadas al comercio. Los autores dan cuenta de 25 años de evaluación, importación y liberación al medio de agentes exóticos de control biológico. En ese tiempo, ni una sola importación provista de permiso ha sido denegada en aplicación de requisitos fitosanitarios. Semejante historial pone de relieve el compromiso de la región con fórmulas de gestión integrada de plagas que reposan en los envíos internacionales de insectos vivos y otros organismos beneficiosos.

Recent epidemiological trends in extrapulmonary TB in Galicia, Spain.

OBJECTIVE: To describe the epidemiological trends and characteristics of extrapulmonary tuberculosis (EPTB) in Galicia, Spain, from 2000 to 2019.METHODS: This was a retrospective cohort study based on data from the Galician TB information system.RESULTS: Of the total number of TB cases (n = 15,871), 5,428 (34.2%) had EPTB. The absolute number of cases and incidence of EPTB decreased dramatically (from 480 cases and 17.8 cases/100,000 in 2000, to 172 and 6.4 cases/100,000 in 2019, respectively), with a mean annual decrease of respectively 64% and 4.7% for absolute cases and incidence rates. The risk for EPTB was higher in men than in women (RR 3.86, 95% CI 3.66-4.07). The most frequent age group was 15-44 years (2,234 patients, 41.2%); overall reductions per age group were 82% (0-14 years), 75% (15-44 years), 44% (45-64 years) and 63% (≥65 years), with statistically significant differences. The most frequently locations were the pleura (1,916 cases; 35.3%) and the lymph nodes (1,504; 27.7%).CONCLUSION: The incidence of EPTB in Galicia has decreased significantly in the last 20 years. The epidemiological characteristics have not changed, except for the number of patients with risk factors. This improvement of EPTB epidemiological trends coincides with the implementation of the programme for the prevention and control of TB, which suggests that it has been very effective in the control of the EPTB.

A quantitative PCR assay for antiviral activity screening of medicinal plants against Herpes simplex 1.

Herpes simplex virus one is one of the most prevalent pathogens worldwide. Strains resistant to current treatment have been reported, so it is necessary to search for new antiviral molecules. The most common method to quantify antiviral activity from natural products is the plaque reduction assay, a technically demanding method. In order to provide a simple alternative to this method, we have established a procedure for viral quantification by qPCR, and coupled with a cytotoxicity evaluation system using resazurin. In this way, it is possible to obtain both the estimation of cytotoxicity and the antiviral activity simultaneously, allowing rapid screening of plant extracts. Ten out of twenty-eight Paraguayan medicinal plant extracts evaluated using this method showed antiviral activity, and the EC50, CC50, and SI values were calculated for each extract. Our experience supports the employment of the described method for a rapid identification of plant extracts with antiviral activity.

Intentional Hydroxychloroquine Overdose Treated with High-Dose Diazepam: an Increasing Concern in the COVID-19 Pandemic.

INTRODUCTION: Recent attention on the possible use of hydroxychloroquine and chloroquine to treat COVID-19 disease has potentially triggered a number of overdoses from hydroxychloroquine. Toxicity from hydroxychloroquine manifests with cardiac conduction abnormalities, seizure activity, and muscle weakness. Recognizing this toxidrome and unique management of this toxicity is important in the COVID-19 pandemic. CASE REPORT: A 27-year-old man with a history of rheumatoid arthritis presented to the emergency department 7 hours after an intentional overdose of hydroxychloroquine. Initial presentation demonstrated proximal muscle weakness. The patient was found to have a QRS complex of 134 ms and QTc of 710 ms. He was treated with early orotracheal intubation and intravenous diazepam boluses. Due to difficulties formulating continuous diazepam infusions, we opted to utilize an intermitted intravenous bolus strategy that achieved similar effects that a continuous infusion would. The patient recovered without residual side effects. DISCUSSION: Hydroxychloroquine toxicity is rare but projected to increase in frequency given its selection as a potential modality to treat COVID-19 disease. It is important for clinicians to recognize the unique effects of hydroxychloroquine poisoning and initiate appropriate emergency maneuvers to improve the outcomes in these patients.

Injury narratives in occupational safety and health prevention in Italy.

BACKGROUND: Storytelling is an effective information source when coupled with technical-scientific evidence. It can promote a structured relationship between evidence-based knowledge and field experience of workplace safety and prevention services (WSPS) inspectors. This is key to identifying the causes of workplace injuries and to set priorities for prevention strategies. AIMS: The main aim was to describe and report how story collection can be used for deriving validated indications for injury prevention. The specific objectives were to report the results of the creation and dissemination on the web of the story collection and the experience of setting up a community of practice (CoP) to develop preventive recommendations. METHODS: WSPS inspectors from local health boards in Piedmont (northwest Italy) were asked to write injury stories. They identified the key elements of their stories and developed a narrative of witness accounts to explore the critical issues identified during the investigation. In sessions with the CoP, the inspectors validated the indications for prevention elaborated in each story to reduce bias and standardize recommendations. RESULTS: Between 2012 and 2017, 60 WSPS inspectors wrote 53 injury stories which were collected and published on the institutional website. Twenty-two stories were selected for discussion during peer review sessions in the CoP and the indications for prevention were transformed as preventive solutions. CONCLUSIONS: Occupational safety and health prevention can benefit from a narrative-based approach that provides a more comprehensive look at health and safety by facilitating knowledge improvement and sharing.

Spatial prediction of risk areas for vector transmission of Trypanosoma cruzi in the State of Paraná, southern Brazil.

After obtaining certification of the absence of transmission of the Trypanosoma cruzi by Triatoma infestans in 2006, other native species of protozoan vectors have been found in human dwellings within municipalities of the State of Paraná, Southern Brazil. However, the spatial distribution of T. cruzi vectors and how climatic and landscape combined variables explain the distribution are still poorly understood. The goal of this study was to predict the potential distribution of T. cruzi vectors as a proxy for Chagas disease transmission risk using Ecological Niche Models (ENMs) based on climatic and landscape variables. We hypothesize that ENM based on both climate and landscape variables are more powerful than climate-only or landscape-only models, and that this will be true independent of vector species. A total of 2,662 records of triatomines of five species were obtained by community-based entomological surveillance from 2007 to 2013. The species with the highest number of specimens was Panstrongylus megistus (73%; n = 1,943), followed by Panstrongylus geniculatus (15.4%; 411), Rhodnius neglectus (6.0%; 159), Triatoma sordida (4.5%; 119) and Rhodnius prolixus (1.1%; 30). Of the total, 71.9% were captured at the intradomicile. T. cruzi infection was observed in 19.7% of the 2,472 examined insects. ENMs were generated based on selected climate and landscape variables with 1 km2 spatial resolution. Zonal statistics were used for classifying the municipalities as to the risk of occurrence of synanthropic triatomines. The integrated analysis of the climate and landscape suitability on triatomines geographical distribution was powerful on generating good predictive models. Moreover, this showed that some municipalities in the northwest, north and northeast of the Paraná state have a higher risk of T. cruzi vector transmission. This occurs because those regions present high climatic and landscape suitability values for occurrence of their vectors. The frequent invasion of houses by infected triatomines clearly indicates a greater risk of transmission of T. cruzi to the inhabitants. More public health attention should be given in the northern areas of the State of Paraná, which presents high climate and landscape suitabilities for the disease vectors. In conclusion, our results-through spatial analysis and predictive maps-showed to be effective in identifying areas of potential distribution and, consequently, in the definition of strategic areas and actions to prevent new cases of Chagas' disease, reinforcing the need for continuous and robust surveillance in these areas.

Different concentration of human cord blood HMGB1 according to delivery and labour: A pilot study.

OBJECTIVE: Oxidative stress is involved in several maternal conditions characterized both by an increase in free radicals synthesis and a parallel decrease in the antioxidant activity. Parturition induces considerable oxidative stress and many inflammatory mediators, among which HMGB1, are involved from the beginning of pregnancy to the birth of the infant. We evaluated serum cord blood HMGB1 levels in a population of neonates to investigate correlation with mode of delivery, as well as the influence of labour. SETTING AND PATIENTS: The study subjects were 325 neonates delivered at University Hospital "G. Martino" of Messina over an 18-month period. Following cord separation, venous blood sampling was performed on umbelical cords. RESULTS: In the cord venous blood, we found HMGB1 values significantly more elevated in spontaneous vaginal group when compared to elective or emergency caesarean section group. Regarding labour, umbilical cord venous blood HMGB1 levels were significantly higher in the spontaneous and induced labour group, compared to non-labouring women. CONCLUSION: These results could highlight a possible role of HMGB1 during birth time related to mode of delivery and labour.

Inflammatory biomarkers and intellectual disability in patients with Down syndrome.

BACKGROUND: Intellectual disability (ID) is part of the Down syndrome (DS) phenotypic spectrum, but the exact molecular pathophysiology of ID in individuals with DS is not yet fully understood, with many research hypotheses still unproven. Basing on previous studies (which suggested a possible role of altered inflammatory response in DS-related ID), we assessed the serum levels of a number of inflammatory biomarkers [serum amyloid A (SAA), C-reactive protein (C-RP), high mobility group box-1 (HMGB1)] in a cohort of individuals with DS and healthy controls. METHODS: In total, 24 children diagnosed with DS and 12 healthy controls were enrolled, and all underwent detailed cognitive assessment. Also, serum SAA, C-RP and HMGB1 levels were measured in all recruited subjects and correlated to the severity of ID in the DS group. RESULTS: Serum SAA, C-RP and HMGB1 values were found to be significantly higher in the DS group compared with the healthy subjects (P = 0.001). In addition, serum HMGB1 levels positively correlated with C-RP and SAA in the DS group but not in the healthy controls. Only serum C-RP levels resulted inversely correlated (P < 0.01) with intelligence quotient (IQ); conversely, significant statistical correlations between serum SAA levels and IQ (as well as between HMGB1 and IQ) have been not found (P > 0.05). CONCLUSIONS: The levels of the determined markers were higher in DS individuals compared with (cognitively) healthy subjects, and CRP showed a negative correlation with IQ in children with DS.

Peptidomic analysis of the anterior temporal lobe and corpus callosum from schizophrenia patients.

Schizophrenia is a complex disorder hypothesized to develop from a combination of genetic, neurodevelopmental, and environmental factors. Molecules that are directly involved in the pathogenesis of schizophrenia and may serve as biomarker candidates can be identified with "omics" approaches such as proteomics and peptidomics. In this context, we performed a peptidomic study in schizophrenia postmortem brains, to our knowledge the first such study in schizophrenia patients. We investigated the anterior temporal lobe (ATL) and corpus callosum (CC) by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) and a label-free ion quantification technique based on data-dependent acquisition (DDA). Results indicated alterations in a specific intracellular neurogranin peptide in both the ATL and CC and a decrease of PepH, a fragment of histone H2B type 1-H intracellular peptide, in the ATL. PepH was tested in serum-deprived Neuro2A cells and showed a protective effect against cell death. Cells were also challenged with lipopolysaccharide (LPS), and PepH was able to prevent the endotoxic effects of LPS. Our data suggest that specific intracellular peptides are altered in schizophrenia patients. The potential biological activity of PepH supports intracellular peptides as novel targets in the study not only of schizophrenia but also of other neuropsychiatric diseases. BIOLOGICAL SIGNIFICANCE: Psychiatric disorders are considerably more difficult to diagnose in their early stages. Usually, by the time the diagnosis is clear and clinical treatment can be started, the disorder is already established and thus of greater severity. Consequently, the scientific community has been searching for biomarker candidates that can aid the early detection of such disorders and for novel therapeutics to improve treatment or at least delay disease progression. Moreover, key molecules involved in the establishment of psychiatric diseases may help the understanding of their pathogenesis and thus drive the development of more effective treatments. The present work screened peptides that might be possible novel targets to control cell machinery in schizophrenia and identified an intracellular peptide with potential cytoprotective activity. To our knowledge, this is the first peptidomic study in schizophrenia patients.

Desempenho da cultura líquida MGIT após implementação em uma rede de laboratórios públicos do estado de São Paulo / Performance of liquid culture MGIT after implementation in a network of public laboratories of Sao Paulo state

A OMS, em 2007, recomendou a implementação da cultura líquida para o diagnóstico da tuberculose (TB) e teste de sensibilidade para países de baixa e média renda. Neste estudo foi avaliado odesempenho da cultura líquida MGIT em condição de rotina após dois anos de implantação em uma rede de laboratórios públicos. Foi efetuada análise retrospectiva de dados da cultura líquida,realizadas em dez laboratórios regionais do Instituto Adolfo Lutz, de janeiro a março de 2010. Foram incluídas amostras submetidas a baciloscopia, cultura líquida MGIT automatizada ou manual eidentificação presuntiva do complexo Mycobacterium tuberculosis (CMTB). Foram detectadas 1.159 culturas positivas. Destas, 113 (9,7%) contaminaram, e 1.046 foram analisadas, sendo 850 (81,3%) CMTB, 116 (11,1%) micobactérias não tuberculosas e 6 (0,6%) Nocardia sp. A taxa de contaminação foi de 2,2% e o acréscimo da cultura para o diagnóstico da TB foi de 29,9%. A média do tempo de detecção da cultura foi de 14,7 dias (DP+/- 11,7 dias). A acurácia da identificação presuntiva foide 91,3%. A cultura líquida MGIT demonstrou ser excelente alternativa para efetuar diagnóstico da TB e das micobacterioses, em razão da rapidez possibilitando uma intervenção rápida e eficaz no tratamento.

In 2007, WHO recommended the implementation of liquid culture for tuberculosis (TB) diagnosis anddrug-susceptibility test in low and middle-income countries. This study evaluated the performanceof MGIT culture in routine condition after two years of its implementation in a public laboratoriesnetwork. This is a retrospective study, which analyzed the data on the liquid culture performed in ten regional laboratories of the Institute Adolfo Lutz, from January to March 2010. The data included clinical samples submitted to microscopy, automated or manual MGIT culture and presumptive M. tuberculosis complex (MTBC) identification by analyzing the cord formation. Culture waspositive in 1,159 samples. Of these, 113 (9.7%) contaminated, and 1,046 were analyzed, of which 850 (81.3%) were identified as MTBC, 116 (11.1%) as non-tuberculous mycobacteria and 6 (0.6%)as Nocardia sp. Contamination rate was 2.2% and the contribution of culture to the TB diagnosis was 29.9%. The detection mean time was 14.7 days (SD+/-11.7 days). The accuracy of the presumptive identification of MTBC was 91.3%. MGIT liquid culture demonstrated to be an excellent alternative for diagnosing TB and mycobacterioses, because of the rapidity of diagnosis, thus allowing an immediate and effective treatment.

Desempenho da cultura líquida MGIT após implementação em uma rede de laboratórios públicos do estado de São Paulo / Performance of liquid culture MGIT after implementation in a network of public laboratories of Sao Paulo state

A OMS, em 2007, recomendou a implementação da cultura líquida para o diagnóstico da tuberculose (TB) e teste de sensibilidade para países de baixa e média renda. Neste estudo foi avaliado o desempenho da cultura líquida MGIT em condição de rotina após dois anosde implantação em uma rede de laboratórios públicos. Foi efetuada análise retrospectiva de dados da cultura líquida, realizadas em dez laboratórios regionais do Instituto Adolfo Lutz, de janeiro a março de 2010. Foram incluídas amostras submetidas a baciloscopia, cultura líquida MGIT automatizada ou manual e identificação presuntiva do complexo Mycobacterium tuberculosis (CMTB). Foram detectadas 1.159 culturas positivas. Destas, 113 (9,7%) contaminaram, e 1.046 foram analisadas, sendo 850 (81,3%) CMTB, 116 (11,1%) micobactérias não tuberculosas e 6 (0,6%) Nocardia sp A taxa de contaminação foi de 2,2% e o acréscimo da cultura para o diagnóstico da TB foi de 29,9%. A média do tempo de detecção da cultura foi de 14,7 dias (DP+/- 11,7 dias). A acurácia da identificação presuntiva foi de 91,3%. A cultura líquida MGIT demonstrou ser excelente alternativa para efetuar diagnóstico da TB e das micobacterioses, em razão da rapidez possibilitando uma intervenção rápida e eficaz no tratamento.

In 2007, WHO recommended the implementation of liquid culture for tuberculosis (TB) diagnosis and drug-susceptibility test in low and middle-income countries. This study evaluated the performance of MGIT culture in routine condition after two years of its implementation in a public laboratories network.This is a retrospective study, which analyzed the data on the liquid culture performed in ten regional laboratories of the Institute Adolfo Lutz, from January to March 2010. The data included clinical samples submitted to microscopy, automated or manual MGIT culture and presumptive M. tuberculosis complex (MTBC) identification by analyzing the cord formation. Culture was positive in 1,159 samples. Of these, 113 (9.7%) contaminated, and 1,046 were analyzed, of which 850 (81.3%) were identified as MTBC, 116 (11.1%) as non-tuberculous mycobacteria and 6 (0.6%) as Nocardia sp. Contamination rate was 2.2% and the contribution of culture to the TB diagnosis was 29.9%. The detection mean time was 14.7 days (SD+/-11.7 days). The accuracy of the presumptive identification of MTBC was 91.3%. MGIT liquid culture demonstrated to be an excellent alternative for diagnosing TB and mycobacterioses, because of the rapidity of diagnosis, thus allowing an immediate and effective treatment.

Peptidomic analysis of the anterior temporal lobe and corpus callosum from schizophrenia patients

Schizophrenia is a complex disorder hypothesized to develop from a combination of genetic, neurodevelopmental, and environmental factors. Molecules that are directly involved in the pathogenesis of schizophrenia and may serve as biomarker candidates can be identified with "omics" approaches such as proteomics and peptidomics. In this context, we performed a peptidomic study in schizophrenia postmortem brains, to our knowledge the first such study in schizophrenia patients. We investigated the anterior temporal lobe (ATL) and corpus callosum (CC) by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) and a label-free ion quantification technique based on data-dependent acquisition (DDA). Results indicated alterations in a specific intracellular neurogranin peptide in both the ATL and CC and a decrease of PepH, a fragment of his tone H2B type 1-H intracellular peptide, in the ATL. PepH was tested in serum-deprived Neuro2A cells and showed a protective effect against cell death. Cells were also challenged with lipopolysaccharide (LPS), and PepH was able to prevent the endotoxic effects of LPS. Our data suggest that specific intracellular peptides are altered in schizophrenia patients. The potential biological activity of PepH supports intracellular peptides as novel targets in the study not only of schizophrenia but also of other neuropsychiatric diseases. Biological significance: Psychiatric disorders are considerably more difficult to diagnose in their early stages. Usually, by the time the diagnosis is clear and clinical treatment can be started, the disorder is already established and thus of greater severity. Consequently, the scientific community has been searching for biomarker candidates that can aid the early detection of such disorders and for novel therapeutics to improve treatment or at least delay disease progression. Moreover, key molecules involved in the establishment of psychiatric diseases may help the understanding of their pathogenesis and thus drive the development of more effective treatments. The present work screened peptides that might be possible novel targets to control cell machinery in schizophrenia and identified an intracellular peptide with potential cytoprotective activity. To our knowledge, this is the first peptidomic study in schizophrenia patients.

Phenotypic and genotypic characterization of two Toxoplasma gondii isolates in free-range chickens from Uberlândia, Brazil.

The aim of this study was to determine the seroprevalence of Toxoplasma gondii infection in free-range chickens from Uberlândia, Minas Gerais state, Brazil, and characterize the genotypic and phenotypic features of two isolates of this parasite, considering the importance of these hosts in the epidemiology of toxoplasmosis. Serum samples from 108 free-range chickens were obtained from ten different districts, and submitted to the modified agglutination test (MAT) for the presence of anti-T. gondii antibodies, and brain and heart tissue samples from infected chickens were processed for mouse bioassay. An overall seroprevalence of 71·3% was found and antibody titres ranged from 16 to 4096. After confirmation of seropositivity by mouse bioassay, the determination of the T. gondii genotypes of two isolates was performed by PCR-RFLP, using primers for the following markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, new SAG2, Apico and CS3. These T. gondii isolates, designated TgChBrUD1and TgChBrUD2, were obtained from heart samples of free-range chickens. The TgChBrUD1 isolate belonged to ToxoDB PCR-RFLP genotype 11 and the TgChBrUD2 isolate belonged to ToxoDB PCR-RFLP genotype 6. Both isolates demonstrated high virulence in a rodent model, with the TgChBrUD1 isolate able to induce brain cysts, in accord with its pattern of multiplication rates in human fibroblast culture. Taken together, these results reveal high prevalence of T. gondii infection in free-range chickens throughout Uberlândia, indicating an important degree of oocyst environmental contamination and the existence of considerable risk for T. gondii transmission to humans by consumption of free-range chicken as a food source.

Tuberculose pulmonar paucibacilar em Centros de Detenção Provisória / Paucibacillary pulmonary tuberculosis in ProvisionalDetention Centers

Cultura de micobactérias proporciona o crescimento de bacilos viáveis, mesmo presentes emescassa quantidade e não detectados pela baciloscopia. Neste estudo foram analisadas as amostrasde escarro que apresentaram baciloscopia negativa e cultura positiva. As amostras foram coletadasde 2008 a 2013, de indivíduos detidos em Centros de Detenção Provisória de Santo André,Mauá e Diadema, Estado de São Paulo. As metodologias utilizadas foram baciloscopia porcoloração Ziehl-Neelsen e cultura pelo Sistema BACTEC MGIT 960 e Ogawa-Kudoh. Dos11.529 exames realizados, 221 (1,9 %) apresentaram baciloscopias negativas e culturas positivas.Dos 221 isolados, 166 (75,1 %) pertenciam ao Complexo Mycobacterium tuberculosis, 21 (9,5 %)micobactérias não membros do Complexo Mycobacterium tuberculosis (MNT), 33 (14,9 %)Mycobaterium sp e uma cultura mista do Complexo M. tuberculosis e M. avium. MNT maisfrequentes foram M. avium (23,8 %) e M. fortuitum (19,0 %). A maioria dos isolados do ComplexoM. tuberculosis (155/166 - 93,4 %) foi sensível aos antimicrobianos. Sete amostras apresentaramresistência à isoniazida e uma apresentou multirresistência à isoniazida e rifampicina. Este estudomostra a importância da realização da cultura em escarros que apresentam baciloscopia negativano diagnóstico da TB e micobacteriose. O tratamento tardio causa a continuidade da transmissãoda doença e agravamento do quadro clínico.

Culture of mycobacteria induces the growth of viable bacillus occurring in small quantity,which are no detectable by bacilloscopy. This study aimed at identifying the mycobacteria isolatesfrom sputum presenting negative bacilloscopy and positive culture. The samples were collectedfrom 2008 to 2013 from criminals of Provisional Detention Centers in Santo André, Mauáand Diadema/SP. Smears were stained by Ziehl-Neelsen staining and the cultures were performedby the BACTEC MGIT 960 system and Ogawa-Kudoh culture medium. Of 11,529 isolates, 221(1.9 %) showed negative bacilloscopy and positive cultures. Of 221 isolates, 166 (75.1 %) belongedto Mycobacterium tuberculosis complex, 21 (9.5 %) were nontuberculous mycobacteria (NTM),33 (14.9 %) Mycobacterium sp, and one identified as a mixed culture of M. tuberculosis andM. avium complex. The most common NTM species were M. avium (23.8 %) and M. fortuitum(19.0 %). Most of the isolates (155/166-93.4 %) were susceptible to antimicrobial agents.Seven samples were resistant to isoniazid, and one presented multiresistance to isoniazid andrifampicin. This study shows the importance in performing sputum culture, when these samplesare negative on bacilloscopy in diagnosing TB and mycobacteriosis. The treatment delay resultsin the maintenance of disease transmission and worsening of clinical symptoms.

Tuberculose pulmonar paucibacilar em Centros de Detenção Provisória / Paucibacillary pulmonary tuberculosis in Provisional Detention Centers

Cultura de micobactérias proporciona o crescimento de bacilos viáveis, mesmo presentes em escassa quantidade e não detectados pela baciloscopia. Neste estudo foram analisadas as amostras de escarro que apresentaram baciloscopia negativa e cultura positiva. As amostras foram coletadas de 2008 a 2013, de indivíduos detidos em Centros de Detenção Provisória de Santo André, Mauá e Diadema, Estado de São Paulo. As metodologias utilizadas foram baciloscopia por coloração Ziehl-Neelsen e cultura pelo Sistema BACTEC MGIT 960 e Ogawa-Kudoh. Dos 11.529 exames realizados, 221 (1,9 %) apresentaram baciloscopias negativas e culturas positivas. Dos 221 isolados, 166 (75,1 %) pertenciam ao Complexo Mycobacterium tuberculosis, 21 (9,5 %) micobactérias não membros do Complexo Mycobacterium tuberculosis (MNT), 33 (14,9 %) Mycobaterium sp e uma cultura mista do Complexo M. tuberculosis e M. avium. MNT mais frequentes foram M. avium (23,8 %) e M. fortuitum (19,0 %). A maioria dos isolados do Complexo M. tuberculosis (155/166 - 93,4 %) foi sensível aos antimicrobianos. Sete amostras apresentaram resistência à isoniazida e uma apresentou multirresistência à isoniazida e rifampicina. Este estudo mostra a importância da realização da cultura em escarros que apresentam baciloscopia negativa no diagnóstico da TB e micobacteriose. O tratamento tardio causa a continuidade da transmissão da doença e agravamento do quadro clínico.

Culture of mycobacteria induces the growth of viable bacillus occurring in small quantity, which are no detectable by bacilloscopy. This study aimed at identifying the mycobacteria isolates from sputum presenting negative bacilloscopy and positive culture. The samples were collected from 2008 to 2013 from criminals of Provisional Detention Centers in Santo André, Mauá and Diadema/SP. Smears were stained by Ziehl-Neelsen staining and the cultures were performed by the BACTEC MGIT 960 system and Ogawa-Kudoh culture medium. Of 11,529 isolates, 221 (1.9 %) showed negative bacilloscopy and positive cultures. Of 221 isolates, 166 (75.1 %) belonged to Mycobacterium tuberculosis complex, 21 (9.5 %) were nontuberculous mycobacteria (NTM), 33 (14.9 %) Mycobacterium sp, and one identified as a mixed culture of M. tuberculosis and M. avium complex. The most common NTM species were M. avium (23.8 %) and M. fortuitum (19.0 %). Most of the isolates (155/166-93.4 %) were susceptible to antimicrobial agents. Seven samples were resistant to isoniazid, and one presented multiresistance to isoniazid and rifampicin. This study shows the importance in performing sputum culture, when these samples are negative on bacilloscopy in diagnosing TB and mycobacteriosis. The treatment delay results in the maintenance of disease transmission and worsening of clinical symptoms.

Trophoblast-macrophage crosstalk on human extravillous under Toxoplasma gondii infection.

INTRODUCTION: The interaction between human extravillous trophoblasts and macrophages has an important role in implantation and placentation. However, any dysfunction in this communication system is associated with pregnancy pitfalls, and a Toxoplasma gondii infection can be a potential problem in this crosstalk. Therefore, the aim of this study was to assess the influence of infected macrophages on cytokine production and the incidence of apoptosis in T. gondii-infected extravillous trophoblast cells. METHODS: HTR-8/SVneo cells were treated with supernatant from macrophages infected or not by T. gondii (conditioned medium) in order to analyze apoptosis and cytokine production in comparison to uninfected control conditions. RESULTS: The IL-6 secretion by HTR-8/SVneo cells increased synergistically by treatment with conditioned medium and T. gondii infection. The apoptosis index of HTR-8/SVneo cells was also upregulated by treatment with conditioned medium and infection. In addition, a low expression of Fas/CD95 and a high soluble FasL release were observed during infection, although no significant change was observed in the proliferation of T. gondii. DISCUSSION: The parasite modulates the high apoptosis index in HTR-8/SVneo cells in order to favor its establishment inside its host cells. On the other hand, the conditioned medium from uninfected macrophages restores the apoptosis rates, although the effect of the infection seems to be stronger. In conclusion, our results showed that T. gondii infection in human extravillous trophoblasts is able to modulate the trophoblast-macrophage crosstalk.

Rapid detection of Mycobacterium tuberculosis complex by real-time PCR in sputum samples and its use in the routine diagnosis in a reference laboratory.

Tuberculosis (TB) is an infectious disease of global distribution, constituting a serious public health problem in Brazil. São Paulo State, located in the south-east of Brazil, notified 16,580 new TB cases in 2013. The Instituto Adolfo Lutz is a public health reference laboratory for TB diagnosis for all the State. Considering that rapid and accurate diagnosis is essential for TB control, the aim of this study was to evaluate the use of an in-house real-time (RT)-PCR assay targeting the mpt64 gene in the routine diagnosis of TB, and to compare this technique with smear microscopy and culture. From August 2012 to October 2013, 715 sputum samples from 657 patients were included in the study. Smear microscopy, culture, phenotypic and PRA-hsp65 identification of mycobacteria, and mpt64 RT-PCR were performed. With respect to confirmed TB cases (n = 62/657; 9.4%), smear microscopy had a sensitivity of 82.3%. Culture and RT-PCR showed the same sensitivity, i.e. 90.3%. Specificity was 99.7, 99.4 and 98.6% for smear microscopy, culture and RT-PCR, respectively. mpt64 RT-PCR showed high sensitivity and specificity for the detection of Mycobacterium tuberculosis complex in sputum samples. This technique can be deployed in laboratories that do not have a rapid test for TB available, enabling the performance of TB diagnosis in up to 5 h.

Short communication: Relationship between urinary neutrophil gelatinase-associated lipocalin and noninfectious pyuria in dogs.

Neutrophil gelatinase-associated lipocalin (NGAL) is a neutrophil-derived protein whose concentration increases in plasma and urine with ongoing renal damage. Urinary leucocytes can be a potential source of urinary NGAL. The aim of this study is to investigate the effects of urinary neutrophil count and other urinary parameters on urinary NGAL values in urine with negative culture. Urinalysis, urine culture, and determination of urinary NGAL were performed on 33 clinically healthy nonproteinuric dogs with negative urinoculture. The median uNGAL concentration in dogs in this study population was 9.74 ng/mL (IQR 1.93-25.43 ng/mL). In samples with WBCs > 5 hpf (mean 15.9, 6-50 leucocytes/hpf), median uNGAL value was significantly higher than that in samples with WBCs < 5 hpf (mean 0.9, 0-3 leucocytes/hpf), (4.96 pg/mL (0.29-11.34) and 23.65 pg/mL (20.04-29.80), resp.; P = 0.0053). The severity of urinary pyuria and the UPC value were correlated with uNGAL concentration. The results of our study show that urinary NGAL concentration is correlated with WBCs number in urinary sediment of dogs with negative urinoculture. The present study suggests that noninfectious pyuria is significantly correlated with urinary NGAL values and might influence uNGAL values.

Antioxidant activity of hyaluronic acid investigated by means of chemiluminescence of equine neutrophil bursts and electron paramagnetic resonance spectroscopy.

Activated neutrophils (PMNs), the ROS/RNS released by PMNs and the derived inflammatory processes are involved in the pathogenesis and progression of human inflammatory airway diseases. Similar diseases are also present in horses which suffer from recurrent airway obstruction (RAO), exercise-induced pulmonary haemorrhage (EIPH) and inflammatory airway diseases (IAD). Hyaluronic acid (HA) plays numerous roles in modulating inflammatory processes. The aim of this study was to examine whether a preparation of HA (MW 900 000 Da) interferes with ROS/RNS during the course of equine PMN respiratory bursts, and to establish the lowest concentration at which it still has antioxidant activity by means of luminol-amplified chemiluminescence (LACL). Electron paramagnetic resonance (EPR) spectroscopy was also used to investigate the direct antiradical activity of HA. The hydroxyl radical was significantly scavenged in a concentration-dependent manner at HA concentrations ranging from 2.5 to 0.16 mg/mL. Superoxide anion, Tempol radical and the ABTS(•+) were significantly inhibited at concentrations ranging from 2.5 to 0.62 mg/mL. The LACL of stimulated equine neutrophils showed that HA induced a statistically significant concentration-effect reduction from 5 mg/mL to 1.25 mg/mL. These findings were confirmed also when l-Arg was added to investigate the inhibition of the resulting peroxynitrite anion. Our findings indicate that, in addition to the human use, HA can also be used to antagonize the oxidative stress generated by free radicals in horses peripheral blood mononuclear cells (PBMCs). In order to achieve therapeutic concentrations, a direct aerosol administration to horses with horse respiratory diseases can be considered, as this route of application is also recommended in human medicine.