A VP3/VP1 gene polymerase chain reaction assay for detection of chicken anemia virus in broiler samples

Desenvolveu-se uma reação em cadeia de polimerase (PCR) para amplificação do altamente conservado gene VP3 e da região 5 do gene VP1, para o diagnóstico do vírus da anemia das galinhas (CAV), diretamente em amostras de campo de órgãos de frangos de corte com suspeita clínica da doença. A comparação entre o PCR VP3/VP1 com isolamento viral in vivo indicou 100 de concordância dos resultados, com 13 amostras positivas e três negativas em ambos os testes. Órgãos de outros 24 lotes de frangos com lesões e história clínica compatível com CAV foram testados com o PCR VP3/VP1 e com um PCR de referência com primers conhecidos para o gene VP1. Dezenove amostras resultaram positivas e uma negativa em ambos os PCR e quatro foram positivas apenas no PCR VP3/VP1. Estes resultados indicam que o PCR VP3/VP1 é um teste de diagnóstico sensível e específico, aplicável como alternativa ao método caro e demorado de isolamento viral in vivo especialmente considerando-se amostras do CAV não adaptáveis a cultivos de células MSB-1.

Detection of chicken anemia virus in the gonads and in the progeny of broiler breeder hens with high neutralizing antibody titers.

Previous evidence for the presence of chicken anemia virus (CAV) in the gonads of immune specific-pathogen-free chickens raised the question whether this occurs also in commercial breeders. The presence of CAV was investigated by nested PCR in the gonads and spleens of hens from two 55- and 59-week-old, CAV-vaccinated (flocks 2 and 3), and two 48- and 31-week-old non-vaccinated broiler breeder flocks (flocks 1 and 4). In addition, lymphoid tissues of 20-day-old embryos from these hens were also investigated for the presence of CAV. CAV was detected in the gonads and of 5/6 and 11/22 of the vaccinated hens and in some hens also in the spleen alone. Embryos from 7/8 and 5/18 of these hens were positive. In the non-vaccinated flocks, CAV was detected in the gonads of 11/34 and 10/10 hens in flocks 1 and 4, respectively. In addition, 11 birds in flock 1 had positive spleens. CAV DNA was detected in 3/11 and 2/10 of their embryos. CAV-positive gonads and embryos were detected in samples from hens with moderate as well as high VN antibody titers. Vaccinated chickens positive for CAV in the gonads and in their embryos had VN titers ranging from >1:512 to <1:2048. In non-vaccinated chickens, the VN titers of CAV positive chickens ranged from 1:128 to 1:4096. These results demonstrate that CAV genome can remain present in the gonads of hens in commercial broiler breeder flocks even in the presence of high neutralizing antibody titers that have been associated with protection against CAV vertical transmission. It also suggests that transmission to the progeny may occur irrespectively of the level of the humoral immune response in the hens.

Survival and prognostic factors in patients with resected epidermoid oesophageal carcinoma.

To determine long-term survival of patients with thoracic oesophageal tumor who underwent resection, and to identify possible associated prognostic factors, 58 patients underwent oesophagectomy alone (group A) and 16 combined with neo-adjuvant chemoradiotherapy (group B). Univariate and multivariate analysis of prognostic factors were performed for age, depth of oesophageal wall tumour penetration, node involvement, type of resection, TNM stage and degree of tumour differentiation. Long-term survival rates at 1-5 years were 81% versus 89%; 56% versus 67%; 30% versus 67%; 12% versus 44%; and 0% versus 33% for group A and B, respectively (P = 0.0543, NS). Univariate analysis revealed only depth of invasion (P = 0.0076) and TNM stage (P = 0.0452) as isolated prognostic factors for long-term survival and multivariate analysis did not demonstrate any independent factor. Despite the small number of cases, neoadjuvant chemoradiotherapy seems to improve prognosis as well as to allow resection in a greater number of cases due to tumor downstaging.

Alternative surgical treatment for complex enterocutaneous fistula.

A retrospective analysis on the clinical-surgical handling of patients with enterocutaneous fistula (ECF) was performed, where an alternative surgical technique was discussed: intestinal bypass. Fistula with draining over 500 ml/24 h, which were present in 13 patients, were classified as high debit. We defined as complex, the fistula with multiple orifices, high defect of the abdominal wall or through the mesh. The population studied consisted of 25 patients, 11 male, in a total of 34 ECF and mean age of 41.9 years. At clinical treatment with TPN for high debit ECF, 2 patients (16.6%) were cured, another 2 died and 8 (66.8%) needed surgical treatment. The surgery cured 7 patients (77.7%) with high debit ECF but 2 (22.3%) died. In the patients with low debit ECF, TPN cured 2 patients (40%) but failed in another 3 (60%). All patients with low debit ECF resolved with surgical treatment.

Efficacy of computed axial tomography in the evaluation of the involvement of the respiratory tract in patients with squamous cell carcinoma of esophagus.

Squamous cell carcinoma of the esophagus is endemic in Rio Grande do Sul, the most southern state of Brazil, where the incidence reaches 27 out of 100,000 inhabitants per year. Palliative treatments, surgical or nonsurgical, impose lower mortality and lower costs. The choice of the best treatment for each patient depends on the neoplasia staging, and the local involvement of the tracheobronchial tree by tumor characterizes incurability. Computed axial tomography (CAT) is indicated, but remarkable contradictions were found in the literature. We studied 134 patients, diagnosed with esophageal squamous cell carcinoma, who underwent computed tomography and fiberoptic bronchoscopy (gold standard) performed with an uniform technique. An accuracy of 85.1% was obtained when computed tomography was compared with bronchoscopy with regard to impingement, displacement and invasion of trachea and bronchi. In conclusion, there is no statistically significant difference between computed tomography and fiberoptic bronchoscopy in the evaluation of tracheobronchial involvement by squamous cell carcinoma of the esophagus (p < 0.05).

Laparoscopic resection of a mesenteric cyst.

Mesenteric cysts are rare intra-abdominal tumors with an incidence around one case per 100,000 hospital admissions. The clinical presentation is variable; patients may be asymptomatic or present with either acute or chronic abdominal pain. Physical examination commonly demonstrates a smooth, round and mobile abdominal mass. Differential diagnosis includes any abdominal cyst or tumor. Laboratory tests are usually helpless. Ultrasonography and CT scans are the best diagnostic tools. The treatment of choice is the total resection of the cyst, which is regularly performed by open surgery. This paper reports a case of a mesenteric cyst successfully resected by laparoscopy, and addresses the possible uses of this approach.

The reovirus protein mu2, encoded by the M1 gene, is an RNA-binding protein.

The reovirus M1, L1, and L2 genes encode proteins found at each vertex of the viral core and are likely to form a structural unit involved in RNA synthesis. Genetic analyses have implicated the M1 gene in viral RNA synthesis and core nucleoside triphosphatase activity, but there have been no direct biochemical studies of mu2 function. Here, we expressed mu2 in vitro and assessed its RNA-binding activity. The expressed mu2 binds both poly(I-C)- and poly(U)-Sepharose, and binding activity is greater in Mn2+ than in Mg2+. Heterologous RNA competes for mu2 binding to reovirus RNA transcripts as effectively as homologous reovirus RNA does, providing no evidence for sequence-specific RNA binding by mu2. Protein mu2 is now the sixth reovirus protein demonstrated to have RNA-binding activity.

Neoplasias associadas ao carcinoma epidermóide do esôfago / Neoplasms associated to esophageal epidermoid carcinoma

INTRODUÇÄO. O carcinoma epidermóide de esôfago (CEE) tem uma importante associaçäo com neoplasias do trato aerodigestivo e, provavelmente, compartilham dos mesmos fatores de risco. Além destes, outras neoplasias podem estar associadas com o carcinoma de esôfago. OBJETIVO. Analisar, retrospectivamente, pacientes com carcinoma epidermóide de esôfago tratados pelo Grupo de Cirurgia do Esôfago, Estômago e Intestino Delgado (GCEEID) do Hospital de Clínicas de Porto Alegre (HCPA), no período de janeiro/88 a junho/95, os quais tinham neoplasias associadas ao CEE. PACIENTES E MÉTODOS. Dentre os 261 pacientes estudados, 19 (7,28 por cento) tinham neoplasia associada ao CEE. Dez pacientes apresentaram tumores sincrônicos e 9, metacrônicos. O sexo predominante foi o masculino, com 17 casos. A média de idade ficou em 62,52 anos no momento do diagnóstico da neoplasia esofágica. RESULTADOS. Os tumores aerodigestivos, na sua totalidade carcinomas escamosos, representaram o tipo histológico predominante da neoplasia associada em 68,42 por cento dos casos. O sítio mais freqüente da neoplasia aerodigestiva associada foi a árvore respiratória (53,8 por cento), seguido da cavidade oral e orofaringe (23 por cento) e laringe (23 por cento). Dos 19 pacientes, 12 eram tabagistas e nove ingeriam bebidas alcoólicas regularmente. Para o tratamento do CEE, optou-se por cirurgia em seis pacientes. A neoplasia associada foi tratada com cirurgia radical em 11 pacientes e radioterapia em cinco. Surpreendentemente, foram diagnosticados quatro casos (21 por cento) de adenocarcinomas gßstricos associados ao CEE, tratados com cirurgia radical em três pacientes. CONCLUSÄO. Os autores ressaltam a importância do estadiamento criterioso dos pacientes com CEE devido a associaçäo significativa com outras neoplasias, principalmente com tumores aerodigestivos. Alertam para o seguimento desses pacientes e discutem a possibilidade de fatores de risco comuns: fumo e álcool. Nesta casuística, encontrou-se associaçäo importante com neoplasias gástricas.

Delayed cervical anastomosis of the esophagus for esophageal carcinoma.

BACKGROUND: The surgical approach for patients with advanced epidermoid esophageal carcinoma should provide an effective palliative effect with morbidity ratio as low as possible. Anastomotic leakage is a frequent complication and may be responsible for both early and late morbidity and, therefore, we assessed the role of delayed cervical esophagovisceral anastomosis technique in relation to the incidence of anastomotic complications. METHODS: Eight patients (Group 1) and 12 patients (Group 2) submitted to one-stage or two-stage operation, respectively, were selected by an intraoperative assessment by the surgeon, considering mainly tissue blood flow of the replacement organ after its placement in the cervical region. RESULTS: In Group 1 anastomotic dehiscence was observed in 37.5% of patients, while in Group 2 no cases of dehiscence occurred (p = 0.049). However, the postoperative mortality rate did not differ between the two groups (12.5% versus 0%, NS). CONCLUSION: When organ viability is uncertain, esophagovisceral anastomosis is best done by two-stage operation, since it decreases the incidence of anastomotic leak.

[Neoplasms associated with esophageal epidermoid carcinoma]. / Neoplasias associadas ao carcinoma epidermóide do esôfago.

INTRODUCTION: The esophageal epidermoid cancer has an important association with aerodigestive tract neoplasms and possibly share the same risk factors. Furthermore, other neoplasms can be associated with esophagus cancer. OBJECTIVE: To analyze retrospectively the patients with esophageal epidermoid cancer (EEC) and associated neoplasms, treated by the Esophagus Stomach and Small Intestine Group of Surgery at Hospital de Clinicas de Porto Alegre from January 1988 to June 1995. PATIENTS AND METHODS: Nineteen (7.28%) of the 261 studied patients had associated neoplasms to the EEC. Ten patients presented synchronic tumours and 9 metachronic ones. The predominant sex was the masculine with 17 cases. The mean age was 62.52 years in the moment of the esophageal cancer diagnostic. RESULTS: The aerodigestive tumours, squamous carcinomas in totality, represented the predominant associated neoplasm histological type in 68.42% of the cases. The most frequent associated aerodigestive tumours site was the respiratory tract (53.8%), followed by the oral cavity and oropharynx (23%) and larynx (23%). In our sample, twelve patients were smokers and 9 were alcohol abusers. In relation to the EEC treatment, surgery was performed in 6 patients. The associated neoplasm was treated with radical surgery in 11 patients and radiotherapy in 5. Surprisingly 4 cases (21%) of gastric adenocarcinoma associated to the EEC were diagnosed, treated with radical surgery in 3 patients. CONCLUSION: The authors call attention to the importance of a criterial staging as well as the follow up in patients with EEC owing to the significant association with others neoplasms, principally with aerodigestive tumours, and discuss the common risk factors possibility: tobacco and alcohol use. Important association with gastric neoplasms were found in this casuistry.

Isolation and identification of chicken infectious anemia virus in Brazil.

Seven chicken infectious anemia virus (CIAV) isolates were obtained from seven broiler flocks with poor performance in two states of Brazil. All isolates induced thymus atrophy, bone-marrow aplasia, and low hematocrit values when inoculated into 1-day-old susceptible chicks. The CIAV isolates were resistant to treatment with chloroform and were able to pass through 50-nm-pore-size filters. CIAV-specific antigens could be demonstrated in tissues of experimentally infected chicks using a monoclonal antibody specific for CIAV. These characteristics of the virus and the virus-induced lesions demonstrate that CIAV is present in Brazil and that the virus is associated with production problems.

Experimental vaccination of rabbits with non-pathogenic strains of Aujeszky's disease virus.

1. The Bartha K and NIA-4 strains of Aujeszky's disease virus (ADV) were non-pathogenic for rabbits vaccinated once or twice by nasal instillation or intramuscular injection. Neutralizing antibodies were detected in 68% of the rabbits two weeks after primary vaccination and in all rabbits at challenge. 2. Challenge doses of virulent ADV greater than 10(5.0) median tissue culture infective doses (TCID50) resulted in the death of most vaccinated and all unvaccinated rabbits with typical signs of Aujeszky's disease within 4 days. ADV was recovered from brain and lung suspensions of vaccinated and unvaccinated rabbits who had died as a result of the challenge. 3. When the challenge dose was reduced to approximately 10(3.0) TCID50, rabbits vaccinated twice survived while all unvaccinated controls died within 3 days.

Experimental vaccination of rabbits with non-pathogenic strains of Aujeszky's disease virus

1. The Bartha K and NIA-4 strains of Aujeszky's disease virus (ADV) were non-pathogenic for rabbits vaccinated once or twice by nasal instillation or intramuscular injection. Neutralizing antibodies were detected in 68% of the rabbits two weeks after primary vaccination and in al rabbits at challenge. 2. Challenge doses of virulent ADV greater than 10**5.0 median tissue culture infective doses (TCID50) resulted in the death of most vaccinated and all unvaccinated rabbits with typical signs of Aujeszky's disease withing 4 days. ADV was recovered from braim and lung suspensions of vaccinated and unvaccinated rabbits who had died as a result of the challenge. 3. When the challenge dose was reduced to approximately 10**3.0 TCID50, rabbits vaccinated twice survived while al unvaccinated controls died within 3 days