Anti-serum validation for use in immunohistochemistry for Trypanosoma cruzi detection

Abstract INTRODUCTION: The detection of Trypanosoma cruzi in tissue samples is important in many situations, such as testing of the reactivation of the infection. The detection of T. cruzi nests in endomyocardial biopsies (EMB) may be useful to evaluate graft rejection. Given their scarcity, such nests are not routinely identified. To increase the diagnosis sensitivity, immunohistochemistry (IHC) may serve as a promising strategy. Here, we validate an antiserum for the detection of T. cruzi infection by IHC. METHODS: We used 1) positive controls (PCs) - 13 EMB, 12 skin biopsies, and 1 heart with T. cruzi nests as sections stained with hematoxylin and eosin (HE); 2) negative controls - a) 10 explant hearts and 10 EMB with no amastigote nests or clinical/laboratory signs of chagasic infection; and b) eight samples with leishmaniasis, toxoplasmosis, or histoplasmosis; and 3) Cases - 31 EMB of chagasic patients with no parasite nests in HE sections but detected positive for T. cruzi DNA by polymerase chain reaction. As a primary antibody, a hyperimmune serum from T. cruzi-infected rabbits was used. RESULTS: IHC results were positive for 21 of 26 PCs (80.8%) and one case of cutaneous leishmaniasis. In 4 of 31 cases, IHC revealed nests (12.9%), which were undetected by conventional histological examination. CONCLUSIONS: This study shows that IHC with the tested antiserum increases the sensitivity of the diagnosis and may be recommended for routine use in EMB analyses of cardiac transplant patients with Chagas disease.

Correlation of nndoscopic and histological featres in adults with suspected ceeliac disease in a referral center of minas gerais, Brazil / Correlação entre os aspectos endoscópicos e histológicos de pacientes adultos com suspeita de doença celíaca em um centro de referência de Minas Gerais, Brasil

Context Clinical presentation of celiac disease is extremely variable and the diagnosis relies on serologic tests, mucosal intestinal biopsy and clinic and serologic response to a gluten-free diet. Objectives To correlate the endoscopic and histological aspects of adult patients with suspicion of celiac disease and to evaluate the interobserver histological agreement. Methods Endoscopic aspects of 80 adult patients were evaluated and correlated with the histological features according the Marsh-Oberhuber classification system. The interobserver histological agreement was based on kappa values. Results The symptoms of the patients varied largely, with prominence for chronic diarrhea, present in 48 (60%) patients. The endoscopic aspects related with the duodenal villous atrophy had been observed in 32 (40%) patients. There were confirmed 46 cases of celiac disease, with prevalence of 57.5%. The sensitivity, specificity, positive predictive value and negative predictive value of the endoscopic markers for celiac disease diagnosis were of 60.9%, 88.2%, 87.5% and 62.5%. There was moderate interobserver histological agreement (kappa = 0.46). Conclusions The endoscopic markers of villous atrophy, although not diagnostic, had assisted in the suspicion and indication of the duodenal biopsies for diagnosis proposal. Histology is sometimes contradictory and new biopsies or opinion of another professional can provide greater diagnostic agreement. .

Contexto A apresentação clínica da doença celíaca é extremamente variável e o diagnóstico se baseia em testes sorológicos, histologia intestinal e respostas clínica e sorológica à dieta sem glúten. Objetivos Correlacionar os aspectos endoscópicos e histológicos de pacientes adultos com suspeita de doença celíaca e avaliar a concordância histológica interobservadores. Métodos Os aspectos endoscópicos de 80 pacientes adultos foram avaliados e correlacionados com os achados histológicos de acordo com a classificação de Marsh-Oberhuber. A concordância histológica foi baseada nos valores kappa. Resultados A sintomatologia clínica foi muito variável com destaque para a diarréia crônica, presente em 48 (60%) pacientes. Os aspectos endoscópicos relacionados à atrofia vilositária duodenal foram observados em 32 (40%) pacientes. Foram confirmados 46 casos de doença celíaca, prevalência de 57.5%. A sensibilidade, a especificidade, o valor preditivo positivo e o valor preditivo negativo dos aspectos endoscópicos para o diagnóstico da doença celíaca foram, respectivamente, 60,9%, 88,2%, 87,5% e 62,5%. A concordância histológica interobservadores foi moderada (kappa = 0,46). Conclusões Os aspectos endoscópicos de atrofia vilositária contribuíram para a suspeita e a indicação das biópsias duodenais com objetivo diagnóstico. A histologia pode ser contraditória e novas biópsias ou a opinião de outro profissional podem propiciar maior concordância diagnóstica. .

Brain schistosomiasis in mice experimentally infected with Schistosoma mansoni

Introduction Human neuroschistosomiasis has been reported in the literature, but the possibility of modeling neuroschistosomiasis in mice is controversial. Methods In two research laboratories in Brazil that maintain the Schistosoma mansoni life cycle in rodents, two mice developed signs of brain disease (hemiplegia and spinning), and both were autopsied. Results S. mansoni eggs, both with and without granuloma formation, were observed in the brain and meninges of both mice by optical microscopy. Conclusions This is the first description of eggs in the brains of symptomatic mice that were experimentally infected with S. mansoni. An investigation of experimental neuroschistosomiasis is now feasible. .

Ischemia-reperfusion histopathology alterations of the rabbit intestinal wall with and without ischemic preconditioning / Alterações histopatológicas da parede intestinal de coelhos na isquemia-reperfusão com e sem precondicionamento isquêmico

PURPOSE: To evaluate the histopathology alterations of the intestinal mucosa of rabbits submitted to mesenteric artery ischemia and reperfusion with and without ischemic preconditioning. METHODS: Two groups of ten male New Zealand white rabbits body (weight 2.2-3.0, average 2.5 kg). For mesenteric ischemia induction in all animals the small bowel and mesentery were cut 30cm and 60cm far from the gastroduodenal pyloric transition before the proximal mesenteric artery occlusion. In the Group 1 animals, the proximal mesenteric artery was occluded for 45 min with an atraumatic vascular clamp, followed by reperfusion for 30 min. In the Group 2 the 45 min ischemic phase was preceded by three cycles of ischemia (2 minutes each) alternated with three cycles of reperfusion (2 minutes each). For istopathology study small bowel biopsies were obtained before ischemia (control), after 45 min of mesenteric ischemia and at 30 min. of mesenteric artery reperfusion. RESULTS: In the Group I animals, the followings histopathology grade results were observed: t1, mean 2,8; t2, mean 3,3. Using the Kruskal-Wallis non-parameter test, differences between t0 and t1 and t0 and t2 were significants (p<0.05), but not significant between t1 and t2 (p>0.05). In the Group 2 animals histopathology grade results were: t1 mean 2,6 and t2, mean 2,1. Differences between t0 and t1, t0 and t2 were significant (p<0.05). It was not observed differences (p>0.05) between results of t1 in both groups but histopathology injury observed in Group 1 t2 biopsies were higher (p<0.05) than observed in the same period (t2) of Group 2 animals. CONCLUSION: Microscopic examination of the biopsies revealed significant evidence of preconditioning protection against small bowel wall ischemia-reperfusion injury.

OBJETIVO: Avaliar as alterações histopatológicas da mucosa intestinal de coelhos submetidos a isquemia-reperfusão com e sem precondicionamento isquêmicol. MÉTODOS: Foram estudados dois grupos de dez coelhos Nova Zelândia machos com pesos variáveis entre 2,2 e 3,0 kg (média de 2,5 kg) de peso corpóreo. Para indução da isquemia, em todos os animais, o intestino delgado e o mesentério foram seccionados 30 cm e 60 cm após a transição pilórica gastroduodenal, antes da oclusão da artéria mesentérica cranial. Nos animais do Grupo 1, a artéria mesentérica proximal foi ocluida por pinçamento atraumático durante 45 min., seguido de reperfusão por 30 min. No Grupo 2, foi realizado precondicionamento por três ciclos de 2 min. de oclusão mesentérica intercalados com três ciclos de 2 min. de reperfusão, seguido de oclusão mantida por 45 min e reperfusão de 30min. como no Grupo I. Para estudo histopatológico, foram obtidas biópsias da parede intestinal antes da isquemia (t0-controle), após 45 min. de isquemia (t1) e após 30 min. de reperfusão (t2). RESULTADOS: No Grupo I foram observados os seguintes graus de lesões: t1, média de 2,8 e t2, média 3,3, Foram significantes as diferenças entre t0 e t1 e t0 e t2, mas não foram significantes as variações entre t1 e t2 (p>0,05). No Grupo 2, obteve-se em t1,média de 2,6 e t2, média 2,1. Foram significantes (p<0,05) as diferenças entre t0 e t1, t0 e t2 e entre t1 e t2. . Não ocorreu diferença significante (p>0,05) entre os resultados de t1 nos dois Grupos, mas foram significantes (p<0,05) as diferenças entre os resultados histopatológicos das biopsias de t2 dos Grupos 1 e 2. CONCLUSÃO: O precondicionamento isquêmico reduziu significantemente a degeneração histopatológica determinada pela reperfusão pós-isquêmica da parede intestinal.

Ischemia-reperfusion histopathology alterations of the rabbit intestinal wall with and without exclusion of the collateral mesenteric circulation supply / Alterações histopatológicas da parede intestinal de coelhos na isquemia-reperfusão com e sem exclusão da circulação mesentérica colateral

PURPOSE: To evaluate the histopathology alterations of the intestinal mucosa of rabbits submitted to different times of mesenteric artery ischemia and reperfusion with and without celiac artery collateral circulation supply. METHODS: Two groups of eight male New Zealand white rabbits (weight 2.2-3.5 kg) were used in this study. In the Group 1 animals, the proximal mesenteric artery was occluded for 60 min with an atraumatic vascular clamp, followed by reperfusion for 60 min. In the Group 2 animals the small bowel and mesentery were cut 30cm and 60cm far from the gastroduodenal pyloric transition before the proximal mesenteric artery occlusion. Small bowel biopsies were obtained before ischemia (control), after 30 min and 60 min of mesenteric ischemia and at 30 and 60 min. of mesenteric artery reperfusion. RESULTS: In the Group I animals, the followings histopathology grade results were observed: t1, mean 0.4 + 0.29; t2, mean 1.9 ± 0.38; t3, 1.9 ± 0.33; t4, 1.2 ± 0.36 and t5, 1.2 ± 0.32. Differences between t0 and t2 and between t3 and t4 were statistically significant (p<0.05). Differences between t2 and t3 and t4 and t5 were not significant (p>0.5). In the Group II animals, it was observed: t1, mean 1.6 ± 0.33; t2, 2.4 ± 0.36; t3, 3.0 ± 0.35; t4 3.4 ± 0.31; t5, 3 ± 031. Differences between t0 and t1, t1 and t2, and t2 and t3 were significant (p<0.05). Differences between histopathology grades results of samples t1 to t5 in Group 1 and 2 were statistically significant (p<0.5). CONCLUSION: Microscopic examination of the biopsies revealed significant evidence of worse small bowel wall ischemia-reperfusion lesions by exclusion of the celiac artery collateral circulation supply.

OBJETIVO: Avaliar as alterações histopatológicas da mucosa intestinal de coelhos submetidos à isquemia-reperfusão com e sem exclusão da circulação mesentérica colateral. MÉTODOS: Foram estudados dois grupos de oito coelhos Nova Zelândia machos com pesos variáveis entre 2,2 e 3,5 kg de peso corpóreo. Nos animais do Grupo 1, a artéria mesentérica proximal foi ocluida por pinçamento atraumático durante 60 min, seguido de reperfusão por 60 min. No Grupo 2 o intestino delgado e o mesentério foram seccionados 30 cm e 60 após a transição pilórica gastroduodenal antes da oclusão da artéria mesentérica cranial. Biópsias da parede intestinal foram obtidas antes da isquemia (controle), após 30 e 60 min. de isquemia. RESULTADOS: No Grupo I foram observados os seguintes graus de lesões: t1,média de 0.4 + 0.29; t2, média 1.9 ± 0.38; t3, 1.9 ± 0.33; t4, 1.2 ± 0.36 e t5, 1.2 ± 0.32. As diferenças entre t0 e t2 e entre t3 e t4 foram significantes (p<0.05). As diferenças entre t2 e t3 e t4 e t5 não foram significantes (p>0.5). No Group II observou-se: t1, média de 1.6 ± 0.33; t2, 2.4 ± 0.36; t3, 3.0 ± 0.35; t4 3.4 ± 0.31; t5, 3 ± 031. As diferenças entre t0 e t1, t1 e t2, e t2 e t3 foram significantes (p<0.05). As diferenças entre os resultados histopatológicos das biopsies de t1 a t5 dos Grupos 1 e 2 foram significantes (p<0.5). CONCLUSÃO: A exclusão da circulação mesentérica colateral agravou significantemente a degeneração histopatológica na isquemia-reperfusão da parede intestinal.

Poluição atmosférica e efeitos respiratórios, cardiovasculares e reprodutivos na saúde humana / Air Pollution and its respiratory, cardiovascular andreproductive effects in the human health

A poluição atmosférica é responsável por vários danos à saúde humana, resultando em aumento da morbidade e/ou mortalidade por inúmeras doenças. A poluição do ar deriva da queima de combustíveis fósseis, sendo as indústrias e os veículos automotores suas principais fontes. Os poluentes atmosféricos podem ser voláteis ou particulados. Destes, o mais danoso é o material particulado com diâmetro aerodinâmico < 2,5 µm (PM 2,5). Neste artigo, são descritos os principais efeitos provocados à saúde pelo PM 2,5, notadamente nos sistemas respiratório, circulatório e reprodutor. No sistema respiratório, os danos mais importantes são desencadeamento ou agravamento de inflamações pulmonares, asma, doença pulmonar obstrutiva crônica e câncer. Os principais eventos cardiovasculares são alterações no ritmo cardíaco, isquemia miocárdica, modificações na coagulação sanguínea e progressão da aterosclerose. Na gravidez, crescimento intrauterino restrito, prematuridade e baixo peso ao nascimento são as consequências mais marcantes. Como indicam vários estudos, a redução da poluição atmosférica, tarefa nada fácil, mas perfeitamente alcançável, reduz a morbidade e a mortalidade por essas doenças, indicando que ações públicas sobre o assunto são necessárias e inadiáveis, pois trazem efeitos palpáveis à vida das pessoas.

Air pollution is responsible for several damages to human health, resulting in morbidity and/or mortality increasing due to a number of diseases. Air pollution comes from fossil fuel burning, and the industries and automotor vehicles are the main sources. Air pollutants can be volatile or particulate. Among these, the most damaging is the particulate matter with aerodynamic diameter < 2,5 µm (PM 2,5). The main PM 2,5 effects for the health are described in this article, particularly regarding the respiratory, circulatory and reproductive systems. In the respiratory system, the most relevant damage is the onset or worsening of pulmonary inflammation, asthma, chronic obstructive pulmonary disease and cancer. The major cardiovascular events are changes in the heart rate, myocardial schemia, changes in blood coagulation and arteriosclerosis progression. In pregnancy, restrict intrauterine growth, prematurity and low birth weight are the most noteworthy consequences. As indicated by several studies, the reduction of air pollution, no easy task but quite achievable, reduces morbidity and mortality from these diseases, indicating that public actions on the matter are necessary and unavoidable , as they bring tangible improvements to people?s lives.

Comparison of HPV genotyping by type-specific PCR and sequencing

Human papillomavirus (HPV) infection is the most common sexually transmitted disease worldwide and there is a strong link between certain high-risk viral types and cervical carcinogenesis. Although there are several typing methods, it is still unclear which test is the best. This study compared the effectiveness of type-specific PCR (TS-PCR) and sequencing, with a focus on their clinical application. A total of 260 cervical samples from HPV-positive patients were tested for types 6, 11, 16, 18, 31, 33 and 35 using TS-PCR and sequencing. The genotype was identified in 36 percent of cases by TS-PCR and in 75 percent by sequencing. Sequencing was four times more likely to identify the viral type in positive samples than TS-PCR (p = 0.00). Despite being more effective for virus genotyping, sequencing was unable to identify viral types in multiple infections. Combining both techniques resulted in highly sensitive detection (87 percent of cases), showing that they are complementary methods. HPV genotyping is an important step in HPV management, helping to identify patients with a higher risk of developing cervical cancer and contributing to the development of type-specific vaccines.

Prenatal toxoplasmosis diagnosis from amniotic fluid by PCR

Toxoplasmosis is one of the most common infections all over the world. Most cases are asymptomatic, except in immunosuppressed individuals and fetuses, which can be seriously damaged. Prenatal diagnosis should be made as soon as possible since treatment of the mother can minimize fetal sequelae. Our aim in this study was to test the polymerase chain reaction technique (PCR) in 86 samples of amniotic fluid from women who seroconverted during pregnancy. DNA was amplified using external primers and, in a second step, internal primers, in a nested PCR system. Samples were also inoculated into mice and the newborn were evaluated by T. gondii serology, skull x-ray, transfontanel ultrasound, fundoscopic examination, lumbar puncture and clinical examination. PCR was positive in seven cases and negative in 79. Among PCR-positive cases, two were negative by inoculation into mice and by clinical evaluation; among PCR-negative ones, three had clinical evidence of toxoplasmosis and one was positive after inoculation into mice. PCR showed values of sensitivity = 62.5 and specificity = 97.4; the values of inoculation into mice where 42.9 and 100, respectively. Although PCR should not be used alone for prenatal diagnosis of congenital toxoplasmosis, it is a promising method and deserves more studies to improve its efficacy.

Comparação dos métodos para diagnóstico da toxoplasmose congênita / A comparasion between methods for the diagnosis of congenital toxoplasmose

Objetivo: avaliar a eficácia da reação em cadeia da polimerase (PCR) no líquido amniótico para a detecção da contaminação fetal pelo Toxoplasma gondii em gestantes com infecção aguda e correlacionar com a técnica de inoculação e a histologia da placenta. Métodos: trinta e sete pacientes foram estudadas prospectivamente e o diagnóstico foi baseado na identificação da infecção aguda materna, seguido pela amniocentese guiada pela ultra-sonografia para obtenção de líquido amniótico para a realização de PCR e inoculação em camundongo. As pacientes foram tratadas com espiramicina durante a gestação; se a infecção fetal era demonstrada, pirimetamina e sulfadiazina eram acrescentadas ao esquema. As placentas foram encaminhadas para exame histológico. As crianças foram acompanhadas durante um período que variou de três a 23 meses para a confirmação ou exclusão da toxoplasmose congênita. Resultados: medidas de associação como sensibilidade, especificidade e valores preditivos foram calculadas para a PCR no líquido amniótico, a inoculação em camundongo e a histologia da placenta, mostrando os seguintes resultados: para a PCR, uma sensibilidade de 66,7 por cento e especificidade de 87,1 por cento; os respectivos valores para a inoculação em camundongo foram 50 e 100 por cento e para a histologia da placenta foram 80 e 66,7 por cento. Conclusões: embora a PCR não deva ser utilizada isoladamente para diagnóstico de toxoplasmose congênita, é um método promissor e necessita de maiores estudos para melhorar sua eficácia.

Whipple's disease. Report of five cases with different clinical features

Whipple's disease (WD) is a rare systemic disease of infectious etiology which involves the small intestine but can virtually affect any organ. We present here five cases (four males and one female) ranging in age from 20 to 59 years. All patients had intestinal involvement associated or not with clinical manifestations linked to this organ. Vegetation in the tricuspid valve was observed in one patient, suggesting endocarditis caused by Tropheryma whippelii, with disappearance of the echocardiographic alterations after treatment. In one of the male patients the initial clinical manifestation was serologically negative spondylitis, with no diarrhea occurring at any time during follow-up. Ocular involvement associated with intestinal malabsorption and significant weight loss were observed in one case. In the other two cases, diarrhea was the major clinical manifestation. All patients were diagnosed by histological examination of the jejunal mucosa and, when indicated, of extraintestinal tissues by light and electron microscopy. After antibiotic treatment, full remission of symptoms occurred in all cases. A control examination of the intestinal mucosa performed after twelve months of treatment with sulfamethoxazole-trimethoprim revealed the disappearance of T. whippelii in four patients. The remaining patient was lost to follow-up

Congenital cytomegalovirus infection in a neonatal intensive care unit in Brazil evaluated by PCR and association with perinatal aspects

Cytomegalovirus (CMV) infection is the most common congenital infection, affecting 0.4 percent to 2.3 percent newborns. Most of them are asymptomatic at birth, but later 10 percent develop handicaps, mainly neurological disturbances. Our aim was to determine the prevalence of CMV shed in urine of newborns from a neonatal intensive care unit using the polymerase chain reaction (PCR) and correlate positive cases to some perinatal aspects. Urine samples obtained at first week of life were processed according to a PCR protocol. Perinatal data were collected retrospectively from medical records. Twenty of the 292 cases (6.8 percent) were CMV-DNA positive. There was no statistical difference between newborns with and without CMV congenital infection concerning birth weight (p=0.11), gestational age (p=0.11), Apgar scores in the first and fifth minutes of life (p=0.99 and 0.16), mother's age (p=0.67) and gestational history. Moreover, CMV congenital infection was neither related to gender (p=0.55) nor to low weight (<2,500g) at birth (p=0.13). This high prevalence of CMV congenital infection (6.8 percent) could be due to the high sensitivity of PCR technique, the low socioeconomic level of studied population or the severe clinical status of these newborns.

Visceral larva migrans and tropical pyomyositis: a case report

Os autores relatam o caso de um jovem com piomiosite tropical grave, eosinofilia e hiperimunoglobulinemia E. O paciente relatou historia de contato com um cao e o teste de ELISA para Toxocara canis revelou-se positivo. O Staphylococcus aureus foi a unica bacteria isolada da secrecao purulenta obtida dos abscessos musculares. Sugere-se que a piomiosite tropical possa ser iniciada pela presenca das larvas desse ou de outros parasitos com tropismo para os musculos. As alteracoes imunologicas e estruturais nos musculos acometidos pelas larvas e a presenca de bacteriemia podem favorecer a instalacao da bacteria e o desenvolvimento da piomiosite