Survival and prognostic factors of patients treated for Stage I to Stage III endometrial carcinoma in a reference cancer center in Southern Brazil.

PURPOSE: To describe two- and five-year survival of patients with Stage I to III endometrial carcinoma and to identify prognostic factors. STUDY DESIGN: Concurrent cohort study. PATIENTS AND METHODS: Seventy-two patients were operated on by the same surgeon and followed up for at least two years. All the histopathological examinations were performed by the same pathologist. Survival was analyzed by the Kaplan-Meier method. Age, body mass index, tumor grade, myometrial invasion, histological type and stage were correlated with death. RESULTS: Overall survival at two and five years was 90.2% and 81.4%, respectively. By bivariate analysis, FIGO stage, myometrial invasion, tumor grade, histology, adnexal and/or lymph node metastasis and age were significant predictors of death (p < 0.05). Multivariate analysis revealed significant associations with death: FIGO Stage III (p = 0.001), histological type other than endometrioid (p = 0.027) and age 70 or more (p = 0.04). CONCLUSION: Endometrial carcinoma Stage III patients, histological types other than endometrioid and age 70 years or more are at significant risk for death.

Herpes simplex virus type 2 and Chlamydia trachomatis in adenocarcinoma of the uterine cervix.

INTRODUCTION: Adenocarcinoma of the uterine cervix (AC) occurs in 15-20% of primary cervical neoplasias. Although some etiologic factors for squamous cell carcinoma are well defined, and its relationship with sexually transmitted disease as human papillomavirus (HPV) is established, we still do not know about the causative factors of most of AC besides HPV infection. OBJECTIVES: To determine the presence of herpes simplex virus type 2 (HSV-2) and Chlamydia trachomatis (CT) DNA in AC specimens, and its correlation with HPV infection. METHODS: 206 paraffin-embedded cases of AC were selected to DNA extraction. The specimens and the DNA were isolated. Samples were first screened for beta-globin DNA sequences, and 67 cases were considered adequate to further analysis. In a previous analysis, DNA of HPV was identified in 79.4% of specimens included in this series (51% HPV 18 and 34% HPV 16). The local ethical committee approved the study. RESULTS: All samples were negative for HSV-2 DNA and CT DNA. CONCLUSIONS: In our series HSV-2 DNA and CT DNA were not found to be integrated to the genome of adenocarcinoma of the uterine cervix and do not seem to be a co-factor for HPV on the etiology of this histologic subtype.

Improving anaesthetists' communication skills.

The attitude, behaviour and communication skills of specialised doctors are increasingly recognised as important and they have been identified as training requirements. We designed a programme to teach communication skills to doctors in a University Department of Anaesthesia and evaluated its effect on patient outcomes such as satisfaction and anxiety. The 20 h programme was based on videotaped reviews of actual pre-operative visits and role-playing. Effects on patient satisfaction and pre-operative anxiety were assessed using a patient questionnaire. In addition, all participating anaesthetists assessed the training. We provide evidence that the training increased patient satisfaction with the pre-operative anaesthetic visit. Training also decreased anxiety associated with specific aspects of anaesthesia and surgery, but the effect was rather small given the intense programme. The anaesthetists agreed that their interpersonal skills increased and they felt better prepared to understand patients' anxieties. Communication skills training can increase patient satisfaction and decrease specific anxieties. The authors conclude that in order to better demonstrate the efficacy of such a training programme, the particular communication skills of anaesthetists rather than indirect patient outcome parameters should be measured.

[Experimental cirrhosis induced by carbon tetrachloride inhalation: adaptation of the technique and evaluation of lipid peroxidation]. / Cirrose experimental induzida pela inalação de tetracloreto de carbono: adaptação da técnica e avaliação da peroxidação lipídica.

BACKGROUND: Long-term administration of carbon tetrachloride is an accepted experimental model to produce hepatic fibrosis. Oxidative stress has been postulated as a major molecular mechanism involved in carbon tetrachloride hepatotoxicity, where the reactive oxygen species play an important role in the pathogenesis of liver fibrosis. AIMS: This study was conducted to evaluate the effectiveness of an experimental model of hepatic cirrhosis induced by carbon tetrachloride inhalation as well as the importance of lipid peroxidation and the characteristics of the ascitic fluid in this model. METHODS: At first the hepatic histologic findings were assessed using the hematoxilineosin technique in different moments of carbon tetrachloride inhalation (5th, 7th, 9th, 12th weeks). Later, at the end of 15 weeks of the study the rats were divided in three groups (control; control + phenobarbital; and carbon tetrachloride + phenobarbital) for lipid peroxidation, ascitic fluid and histologic characteristics evaluation. For the lipid peroxidation analysis, thiobarbituric acid and QL techniques were used. Cytologic and bacteriologic parameters were analysed in the ascitic fluid. RESULTS: Cirrhosis was established in 100% of carbon tetrachloride rats between the 12th and 15th weeks with an elevation in the lipid peroxidation carbon tetrachloride rats' livers. Ascitic fluid infection was observed in one of seven rats who has developed ascites. CONCLUSIONS: The carbon tetrachloride inhalation method developed in this study is effective in cirrhosis induction and ascites formation, and the carbon tetrachloride cirrhosis physiopathogenesis is probably related to the oxidative stress installation.

Expression of alpha4beta1 and alphavbeta3 integrins in the endometrium of women using the T200 copper intrauterine device.

OBJECTIVE: To investigate the expression of alpha4 and beta3 integrin subunit levels in the endometrium of healthy women and copper intrauterine device (IUD) T200 users. DESIGN: Case control study. SETTING: An academic teaching hospital and a primary care clinic. PATIENT(S): Thirteen copper IUD users and 13 normal fertile women. INTERVENTION(S): Timed endometrial biopsies during the mid-secretory phase (days 20 to 24). MAIN OUTCOME MEASURE(S): Histologic dating of endometrium and immunohistochemical staining intensity of alpha4 and beta3, using the semiquantitative immunohistochemical score (HSCORE). RESULT(S): All endometrial biopsies consistent with menstrual dates were examined for integrin expression (beta3 and alpha4). No difference in alpha4 integrin expression was found between IUD users and controls in both luminal and glandular epithelium. In fertile controls, alphavbeta3 staining was present in 100% and 38.4% of glandular and luminal epithelium, respectively. In contrast, only 61.5% of the IUD users had any alphavbeta3 staining in the glandular epithelium and only 53.9% in the luminal epithelium. The intensity of immunoreactivity between the two groups (mean HSCORE) did not differ significantly. CONCLUSION(S): Proportionately, significantly fewer women using copper IUD had positive alphavbeta3 immunoreactivity in the glandular epithelium of mid-secretory endometrium.

The effects of allopurinol in hepatic ischemia and reperfusion: experimental study in rats.

BACKGROUND/AIMS: Some studies have shown that postischemic hepatic dysfunction is mainly due to oxygen free radicals that are generated by xanthine oxidase. The present study was undertaken to determine the effect of allopurinol, an inhibitor of xanthine oxidase, on oxidative stress, liver injury and histologic alterations induced by hepatic ischemia-reperfusion in rats. METHODS: One hundred and sixty Wistar rats were used and divided into three groups. Group 1: sham operation; group 2: 50 min of ischemia followed by 1 h of reperfusion, and group 3: pretreatment with allopurinol and 50 min of ischemia followed by 1 h of reperfusion. The effect of allopurinol was evaluated by plasma levels of alanine aminotransferase and aspartate aminotransferase, histopathologic studies, and lipid peroxidation measured by the thiobarbituric acid reactive substances method and chemiluminescence initiated by tert-butyl hydroperoxide technique. RESULTS: Ischemia followed by reperfusion promoted an increase in lipid peroxidation of the hepatic cells when compared to the sham-operated group (p<0.05). This increase was attenuated in the group treated with allopurinol (p< 0.05). Allopurinol also showed a protective effect on hepatocellular necrosis (p<0.05), and the plasma levels of liver enzymes returned earlier to the normal range in rats pretreated with allopurinol in comparison to those that did not receive the drug (p<0.05). CONCLUSIONS: Allopurinol exerted a protective effect on hepatic ischemia and reperfusion in rats. The administration of this drug prior to liver operations should be considered to be submitted to trials in humans.

Neurotrophin 4/5 immunoassay: identification of sources of errors for the quantification of neurotrophins.

Neurotrophin 4/5 (NT4/5) is the least understood member of the mammalian neurotrophin family. Precise and reliable determinations of endogenous NT4/5 levels are essential to understand its physiology. Immunoassay has been used for neurotrophin quantification for over three decades. However, this apparently simple task has proved elusive: conflicting results have long been recognized for nerve growth factor (NGF; up to 10000-fold variations in serum values have been reported in the literature) and more recently, for brain-derived neurotrophic factor (as much as 50-fold reported in rat hippocampus). Reasons for these variations have been extensively investigated by researchers, but rarely explained. During the development of our NT4/5 immunoassay, we discovered that false positive reactions resulted when tissues were extracted and assayed under certain conditions. In this study, we examined the major factors that adversely affect the quantification of NT4/5. Tissue samples from Sprague-Dawley rats were dissected and extracted in a range of buffers. The assay was performed on 96 well vinyl plates using sheep anti-NT4/5 immunoglobulin (Ig) as the capture (first) antibody, and a monoclonal anti-NT4/5 as the detector (second) antibody, followed by anti-mouse IgG (third) conjugated with peroxidase or alkaline phosphatase from several manufacturers. Our results show that: (1) tissue extraction at high or low pH, a method previously found to increase the measurable amount of NGF, produced greater false positive results for NT4/5 when compared with extraction at neutral pH; (2) the most significant source of error derived from the use of conjugated antibodies capable of reacting with molecules within tissue extracts which bind to the plate, even after thorough blocking; and (3) quantification is also significantly affected by both the standards used and the ability of the antibodies to react with these standards. Our findings indicate that the precise determination of neurotrophin levels requires quality reagents and the optimization of extraction conditions for each neurotrophin. The use of a two - rather than a three - antibody assay system avoids most of the interactions which give rise to false positive reactions.

Effects of ischemia and reperfusion on oxidative stress in hepatic cirrhosis induced by carbon tetrachloride in rats.

The present study was undertaken to determine the effect of ischemia and reperfusion on oxidative stress in hepatic cirrhosis induced by carbon tetrachloride (CCl4) in rats by the evaluation of lipid peroxidation products (LPO). Cirrhosis of the liver was induced by CCl4 administration. This drug was dissolved in mineral oil and the control group received only mineral oil intraperitoneally. Forty-five minutes of ischemia followed by one hour of reperfusion were performed. LPO products were evaluated by the thiobarbituric acid reactive substances method (TBARS) and chemiluminescence initiated by tert-butyl hydroperoxide technique (CL). The liver was submitted to histologic evaluation to check whether cirrhosis was present. The results demonstrated that ischemia-reperfusion caused an increase of LPO products in cirrhotic rats when compared to the control group (p < 0.05). Hepatic cirrhosis was present in all animals treated with CCl4 and no significant histologic alterations were observed in the control group. According to this study, we can conclude that the effect of ischemia and reperfusion in a rat model of hepatic cirrhosis caused a significant increase of the hepatic-levels of LPO products when compared to the noncirrhotic livers.

Protective effect of allopurinol in the renal ischemia--reperfusion in uninephrectomized rats.

The effect of allopurinol (an inhibitor of xanthine oxidase) on oxidative stress, renal dysfunction, and histologic alterations was evaluated during the renal ischemia--reperfusion in uninephrectomized rats. Renal malondialdehyde and serum creatinine levels significantly increased after renal ischemia--reperfusion. However, the pretreatment with allopurinol demonstrated a protector effect in these parameters. Renal ischemia--reperfusion provoked a significant renal damage in the operated group. Tubular atrophy and interstitial fibrosis were attenuated by allopurinol when given prior to the surgery. In our study, allopurinol had a strong tendency to exert a beneficial effect during renal ischemia--reperfusion in uninephrectomized rats.

The role of colchicine in prevention of hepatic cirrhosis induced by carbon tetrachloride.

BACKGROUND/AIMS: The present study was undertaken to determine whether colchicine has a beneficial effect in the prevention of hepatic cirrhosis when it is given simultaneously with CCl4. METHODOLOGY: Wistar rats were employed as experimental animals and divided into 6 groups: Group I received saline solution, Group II, saline solution and mineral oil; Group III, colchicine (10 micrograms/100 g) and mineral oil; Group IV, colchicine (10 micrograms/100 g) and CCl4; Group V, colchicine (5 micrograms/100 g) and CCl4; and, Group VI received saline solution and CCl4. The effect of colchicine was evaluated by liver function tests, serum total proteins, electrolytes and histological evaluation. RESULTS: The results demonstrated higher values of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and total bilirubin in groups IV and V when compared with group VI (p < 0.05). No difference between group VI and groups IV and V was observed in histological evaluation, serum total proteins and electrolytes (p < 0.05). CONCLUSIONS: Colchicine, as given in this study, did not have any protective effect in the prevention of cirrhosis induced by carbon tetrachloride.

Elevated nerve growth factor levels in the synovial fluid of patients with inflammatory joint disease.

A novel pH shock extraction procedure was used to measure nerve growth factor (NGF) levels in both normal and inflamed synovial fluids using a sensitive and specific two-site enzyme linked immunosorbant assay. To date no data is available on NGF levels in normal synovial fluids. Synovial fluids were taken from 5 normal volunteers, 12 patients with rheumatoid arthritis and 10 patients with other inflammatory arthropathies. The mean +/- SEM NGF concentration in normal synovial fluids was 95 +/- 33.2 pg/ml (range 39.1-143.1 pg/ml), whereas the mean NGF concentration in the synovial fluids taken from patients with rheumatoid arthritis was 532.5 +/- 123.8 pg/ml (range 152-1686 pg/ml). The mean NGF concentration in patients with other inflammatory arthropathies was also raised (430.6 +/- 90 pg/ml; range 89-1071 pg/ml). The NGF concentrations were significantly higher in the synovial fluids from both inflamed groups (ANOVA p < 0.05) compared to normals. Raised levels of NGF in synovial fluid may contribute directly to joint inflammation via activation of inflammatory cells.

Left ventricular function and endomyocardial biopsy in early and advanced dilated cardiomyopathy.

We evaluated left ventricular function and endomyocardial biopsy in 20 patients with early and advanced dilated cardiomyopathy, with the purpose of assessing the correlation between histologic variables and systolic and diastolic filling indexes. Group 1 included 10 patients with no clinical history of heart failure and left ventricular ejection fraction > or = 45% and group 2, 10 patients with a clinical history of heart failure and left ventricular ejection fraction <45%. Group 1 showed lower left ventricular end-systolic and end-diastolic volumes indexes (49+/-14 versus 86+/-23 ml/m2, P<0.001; 98+/-25 versus 127+/-35 ml/m2, P=0.049), higher left ventricular ejection fraction (50+/-4 versus 32+/-4%, P<0.001) and lower coefficient of variation of percentage shortening of left ventricular transverse hemiaxes (0.3+/-0.1 versus 0.5+/-0.1, P=0.001) compared with group 2. Group 1 had higher A wave peak velocity (78+/-18 versus 60+/-20 cm/s, P=0.048), lower E/A ratio (0.9+/-0.3 versus 1.5+/-0.6, P=0.02) and slower E wave deceleration time (204+/-51 versus 155+/-50 ms, P=0.047) compared with group 2. Semiquantitative histologic scores did not differ significantly between groups. There was no significant correlation between histologic variables and left ventricular systolic and diastolic indexes. Thus, dilated cardiomyopathy shows borderline to severe left ventricular systolic impairment and distinct left ventricular diastolic filling abnormalities, according to the clinical stage. This study suggests a marked dissociation between histologic findings and functional abnormalities in early and advanced dilated cardiomyopathy.

Neurotrophic factors are required by mature sympathetic neurons for survival, transmission and connectivity.

1. Two neuronal growth factors, nerve growth factor (NGF) and neurotrophin 3 (NT3), have been studied for their action on the developing and mature sympathetic nervous system. 2. Antibodies to each factor have proved useful as reagents for the detection and quantification of NGF and NT3. They have also proved valuable in uncovering the functional roles of each factor by their ability to neutralize the endogenous molecules. 3. Nerve growth factor acts on postnatal neurons to control neurotransmission, connectivity and survival. Like NGF, NT3 is synthesized by effector tissues and is retrogradely transported by post-ganglionic neurons to prevent cell death. However, the two factors have been shown to have quite distinct functions in mature neurons, indicating the existence of different signalling pathways. This differential action extends to secondary influences on satellite glia. 4. Pathological consequences result from excessive growth factor synthesis leading, in the hypertensive rat, to hyperinnervation and elevated blood pressure. Satellite glial cell synthesis of the factors and their receptors following peripheral nerve damage appears to be responsible for the establishment of inappropriate neuronal connections between sympathetic nerve terminals and sensory somata. 5. It is concluded that these potent factors control, by both coincident and independent mechanisms, sympathetic neuronal function throughout the life of the animal.

[Protective effect of colchicine on tissue damage caused by free radicals in hepatic cirrhosis: an experimental study in rats]. / Efeito protetor da colchicina no dano tecidual causado por radicais livres na cirrose hepática: um estudo experimental em ratos.

The present study was undertaken to evaluate the effect of colchicine on oxidative stress in cirrhosis assessed by lipid peroxidation products. Wistar rats were used and induced hepatic cirrhosis by carbon tetrachloride. After the cirrhosis-induced period colchicine was administrated daily during 90 days. Lipid peroxidation was evaluated by the thiobarbituric acid reactive substances method (TBARS) and chemiluminescence initiated by tert-butyl hydroperoxide. The liver was submitted to histological evaluation to check whether cirrhosis was present. The results demonstrated an higher increase in lipid peroxide levels in cirrhotic tissue when compared with normal tissue and it was decreased by colchicine treatment (P < 0.05). Observing this study, we can conclude that hepatic cirrhosis produce an higher oxidative stress than normal liver and it can be decreased by colchicine treatment.

Detection of increased tissue concentrations of nerve growth factor with an improved extraction procedure.

Nerve growth factor (NGF) is a protein essential for the survival and normal function of sympathetic neurons. Two-site immunoassays have been developed over the past decade in several laboratories and used to estimate its endogenous concentrations in a variety of effector tissues. However, levels appear restricted to a narrow range, display only a poor correlation with innervation density, and show obvious inter- and intralaboratory variations, the origins of which are unclear. This led us to examine alternative extraction procedures for NGF before quantification. In particular, we have found treatment of tissue extracts with high and low pH in the presence of detergent results in the detection of higher NGF concentrations in immunoassays using either polyclonal or commercially available monoclonal antibodies. These increases were tissue-specific (sciatic nerve, mesenteric arteries, and thoracic aorta > heart and brain > sympathetic ganglia > abdominal aorta) and as much as 10 times greater than the amounts detected by traditional procedures. The method should also prove useful for the assay of other members of the neurotrophin family when appropriate antibodies become available.

The influence of the renin angiotensin system on abnormal expression of nerve growth factor in the spontaneously hypertensive rat.

1. The levels of the neurotrophic factor, nerve growth factor (NGF) in the mesenteric vascular bed of the spontaneously hypertensive rat (SHR) were greater than those in the corresponding vascular bed of normotensive Wistar-Kyoto rats (WKY). 2. Administration of angiotensin II (200 ng/kg per min, by minipump) for 2 weeks to juvenile WKY rats increased the levels of NGF in the mesenteric vasculature to those seen in untreated SHR. 3. Administration of the angiotensin II receptor antagonists losartan (30 mg/kg per day, p.o.) or PD144277 (10 mg/kg per day, p.o.) to juvenile SHR for 4 weeks reduced the levels of NGF such that they were indistinguishable from the values obtained for normotensive WKY rats. 4. The results confirm the elevated level of NGF in the mesenteric vasculature of the SHR and suggest that angiotensin II may play a role in regulating the abnormal concentrations of the protein in this tissue.

The regulation of nerve growth factor synthesis and delivery to peripheral neurons.

Recent experiments have provided new insights into the way in which the synthesis of nerve growth factor (NGF) is regulated. In particular, a variety of agents have been found to influence the cellular concentration of mRNANGF and NGF both in vitro and in vivo. However, no clear mechanism has been found to indicate the existence of a feedback regulation of NGF synthesis by effector tissue innervation. We have argued in this review that some form of feedback control is likely to exist between the innervation and the cells producing the factor. One such possibility, the regulation of the availability of NGF by control of its secretion, is discussed.

An improved procedure for the immunohistochemical localization of nerve growth factor-like immunoreactivity.

Nerve growth factor (NGF) is a survival factor required by a number of neuronal populations including most post-ganglionic sympathetic neurones. NGF has been detected and quantified in many tissues but there is little information regarding its cellular localization. Although it has been argued that histological detection has proven difficult due to the low levels of NGF present, other factors may contribute to prevent its identification. In the present study, we report a method for the histological detection of NGF-like immunoreactivity in the rat superior cervical ganglia (SCG). Adult Wistar-Kyoto rats were perfused briefly with either a high or low pH buffer prior to fixation and routine immunohistochemistry. Polyclonal antibodies to native mouse NGF used in the present study recognized mouse NGF but not recombinant human neurotrophin 3 (rhNT3) or brain-derived neurotrophic factor (rhBDNF) by immunoblot analysis. NGF-like immunoreactivity was localized to most sympathetic neurones. Immunoreactivity was detected in the cytoplasm with dense labelling around nuclei. No stain was seen in sections incubated with normal sheep IgG or from animals perfused with phosphate buffer (pH 7.4) prior to fixation. In addition, axotomy resulted in the disappearance of NGF immunoreactivity which was confirmed by biochemical quantification. Finally, no NGF immunoreactivity was found in neurones of rats treated systemically with NGF antiserum 3 days earlier. Possible mechanisms underlying the improvement of NGF immunohistochemistry by pH manipulation before fixation are discussed.

Ovalbumin-like immunoreactivity detected in chicken sensory neurons by antibodies to aldehyde-treated ovalbumin.

A method has been developed to raise an antiserum against ovalbumin that can detect this antigen immunohistochemically in chicken sensory ganglia. Ovalbumin-like immunoreactivity has been identified in a subpopulation of chicken dorsal root ganglion neurons by the generation of antibodies to aldehyde-conjugated ovalbumin but not by the antibodies to native ovalbumin, although both antibodies recognize the much higher concentrations of ovalbumin in sections of the oviduct. Biochemical analysis demonstrated that the antigen is more readily detectable in fixed tissue extracts than in fresh tissue extracts. Sensitive immunoblot analysis combined with affinity purification of the antigen, has confirmed that the antigen is of the same molecular weight as ovalbumin. Furthermore, the immunoreactive material elutes at a position identical to native ovalbumin on a molecular sieve column. These findings argue that molecules sensitive to aldehyde fixation may be more readily detected by the use of antisera prepared against aldehyde-modified antigens. The function of the ovalbumin-like antigen in these neurons is unknown.