Schistosoma mansoni granulomas in the skeletal striated muscles in the murine model of neuroschistosomiasis: histological findings.

Schistosomiasis mansoni presents many clinical manifestations during migration of schistosomes in their hosts, including diarrhea, hepatomegaly, splenomegaly, liver abscesses, skinlesions, brain tumors and myeloradiculopathy. No lesions have been reported in skeletal striated muscles due to schistosomiasis mansoni in the literature. This short communication reports the histopathological findings on skeletal musculature in a murine model of neuroeschistosomiasis mansoni. Lesions were found in the tongue, masseter muscle, buccinator muscle, digastric muscle and temporalis muscle. Worm recovery was carried out to confirm the infection. We describe here, for the first time in the literature, injuries in the skeletal musculature due to Schistosoma mansoni nfection.

Characterisation of ocular involvement in an experimental model of neuroschistosomiasis mansoni.

The Global Burden of Disease Study 2010 listed schistosomiasis among the leading 100 causes of death in Brazil, responsible for 3.6% of the estimated total of deaths globally. Eye and adnexa are very rarely affected by schistosomiasis mansoni, with limited documentation of ocular pathology in this setting. This short communication reports ocular histolopathological findings in a murine model of neuroschistosomiasis mansoni. Lesions were found in the bulbar conjunctiva, lacrimal gland, choroid and corneoscleral limbus.

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% of cases by TS-PCR and in 75% 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% 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.

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

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.

Bracken fern-induced malignant tumors in rats: absence of mutations in p53, H-ras and K-ras and no microsatellite instability.

Bracken fern (genus Pteridium) has been shown to induce tumors in domestic and experimental animals. Epidemiological studies have also shown an association between human exposure to bracken toxins and increased risk for the development of upper gastrointestinal tract tumors. Our aim in this study was to investigate possible genomic alterations in bracken fern-induced tumors of experimental animals searching for molecular markers that might be used for human epidemiological studies. Using human colorectal carcinogenesis as a molecular model, we examined eight malignant bracken fern-induced tumors of rats for mutations in the genes associated with the "classic pathway" of colorectal cancer, i.e. p53 and ras, and also in the "mutator pathway" by evaluating microsatellite instability. Exons 5-9 of the p53 gene and exons 1 and 2 of the K-ras and H-ras genes were examined by DNA sequencing and no mutations were found in any of the eight tumors. Amplification of five previously validated microsatellite loci (one with mono-, three with di- and one with tetra-nucleotide repeat motifs) in the malignant tumors and in the surrounding normal tissue did not reveal any instability. The involvement of epigenetic alterations or of mutations in other tumor suppressor genes or oncogenes should be further investigated in the search for human epidemiological markers.

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.

Correlation of endoscopic and histological features in adults with suspected celiac disease in a referral center of Minas Gerais, Brazil.

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.

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.

Mitochondrial cardioencephalomyopathy due to a novel SCO2 mutation in a Brazilian patient: case report and literature review.

OBJECTIVES: To review all patients with SCO2 mutations and to describe a Brazilian patient with cardioencephalomyopathy carrying compound heterozygous mutations in SCO2, one being the known pathogenic p.E140K mutation and the other a novel 12-base pair (bp) deletion at nucleotides 1519 through 1530 (c.1519_1530del). DESIGN: Case report and literature review. SETTING: University hospital. PATIENT: Infant girl presenting with an encephalomyopathy, inspiratory stridor, ventilator failure, progressive hypotonia, and weakness, leading to death. MAIN OUTCOME MEASURES: Clinical features, neuroimaging findings, muscle biopsy with histochemical analysis, and genetic studies. RESULTS: This infant girl was the first child of healthy, nonconsanguineous parents. She developed progressive muscular hypotonia and ventilatory failure. At the end of the first month of life, she developed cardiomegaly and signs of cardiac failure. Routine blood tests showed lactic acidosis and mild elevation of the creatine kinase level. Brain magnetic resonance imaging showed increased T2 and fluid-attenuated inversion recovery signals in the putamen bilaterally. Nerve conduction studies showed severe axonal sensorimotor neuropathy. Muscle biopsy revealed a neurogenic pattern with mitochondrial proliferation and total absence of cytochrome- c oxidase histochemical stain. Sequencing of SCO2 showed that the patient had compound heterozygote SCO2 mutations: the previously described c.1541G>A (p.E140K) mutation and a novel 12-bp deletion at nucleotides 1519 through 1530 (c.1519_1530del). The patient died at age 45 days. CONCLUSIONS: Our findings and the literature review indicate that it is important to consider the diagnosis of mitochondrial disease in newborns with hypotonia and cardiomyopathy. In our case, the accurate diagnosis of SCO2 mutations is particularly important for genetic counseling.

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.

Clinical and molecular characterization of McArdle's disease in Brazilian patients.

McArdle's disease, a glycogen storage disease type V, is caused by a deficiency of the enzyme myophosphorylase, encoded by the PYGM gene. Worldwide distribution of mutations has revealed interesting data about the prevalence of mutations and population migrations. Currently, more than 100 mutations in the PYGM gene have been described, with some recurrent mutations in the different populations. However, no molecular studies of McArdle's disease were reported in Brazilian patients. Here, we describe the clinical phenotype and genotype of 10 patients from 8 unrelated Brazilian families. Among the 10 patients (3 females, 7 males), eight presented with the typical phenotype, with exercise intolerance, cramps, and myalgia; one patient showed permanent muscle weakness; and one patient showed a mild phenotype. Molecular analysis identified 5 different mutations in the 8 families, both in homozygosis or compound heterozygosis state. Four of them had already been described (p.R50X, p.T692kfs30, p.K609K, and p.G455R), and one, pI513V, is a novel heterozygous mutation. The common nonsense p.R50X mutation was found in 6 of the 8 families, being therefore the commonest mutation in the Brazilian population as well. Other mutations previously reported in European patients were also found in the patients in this study, which was expected considering the European ancestry of the Brazilian population.

Early polymerase chain reaction detection of Chagas disease reactivation in heart transplant patients. / Detecção precoce da reação em cadeia da polimerase da reativação da doença de Chagas em pacientes transplantados cardíacos.

FUNDO: O transplante cardíaco é uma valiosa opção terapêutica para pacientes com doença de Chagas com cardiomiopatia grave. Durante o seguimento dos pacientes, o diagnóstico diferencial entre a rejeição ao transplante cardíaco e a reativação da infecção por doença de Chagas continua sendo uma tarefa desafiadora, o que dificulta a rápida implementação do tratamento adequado. Aqui investigamos se as estratégias de reação em cadeia da polimerase (PCR) poderiam facilitar a detecção precoce do Trypanosoma cruzi (T cruzi) em biópsias endomiocárdicas transplantadas (EMBs). MÉTODOS: Neste estudo, foram analisadas 500 amostras EMB obtidas de 58 pacientes submetidos a transplante cardíaco chagásico, utilizando-se métodos de PCR direcionados aos marcadores nuclear (rDNA 24Sα) e cinetoplasto (kDNA), e compararam a eficiência dessas abordagens com a de outros testes rotineiramente utilizados. RESULTADOS: O DNA do T cruzi foi detectado em 112 espécimes EMB derivados de 39 pacientes (67,2%). O primeiro resultado positivo ocorreu em uma mediana de 1,0 mês após o transplante. Análises histopatológicas convencionais, esfregaços de sangue e hemoculturas mostraram menor sensibilidade e maior tempo médio para o primeiro resultado positivo. O seguimento dos pacientes revelou que 31 dos 39 casos PCR-positivos apresentaram reativação clínica da doença de Chagas em diferentes momentos após o transplante. As técnicas de PCR mostraram sensibilidade considerável (0,82) e especificidade (0,60), com área sob as curvas ROC (Receiver Operating Characteristic) de 0,708 (p = 0,001). Além disso, técnicas de PCR anteciparam os sinais clínicos de reativação da doença de Chagas em até 36 meses, com tempo mediano de 6 meses e média de 9,1 meses. CONCLUSÕES: Encontramos uma boa associação entre o diagnóstico de PCR e os sinais clínicos da doença, indicando que as abordagens de PCR aqui utilizadas são adequadas para o diagnóstico precoce da reativação da doença de Chagas, com alto potencial para auxiliar os médicos nas decisões de tratamento. Para este propósito, um algoritmo é proposto para vigilância baseada nos testes moleculares.

Terminal ischemic cardiomyopathy associated with complication of stenting in the treatment of acute myocardial infarction.

A serious complication such as dissection of the left main coronary artery, with significant reduction in coronary blood flow by the true light, requires quick action. Therefore, the immediate choice of stent with appropriate length and size to treat the complication is necessary.

Ischemia-reperfusion histopathology alterations of the rabbit intestinal wall with and without ischemic preconditioning.

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.