Trypanosoma cruzi: identification and characterization of a novel ribosomal protein L27 (TcrL27) that cross-reacts with an affinity-purified anti-Sm antibody

Small nuclear ribonucleoproteins (snRNPs)are involved in trans-splicing processing of pre-mRNA in Trypanosoma cruzi. To clone T. cruzi snRNPs we screened an epimastigote cDNA library with a purified antibody raised against the Sm-binding site of a yeast sequence. A clone was obtained containing a 507 bp-insert with an ORF of 399 bp and coding for a protein of 133 amino acids. Sequence analysis revealed high identity with the L27 ribosomal proteins from different species including: Canis familiaris, Homo sapiens, Schizosaccharomyces pombe and Saccharomyces cerevisiae. This protein has not been previously described in the literature and seems to be a new ribosomal protein in T. cruzi and was given the code TcrL27. To express this recombinant T. cruzi L27 ribosomal protein in E. coli, the insert was subcloned into the pET32a vector and a 26 kDa recombinant protein was purified. Immunoblotting studies demonstrated that this purified recombinant protein was recognized by the same anti-Sm serum used in the library screening as well as by chagasic and systemic lupus erythemathosus (SLE) sera. Our results suggest that the T. cruzi L27 ribosomal protein may be involved in autoimmunity of Chagas disease.

TOXIC DILATATION OF THE COLON IN CHAGAS-DISEASE

Toxic megacolon occurs in colitis of differing aetiology. This report describes 15 patients with chagasic megacolon with this complication. The clinical signs and symptoms in all patients were pain and progressive abdominal distension accompanied by fever, severe toxaemia and shock. Seven patients developed this clinical pattern after manual removal of faeces. The remaining patients had pain and abdominal distension followed by signs of severe toxaemia when first examined. Nine patients underwent total colectomy with ileostomy (one death), four partial colectomy (all died) and two received medical treatment (both died). At autopsy, three of the four patients undergoing partial colectomy had residual colitis and enteritis. The surgical procedure of choice for this complication of chagasic megacolon is total colectomy.

Sympathetic hyperactivity, respiratory failure, pruritus, and anesthesia after unintentional epidural injection of potassium chloride: Case report

Background and Objectives. A combination of epidural and general anesthesia has been widely used to attenuate the surgical stress response and to provide postoperative analgesia. This case report illustrates the use of this anesthetic technique. Analgesia was induced with local anesthetic in the immediate postoperative period using unintentional 19.1% potassium chloride (KCI) as diluent. Methods. An ASA I male patient was scheduled for surgical correction of idiopathic megaesophagus under continuous epidural anesthesia combined with general anesthesia. In the postoperative period, while preparing 10 mt 0.125% bupivacaine to be administered through the epidural catheter for pain control, 5 mt 19.1% KCI was unintentionally used as diluent, resulting in a 9.55% potassium solution concentration. Results. The patient developed warmness of the lower limbs, tachycardia, hypertension, intense pruritus on the chest, agitation, exacerbation of sensory and motor blocks, and respiratory failure secondary to pulmonary edema, requiring ventilatory support. Total recovery was observed after 24 hours. Conclusions. Epidurally injected potassium leads to severe clinical manifestations caused by autonomic dysfunction, spinal cord irritation, and possible release of histamine. Despite continuous recommendations, ampule misidentification still happens in hospitals, frequently leading to serious accidents.

CLINICAL STUDY of NATURAL INFECTION BY Trypanosoma cruzi IN DOGS RESIDING IN A RURAL AREA IN MATO GROSSO do SUL STATE, BRAZIL

This study was carried out to describe the clinical characteristics of natural infection caused by Trypanosoma cruzi in dogs that reside in a rural area of Mato Grosso do Sul State, Brazil. Conventional and nonconventional diagnostic methods were used for screening T. cruzi infection in 75 dogs that lived in the area. Cardiovascular tests and biochemical examination of sera were also performed in four confirmed positive dogs. The following techniques were employed: indirect immunofluorescence test (IFAT), enzyme-linked immunosorbent assay with T. cruzi epimastigote antigens (EAE-ELISA) and enzyme-linked immunosorbent assay with T. cruzi excreted-secreted trypomastigote antigens (TESA-ELISA) with antibodies detected in 45.33% (n = 34), 24.0% (n = 18) and 12.0% (n = 9) of the dogs, respectively. The current prevalence of the infection was confirmed as 10.7% (n = 8) by immunoblotting test with T. cruzi excreted-secreted antigens (TESA-blot). The test that showed the best concordance index (Kappa; 0.93), sensitivity (100%) and specificity (98.5%) was TESA-ELISA, that when associated with IFAT had the same results as those obtained by TESA-blot (10.7%). Three out of the four chagasic animals showed enlarged cardiac silhouette on X-ray and an increase of the P-wave duration and QRS complex in electrocardiogram. Two dogs presented conduction disturbances, right bundle branch block in one dog and first-degree atrioventricular block and sinus arrest in another. The ecodopplercardiography presented left-ventricular-wall thickness increased during diastole, decrease of the shortening fraction and inversion in the speed peaks of the E and A waves, indicating the presence of systolic and diastolic disorders. The four animals showed enzymatic activities of creatine kinase (221-404 U/L), MB fraction of creatine kinase (189-304 U/L), elevated total proteins (7.6-10.2 g/dL) and total globulins (4.6-7.7g/dL) and reduction of albumin/globulin ratio, which suggested a myocardial injury and continuous antigenic stimulus.

Dilatacao pneumatica no megaesofago avancado. Resultados do sequimento tardio

Six patients with advanced Chagas' megaesophagus with poor condition for surgical indication due to severe malnutrition and concomitant diseases were submitted to pneumatic dilatation with the aid of the endoscope. The method consists in leading the tip of the pneumatic dilatator into the cardia with the tip of the endoscope. Insufflation of the balloon must be slow and progressive with visualization of the mucosa of the cardia after insufflating of 2 pounds of air pressure. An excellent symptomatic relief was seen in the five-year follow-up but without improvement in the degree of the esophageal dilatation.

Chagas disease and triatomine biology (Heteroptera, Triatominae), with emphasis on aspects of spermatogenesis

A century after the discovery of Chagas disease, it is still one of the most important parasitic diseases affecting humans. The subfamily Triatominae is important in medical health, because these insects are vectors of Trypanosoma cruzi, the etiologic agent of Chagas disease. These insects are also of important cytological relevance because they have particular cell characteristics, such as persistence of nucleolar material in spermatogenesis. The germ cells of the animal kingdom have chromatoid bodies (CBs) in their cytoplasm that can originate from nucleolar material that is fragmented in the early stages of spermatogenesis and plays an important role in cellular communication between the spermatids during spermiogenesis. Currently, there are few studies on the function and formation of the CB in nucleologenesis, especially with emphasis on the ultrastructure of the cells involved in spermatogenesis of insects. Considering the importance of knowledge about the triatomine fauna, we conducted a study of the biogeography and reports of these insects and a survey of patients with Chagas disease in the northwestern region of São Paulo State. Data collected from 1995 to 2009 indicated 700 individuals with Chagas disease, demonstrating a range of 0 to 40 years, which shows that the disease may be active in this region. Moreover, of the 1150 patients treated for cardiomyopathy, 44% were chagasic. Regarding the triatomines noted and captured in the period from 2004 to 2009, the species were Triatoma sordida and Rhodnius neglectus, with T. sordida being the most abundant. In addition, some triatomines were infected by T. cruzi in various developmental stages. We also analyzed the nucleolar cycle and fibrillarin nucleolar protein expression in CB of spermatogenic cells of T. infestans and T. sordida, using histological, ultrastructural and immunocytochemical techniques. The results revealed fibrillarin nucleolar protein expression in the nucleus and in some cytoplasmic spots of germ cells during spermatogenesis in triatomines. These data suggest that fibrillarin could be a constituent of CB, which was most likely derived from nucleolar fragmentation. This is the first time that fibrillarin protein expression has been shown in CB during spermatogenesis progression in triatomines. Knowledge about the biology of triatomines was deepened in this study and, in particular, the structural and ultrastructural aspects of spermatogenesis in triatomines. This study showed that the disease may be active in the northwestern region of São Paulo and expanded our knowledge of the biology of triatomines, the main vectors of Chagas disease. © FUNPEC-RP.

Biologic and genetics aspects of chagas disease at endemic areas

The etiologic agent of Chagas Disease is the Trypanosoma cruzi, transmitted through blood-sucking insect vectors of the Triatominae subfamily, representing one of the most serious public health concerns in Latin America. There are geographic variations in the prevalence of clinical forms and morbidity of Chagas disease, likely due to genetic variation of the T. cruzi and the host genetic and environmental features. Increasing evidence has supported that inflammatory cytokines and chemokines are responsible for the generation of the inflammatory infiltrate and tissue damage. Moreover, genetic polymorphisms, protein expression levels, and genomic imbalances are associated with disease progression. This paper discusses these key aspects. Large surveys were carried out in Brazil and served as baseline for definition of the control measures adopted. However, Chagas disease is still active, and aspects such as host-parasite interactions, genetic mechanisms of cellular interaction, genetic variability, and tropism need further investigations in the attempt to eradicate the disease. Copyright 2012 Marilanda Ferreira Bellini et al.

Avaliação clínica da cardiomiopatia chagásica crônica em cães

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação da técnica de hemocultura pela reação em cadeia da polimerase (PCR) utilizando os iniciadores P35/P36 e TCZ1/TCZ2 para a detecção de Trypanosoma cruzi em cães e gatos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo clínico da infecção natural por T. cruzi em cães residentes em uma área rural de Mato Grosso do Sul, Brasil

Pós-graduação em Medicina Veterinária - FCAV

Expressão de RNAm TLR 2, TLR 4 e citocinas em infecção experiemental por cepas Trypanosoma cruzi de diferentes origens

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Doença de chagas e biologia de triatomíneos (Heteroptera, Triatominae), com ênfase aos aspectos da espermatogênese

Pós-graduação em Genética - IBILCE

Epidemiological features of esophageal cancer. Squamous cell carcinoma versus adenocarcinoma

PURPOSE: To analyze the epidemiological features of patients with esophageal cancer according to the histopathological types: squamous cell carcinoma or adenocarcinoma. METHODS: A total of 100 patients with esophageal cancer, being 50 squamous cell carcinomas and 50 adenocarcinomas were analyzed for demographics, nutritional factors, lifestyle habits, benign pathological conditions associated, like Barrett's esophagus and megaesophagus, tumor stage and survival rates. The nutritional factors evaluated included body mass index, percent weight loss, hemoglobin and albumin serum levels. RESULTS: Esophageal cancer occurred more often in men over 50 years-old in both histological groups. No significant differences on age and gender were found between the histological groups. Squamous cell carcinoma was significantly more frequent in blacks than adenocarcinoma. Alcohol consumption and smoking were significantly associated with squamous cell carcinoma. Higher values of body mass index were seen in patients with adenocarcinoma. Barrett's esophagus was found in nine patients (18%) with adenocarcinoma, and megaesophagus in two patients (4%) with squamous cell carcinoma. The majority of patients were on stages III and IV in both histological groups. The mean survival rates were 7.7 ± 9.5 months for patients with squamous cell carcinoma and 8.0 ± 10.9 months for patients with adenocarcinoma. No significant differences on tumor stage and survival rates were detected between the histological groups. CONCLUSION: Epidemiological features are distinct for the histopathological types of esophageal cancer. Squamous cell carcinoma is associated with black race, alcohol and smoking, while adenocarcinoma is related to higher body mass index, white race and Barrett's esophagus.

Anticorpos naturais e infecção: anticorpos naturais na doença de Chagas e na paracoccidioidomicose

Using ELISA technique, natural antibodies against self and non self antigens were determined in 80 patients chronically intected by T. cruzi and 40 individuals suffering from a deep mycosis frequentely found in Latin Amarica (Paracoccidioidomycosis - PCM). Two forms of PCM were investigated: adult forms and juvenil type of disease. Eighty percent (80%) of the former group had significantly elevated anti-laminin antibody levels (M=4.7,SD±1.8) compared with healthy controls and different specificities of antibody were associated with anti-laminin in pathological sera. A notable binding to cytoskeletal proteins was observed, specially with band 3 and their peptides derivates, such as 62 kDa peptide. By means of Protein A chromatography we were able to show that natural anti-Gal antibodies may be bound by their Fab region to other immunoglobulins and/or to Protein A by alternative sites of binding. The finding of lgG anti-Gal antibodies in circulating immune complexes isolated from chagasic sera supported the first alternative. However, it is possible that some of lgG anti-Gal antibodies, belong to VH111 subgroup of immunoglobulins, that bind directly to Protein A. Among the 40 sera from PCM examined, the majority was considered as not exhibiting a signilicantly higher binding than normal sera to antigens tested. However thirty percent (30%) of the chronic patients had an increased levels of natural antibodies at least for one specificity such as actyn, myosin and Gala1,3Gal epitopes. ln juvenil type of PCM the mean value found for actyn was also increased 2,42 (range 1,0 to 5,3). Utilizing the polyethylene glicol precipitation the presence of circulating immune complexes was investigated in PCM sera. Specific antibodies for soluble antigens from P. brasiliensis and natural antibodies against myoglobin, myosin and Gala1,3 Gal epitopes were characterized

Modelos para dados grupados e sensurados aplicados à área biológica: comparação usando fator de Bayes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)