Clemastine/tamoxifen hybrids as easily accessible antileishmanial drug leads.

A library of hybrid molecules is developed based on the common chemical features shared by clemastine and tamoxifen both of which are well known for their antileishmanial activities. In the initial screening against Leishmania major and L. amazonensis promastigotes, as well as cytotoxicity assays using HepG2 cells, several hybrids showed submicromolar activity against the parasite and no toxicity against human cells. The compounds with an EC50 < 2 µM against promastigotes of both species and a selectivity index >10 were further characterized against intracellular amastigotes as well as promastigotes of species that cause both visceral and cutaneous leishmaniasis, such as L. infantum and L. braziliensis, respectively. These sequential screenings revealed the high pan-activity of this class of molecules against these species, with several compounds displaying an EC50 ≤ 2 µM against both promastigotes and intracellular amastigotes. Two of them were identified as the potential templates for lead optimization of this series having shown the highest activities against all species in both stages of parasite. The present findings can serve as a good starting point in the search for novel antileishmanial compounds that are easy to access and highly active.

Identification of equine herpesvirus type 1 as cause of abortion in mares in Southern Brazil / Identificação do herpesvirus equino tipo 1 como causa de abortos em éguas no sul do Brasil

Foi realizado um estudo retrospectivo dos casos suspeitos de aborto por herpesvírus equino observados na região sul do Rio Grande do Sul entre 1978 e 2016. Foram revisados os protocolos de necropsia do Laboratório Regional de Diagnóstico da Faculdade de Veterinária da Universidade Federal de Pelotas resgatando-se os dados epidemiológicos, lesões macroscópicas e histológicas de cada caso. Foram observados dois surtos da enfermidade com prevalência entre 5,7% e 50% nos diferentes estabelecimentos, e dois casos individuais. Em todos os casos foram enviados fragmentos de órgãos fetais formolizados. Histologicamente, em todos os casos foram observados focos de necrose no fígado, pulmão e baço e presença de corpúsculos de inclusão acidofílico em hepatócitos, células epiteliais pulmonares e leucócitos. A imuno-histoquímica utilizando anticorpo policlonal comercial para herpesvirus equino-1 (EHV-1) revelou marcação positiva em todos os casos. Além disso, foi extraído DNA dos tecidos emblocados em parafina dos casos e submetidos à técnica de nested-PCR seguida de sequenciamento genômico dos amplicons em duas amostras. Estes achados indicam, que EHV-1 deve ser considerado como diagnóstico diferencial em casos de aborto em equinos no sul do Rio Grande do Sul.(AU)

Assessing epidemiology of leprosy and socio-economic distribution of cases.

Leprosy still represents a serious health problem in a number of countries, including Brazil. Although leprosy has been associated with poverty for a long time, it is still difficult to accurately define this relationship. Here, we evaluated in an endemic municipality the progress from 1995 to 2015 of epidemiological indicators to establish if there were any strong associations between social indicators and the occurrence of leprosy. An ecological study was conducted using the SINAN database (Brazilian leprosy-national notifiable diseases information system) in combination with georeferencing of leprosy cases. The georeferencing used the ArcGis programme and occurrence of cases was evaluated in relation to the Health Vulnerability Index (HVI), an indicator that categorises socio-economic and sanitation factors. The data identified a marked decrease in the overall prevalence of leprosy, a reduction in the new case-detection rate and a reduction in the number of cases with grade 2 disabilities (albeit with transient peaks in 2007 and 2015). Logistic regression analysis showed association of detection rates with elevated HVI. Thus, while the epidemiological indicators point to the elimination of leprosy, there is evidence of hidden cases and an association between higher rates of leprosy detection and greater social vulnerability remain.

Serologic evidence for Schmallenberg virus circulation at high altitude, Central Portugal, 2015-2016.

A few years after the initial 2011 large scale Schmallenberg virus (SBV) epidemic that affected Europe, a subsequent decrease in new SBV infections was observed presumably associated with natural substitution of previously exposed animals. In the present work, a 2-year prospective serosurvey was performed to evaluate SBV circulation in a population of sheep living at high altitude in the central region of Portugal and with restricted movement. Blood from a representative sample of 168 autochthonous sheep was collected in 2015 and again in 2016, and tested for the presence of anti-SBV IgG by ELISA. Of the 2015 sample collection, seven animals tested positive for anti-SBV IgG, corresponding to a seroprevalence of 4.2% while of the 2016 sample collection, 10 presented SBV antibodies, showing a seroprevalence of 6.0% (p = 0.619). Results show that SBV is endemic in sheep of central Portugal, even in herds at high altitude locations. When comparing anti-SBV seroprevalences of 2015/2016 found in this study, to one detected in 2014 in the same region, a steep decrease could be observed (p < 0.001). This is in accordance with what has been documented in Western European countries, where a decrease in the number of SBV-infected sheep has been found, a fact which may pose a new threat for SBV re-emergence.

Mesenteric lymph node granulomatous lesions in naturally infected wild boar (Sus scrofa) in Portugal--Histological, immunohistochemical and molecular aspects.

Several studies have demonstrated that wildlife reservoirs of mycobacteria are responsible for the maintenance and spreading of the infection to livestock and wildlife counterparts. Recent data report the role of wild boar (Sus scrofa) as a reservoir for Mycobacterium bovis. This study was conducted to evaluate the chronic inflammatory response in the mesenteric lymph nodes (MLN) of wild boar with granulomatous lymphadenitis (n=30). Morphological parameters of the lesions were recorded. The expression of CD3 and CD79α molecules was evaluated by immunohistochemistry. Molecular genotyping and culture to identify mycobacteria were performed. The lesions consisted mainly of stage III and stage IV granulomas. CD3 and CD79α positive cells were observed in 15 (50%) and in 11 (36.6%) MLN, respectively. In these lesions, higher percentages of T lymphocytes were found and a limited number of animals exhibited a tendency for an increased percentage of B lymphocytes. Our results suggest that there are similar percentages and distribution patterns of CD3 and CD79α in the lesions, regardless of the presence of Mycobacterium avium subsp. paratuberculosis (Map), M. bovis or Map-M. bovis co-infection, and confirm that wild boar is both susceptible and could be an important Map and M. bovis wild reservoir in the study area.

New Insights into Mycobacterium bovis Prevalence in Wild Mammals in Portugal.

A survey to determine the prevalence of Mycobacterium bovis in wild mammals in Portugal was conducted by testing samples from hunted animals and those found dead between 2009 and 2013. In this study, we investigated 2116 wild mammals. Post-mortem examinations were performed, and tissues were collected from wild mammals representing 8 families and 11 different species, with a total of 393 animals analysed. Cultures were performed, and acid-fast isolates were identified by PCR. Tissues were also screened for Mycobacterium bovis by directly extracting DNA and testing for the Mycobacterium bovis-specific sequences. Mycobacterium bovis prevalence was 26.9% (95% CI: 22.8-31.5%). Mycobacterium bovis was recorded in 106 of the 393 studied species: prevalence by species were 26.9% (95% CI: 16.8-40.2%) in red foxes, 20.0% (95% CI: 7.0-45.2%) in Egyptian mongooses, 21.4% (95% CI: 16.2-27.7%) in wild boar and 38.3% (95% CI: 29.9-47.4%) in red deer. Mycobacterium bovis infection was detected in six of eight taxonomic families. For some species, the small sample sizes obtained were a reflection of their restricted range and low abundance, making estimates of infection prevalence very difficult (1 beech marten of 4; 1 Eurasian otter of 3; 2 common genet of 3). Infection was not detected in European badgers, hedgehog, wild rabbits and hare. The results of this study confirm the presence of Mycobacterium bovis infection in wild carnivores in Portugal.

Instantaneous quantification of skeletal muscle activation, power production, and fatigue during cycle ergometry.

A rapid switch from hyperbolic to isokinetic cycling allows the velocity-specific decline in maximal power to be measured, i.e., fatigue. We reasoned that, should the baseline relationship between isokinetic power (Piso) and electromyography (EMG) be reproducible, then contributions to fatigue may be isolated from 1) the decline in muscle activation (muscle activation fatigue); and 2) the decline in Piso at a given activation (muscle fatigue). We hypothesized that the EMG-Piso relationship is linear, velocity dependent, and reliable for instantaneous fatigue assessment at intolerance during and following whole body exercise. Healthy participants (n = 13) completed short (5 s) variable-effort isokinetic bouts at 50, 70, and 100 rpm to characterize baseline EMG-Piso. Repeated ramp incremental exercise tests were terminated with maximal isokinetic cycling (5 s) at 70 rpm. Individual baseline EMG-Piso relationships were linear (r(2) = 0.95 ± 0.04) and velocity dependent (analysis of covariance). Piso at intolerance (two legs, 335 ± 88 W) was ∼45% less than baseline [630 ± 156 W, confidence interval of the difference (CIDifference) 211, 380 W, P < 0.05]. Following intolerance, Piso recovered rapidly (F = 44.1; P < 0.05; η(2) = 0.79): power was reduced (P < 0.05) vs. baseline only at 0-min (CIDifference 80, 201 W) and 1-min recovery (CIDifference 13, 80 W). Activation fatigue and muscle fatigue (one leg) were 97 ± 55 and 60 ± 50 W, respectively. Mean bias ± limits of agreement for reproducibility were as follows: baseline Piso 1 ± 30 W; Piso at 0-min recovery 3 ± 35 W; and EMG at Piso 3 ± 14%. EMG power is linear, velocity dependent, and reproducible. Deviation from this relationship at the limit of tolerance can quantify the "activation" and "muscle" related components of fatigue during cycling.

An evaluation of cattle farmers' knowledge of bovine brucellosis in northeast Portugal.

Little information is available regarding the connection between the risk of brucellosis infection in cattle and the lack of training and education of cattle producers. A total of 154 cattle farmers from the Vila Real (northern Portugal) municipality were interviewed in person to evaluate their knowledge of bovine brucellosis. Basic knowledge of the zoonotic characteristics and clinical signs of brucellosis infection and cattle management was obtained from 78.6%, 68.8% and 79.9% of the respondents, respectively. The respondents with infected animals in their herds (odds ratio (OR) 5.5; 95% confidence interval 1.6, 19.5) were more likely to have greater knowledge about bovine brucellosis. The study also revealed a relationship (p<0.01) between the use of breeding males and farms that were already infected with brucellosis. Moreover, the knowledge that brucellosis is a zoonotic disease was also influenced by the number of farms already infected with brucellosis (p<0.01). Conversely, the number of respondents who were unaware that bovine brucellosis is a zoonotic disease (25.3%) and a foodborne pathogen (21.4%), and the fact that over half (54.5%) of the respondents believed that bovine brucellosis was a treatable infectious disease was associated with the absence of veterinary assistance on the farm (60.4%). Because the eradication of bovine brucellosis has multiple factors, the success of the national eradication program cannot be based only on the sanitary management of infected herds. Successful eradication will only occur with adequate training programs for farmers, including farm biosecurity, legal fulfillment and veterinary public health programs (in which the role of the veterinarian is fundamental).

New insights into the chemistry of fac-[Ru(CO)3]²âº fragments in biologically relevant conditions: the CO releasing activity of [Ru(CO)3Cl2(1,3-thiazole)], and the X-ray crystal structure of its adduct with lysozyme.

Complexes of the general formula fac-[Ru(CO)(3)L(3)](2+), namely CORM-2 and CORM-3, have been successfully used as experimental CO releasing molecules (CO-RMs) but their mechanism of action and delivery of CO remain unclear. The well characterized complex [Ru(CO)(3)Cl(2)(1,3-thiazole)] (1) is now studied as a potential model CO-RM of the same family of complexes using LC-MS, FTIR, and UV-vis spectroscopy, together with X-ray crystallography. The chemistry of [Ru(CO)(3)Cl(2)(1,3-thiazole)] is very similar to that of CORM-3: it only releases residual amounts of CO to the headspace of a solution in PBS7.4 and produces marginal increase of COHb after long incubation in whole blood. 1 also reacts with lysozyme to form Ru adducts. The crystallographic model of the lysozyme-Ru adducts shows only mono-carbonyl Ru species. [Ru(H(2)O)(4)(CO)] is found covalently bound to a histidine (His15) and to two aspartates (Asp18 and Asp119) at the protein surface. The CO release silence of both 1 and CORM-3 and their rapid formation of protein-Ru(CO)(x)(H(2)O)(y) (x=1,2) adducts, support our hypothesis that fac-[Ru(CO)(3)L(3)] CO-RMs deliver CO in vivo through the decay of their adducts with plasma proteins.

Diagnosis of Mycobacterium avium complex in granulomatous lymphadenitis in slaughtered domestic pigs.

The aim of this study was to compare two diagnostic methods for the detection of Mycobacterium avium complex (MAC) infection in lymph nodes with granulomatous lymphadenitis from slaughtered domestic pigs. Fifty affected lymph nodes were collected from 50 pigs and examined microscopically and by polymerase chain reaction (PCR). Microscopically, granulomatous lesions were observed in 92% of the samples, consisting mostly of central necrosis (78%) with dystrophic calcification (46%) and associated with inflammatory infiltration by epithelioid giant cells, lymphocytes, neutrophils (92%), eosinophils (60%) and Langhans-type cells (70%). In 64% of the lesions, a capsule of connective tissue was found. Acid-fast bacilli were observed in all cases. PCR detected DNA from Mycobacterium spp. in 82% (41/50) of the lymph nodes. MAC was confirmed in 58% (24/41) and M. avium avium/silvaticum subspecies in 39% (16/41). The results of this study suggest that combined histopathology and PCR of lymph nodes are useful in the diagnosis of granulomatous lymphadenitis in slaughtered pigs.

Antimicrobial efficacy of alcohol-based hand gels with a 30-s application.

AIMS: The objective of this study was to assess the antimicrobial efficacy of alcohol-based hand gels according to European Norm 1500 (EN 1500). METHODS AND RESULTS: We assessed the antimicrobial efficacy of 12 alcohol-based hand gels produced in Brazil, containing 70% w/w or v/v ethyl alcohol as the active ingredient, according to EN 1500, with a 30-s application. In addition, 70% w/w ethyl alcohol and three alcohol-based hand rubs commonly used in Europe and effective according to EN 1500 were also tested. Eight of 12 (67%) alcohol-based hand gels produced in Brazil failed by EN 1500. In contrast, 70% w/w ethyl alcohol and European alcohol-based hand rubs were approved by EN 1500. CONCLUSIONS: In this study, the majority of Brazilian alcohol-based hand gels showed limited efficacy on hand hygiene within 30 s. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings of this study may be used as an important argument to motivate Brazilian manufacturers to improve the antimicrobial efficacy of alcohol-based hand gels, because it is prudent to suppose that alcohol-based hand gels can be recommended for use in healthcare settings only if they show antimicrobial activity at least similar to that of alcohol-based liquid preparations, including the traditional 70% w/w ethyl alcohol.

Effects of gastroesophageal reflux on esophageal motility.

UNLABELLED: Esophageal characteristics during swallowing in individuals with reflux are not well known. This study aimed to compare manometric esophageal characteristics between a group of individuals with gastroesophageal reflux and a group without these symptoms. METHODS: A stationary esophageal manometry and a questionnaire of gastrointestinal symptoms were performed on 41 individuals, of whom 13 (31.7%) complained of gastroesophageal reflux. RESULTS: The following data were obtained in cases of non-reflux vs. reflux: mean wave amplitude, 53.4 +/- 17.9 mm Hg vs. 48.6 +/- 11.8 mm Hg; similar mean upstroke; and lower esophageal sphincter pressure of 16.7 +/- 6.4 mm Hg vs. 17.9 +/- 4.9 mm Hg (p = 0.5). The relaxing degree and duration were similar. In the analysis according to the severity of the symptoms, we verified the following differences between cases of slight and severe reflux: amplitude in proximal esophagus, 40.75 +/- 13.1 mm Hg sec vs. 23.0 +/- 9.2 mm Hg sec (p = 0.02); mean average upstroke, 36.6 +/- 8.2 mm Hg sec vs. 24.8 +/- 7.3 mm Hg sec (p = 0.02); mean maximum upstroke, 61.4 +/- 10.4 mm Hg sec vs. 47.0 +/- 12.4mm Hg sec (p = 0.04); and relaxing duration, 5.36 +/- 2.0 sec vs. 8.76 +/- 1.6 sec (p = 0.01). CONCLUSIONS: 1. No differences in the manometric characteristics of the esophagus were found between individuals with and without reflux. 2. The wave amplitude and the upstroke were significantly higher in the proximal esophagus of patients with less severe complaints. 3. The mean and maximum upstroke of the esophageal wave were lower in individuals with severe reflux. 4. The relaxation duration was higher in patients with severe complaints.

Análise de custo-benefício da vacinação com Rev.1 da brucelose ovina e caprina no norte de Portugal entre 2000 e 2005 / Cost-benefit analysis of sheep and goat brucellosis vaccination with Rev.1 in the north of Portugal from 2000 to 2005

In the North of Portugal, a mass vaccination programme of small ruminants was conducted from 2001 to 2004. A study of cost-benefit was carried out for the 2000/2005 period to ascertain the economic benefits of this strategy. In order to estimate the cost of the zoonosis, the compensation costs paid to farmers for culled animals in the Brucellosis Eradication Campaign, data from vaccine Rev. 1 costs, and costs of people internment due to brucellosis were studied. An increase in the cost was observed from 2000 to 2001 (aproximately US$ 110,000), essentially due to compensation for culled animals, but also from vaccination and human internment costs. However, a progressive decrease was observed from 2001 to 2004 (from more than US$1,200,000 in 2001 to US$180,000 in 2004), roughly US$ 1,020,000 less. As the main conclusion, the annual cost of this zoonosis decreased by an average of almost US$600,000 per year, and a total of more than three million dollars were saved from 2000 to 2005. The results of this study suggest that mass vaccination with Rev. 1 reduced overall costs and was effective in reducing animal and human brucellosis costs.

No norte de Portugal, procedeu-se à vacinação massiva contra a brucelose em pequenos ruminantes entre 2001 e 2004. Neste estudo, efetuou-se uma análise de custo benefício entre 2000 e 2005 para apurar os benefícios econômicos dessa estratégia. Para estimar o custo da zoonose, estudaram-se três variáveis: a indenização paga aos proprietários dos animais eliminados durante a campanha de erradicação da brucelose, o custo da vacina Rev. 1 e os custos de internamento das pessoas infectadas por brucelose nas unidades de Serviços de Saúde. Durante os anos em estudo, observou-se um aumento de custo de aproximadamente US$110.000, entre 2000 e 2001, principalmente, devido à indenização dos animais eliminados e à aplicação da vacina e os custos por internamento de pessoas infectadas, seguindo-se uma redução acentuada entre 2001 e 2004 (de US$1.200.000 para US$180.000 em 2004), o que perfez US$1.020.000 poupados. O custo anual da zoonose, em relação às variáveis estudadas, diminuiu em média US$600.000 por ano, sendo poupados mais de US$ 3.000.000 entre 2000 e 2005. Estes resultados sugerem que a vacinação com Rev. 1 reduziu os custos totais da infecção humana e animal relativamente às variáveis estudadas.

High levels of leptin modulate esophageal motor characteristics in type 2 diabetic patients.

UNLABELLED: Leptin regulates gastric and intestinal motility, but its effect on oesophageal motility is unknown. We analyzed oesophageal manometric characteristics in diabetics with elevated leptin. METHODS: Fasting blood leptin levels were measured in 32 type 2 individuals aged from 39-81 years. An oesophageal stationary manometry was then performed. Each manometric door (P) registered one third of the oesophageal activity. Results are presented as mean +/- SD. RESULTS: Twenty-one subjects had elevated leptin (HLL) while 11 displayed normal levels (NLL). Peristaltic wave distributions (%) in NLL vs. HLL were 79.4 +/- 26.3 vs. 88.6 +/- 8.3 (p = 0.2). Simultaneous and retrograde waves showed similar trends. Non-transmitted waves were 16.1 +/- 26.5 vs. 4.6 +/- 4.5% (p < 0.05). Amplitudes in NLL vs. HLL (in mm Hg) were P1: 30.2 +/- 10.8 vs. 33.2 +/- 11.7 (p = 0.4), P2: 38.4 +/- 14.4 vs. 58.0 +/- 21.2 (p = 0.01), P3: 42.4 +/- 14.4 vs. 64.7 +/-2 8.3 (p < 0.006), and average amplitudes: 37.1 +/- 12.1 vs. 52.1 +/- 17.6 (p = 0.01). Wave average upstroke (in mm Hgs) was P1: 25.6 +/- 19.1 vs. 23.3 +/- 10.1 (p = 0.6), P2: 26.8 +/- 10.7 vs. 36.2 +/- 11.6 (p < 0.03), and P3: 25.5 +/- 9.1 vs. 34.1 +/- 16.3, (p < 0.06). Wave maximum upstroke was P1: 39.0 +/- 18.6 vs. 40.5 +/- 13.8, (p = 0.8), P2: 45.5 +/- 15.5 vs. 63.8 +/- 19.2 (p = 0.01), P3: 46.6 +/- 17.8 vs. 65.0 +/- 29.1 (p <0.03). Wave duration in distal oesophagus was 4.5 +/- 0.7 vs. 5.5 +/- 1.1 s (p = 0.01), and velocity 3.3 +/- 3.3 vs. 2.96 +/- 3.7 cm/s (p = 0.6). CONCLUSION: 1--Non-transmitted waves were slightly higher in NLL. 2--In medium and distal oesophagus, the wave amplitude, medium and maximum upstroke, and duration in distal oesophagus were increased in HLL.

Comparative evaluation of PCR in Ziehl-Neelsen stained smears and PCR in tissues for diagnosis of Mycobacterium avium subsp. paratuberculosis.

Thirty six tissues from sheep, previously diagnosed with paratuberculosis, were tested by PCR in positive Ziehl-Neelsen staining smears of tissues, and PCR in tissues targeting IS900 specific for Mycobacterium avium subsp. paratuberculosis. DNA amplification was achieved in 33.3% Ziehl-Neelsen smears, and in 61.1% tissue samples. Combination of both techniques found 66.7% samples as positive. Combination of techniques would, therefore, increase the sensitivity of diagnosis.

Complex inguinal hernia repairs.

INTRODUCTION: Complex inguinal hernia treatment is a challenge for general surgeons. The gold standard for the repair of inguinal hernias is the Lichtenstein repair (anterior approach). However, when multiple recurrent hernias or giant hernias are present, it is necessary to choose different approaches because the incidence of poor results increases. There are many preperitoneal approaches described in the literature. For example: (a) open procedure-Nyhus and Stoppa (b) laparoscopic technique-transabdominal pre-peritoneal (TAPP) and totally extraperitoneal (TEP). In this study, we show how we repair complicated cases using open access in huge unilateral or bilateral, recurrent, or multiple recurrent inguinal hernias. METHODS: The present study includes the period from November 1993 through December 2007. One hundred and eighty-eight patients, divided into 121 with unilateral hernias and 67 with bilateral hernias, totaling 255 inguinal hernia repairs, were treated by the Nyhus or Stoppa preperitoneal approach, depending on whether they were unilateral or bilateral. We used progressive preoperative pneumoperitoneum for oversize inguinal hernias in all patients. RESULTS: Orchiectomy was necessary on only two occasions. Despite the repair complexity involved, we had only two known recurrences. The mortality was zero and the morbidity was acceptable. CONCLUSIONS: We conclude that an accurate open preperitoneal approach using mesh prosthesis for complex inguinal hernias is safe, with very low recurrent rates and low morbidity. Progressive preoperative pneumoperitoneum for giant hernias was shown to be an important factor in accomplishing good intraoperative and immediate postoperative results.

Ocorrência de Actinobacillus pleuropneumoniae biótipo 1 em pulmões de suínos com pleuropneumonia no Norte de Portugal / Occurrence of Actinobacillus pleuropneumoniae biotype 1 in swine with pleuropneumonia in the North of Portugal

The isolation and identification of Actinobacillus pleuropneumoniae in swine lungs with pleuropneumonia in the North of Portugal were reported. A total of 127 swine lungs with and without lesions were examined. The system of lesions classification was based on a semi-quantitative method. Diagnosis was made by isolation and identification of the etiological agent in typical lesions. The occurrence of observed lesions was 75.6 percent and the occurrence of isolation of A. pleuropneumoniae was 19.7 percent. In 25 out of 96 (26.0 percent) lung samples with lesions of pleuropneumonia, A. pleuropneumoniae was isolated.