Angiotensin II-induced JNK activation is mediated by NAD(P)H oxidase in isolated rat pancreatic islets.

Angiotensin II (AII), the active component of the renin angiotensin system (RAS), plays a vital role in the regulation of physiological processes of the cardiovascular system, but also has autocrine and paracrine actions in various tissues and organs. Many studies have shown the existence of RAS in the pancreas of humans and rodents. The aim of this study was to evaluate potential signaling pathways mediated by AII in isolated pancreatic islets of rats. Phosphorylation of MAPKs (ERK1/2, JNK and p38MAPK), and the interaction between proteins JAK/STAT were evaluated. AII increased JAK2/STAT1 (42%) and JAK2/STAT3 (100%) interaction without altering the total content of JAK2. Analyzing the activation of MAPKs (ERK1/2, JNK and p38MAPK) in isolated pancreatic islets from rats we observed that AII rapidly (3 min) promoted a significant increase in the phosphorylation degree of these proteins after incubation with the hormone. Curiously JNK protein phosphorylation was inhibited by DPI, suggesting the involvement of NAD(P)H oxidase in the activation of protein.

Allogeneic hematopoietic stem cell transplantation from an alternative stem cell source in Fanconi anemia patients: analysis of 47 patients from a single institution.

We transplanted 47 patients with Fanconi anemia using an alternative source of hematopoietic cells. The patients were assigned to the following groups: group 1, unrelated bone marrow (N = 15); group 2, unrelated cord blood (N = 17), and group 3, related non-sibling bone marrow (N = 15). Twenty-four patients (51%) had complete engraftment, which was not influenced by gender (P = 0.87), age (P = 0.45), dose of cyclophosphamide (P = 0.80), nucleated cell dose infused (P = 0.60), or use of anti-T serotherapy (P = 0.20). Favorable factors for superior engraftment were full HLA compatibility (independent of the source of cells; P = 0.007) and use of a fludarabine-based conditioning regimen (P = 0.046). Unfavorable factors were > or = 25 transfusions pre-transplant (P = 0.011) and degree of HLA disparity (P = 0.007). Intensity of mucositis (P = 0.50) and use of androgen prior to transplant had no influence on survival (P = 0.80). Acute graft-versus-host disease (GVHD) grade II-IV and chronic GVHD were diagnosed in 47 and 23% of available patients, respectively, and infections prevailed as the main cause of death, associated or not with GVHD. Eighteen patients are alive, the Kaplan-Meyer overall survival is 38% at approximately 8 years, and the best results were obtained with related non-sibling bone marrow patients. Three recommendations emerged from the present study: fludarabine as part of conditioning, transplant in patients with < 25 transfusions and avoidance of HLA disparity. In addition, an extended family search (even when consanguinity is not present) seeking for a related non-sibling donor is highly recommended.

Allogeneic hematopoietic stem cell transplantation from an alternative stem cell source in Fanconi anemia patients: analysis of 47 patients from a single institution

We transplanted 47 patients with Fanconi anemia using an alternative source of hematopoietic cells. The patients were assigned to the following groups: group 1, unrelated bone marrow (N = 15); group 2, unrelated cord blood (N = 17), and group 3, related non-sibling bone marrow (N = 15). Twenty-four patients (51 percent) had complete engraftment, which was not influenced by gender (P = 0.87), age (P = 0.45), dose of cyclophosphamide (P = 0.80), nucleated cell dose infused (P = 0.60), or use of anti-T serotherapy (P = 0.20). Favorable factors for superior engraftment were full HLA compatibility (independent of the source of cells; P = 0.007) and use of a fludarabine-based conditioning regimen (P = 0.046). Unfavorable factors were > or = 25 transfusions pre-transplant (P = 0.011) and degree of HLA disparity (P = 0.007). Intensity of mucositis (P = 0.50) and use of androgen prior to transplant had no influence on survival (P = 0.80). Acute graft-versus-host disease (GVHD) grade II-IV and chronic GVHD were diagnosed in 47 and 23 percent of available patients, respectively, and infections prevailed as the main cause of death, associated or not with GVHD. Eighteen patients are alive, the Kaplan-Meyer overall survival is 38 percent at ~8 years, and the best results were obtained with related non-sibling bone marrow patients. Three recommendations emerged from the present study: fludarabine as part of conditioning, transplant in patients with <25 transfusions and avoidance of HLA disparity. In addition, an extended family search (even when consanguinity is not present) seeking for a related non-sibling donor is highly recommended.

Induction of apoptosis in monocytes by Mycobacterium leprae in vitro: a possible role for tumour necrosis factor-alpha.

A diverse range of infectious organisms, including mycobacteria, have been reported to induce cell death in vivo and in vitro. Although morphological features of apoptosis have been identified in leprosy lesions, it has not yet been determined whether Mycobacterium leprae modulates programmed cell death. For that purpose, peripheral blood mononuclear cells obtained from leprosy patients were stimulated with different concentrations of this pathogen. Following analysis by flow cytometry on 7AAD/CD14+ cells, it was observed that M. leprae induced apoptosis of monocyte-derived macrophages in a dose-dependent manner in both leprosy patients and healthy individuals, but still with lower efficiency as compared to M. tuberculosis. Expression of tumour necrosis factor-alpha (TNF-alpha), Bax-alpha, Bak mRNA and TNF-alpha protein was also detected in these cultures; in addition, an enhancement in the rate of apoptotic cells (and of TNF-alpha release) was noted when interferon-gamma was added to the wells. On the other hand, incubation of the cells with pentoxifylline impaired mycobacterium-induced cell death, the secretion of TNF-alpha, and gene expression in vitro. In addition, diminished bacterial entry decreased both TNF-alpha levels and the death of CD14+ cells, albeit to a different extent. When investigating leprosy reactions, an enhanced rate of spontaneous apoptosis was detected as compared to the unreactive lepromatous patients. The results demonstrated that M. leprae can lead to apoptosis of macrophages through a mechanism that could be at least partially related to the expression of pro-apoptotic members of the Bcl-2 protein family and of TNF-alpha. Moreover, while phagocytosis may be necessary, it seems not to be crucial to the induction of cell death by the mycobacteria.

Induction of apoptosis in monocytes by Mycobacterium leprae in vitro: a possible role for tumour necrosis factor-alpha

A diverse range of infectious organisms, including mycobacteria, have been reported to induce cell death in vivo and in vitro. Although morphological features of apoptosis have been identified in leprosy lesions, it has not yet been determined whether Mycobacterium leprae modulates programmed cell death. For that purpose, peripheral blood mononuclear cells obtained from leprosy patients were stimulated with different concentrations of this pathogen. Following analysis by flow cytometry on 7AAD/CD14+ cells, it was observed that M. leprae induced apoptosis of monocyte-derived macrophages in a dose-dependent manner in both leprosy patients and healthy individuals, but still with lower efficiency as compared to M. tuberculosis. Expression of tumour necrosis factor-alpha (TNF-alpha), Bax-alpha, Bak mRNA and TNF-alpha protein was also detected in these cultures; in addition, an enhancement in the rate of apoptotic cells (and of TNF-alpha release) was noted when interferon-gamma was added to the wells. On the other hand, incubation of the cells with pentoxifylline impaired mycobacterium-induced cell death, the secretion of TNF-alpha, and gene expression in vitro. In addition, diminished bacterial entry decreased both TNF-alpha levels and the death of CD14+ cells, albeit to a different extent. When investigating leprosy reactions, an enhanced rate of spontaneous apoptosis was detected as compared to the unreactive lepromatous patients. The results demonstrated that M. leprae can lead to apoptosis of macrophages through a mechanism that could be at least partially related to the expression of pro-apoptotic members of the Bcl-2 protein family and of TNF-alpha. Moreover, while phagocytosis may be necessary, it seems not to be crucial to the induction of cell death by the mycobacteria.

Management of erythema nodosum leprosum by thalidomide: thalidomide analogues inhibit M. leprae-induced TNFalpha production in vitro.

Thalidomide is being successfully used for the treatment of erythema nodosum leprosum (ENL), among other disorders with inflammatory and immunological bases. Although the active molecules responsible for the diverse therapeutic activities of the drug and the sequence of reactions triggered inside the cells remain unclear, it was demonstrated that thalidomide (THAL) inhibits TNFalpha mRNA expression and protein production by stimulated monocytes and activated T lymphocytes. Patients treated with THAL experienced a reduction in serum TNFalpha levels and it diminished cytokine gene expression at the lesion site, with a concomitant abrogation of clinical symptoms. It has been reported that thalidomide as well as some its analogues decrease M. leprae-induced TNFalpha and IL-12 mRNA in vitro. THAL also reduced monocyte apoptosis in the cultures. The present data further support thalidomide's effects on TNFa synthesis and the growing need to search for new specific TNFalpha inhibitors (non-teratogenic compounds) that might be potentially used in clinical disorders such as leprosy.

Neck lymph node metastases to the posterior triangle apex: evaluation of clinical and histopathological risk factors.

BACKGROUND: Dissection of posterior triangle apex (APEX) is a surgical step in supraomohyoid and lateral neck dissections. The prevalence of lymphatic metastases at this site and the clinicohistopathologic conditions that influence their occurrence have not been established. We have evaluated the prevalence and the risk factors for cervical metastases in lymph nodes of the APEX. METHODS: Sixty-two neck dissections were performed in 51 patients with squamous cell carcinoma of the oropharynx, hypopharynx, oral cavity, glottic larynx, and supraglottic larynx or with primary occult tumor. We correlated the presence of positive metastases in the APEX with the neck level involved either clinically (CLIN) or histopathologically (H/P) and with the number of CLIN- or H/P-positive neck levels with metastases. The prevalence of metastases in the APEX in elective (N0) and therapeutic (N+) neck dissections was also compared. This prevalence was also compared with that for each neck level. The histopathologic comparisons between the APEX and the neck levels were calculated for N0, N+, and all neck dissections. The primary site of tumor was correlated with the presence of H/P-positive nodes in the APEX. RESULTS: The overall prevalence of lymphatic metastases in the APEX was 6.5%. The prevalence in N0 neck dissections was 2.3% and in N+ neck dissections it was 16.7%. The prevalence of lymphatic metastases in the APEX for primary tumors of pharynx was 23.1%, for the oral cavity it was 3.6%, and it was 0% for other sites. Metastases in the APEX were not influenced by the neck level with CLIN or H/P metastases in N+ necks. The number of CLIN- or H/P-positive neck levels had no influence on histopathologic metastases in the APEX. Factors that influenced metastases in the APEX were positive histopathologic metastases at level II for N0 neck dissections and positive histopathologic metastases at level II or III for all neck dissections. All the comparisons were analyzed using Fisher's or Poisson's test. CONCLUSIONS: The prevalence of histopathologic metastases in the APEX in N+ necks is 7.3 times greater than that of N0 necks and for primary tumors of pharynx it was 6.4 times greater than for the oral cavity and significantly greater than for the larynx. Histopathologic metastases at level II for clinically N0 necks and histopathologic metastases to level II or III for all neck dissections are risk factors for metastases in the APEX. The number of positive levels did not influence the prevalence of metastases in the APEX. There are no isolated metastases in the APEX of the posterior triangle.

[Intrauterine and perinatal mortality: comparative analysis of 3,904 necropsies, Hospital de Clínicas, Curitiba, from 1960 to 1995] / Mortalidade intra-uterina e perinatal: análise comparativa de 3.904 necropsias do Hospital de Clínicas de Curitiba no período de 1960 a 1995.

OBJECTIVE: To analyze the influence of the perinatal, fetal or maternal pathological processes in the induction of intra or extrauterine death as a guideline to a better perinatal medical assistance.METHODS: The authors studied retrospectively 3,094 consecutive necropsy cases carried out in stillborn fetuses (NM) and newborns (ON) in the period between 1960 and 1995 in the SAP of HC, Curitiba. The data analyzed included sex, time of gestation and causes of death of fetus and newborns. All maternal conditions that might have contributed to intra or extrauterine fetal death were also studied.RESULTS: The prevalence of intrauterine death due to maternal diseases was two times higher than extrauterine. In contrast, the fetal diseases were responsible mainly for extrauterine deaths. Primary placental diseases were responsible for 30% of the deaths with low time of gestation and 40% of them in the end of gestation. The same group of diseases was responsible for only 15-9% of the extrauterine deaths. Diseases of labor contributed to 12-20% of deaths after birth while only 7 to 17% of intrauterine deaths.CONCLUSION: Necropsy studies contribute to the identification of pathologic processes which affect the patients of any medical center. If a pregnancy is associated with a certain disease, the epidemiologic data of this study can help to identify the period of major risk of death of the fetus or newborn.

Panorama global da bacteriologia das amígdalas palatinas de crianças / Children palatine tonsils bacteriology: a comprehensive view

A microbiologia das amígdalas continua sendo assunto de muita discussäo. Este trabalho investiga a flora amigdaliana abrangendo-a em praticamente todas as situaçöes patogênicas e também normais. Foram colhidos swabs da superfície de amígdalas de 132 crianças normais (sem AR) em meses quentes (Q> (61 crianças) e frios (F) (71 crianças), de 64 crianças na fase aguda de amigdalite (AA) e de 76 com amigdalite recorrente (com AR, fora da fase aguda). Neste último grupo ainda foi realizado o estudo do core (interior) das amígdalas. Imediatamente após a coleta, o material era introduzido em meio específico e a cultura era realizada no máximo em 8 horas, seguindo padröes da microbiologia clínica. Como resultados, obtivemos uma alta prevalência de Streptococcus viridans nos três grupos. Entre o grupo Q e F foi notado que realmente existem diferenças sazonais, sendo Streptococcus viridans, Neisseria sp e enterobactérias mais freqüentes no grupo F. Comparando-se o grupo com AR e o sem AR vimos que Neisseria sp e enterobactérias säo mais freqüentes no sem AR. No grupo AA, Neisseria sp foi mais freqüente que no grupo normal (sem AR). Este trabalho é importante, pois traz a microbiologia das amígdaIas estudada de forma ampla, melhorando assim a interpretaçäo dos swabs e direcionando a um correto tratamento das afecçöes amigdalianas

Angiotensin-converting enzyme inhibitor increases insulin-induced pp185 phosphorylation in liver and muscle of obese rats.

The clinical use of angiotensin-converting enzyme (ACE) inhibitors has been associated with increased insulin sensitivity. However, the molecular mechanism is unknown. The authors examined the early steps in insulin action, i.e., the phosphorylation status of the insulin receptor and of the pp185 in liver and muscle of obese rats treated acutely with captopril, using immunoblotting with antiphosphotyrosine antibodies. Following treatment with captopril there was an improvement in insulin-induced insulin receptor and pp185 phosphorylation in the liver and muscle of obese rats. This finding contribute to an explanation of the mechanism by which ACE inhibitors appear to improve insulin sensitivity.

Comparative bacteriology of the surface of normal and pathological palatine tonsils in children.

There are only few studies on the normal bacteriology of tonsils. The purpose of this study was to acquire knowledge about the normal microflora: patients without recurrent tonsillitis (RT) and without tonsil hypertrophy (TH) and to compare these results with the pathological microflora: patients who have recurrent tonsillitis and/or tonsillar hypertrophy. We did 132 cultures of tonsil surface obtained from normal children and 96 cultures from pathological surfaces during the summer and in the winter. Comparing normal and pathological groups, we found Neisseria spp and Enterobacteria spp more frequently in the normal group. There are differences in the surface microflora of tonsils from normal persons and from individuals with tonsillitis.

Effect of glucagon on insulin receptor substrate-1 (IRS-1) phosphorylation and association with phosphatidylinositol 3-kinase (PI 3-kinase).

In the present study we have examined the levels and phosphorylation state of the insulin receptor and insulin receptor substrate 1 (IRS-1) as well as the association between IRS-1 and phosphatidylinositol 3-kinase (PI 3-kinase) in the liver and muscle of rats treated with glucagon. There was a decrease in the insulin-stimulated receptor and IRS-1 phosphorylation levels which was paralleled by a reduced association between IRS-1 and PI 3-kinase in vivo in the liver and muscle of glucagon-treated rats. These observations suggest that glucagon, probably acting through cAMP, may impair insulin signaling in the three early steps in insulin action after binding.

Modulation of early steps in insulin action in the liver and muscle of epinephrine treated rats.

Epinephrine is known to produce insulin resistance, but the exact molecular mechanism involved is unknown. In the present study we have examined the levels and phosphorylation state of the insulin receptor and of insulin receptor substrate 1 (IRS-1), as well as the association between IRS-1 and phosphatidylinositol 3-kinase (PI 3-kinase) in the liver and muscle of rats treated with epinephrine. The results demonstrate a decrease in insulin-stimulated receptor and IRS-1 phosphorylation levels which was accompanied by a reduction in the association of IRS-1 with PI 3-kinasein vivo in liver and muscle of epinephrine treated rats. These data suggest that molecular post-receptor defects may explain some aspects of the insulin resistance induced by catecholamines.

[Confusional state and infantile migraine: report of a case]. / Estado confusional e enxaqueca infantil: relato de um caso.

The case of a male patient having had episodes of confusion and motor deficits at age 5, 9 and 11, admitted to the hospital on these three occasions is reported. All investigations showed negative results, except for the EEG. A clue for the clinical diagnosis of confusional state due to migraine was the past history of the patient, who suffered several migranous attacks from the age of 9. A review of the pathophysiology is made, based on the available literature.