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The microbiome is defined as the sum of microbes, their genomes, and interactions in a given ecological niche. Atopic dermatitis is a multifactorial chronic inflammatory skin disease leading to dryness and itchiness of the skin. It is often associated with comorbidities such as allergic rhinoconjunctivitis and asthma. Today, culture-free techniques have been established to define microbes and their genomes that may be both detrimental and beneficial for their host. There are signs that microbes, both on skin and in the gut, may influence the course of atopic dermatitis. Antiseptic treatment has been used for decades, however now, with the help of traditional culture-based methods and modern metagenomics, we are beginning to understand that targeted treatment of dysbiosis may possibly become part of an integrated therapy plan in the future.

Background and Study Aims: Acute pancreatitis has no obvious cause after clinical, laboratory and radiologic investigation in 10%-30% of patients, and the diagnosis of idiopathic pancreatitis is given. This study investigated the role of linear EUS for identification of possible causes for acute pancreatitis when other investigative methods failed., Patients and Methods: Between June 2012 and March 2017, 35 patients [25 women; mean age: 51.9 + 17.8 years] with idiopathic acute pancreatitis underwent linear EUS for investigation. All of these cases were contacted for a follow-up telephone interview to compare the EUS findings with the final diagnosis and outcome., Results: Pancreaticobiliary abnormalities were identified in 19 of 35 (54.3%) patients. Ten (28.6%) patients had microlithiasis or biliary sludge. Microlithiasis and choledocholithiasis were identified in 8 (22.8%) and a single (2.8%) patient, respectively. Two patients presented gallbladder biliary sludge, one of them with microlithiasis. Chronic pancreatitis was found on EUS in 6 (17.1%) patients, and pseudotumoral masses confirmed by EUS-FNA as autoimmune pancreatitis were detected in other 3 (8.6%) cases. Linear EUS was normal in 13 (37.1%) patients, and demonstrated findings of recent acute pancreatitis but no other etiological factor in 3 (8.6%) cases. After a mean follow-up of 33.3 months, no case with a normal EUS evaluation presented a new episode of pancreatitis, 1 of 9 cases with microlithiasis presented an episode of recurrent pancreatitis due to choledocolithiasis after cholecystectomy, and 3 of 9 cases with chronic pancreatitis presented recurrent episodes, including the 2 cases of autoimmune pancreatitis., Conclusions: Linear EUS provides diagnostic information in approximately a half of patients with idiopathic acute pancreatitis. Exclusion of pancreaticobiliary abnormalities on EUS has an important prognostic value for absence of new episodes of acute pancreatitis., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Objectives: To evaluate the diagnostic yield of the cell block (CB) technique with immunohistochemistry in patients with mesenchymal neoplasms of the gastrointestinal tract collected by endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA)., Methods: Tissue samples from consecutive patients with subepithelial lesions collected by EUS-FNA, without analysis by on-site cytopathology, were evaluated by the same pathologist only using CBs in AAF fixative. Sections were stained with haematoxylin-eosin and underwent complementary immunohistochemical staining for SMA, CD117, DOG-1 and S100 in the presence of mesenchymal neoplasms. Specimens were defined as diagnostic when sufficient tissue was present for histopathological evaluation and immunohistochemistry analysis. If they were insufficient for complete evaluation, the specimens were considered nondiagnostic., Results: Between September 2012 and December 2016, a total of 158 patients (median age: 57 years, 64.5% women) underwent EUS-FNA with an average of three needle passes for every lesion. The median lesion size was 17 mm. There were 113 mesenchymal neoplasms confirmed by immunohistochemistry (66 leiomyomas, 44 GISTs, two schwannomas, one leiomyosarcoma). The overall diagnostic yield of CBs was 84.17%. However, diagnosis was obtained in 98.5% (133/135) of the cases after exclusion of 23 cases in which EUS-FNA sampling was insufficient or without tumoural tissue. Only two mesenchymal neoplasms were not confirmed by CBs even after immunohistochemistry., Conclusions: CBs collected by EUS-FNA and analysed by immunohistochemistry showed a high diagnostic yield in patients with mesenchymal neoplasms, even without on-site cytopathology., (© 2018 John Wiley & Sons Ltd.)

Background: Tissue diagnosis is required for gastric subepithelial lesions for differential diagnosis of GISTs. However, there has not been consensus about the best needle for EUS-guided sampling of these lesions., Aim: To evaluate the diagnostic yield of EUS-FNA for gastric subepithelial lesions of the proper muscle layer with large-bore 19 gauge needles., Methods: A prospectively maintained database was retrospectively reviewed to identify consecutive patients who underwent EUS-FNA with 19 and 22 gauge needles for gastric subepithelial lesions of the fourth endosonographic layer in a tertiary care referral center. EUS-FNA was performed by the same endosonographer, using the fanning technique, without on-site cytopathologist. Specimens were analysed through cell blocks by the same pathologist. Procedure results were categorized into diagnostic, defined as enough material for histopathology and immunohistochemistry, or nondiagnostic., Results: Eighty-nine patients (mean age: 59 years, 77% women) underwent 92 EUS-FNA with 19 (75) or 22 (17) gauge needles. Mean lesion size was 22.6 mm. Overall diagnostic yield was 88%. The diagnostic yield of 19 gauge was higher than that of 22 gauge needle (92%x70.6%; p=0.0410), and similar for lesions >2 cm and <2 cm (93.7%x90.7%; p=0.9563). The best performance for 19 gauge needles was obtained performing <3 needle passes. Complication rate was 2.8%., Conclusions: Diagnostic yield of EUS-FNA with 19 gauge needles is 92% for gastric subepithelial lesions of the proper muscle layer. It is safe and highly valuable for differentiation between GIST and leiomyoma, no matter the size of the lesion.

Competing Interests: Competing interests: None

This study describes the performance of 2 new mouse anti-HER2 monoclonal antibodies (Abs), clones 33F and 410G, in evaluating HER2 overexpression in a series of 123 invasive breast carcinoma cases. In-house immunohistochemistry (IHC) was performed and the results were compared with those for the SP3 and A0485 anti-HER2 Abs. Chromogenic in situ hybridization was used to detect ERBB2 amplification and its concordance with IHC was analyzed. Comparison of IHC results for 33F with SP3 and A0485 yielded concordance rates (K) of 0.81 and 0.75, respectively; the same concordance rates were found when comparing results for 410G with SP3 and A0485. Compared with SP3 and A0485, 33F and 410G specificities were 98.6% and 98.6%, and 100% and 100%, respectively, whereas the sensitivities were 80% and 74.1%, and 78% and 72.2%, respectively. The K values between 33F and 410G HER2+ expression and chromogenic in situ hybridization-positive amplification were 1 and 0.96, respectively. These concordance rates were reproduced in another production batch (K=0.96 and K=0.96). Together, these results show that the tested monoclonal Abs would be well suited for detecting HER2 protein overexpression by IHC.

Changes in frequency and intensity of drought events are anticipated in many areas of the world. In pasture, drought effects on soil nitrogen (N) cycling are spatially and temporally heterogeneous due to N redistribution by grazers. We studied soil N cycling responses to simulated summer drought and N deposition by grazers in a 3-year field experiment replicated in two grasslands differing in climate and management. Cattle urine and NH4NO3 application increased soil NH4(+) and NO3(-) concentrations, and more so under drought due to reduced plant uptake and reduced nitrification and denitrification. Drought effects were, however, reflected to a minor extent only in potential nitrification, denitrifying enzyme activity (DEA), and the abundance of functional genes characteristic of nitrifying (bacterial and archaeal amoA) and denitrifying (narG, nirS, nirK, nosZ) micro-organisms. N2O emissions, however, were much reduced under drought, suggesting that this effect was driven by environmental limitations rather than by changes in the activity potential or the size of the respective microbial communities. Cattle urine stimulated nitrification and, to a lesser extent, also DEA, but more so in the absence of drought. In contrast, NH4NO3 reduced the activity of nitrifiers and denitrifiers due to top-soil acidification. In summary, our data demonstrate that complex interactions between drought, mineral N availability, soil acidification, and plant nutrient uptake control soil N cycling and associated N2O emissions. These interactive effects differed between processes of the soil N cycle, suggesting that the spatial heterogeneity in pastures needs to be taken into account when predicting changes in N cycling and associated N2O emissions in a changing climate.

Introduction: Considering the high prevalence of liver tumors and the impact on patient survival, a greater understanding of the biological behavior of those tumors if of great importance. The multidrug resistance gene (MDR1) may present as single nucleotide polymorphism (SNP) which can affect the expression and activity of P-glycoprotein (Pgp), and high expression of Pgp has been associated with a worse prognosis in affected patients., Objective: To correlate the C3435T polymorphism in the MDR1 gene with the immunohistochemical expression of Pgp., Material and Methods: A total of 67 samples from patients with diagnosis of hepatocellular carcinoma (HCC), collected in the period from 2000 to 2009, were analyzed. The polymorphism in the MDR1 gene was determined by the technique of allele-specific real time PCR using TaqMan assay, and the expression of protein Pgp was evaluated by immunohistochemistry., Results: Among the samples evaluated, 56 (83.6%) were from male patients and 11 (16.4%) from females. Mean age was 60.6 years (± 8.8), ranging from 37 to 85 years. The etiology of the HCC was related to hepatitis C virus infection (HCV) in 31 (46.3%) of cases, followed by hepatitis C virus infection + alcohol in 24 cases (35.8%), alcohol in 4 cases (6)%, hepatitis B virus (HBV) in 4 cases (6%) and other factors in 4 cases (6%). Liver transplantation was performed in 48 cases (71.6%) and hepatectomia in 19 cases (28.4%). The genotypes CC, CT and TT showed frequencies of 25.4%, 41.8% and 32.8%, respectively, and the allele frequencies were 46.3% for allele C and 53.7% for allele T. The expression of Pgp in over 75% of the cells was significantly more frequent in tumor tissue. On the other hand, a low expression of Pgp, in less than 25% of the cells, was significantly more frequent in non-tumor tissue. The Pgp expression in more than 50% of tumor cells of individuals with genotypes CC, CT and TT was 15.7%, 51.0% and 33.3%, respectively, and was significantly higher when in the presence of allele T (p = 0.002)., Conclusion: The presence of the polymorphic allele T is related to increased expression of Pgp protein in patients with HCC.

Objectives: • To compare the influence of a 4-week course of empirical antimicrobial therapy or observation on the prostate-specific antigen (PSA) levels of asymptomatic patients with a raised baseline PSA. • To identify whether a decrease in PSA can predict the risk of prostate cancer (PCa) detection on prostate biopsy., Patients and Methods: • Patients were referred to our ambulatory centre because of a raised PSA level (>2.5 ng/mL) with a normal digital rectal examination. A 12-core prostate biopsy was indicated in these patients and they were offered antibiotic treatment with levofloxacin 500 mg daily for 30 days. • Patients who did not agree to use antibiotics but who still showed interest in participating underwent simple observation, serving as controls. • Total and free PSA levels at baseline and after 45 days were measured. Variation in PSA level was calculated. • All patients underwent a 12-core prostate biopsy 6 weeks after the initial visit., Results: • In all, 245 men were enrolled, but 43 were lost due to follow-up. A total of 145 patients who used antibiotics and 57 controls were included in the analysis. • The median baseline PSA levels were 7.6 and 7.7 ng/mL in the antibiotic and control groups, respectively, with median follow-up levels of 6.8 and 7.0 ng/mL. The follow-up PSA level was significantly lower than the initial PSA level (P = 0.009). • Mean absolute and percentage variation in PSA level were similar in both groups (P = 0.828 and 0.128, respectively). • The overall PCa detection rate was 15.8%, and did not differ among the groups (P = 0.203). Regarding the percentage variation in PSA level, patients diagnosed with PCa tended to have their PSA level increased (22.4 vs -5.3%; P = 0.001). Indeed, a decrease of 20% in PSA was not predictive of a negative prostate biopsy (P = 0.41). • The area under the receiver operating characteristic curve for percentage PSA variation as a predictor of PCa was 0.660., Conclusions: • PSA levels tend to fall when repeated after 45 days, regardless of antibiotic use. • Despite being associated with the chance of PCa, no percentage PSA variation threshold value exhibits satisfactory discriminatory properties., (© 2010 THE AUTHORS. BJU INTERNATIONAL © 2010 BJU INTERNATIONAL.)

Objectives: To identify the most cited articles in general surgery published by Brazilian authors., Introduction: There are several ways for the international community to recognize the quality of a scientific article. Although controversial, the most widely used and reliable methodology to identify the importance of an article is citation analysis., Methods: A search using the Institute for Scientific Information citation database (Science Citation Index Expanded) was performed to identify highly cited Brazilian papers published in twenty-six highly cited general surgery journals, selected based on their elevated impact factors, from 1970 to 2009. Further analysis was done on the 65 most-cited papers., Results: We identified 1,713 Brazilian articles, from which nine papers emerged as classics (more than 100 citations received). For the Brazilian contributions, a total increase of about 21-fold was evident between 1970 and 2009. Although several topics were covered, articles covering trauma, oncology and organ transplantation were the most cited. The majority of classic studies were done with international cooperation., Conclusions: This study identified the most influential Brazilian articles published in internationally renowned general surgery journals.

Background: Well differentiated liposarcomas (WDLPS) and dedifferentiated liposarcomas (DDLPS) have been shown to have supernumerary chromosomes with amplified sequences of the MDM2 and CDK4 genes. MDM2 and CDK4 protein overexpression have also been identified in these tumours., Aim: To investigate whether immunohistochemistry (IHC) for MDM2 and CDK4 can be used to diagnose WDLPS and DDLPS., Methods: IHC for MDM2/CDK4 was carried out on a series of 129 paraffin-embedded lipomatous and non-lipomatous soft tissue tumours. The cases were divided into four groups: WDLPS (n = 19), DDLPS (n = 10), benign adipocytic tumours (BAT) (n = 17), and other mesenquimal tumours (OMT) (n = 83). IHC results were compared in each group and the diagnostic efficacy of the test in identifying WDLPS and DDLPS among the other soft tissue tumours was determined. A percentage of tumour cell positivity was evaluated to better characterise the pattern of tumour immunostaining., Results: Sensitivity and specificity of positive MDM2 and CDK4 immunostainings to identify WDLPS among BAT was 100% and 58.8%, and 68.4% and 88.2%, respectively. When distinguishing DDLPS from OMT, sensitivity and specificity of MDM2 and CDK4 were 90% and 65%, and 70% and 96.3%, respectively. The highest specificity was achieved when a case was considered positive with strong and diffuse immunoreactivity in more than 30% of the neoplastic cells (94.1% and 100%, and 77.1% and 98.8%, respectively)., Conclusion: Detection of MDM2/CDK4 protein overexpression by IHC can be used by pathologists to diagnose WDLPS and DDLPS. Considering a strong and diffuse immunostaining pattern in most of the neoplastic cells achieves the best results in identifying these tumours.

Objective: To evaluate the outcomes after the implementation of a multimodal protocol (ACERTO protocol) with patients undergoing colorectal operations., Methods: Fifty-three patients (37 M and 16 F; 57 [18-82] years old) submitted to various colorectal operations were prospectively studied in two different periods of time: from January 2004 through July 2005 (n=25, conventional group) and from August 2005 through June 2008 (n=28; ACERTO group). The patients received either the traditional perioperative management (including mechanical bowel cleansing) or a multidisciplinary protocol of perioperative care (without mechanical bowel cleansing) established by the ACERTO protocol. We looked at morbidity and mortality rates and length of hospital stay for comparisons between the groups., Results: Mortality was 3.8% (2 patients) without difference between groups. Preoperative fasting and postoperative feeding were shortened and intravenous fluids were diminished in patients operated under the ACERTO protocol (p<0.05). Postoperative morbidity (36% vs. 28.6%; p=0.56) and the incidence of anastomotic leak (12 vs. 10.7%; p=1.00) were similar. The number of complications per patient with any complication was lower in the ACERTO group (p=0.01). Changing protocols reduced the length of hospital stay by 4.5 days (12 [4-43] vs. 7.5 [3-47] days, p= 0.04)., Conclusion: The multidisciplinary routines of the ACERTO protocol are safe and enhanced recovery in colorectal surgery by reducing both hospitalization and the severity of postoperative morbidity.

Purpose: To evaluate the effect of black tea on esophageal carcinogenesis induced by the oral administration of diethylnitrosamine (DEN)., Methods: A population of 120 female mice (Mus musculus, strain CF1) were studied for 160 days. The animals were assigned to two control groups and three treatment groups. The control groups received water or tea throughout the study period, while the three experimental groups received DEN weekly, for three consecutive days, and water, tea, or both, in the other days of the week. On completion of the 160-day period, the animals were killed and their esophagi promptly examined macroscopically and subsequently submitted to histopathology (using the hematoxylin-eosin technique)., Results: In the comparative analysis between the treatment groups, tumor incidence (macroscopy) was significantly lower in those animals that received black tea besides the carcinogen. As regards the histopathologic changes, there was a greater number of low grade epithelial lesions in the same groups (p < 0.001)., Conclusion: The animals that received black tea had a lower incidence of effects related to the carcinogen's action, thus indicating that, in this model, the infusion had a significant chemoprophylactic effect on experimental diethylnitrosamine-induced carcinogenesis.

Objective: To evaluate angiogenesis in non-small cell lung cancer (NSCLC) through immunohistochemistry with CD34 and computerized image analysis comparing the microvessel area in needle biopsies and surgical specimens., Study Design: Core biopsies and surgical specimens from 28 patients with NSCLC were reviewed. Microvessels were highlighted by immunohistochemistry with anti-CD34. Tumor microvessel area was measured in digital photographs from hot spots of all samples., Results: Average microvessel area among core samples was 5,093.6 microm2 (range, 233.4-17,916.8) and among surgical specimens was 3,599.3 microm2 (range, 376.9-9,514.0). There was strong correlation between overall microvascular area in biopsies and surgical specimens (r = 0.7; p = 0.0001). Mean area of core specimens was used to divide cases into groups of low and high vascular areas. A slightly stronger correlation was observed between biopsies and surgical specimens with low vascular areas (r = 0.84; p = 0.0001)., Conclusion: There is strong correlation of microvessel area in core biopsies and respective surgical specimens in NSCLC. These data indicate that core specimens could be used to assess the extent of angiogenesis in NSCLC in the pretreatment phase.

We report a case of xanthogranulomatous cystitis (XC) in a 76-year-old man who presented with painless hematuria and a bladder mass on imaging studies. Xanthogranuloma is a chronic inflammatory condition that most commonly involves the kidney. XC is a rare condition of still unknown aetiology with only about 20 cases reported to date. The gold standard treatment is surgical resection. Consideration should be given to this entity in the differential diagnosis of urinary bladder masses.

Background: Phenobarbital has been used in experimental models because it is an important agent of carcinogenesis promotion in the liver of rats, and it is also non-genotoxic, organ-specific and dose-dependent., Aim: To evaluate the effects of the daily administration of phenobarbital in old rats treated with phenobarbital since their birth up to 24 months of age, in the absence of concomitant administration of chemical agents, which initiate carcinogenesis., Patients and Methods: A control group of male Wistar rats was fed with a basic diet and a second group was fed with the same basic diet added of 0.05% of phenobarbital, for a period of 24 months. Medium and right liver fragments were submitted to the histological processing and they were stained by hematoxiciline and eosin and were immunohystochemically colored to glutathione S-transferase placentary form., Results: Glutathione S-transferase placentary positive zones were detected in both groups and the images were analyzed concerning their number and surface extension through the technique of histometry analyses., Conclusion: Chronic use of phenobarbital did not modify the number of glutathione S-transferase placentary form positive areas. Although, data indicates that glutathione S-transferase placentary form positive areas media size are increased, probably because there are an increase in their evolution capacity and irreversibility.

Objective: To describe the reproducibility of the sentinel lymph node technique in patients with prostate cancer and verify if there is improved accuracy over modified lymphadenectomy., Material and Methods: Twenty-three patients with biopsy proven prostate cancer were enrolled in this study. Lymphoscintigraphy was performed after the transrectal administration of Tc sulfur colloid guided by ultrasound, with one injection in each prostate lobe. Images were obtained 15 and 180 min after injection. Sentinel lymph node was harvested during surgery using a gamma probe, followed by extended lymphadenectomy., Results: The mean age of the patients in this study was 66 years. An average of 3.36 sentinel lymph nodes was found for each patient. Radioactive lymph nodes were identified by the gamma probe in 21 out of 23 patients. In one of the patients there was no radiopharmaceutical migration from the injection site and in another the sentinel lymph node was visualized by lymphoscintigraphy but was not found during surgery. Three patients had lymph node metastasis; in one of these patients the sentinel lymph node was the only positive node and was found outside the modified lymphadenectomy region (obturator fossa and the external iliac)., Conclusion: Sentinel lymph node biopsy in prostate cancer adds important information to the staging of patients, not always attained through the lymphadenectomy restricted to the obturator fossa and external iliac. Such information is essential for the choice of the best treatment to be applied.

Background: Barrett's esophagus is the most serious complication of the gastroesophageal reflux disease and presents a malignant potential. The expression of the tumoral marker p53 increases with the dysplasia-adenocarcinoma sequence., Aims: To evaluate the p53 expression in Barrett's esophagus with or without dysplasia according to the two positive immunostaining criteria., Materials and Methods: The material was constituted by endoscopic biopsy specimens from 42 patients with Barrett's esophagus. Section of formalin-fixed and paraffin-embedded biopsies were stained with hematoxylin-eosin, PAS-alcian blue and evaluated the p53 immunohistochemical expression. Two p53 immunostaining criteria were utilized: 1. The staining of, at least, half of the nuclei, and 2. The staining of any nucleus. The diagnosis of dysplasia was confirmed by the agreement between three pathologists., Results: The total number of tissue specimens was 229, with an average of 5.4 specimens per patient. Dysplasia, with agreement for all pathologists examining the same set of slides, was detected in six (14.3%) cases. According to the two different p53 immunostaining criteria, the protein was detected in non-dysplastic Barrett's metaplasia, respectively, in 5 (13.9%) and 14 (38.9%) patients. Specifically in the six dysplastic cases, p53 was detected, according to the immunostaining criteria, in one (16.7%) and four (66.7%) cases, respectively., Conclusions: In this group, p53 immunohistochemical expression, regardless of positive criteria take into account, was not useful for detecting dysplasia in Barrett's esophagus.

Objective: Few studies have evaluated p53 accumulation in the squamous mucosa contiguous (SMC) to Barrett's esophagus (BE) and in the new squamous epithelium after endoscopic ablation. We evaluated the p53 expression in BE, in the SMC, and in the new squamous mucosa generated after ablation by argon plasma coagulation (APC)., Material and Methods: Endoscopic biopsy specimens from 37 BE patients, before and after ablation by APC, were analyzed. The p53 immunostaining criterion used was the staining of at least half of the nuclei., Results: p53 was detected in BE in 5 (13.5%) cases. In all these cases, SMC was p53(+). In addition, SMC was p53(-) in all cases of p53(-) BE (p <0.001). In the 5 cases with p53(+) BE and SMC, the new squamous mucosa continued to be p53(+). However, in the 32 cases with p53(-) SMC, the new squamous mucosa was also p53(-) (p <0.001). No case with p53(+) SMC turned out to be p53(-) after ablation. Similarly, no case with p53(-) BE and SMC before eradication became p53(+) after ablation (p < 0.001)., Conclusions: p53 was highly prevalent in the contiguous squamous mucosa when it is present in BE. After ablation, none of the cases lost p53 expression, and none of the negative cases turned out to be positive.

Background: Barrett's esophagus is a well-known pre-malignant condition. Pathologic interpretation of biopsy specimens guides endoscopic surveillance as well as the therapeutic approach that will be carried out. However, the predictive value of histopathologic diagnosis can be questioned due to its poor intra- and interobserver reproducibility., Aims: To assess intra- and interobserver variability in the diagnosis of Barrett's dysplasia., Material and Methods: Three-micrometer thick sections from biopsy specimens from 42 patients with Barrett's esophagus were stained with hematoxylin-eosin and PAS-alcian blue. The reading of the slides was carried out blindly in a light microscope. Intra and interobserver variability in the interpretation of the slides was determined by kappa statistics., Results: The number of tissue specimens was 229, with average of 5.45 (1 to 18) fragments for patient. Low grade dysplasia was diagnosed by pathologists in 21.4% to 52.4% of the cases. The intra-observer agreement for the diagnosis of low grade dysplasia was slight (kappa = 0.30). The interobserver agreement for the diagnosis of low grade dysplasia was poor, with kappa scores between 0.05 and 0.16. The diagnosis of dysplasia, with agreement for all pathologists examining the same set of slides, was 14.3%., Conclusions: Pathologic interpretation of Barrett's dysplasia may be subject to marked intra- and interobserver variabiliaty. Interpretation of low grade dysplasia, as high grade dysplasia, should also be considered for review by two or more pathologists.

Objective: To accomplish the tumor vascular area measure and the microvessel count, shown by immunohistochemistry, in non small cell lung cancer (NSCLC) microscopic images, and verify the correlation between these measurements and patients survival., Design: cross-sectional and observational., Material and Methods: The material was deriving from 107 NSCLC surgical specimens in Pavilhão Pereira Filho Santa Casa, Porto Alegre. Immunohistochemistry with anti-CD 34 monoclonal antibody (QB-End10; DAKO Corporation) was performed on paraffin-embedded tissue sections. The tumoral vascular area and the microvessel count were obtained by Image Pro Plus 3.0 program., Results: The 5-year survival rate of high vascular area group (the cut-off was the mean of areas) was 21.7 months +/- 2.5, significantly lower (P < 0.0001) than the low vascular area cases, 38.9 months +/- 3.0. The correlation coefficient between microvessel area and microvessel count was 0.5 (P = 0.001). The tumoral vascular area shown higher correlation to survival (correlation coefficient: 0.48 / P = 0.001) than the tumoral microvessel count (correlation coefficient: 0.2 / P = 0.03). There was not any significant correlation between the microvessel areas and the histological type, the TNM, the lymph nodes status and the clinical staging., Conclusion: The tumoral vascular area measurement, with computed assistance, can be used as a prognostic marker in NSCLC.

In an attempt to correlate the TP53 mutation pattern of squamous cell carcinomas of the esophagus (ESCC) and life style factors of patients from the high risk area Rio Grande do Sul, Brazil, 135 ESCC were analyzed, after prescreening by p53 immunohistochemistry, by SSCP and DNA sequencing of TP53, exon 5-9. Forty-nine somatic TP53 mutations (and 1 case with p53 polymorphism) were identified as missense (n = 39), frameshift (n = 6), silent (n = 1), amber (n = 1) or intron border mutations (n = 2) that cause splicing aberrations. They were preferentially found in exon 5 (36.7%) and exon 8 (32.7%). Several mutations were located in the mutation hot spot codons 248, 273 and 282, mainly at CpG sites. Transition mutations were observed in 53.1% (among them 50% G > A), transversion mutations in 34.7% (among them 47.1% G > T) and frameshifts in 12.2%, the latter 2 mainly in smokers and alcohol drinkers. Transitions were more prevalent in females than in males (p < 0.05). TP53 mutations, mainly transversions, were more frequently found in heavy smokers (p = 0.03), with the same tendency after chronic alcohol consumption. Comparison with the worldwide IARC database disclosed differences in the TP53 mutation pattern of the Brazilian tumors, with a higher accumulation of TP53 mutations in exon 8 and a higher prevalence of transition mutations. Mutations at the reported hot spot codon 176 were missing. Although difficult because of the documented coexposure to various life style risk factors in most patients of this series, the hypothesis is proposed that besides smoking and alcohol drinking the commonly consumed hot mate tea in this high risk area for ESCC is responsible for this different pattern of TP53 mutations because of chronic hyperthermic irritation and inflammation in the esophagus with an endogenous formation of radicals or carcinogenic factors that lead to a higher prevalence of transition mutations., (Copyright 2001 Wiley-Liss, Inc.)

Eleven cases of involvement of the genital tract in paracoccidioidomycosis were collected in a retrospective study of the clinical records of 683 patients seen in Porto Alegre, Rio Grande do Sul, Brazil. These cases are herein summarily reported. Eighteen similar cases were gathered in review of the Brazilian literature. Obtained data are discussed.

Although early age is an important risk factor for the occurrence of acute disseminated histoplasmosis in non immunosuppressed infants, cases of this condition have been rarely reported, even in highly endemic areas. This is a reason to add a new case, the first one diagnosed in the state of Rio Grande do Sul, and, also, to review and comment on the Brazilian literature of histoplasmosis in patients under three years of age.

Unlabelled: As the survival rate of children with malignancies has increased over past decades, the follow up of adult long-term survivors (LTS) of childhood cancer should focus on late effects of disease and treatment. Gonadal function was therefore studied in 54 LTS (aged 17-29 years; 33 male, 21 female) 2-18 years after treatment for malignancies during childhood or adolescence. To analyse the sensitivity of different diagnostic methods, tests of endocrine function (n = 52), spermiograms (n = 14), gynaecological status (n = 20) and ultrasonography of the gonads (n = 53) were compared with the results of equivalent tests in 23 age-matched normal controls (12 male, 11 female). There were no differences between male and female LTS concerning age at diagnosis, gonadal dose of irradiation (XRT) and doses of applied chemotherapeutic agents. Whereas male LTS had elevated levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) before (P < 0.05; P < 0.001) and after (P < 0.01; P < 0.001) stimulation with gonadotropin releasing hormone, female LTS exhibited normal endocrine function. Accordingly, male patients exhibited lower testicular volumes than normal controls, as measured with a Prader orchidometer (P < 0.01) or by ultrasonography (P < 0.001). Gynaecological status and ultrasonography of the gonads were normal in female LTS and controls. Whereas all spermiograms of normal controls (n = 8) showed a normal sperm cell density (SCD), only 2 of 14 male LTS exhibited a normal SCD (P < 0.001). Azoospermic LTS (n = 9) had been treated more often with alkylating agents and had received higher (P < 0.05) gonadal doses of XRT. All male LTS with testicular volumes below the normal range (< 13 ml) and basal FSH levels above the normal range (> 10 IU/l) exhibited azoospermia, whereas LTS with normal values for testicular volume and basal FSH had a normal SCD., Conclusion: A sex-specific susceptibility for gonadal damage after treatment for malignancies might be responsible, in part, for the impaired gonadal function of male LTS. Therapy with alkylating agents and/or high gonadal doses of XRT were important risk factors for azoospermia. A simple method to estimate potential fertility in individual LTS is to measure testicular volume, using a Prader orchidometer, and basal FSH serum levels.

In addition to hyperproliferation of keratinocytes, psoriasis is characterized by pronounced leukocytic infiltration. In contrast to the epidermal localization of neutrophils and T lymphocytes, macrophages are almost exclusively restricted to the dermal compartment. By immunohistologic analysis, these dermal macrophages were mainly encountered in the papillary dermis and arranged along the rete ridges in close proximity to proliferating keratinocytes. Monocyte chemoattractant protein (MCP-1) anti-sense RNA probes yielded abundant signals over the proliferating basal keratinocytes of the tips of the rete ridges, and, to a lesser extent, in cells in the papillae. Thus, the strongest MCP-1 message in psoriatic lesions is found above the dermal-epidermal junction and this may explain the characteristic sub-basal distribution of dermal macrophages. These results suggest that MCP-1 is important in regulating the interaction between proliferating keratinocytes and dermal macrophages in psoriasis pathogenesis.

The evaluation of disease activity in systemic lupus erythematosus (SLE) is important for selection of the appropriate therapeutic regimen. In addition to the clinical picture, various laboratory parameters are taken into account. However, no validated criteria for the evaluation of the disease activity in SLE have yet been established. Recently, serum levels of soluble interleukin-2 receptor (sIL-2R) have been proposed as a potential parameter for disease activity in SLE. However, the studies reported on this subject so far have focused mainly on certain subsets of the disease, and the evaluation of the disease activity was based on a very limited number of parameters. In the present study, we determined serum levels of sIL-2R in 23 patients with SLE and 30 patients with discoid LE (DLE). Evaluation of disease activity in SLE was based on a comprehensive scale which considered numerous clinical signs and laboratory parameters. In SLE, serum levels of sIL-2R showed a better correlation with disease activity than all the other parameters investigated, including proteinuria, erythrocyte sedimentation rate, serum globulin concentration, titre of antibodies against double-stranded DNA, serum albumin concentration, serum complement levels and white blood cell count. For the first time, we report on elevated serum levels of sIL-2R in DLE, which also correlated with disease activity.

We report the case of an 80-year-old woman who developed pemphigus foliaceus while taking penicillamine for rheumatoid arthritis. On the basis of this new case, we discuss fresh insights into the pathogenesis of drug-induced pemphigus. In addition, having reviewed the literature, we give a list of drugs that have been reported to induce this disease.

Intercellular adhesion molecule-1 (ICAM-1, CD54), a molecule bound to the cell surface membrane, mediates various cell-cell interactions in inflammation and immunosurveillance. By means of a new specific enzyme-linked immunosorbent assay (ELISA) for soluble ICAM-1, free circulating ICAM-1 was measured in serum from five healthy volunteers, 10 melanoma patients at different stages of their disease, and eight patients receiving high-dose interleukin-2 (IL-2) for metastatic melanoma. No correlation between the concentration of circulating ICAM-1 and the tumor burden could be detected. In melanoma patients receiving high-dose IL-2, we observed an increase of circulating ICAM-1 of up to 200%, compared to the concentration prior to therapy, ranging between 4 and 13 ng/ml. The increase in circulating ICAM-1 was associated with the induction of tumor necrosis factor-alpha and interferon-gamma.

ICAM-1-mediated cell-cell adhesion is essential for various immunologic functions, including non-MHC-restricted cytotoxicity. Shedding of ICAM-1 occurred after stimulation of melanoma cells with TNF alpha and IFN gamma. Soluble forms of ICAM-1 inhibited the conjugate formation of NK clones with K562 and of LAK cells with melanoma cells. Furthermore, the non-MHC-restricted cytotoxicity mediated by NK clones or LAK cells could be abrogated by soluble ICAM-1.

In bullous pemphigoid, autoantibodies alone are not capable of blister formation. In addition, they need complement and various inflammatory cells. The role of mononuclear cells, however, is poorly understood. Therefore, we studied the inflammatory infiltrate in 21 patients with bullous pemphigoid and 11 controls (normal skin, n = 5; eczematous skin, n = 6) using a panel of monoclonal antibodies. In bullous pemphigoid, 60% of the mononuclear infiltrate consists of CD3+ T-lymphocytes, while 25% represents monocytes/macrophages (MOMA); B-lymphocytes are absent. In the area of the basement zone (BMZ), which was studied separately, MOMA predominate. In 30% of bullous pemphigoid biopsies we found a close, almost linear attachment of MOMA to the BMZ. Along the BMZ, lymphocytes belong to the CD3+, CD4+ T-helper subset. CD8+ T-cytotoxic-suppressor cells, however, are rarely encountered there, and other cytotoxic lymphocytes are not found at all. All lymphocytes along the BMZ are memory cells. In small, early bullous pemphigoid lesions, MOMA are frequently the only effector cells. Eosinophilic granulocytes are rare in these lesions. Our findings suggest that mononuclear cells, and particularly MOMA, may be more important than believed hitherto in the inflammatory process resulting in separation of the BMZ.

Concentrations of annexins I to VI were quantitatively determined in extracts of placenta and different human cell types. They were detectable in all extracts studied, but lymphocytes/monocytes, endothelial cells and fibroblasts had very high annexin contents. The results indicate cell type specific annexin-repertoires. Annexins are intracellular proteins lacking signal sequences but which are detectable in trace amounts in plasma of healthy humans. The majority of plasma samples drawn from 14 patients suffering from myocardial infarction had elevated annexin III, IV and V concentrations. Shortly after infarction increased annexin levels were detected, reaching maximal values 24 to 48 h later. In the course of the following days annexin concentrations returned towards normal plasma levels.

Annexins belong to a family of proteins characterized by calcium-dependent binding to the cytoskeleton and phospholipid surfaces. Basing on these properties annexins are discussed to be involved in the regulation of cytodynamic, anticoagulatory and antiinflammatory processes. Since autoantibodies against annexin I had been detected in patients suffering from inflammatory or autoimmune diseases, an impact on the pathophysiological outcome was assumed. Therefore we developed solid phase, enzyme-linked immunoassays for the quantitative determination of autoantibodies directed against six members of the annexin family. Some preliminary results obtained from sera of patients with malignant melanoma show a quite frequent presence of such autoantibodies. These data suggest that autoantibodies are generated against all annexins. Furthermore, in the individual patient autoantibodies of the IgG-type are monospecific, while about 1/4 of the IgM-type are directed against several annexins. These observations imply that for investigation of anti-annexin autoantibodies in inflammatory and autoimmune diseases as well as cancer all members of the annexin family have to be taken into consideration.

The interleukin-2 receptor (IL-2R) is mainly expressed on activated T cells. Depending on its rate of synthesis, a portion is released from the cell surface as soluble IL-2R (sIL-2R). Since the role of mononuclear cells in the pathology of bullous pemphigoid (BP) is not well understood, we determined the sIL-2R in both blister fluid and serum of 15 BP patients with generalized disease before initiating systemic treatment. In addition, we obtained both lesional and perilesional skin biopsies and examined the mononuclear infiltrate with a panel of monoclonal antibodies. In BP blisters, sIL-2R levels were significantly increased (2070 +/- 350 U/ml), (+/- SEM) compared with serum samples taken at the time of blister puncture (1340 +/- 290 U/ml). In six patients with blisters due to second-degree burns or friction and in five suction blister volunteers, sIL-2R levels were normal in both blisters and serum. In BP, elevated serum levels decreased to normal during therapy, correlating with disease activity. The immunohistology showed that 30% of mononuclear cells in the dermal infiltrate of lesional skin expressed the IL-2R, whereas only 15% were positive in perilesional skin. IL-2R-positive cells are the most likely source of the shed receptor in BP blisters. Our results indicate the presence of activated T cells in lesions and peripheral blood of BP and thus underline the importance of cell-mediated immune mechanisms in the pathology of this disease.

ICAM-1-mediated cell-cell adhesion is essential for various immunologic functions, including non-MHC-restricted cytotoxicity. The present study was designed to establish whether shedding of ICAM-1 from melanoma cells occurred and to characterize the effects of soluble ICAM-1 on some cell adhesion-dependent functions. The shed soluble ICAM-1 molecule was detected and quantified by a specific ELISA. Shedding of ICAM-1 could be induced by IFN-gamma and TNF-alpha alone, or more effectively, by a combination of the two cytokines together. The use of purified soluble ICAM-1 enabled us to test for the functional significance of the ICAM-1 shedding from tumor cells. Conjugate formation between the cloned NK cell line CNK6 and the erythromyeloid cell line K562, as well as between lymphokine-activated killer cells and the melanoma cell line M26, could be inhibited by purified soluble ICAM-1 and cell-free supernatants from melanoma cell cultures containing shed ICAM-1. Furthermore, the non-MHC-restricted cytotoxicity mediated by NK and lymphokine-activated killer cells could be abrogated either by purified soluble ICAM-1 or by melanoma cell culture supernatants containing shed ICAM-1. Thus, shedding of ICAM-1 may be one of the mechanisms by which neoplastic cells escape immunosurveillance.