Intestinal cell migration damage induced by enteropathogenic Escherichia coli strains.

Epithelial cell migration is an essential response to enteric pathogens such as enteropathogenic Escherichia coli (EPEC). This study aimed to investigate the effects of EPEC infection on intestinal epithelial cell migration in vitro, as well as the involvement of type III secretion system (T3SS) and Rho GTPases. Crypt intestinal epithelial cells (IEC-6) were infected with EPEC strains (E2348/69, ΔescF, and the LDI001 strain isolated from a malnourished Brazilian child) and commensal E. coli HS. Wound migration and cell death assays were performed at different time-points. Transcription and expression of Rho GTPases were evaluated using real-time PCR and western blotting. Overall, EPEC E2348/69 reduced migration and increased apoptosis and necrosis levels compared to EPEC LDI001 and E. coli HS strains. Moreover, EPEC LDI001 impaired cell migration at a higher level than E. coli HS and increased necrosis after 24 hours compared to the control group. The different profiles of virulence genes between the two wild-type EPEC strains, characterized by the absence of espL and nleE genes in the LDI001, might explain the phenotypic results, playing significant roles on cell migration impairment and cell death-related events. Moreover, the type III secretion system is determinant for the inhibition of intestinal epithelial cell migration by EPEC 2348/69, as its deletion prevented the effect. Active Rac1 concentrations were increased in E2348/69 and LDI001-infected cells, while the T3SS-deficient strain did not demonstrate this activation. This study contributes with valuable insight to characterize the mechanisms involved in the impairment of intestinal cell migration induced by EPEC.

Intestinal cell migration damage induced by enteropathogenic Escherichia coli strains

Epithelial cell migration is an essential response to enteric pathogens such as enteropathogenic Escherichia coli (EPEC). This study aimed to investigate the effects of EPEC infection on intestinal epithelial cell migration in vitro, as well as the involvement of type III secretion system (T3SS) and Rho GTPases. Crypt intestinal epithelial cells (IEC-6) were infected with EPEC strains (E2348/69, ΔescF, and the LDI001 strain isolated from a malnourished Brazilian child) and commensal E. coli HS. Wound migration and cell death assays were performed at different time-points. Transcription and expression of Rho GTPases were evaluated using real-time PCR and western blotting. Overall, EPEC E2348/69 reduced migration and increased apoptosis and necrosis levels compared to EPEC LDI001 and E. coli HS strains. Moreover, EPEC LDI001 impaired cell migration at a higher level than E. coli HS and increased necrosis after 24 hours compared to the control group. The different profiles of virulence genes between the two wild-type EPEC strains, characterized by the absence of espL and nleE genes in the LDI001, might explain the phenotypic results, playing significant roles on cell migration impairment and cell death-related events. Moreover, the type III secretion system is determinant for the inhibition of intestinal epithelial cell migration by EPEC 2348/69, as its deletion prevented the effect. Active Rac1 concentrations were increased in E2348/69 and LDI001-infected cells, while the T3SS-deficient strain did not demonstrate this activation. This study contributes with valuable insight to characterize the mechanisms involved in the impairment of intestinal cell migration induced by EPEC.

[New aspects in reconstructive nasal surgery]. / Neue Aspekte bei der rekonstruktiven Nasenchirurgie.

Developments in surgical procedures have led to improved results in the field of reconstructive nasal surgery. In this article, the authors focus on the forehead flap technique in one of its smallest forms to cover three-layered alar defects. The example of a complete nasal reconstruction shows how, despite the use of a proven surgical concept, serious complications could occur. In the case shown, a satisfactory outcome was achieved by starting anew with the forehead flap in one of the largest forms-the expanded lateral forehead flap. The authors ascribe particular importance to preoperative analysis with standardized series of photographs. Such series should be made before each surgical step. These are indispensable for structured planning and, thus, are prerequisite for an excellent outcome.

Outcome after elective neck dissection and observation for the treatment of the clinically node-negative neck (cN0) in squamous cell carcinoma of the oropharynx.

Optimal elective neck treatment in node-negative (cN0) oropharyngeal squamous cell carcinoma (OPSCC) patients is still controversially discussed. Retrospective chart review of 49 cT1-3 cN0 cM0 OPSCC patients, who had undergone surgical resection of the primary and either elective neck dissection (END) (n = 32) or observation (OBS) (n = 17) of the neck was performed. For systematic review of literature, Pubmed and EMBASE were searched for clinical studies including data on both END and OBS of the neck in cN0 OPSCC patients. Estimated 5-year overall survival (OS) rate was 82 % for END and 76 % for OBS [hazard ratio (HR) = 1.01]. Estimated 5-year disease-free survival (DFS) rate was 78 % for END and 67 % for OBS (HR = 1.79); 5-year DSS rate was 97 % (END) and 81 % (OBS) (HR = 2.22). None of the primary outcome variables (OS, DFS, DSS) revealed statistically significant effects for the treatment assignments. Hazard ratios implied an advantage for END. Systematic review of literature yielded only retrospective chart reviews and no data meeting our selection criteria for further data analysis. Due to lack of high-level evidence, the decision for END in cN0 OPSCC remains a diagnostic and therapeutic challenge. The demonstrated clinical equipoise would provide a solid basis for a multicentric, randomized trial.

Quantificação de Mycobacterium leprae em amostras de secreção nasal de casos paucibacilares e multibacilares por PCR quantitativo

A quantificação do bacilo realizada pelo exame baciloscópico e histopatológico apresenta sensibilidade limitada. Portanto, o emprego de uso de técnicas moleculares permite o diagnóstico direto do material clínico com elevada especificidade e sensibilidade. A PCR em tempo real (qPCR) é um ensaio sensível e específico que permite a quantificação do número de bacilos a partir de diversas amostras, além de poder ser utilizada no diagnóstico diferencial de muitos patógenos. Pacientes multibacilares hansenianos liberam o bacilo Mycobacterium leprae através da secreção nasal, sendo a coleta deste realizada por procedimento não invasivo. Até o momento, nenhum estudo avaliou a sensibilidade e especificidade da qPCR para o diagnóstico da hanseníase utilizando amostras de secreção nasal.

Flow cytometric assay in peripheral blood of dogs--reference values for leukocytes from Brazilian beagles.

Use of domestic reference values in the flow cytometry analysis is known to improve its accuracy by integrating local variations as gender, race and age. Up to date application of flow cytometry in veterinary medicine has been limited to describe the percentual values just for peripheral lymphocytes subsets of blood. We now report establishment of reference values for a wide range of proportional and absolute numbers of peripheral blood leukocytes, including T cells subsets, B cells, monocytes and eosinophils, applicable to the healthy population of Beagles in Brazil and other regions with similar demographic characteristics. Normal reference values were also established to estimate the gender-related differences. This information will provide clinical aid in the evaluation of immunologic status as well as standard values for experimental animals of dogs from Brazil and other similar regions.

Ultrasound-guided piezoelectric extracorporeal shock wave lithotripsy of parotid gland calculi.

PURPOSE: The introduction of piezoelectric extracorporeal shock wave lithotripsy (ESWL) has changed therapy for salivary calculi. This method seems especially suitable for treating calculi in the parotid gland. The purpose of this study was to evaluate ESWL in patients with such calculi. METHODS: From November 1990 to November 1999, all patients with sialolithiasis of the parotid gland were treated with piezoelectric ESWL. Three different lithotriptors were used over the 9-year study period. Results were analyzed according to both the patients' clinical status and follow-up sonograms. RESULTS: In total, 42 patients (21 women, 21 men; mean age, 59 years) were treated with ESWL. The mean follow-up period for all patients was 63 months (range, 7-96 months). After ESWL had been performed, 71% of the patients were completely free of symptoms, and 21% had marked improvement of their symptoms. Sixty-seven percent were completely free of calculi, and 27% had a marked reduction in the size of their calculi. Adverse effects of ESWL included temporary glandular swelling (4 patients), blood-tinged salivary secretions (9 patients), petechiae on the skin surface (3 patients), and parotid abscess (1 patient). CONCLUSIONS: ESWL is an outpatient procedure that can be performed without anesthesia and with scarcely any discomfort for patients. Conventional surgical procedures such as subtotal parotidectomy may be almost entirely replaced by ESWL because of the excellent treatment results and a very low rate of complications associated with ESWL. ESWL should be considered the treatment of choice for parotid calculi.

[B-image sonography in lymph node staging of the head and neck area. A comparison with palpation, computerized and magnetic resonance tomography]. / Die B-Bildsonographie beim Lymphknotenstaging im Kopf-Hals-Bereich. Ein Vergleich mit der Palpation, Computer- und Magnetresonanztomographie.

The results of palpation, CT and MRI and high resolution ultrasound were compared in 100 patients with malignancy of the head and neck. Ultrasound detected far more lymph nodes than the other methods. These diagnostic findings were compared with the operative and histological results in 62 patients. Ultrasound proved superior to the other methods: a lymph node metastasis was missed in only 2 patients by sonography, whereas in 20 patients metastases were overlooked by CT and MRI scans, and in 27 patients by palpation. High resolution ultrasound is at present the most reliable method for the detection of lymph nodes in the head and neck.