Alteplase and DNase for the treatment of pleural empyema in rats.

BACKGROUND AND OBJECTIVES: Adjunctive intrapleural fibrinolytic is an option to treat empyema at fibrinopurulent stage, but there is controversy about which should be use. Our objective is to evaluate the action of alteplase and/or desoxyribonuclease at physical and chemical properties in vitro pus derived from an experimental induced empyema in rats. METHODS: Streptococcus pneumoniae was introduced into the pleural cavity by thoracentesis through pleural pressure monitor. Animals were euthanized after 24 h, with macroscopic thoracic evaluation and measurement of amount of intrapleural liquid that was posteriorly stored at -80 °C. Selected samples were randomly distributed into four groups, then thawed at room temperature before exposure to one of the following: G1 = alteplase (n = 12), G2 = DNase (n = 12), G3 = alteplase + DNase (n = 12), or G4 = saline (n = 6). The mean molecular size in the fluid portion of the empyema was evaluated using dynamic light scattering; viscosity of the empyema fluid was measured using the drip method. RESULTS: Macroscopic showed purulent liquid, with fibrin and septation, with mean volume of 4.16 ml (0.5-8 ml). All samples were culture-positive for Streptococcus pneumoniae. Comparing with control, all experimental groups presented reduction of larger than 135 nm molecular size, but there was only significant difference with alteplase (p = 0,02). Viscosity reduced at all experimental groups, but increased at control. DNase group presented negative median (-5 mPa/s) of viscosity, and differed significantly from that observed in the control group (p = 0.04). CONCLUSIONS: Alteplase, DNase and alteplase + DNase changed significantly physical and chemical properties of experimental empyema at fibrinopurulent phase: alteplase reduced molecular size larger than 135 nm and DNase reduced viscosity.

CD117 Expression in Squamous Cell Carcinoma of the Oesophagus.

BACKGROUND/AIM: Identification of changes in specific genes may help new attempts in finding targeted therapy for oesophageal cancer which still has a very poor prognosis. The aim of this study was to investigate CD117 expression in squamous cell carcinoma of the oesophagus (SCCO). MATERIALS AND METHODS: A preliminary study was performed for CD177 immunoreactivity using a monoclonal antibody against CD117 on 27 SCCO specimens from patients who underwent surgical resection. Specimens of oesophageal mucosa obtained from 10 healthy individuals were studied as a control group. RESULTS: Most patients had TNM American Joint Committee on Cancer stage IIb or III SCCO and mean overall survival was 21 (range=2-72) months. Cytoplasmic membrane CD117 immunoreactivity was demonstrated in only four (15%) out of 27 tumours and in none of the controls. CONCLUSION: Although immunohistochemical expression of CD117 was higher than previously demonstrated, the lack of expression does not warranty further use in targeted therapy of SCCO.

Accuracy of complement activation product levels to detect infected pleural effusion in rats.

BACKGROUND: Pleural empyema is a well-known complication of pneumonia. If treatment is delayed, empyema may increase morbidity and mortality in affected patients. Therefore, the identification of empyema biomarkers in parapneumonic pleural effusion is desirable. Previous research has suggested complement activation products as candidate empyema markers. OBJECTIVE: To compare the levels of complement activation products C3a, C5a, and C5b9 in pleural effusion induced by Staphylococcus aureus (SA), Streptococcus pneumoniae (SP), or turpentine (control). METHODS: Thirty-nine male Wistar rats (mean weight 414 g; 290-546 g) were allocated as follows: 17 animals in the SA group, 12 in the SP group, and 10 in the control group. Bacteria or turpentine were injected into the pleural space. After 12 hr, intrapleural fluid was collected using ultrasound-guided thoracentesis. Levels of complement activation products were determined using ELISA kits. RESULTS: Two SA and one SP animals died before 12 hr. Mean levels were as follows: C3a: 1066.82 µg/ml (937.29-1196.35 µg/ml) in SA, 1188.28 µg/ml (1095.65-1280.92 µg/ml) in SP, and 679.13 µg/ml (601.29-756.98 µg/ml) in controls (P < 0.001); C5a: 55.727 ng/ml (41.22-70.23 ng/ml) in SA, 520.107 ng/ml (278.92-761.3 ng/ml) in SP, and 5.268 ng/ml (1.68-8.85 ng/ml) in controls (P < 0.001); C5b9: 15.02 ng/ml (13.1-16.94 ng/ml) in SA, 16.63 ng/ml (14.37-18.9 ng/ml) in SP, and 14.05 ng/ml (9.8-18.29 ng/ml) in controls (P = 0.692). ROC analysis revealed an area under the curve of 0.987 (95% CI: 0.953-1) for C3a; 1 (1-1) for C5a; and 0.757 for C5b9 (0.523-0.990). CONCLUSIONS: In the present rat model, complement activation fragments C3a and C5a accurately detected infected pleural effusion. Pediatr Pulmonol. 2017;52:757-762. © 2017 Wiley Periodicals, Inc.

Cell therapy in the treatment of bronchiolitis obliterans in a murine model.

OBJECTIVE: To evaluate the importance of stem cells derived from adipose tissue in reducing graft inflammation in a murine model of allogeneic heterotopic tracheal transplant. METHODS: We performed a heterotopic tracheal allografting in dorsal subcutaneous pouch and systemically injected 5x105 mesenchymal stem cells derived from adipose tissue. The animals were divided into two groups according to the time of sacrifice: T7 and T21. We also carried out histological analysis and digital morphometry. RESULTS: The T7 animals treated with cell therapy had median obstructed graft area of 0 versus 0.54 of controls (p = 0.635). The treated T21 subjects had median obstructed graft area of 0.25 versus 0 in controls (p = 0.041). CONCLUSION: The systemically injected cell therapy in experimental murine model of bronchiolitis obliterans did not reduce the severity of the allograft inflammation in a statistically significant way in seven days; Conversely, in 21 days, it increased the allograft inflammatory process.

Cell therapy in the treatment of bronchiolitis obliterans in a murine model / Terapia celular no tratamento da bronquiolite obliterante em modelo murino

OBJECTIVE: To evaluate the importance of stem cells derived from adipose tissue in reducing graft inflammation in a murine model of allogeneic heterotopic tracheal transplant. METHODS:We performed a heterotopic tracheal allografting in dorsal subcutaneous pouch and systemically injected 5x105 mesenchymal stem cells derived from adipose tissue. The animals were divided into two groups according to the time of sacrifice: T7 and T21. We also carried out histological analysis and digital morphometry. RESULTS:The T7 animals treated with cell therapy had median obstructed graft area of 0 versus 0.54 of controls (p = 0.635). The treated T21 subjects had median obstructed graft area of 0.25 versus 0 in controls (p = 0.041). CONCLUSION:The systemically injected cell therapy in experimental murine model of bronchiolitis obliterans did not reduce the severity of the allograft inflammation in a statistically significant way in seven days; Conversely, in 21 days, it increased the allograft inflammatory process.

OBJETIVO: Avaliar a importância das células-tronco derivadas de tecido adiposo na redução do processo inflamatório no enxerto em modelo murino de transplante traqueal heterotópico alogênico. MÉTODOS:Foi realizado alotransplante traqueal heterotópico em bolsa dorsal subcutânea e injetado 5x105 células-tronco mesenquimais, derivadas de tecido adiposo, sistemicamente. Os animais foram distribuídos em dois grupos, conforme o tempo de sacrifício: T7 e T21. Procedida a análise em HE e morfometria digital. RESULTADOS:Os T7 tratados com terapia celular apresentaram mediana de área obstruída do enxerto de 0 contra 0,54 dos controles (p=0,635). Os T21 tratados apresentaram mediana de área obstruída da luz do enxerto de 0,25 nos tratados e 0 nos controles (p=0,041). CONCLUSÃO: A terapia celular injetada sistemicamente em modelo experimental murino de bronquiolite obliterante não reduziu a gravidade do processo inflamatório no aloenxerto de forma estatisticamente significativa em sete dias; de modo contrário, em 21 dias, aumentou o processo inflamatório no aloenxerto.

Gastrin-releasing peptide receptor expression in lung cancer.

CONTEXT: Gastrin-releasing peptide receptors (GRPRs) activate mitogen-activated protein kinase signaling pathway primarily through epidermal growth factor receptor activation and are under investigation as a molecular target because they are overexpressed in several solid tumors. OBJECTIVE: To determine GRPR expression in both non-small cell lung carcinoma and small cell lung carcinoma, comparing results with clinical stages and demographic data. DESIGN: We analyzed the immunohistochemical expression of GRPR in 200 non-small cell lung carcinoma and 38 small cell lung carcinoma archival cases from 2004 to 2008. RESULTS: Non-small cell lung carcinoma cases tended to be higher GRPR expressers at a rate of 62.5% (weak, moderate, and strong expression in 41.5%, 13.5%, and 7.5%, respectively), compared with 52.62% in small cell lung carcinoma cases (weak, moderate, and strong expression in 34.21%, 15.78%, and 2.63%, respectively; P = .30). In non-small cell lung carcinoma there was a trend for higher percentages of strong expression in adenocarcinoma cases (10%; P = .67), and in patients with advanced stages (III and IV; 9.43% and 6.9%; P = .01). CONCLUSIONS: To the best of our knowledge, this is the first study to demonstrate GRPR tissue expression in a large population of patients with lung cancer. Although GRPR expression was similar in small cell and non-small cell carcinoma, the expression was more pronounced in an advanced-stage lung cancer, particularly in adenocarcinoma cases, and may represent a potential target for the development of new treatment approaches in this population.

Effects of methylprednisolone on inflammatory activity and oxidative stress in the lungs of brain-dead rats.

OBJECTIVE: To evaluate the effects that early and late systemic administration of methylprednisolone have on lungs in a rat model of brain death. METHODS: Twenty-four male Wistar rats were anesthetized and randomly divided into four groups (n = 6 per group): sham-operated (sham); brain death only (BD); brain death plus methylprednisolone (30 mg/kg i.v.) after 5 min (MP5); and brain death plus methylprednisolone (30 mg/kg i.v.) after 60 min (MP60). In the BD, MP5, and MP60 group rats, we induced brain death by inflating a balloon catheter in the extradural space. All of the animals were observed and ventilated for 120 min. We determined hemodynamic and arterial blood gas variables; wet/dry weight ratio; histological score; levels of thiobarbituric acid reactive substances (TBARS); superoxide dismutase (SOD) activity; and catalase activity. In BAL fluid, we determined differential white cell counts, total protein, and lactate dehydrogenase levels. Myeloperoxidase activity, lipid peroxidation, and TNF-α levels were assessed in lung tissue. RESULTS: No significant differences were found among the groups in terms of hemodynamics, arterial blood gases, wet/dry weight ratio, BAL fluid analysis, or histological score-nor in terms of SOD, myeloperoxidase, and catalase activity. The levels of TBARS were significantly higher in the MP5 and MP60 groups than in the sham and BD groups (p < 0.001). The levels of TNF-α were significantly lower in the MP5 and MP60 groups than in the BD group (p < 0.001). CONCLUSIONS: In this model of brain death, the early and late administration of methylprednisolone had similar effects on inflammatory activity and lipid peroxidation in lung tissue.

Efeitos da metilprednisolona na atividade inflamatória e estresse oxidativo nos pulmões de ratos com morte cerebral / Effects of methylprednisolone on inflammatory activity and oxidative stress in the lungs of brain-dead rats

OBJETIVO: Avaliar os efeitos da administração sistêmica precoce e tardia de metilprednisolona nos pulmões em um modelo de morte encefálica em ratos. MÉTODOS: Vinte e quatro ratos Wistar machos foram anestesiados e randomizados em quatro grupos (n = 6 por grupo): sham, somente morte encefálica (ME), metilprednisolona i.v. (30 mg/kg) administrada 5 min após a morte encefálica (MP5) e 60 min após a morte encefálica (MP60). Os grupos ME, MP5 e MP60 foram submetidos à morte encefálica por insuflação de um balão no espaço extradural. Todos os animais foram observados e ventilados durante 120 min. Foram determinadas variáveis hemodinâmicas e gasométricas, relação peso úmido/seco, escore histológico, thiobarbituric acid reactive substances (TBARS, substâncias reativas ao ácido tiobarbitúrico), atividade de superóxido dismutase (SOD) e de catalase, assim como contagem diferencial de células brancas, proteína total e nível de desidrogenase lática no LBA. A atividade da mieloperoxidase, peroxidação lipídica e níveis de TNF-α foram avaliados no tecido pulmonar. RESULTADOS: Não foram observadas diferenças significativas nas variáveis hemodinâmicas e gasométricas, relação peso úmido/seco, análises do LBA, escore histológico, SOD, mieloperoxidase e catalase entre os grupos. Os níveis de TBARS foram significativamente maiores nos grupos MP5 e MP60 do que nos grupos sham e ME (p < 0,001). Os níveis de TNF-α foram significativamente menores nos grupos MP5 e MP60 do que no grupo ME (p < 0,001). CONCLUSÕES: Neste modelo de morte cerebral, a administração precoce e tardia de metilprednisolona apresentou efeitos semelhantes sobre a inflamação e a peroxidação lipídica no tecido pulmonar.

OBJECTIVE: To evaluate the effects that early and late systemic administration of methylprednisolone have on lungs in a rat model of brain death. METHODS: Twenty-four male Wistar rats were anesthetized and randomly divided into four groups (n = 6 per group): sham-operated (sham); brain death only (BD); brain death plus methylprednisolone (30 mg/kg i.v.) after 5 min (MP5); and brain death plus methylprednisolone (30 mg/kg i.v.) after 60 min (MP60). In the BD, MP5, and MP60 group rats, we induced brain death by inflating a balloon catheter in the extradural space. All of the animals were observed and ventilated for 120 min. We determined hemodynamic and arterial blood gas variables; wet/dry weight ratio; histological score; levels of thiobarbituric acid reactive substances (TBARS); superoxide dismutase (SOD) activity; and catalase activity. In BAL fluid, we determined differential white cell counts, total protein, and lactate dehydrogenase levels. Myeloperoxidase activity, lipid peroxidation, and TNF-α levels were assessed in lung tissue. RESULTS: No significant differences were found among the groups in terms of hemodynamics, arterial blood gases, wet/dry weight ratio, BAL fluid analysis, or histological score-nor in terms of SOD, myeloperoxidase, and catalase activity. The levels of TBARS were significantly higher in the MP5 and MP60 groups than in the sham and BD groups (p < 0.001). The levels of TNF-α were significantly lower in the MP5 and MP60 groups than in the BD group (p < 0.001). CONCLUSIONS: In this model of brain death, the early and late administration of methylprednisolone had similar effects on inflammatory activity and lipid peroxidation in lung tissue.

Measurement of mid-arm muscle circumference and prognosis in stage IV non-small cell lung cancer patients.

Overall survival (OS) varies widely in patients with stage IV non-small cell lung cancer (NSCLC). Strong prognostic factors are still needed to improve decision-making regarding standard treatment options, to stratify patients for inclusion in innovative therapeutic trials and to identify patients who would be best treated with palliative care rather than with systemic chemotherapy. Mid-arm muscle circumference (MAMC) is a bedside anthropometric measurement that estimates somatic protein reserve, an early indicator of nutritional depletion. This measurement is simple, non-invasive, objective and inexpensive to perform. We evaluated MAMC as a potential prognostic factor in patients with stage IV NSCLC. A total of 56 non-selected consecutive patients with stage IV NSCLC were evaluated. The MAMC measurement results for these patients were expressed as a percentage of the expected reference values, adjusted for gender and age. Patients were categorized as normal (MAMC ≥90%) or depleted (MAMC <90%). The mean age of patients was 63 years (range 47-80), and the mean MAMC was 89 (range 66-122), with 55% of patients classified as depleted. The median OS was 6.2 months (95% CI, 5.1-7.3). In the subgroup with normal MAMC, the median OS was 10.2 months (95% CI, 9.2-11.1). In patients classified as depleted, the median OS was 5.0 months (95% CI, 4.2-5.8). The difference in OS between these two subgroups was highly significant (p<0.001 by the log-rank test; HR=0.21; 95% CI, 0.09-0.5 for patients with normal MAMC). In a multivariate analysis with Karnofsky status, age and gender as covariates, the difference in OS between the MAMC groups remained statistically significant (p<0.001, according to the Cox proportional hazards model). MAMC is a strong independent prognostic factor in stage IV NSCLC patients. Patients with MAMC <90% of the expected value had poor OS.

Validation of cofilin-1 as a biomarker in non-small cell lung cancer: application of quantitative method in a retrospective cohort.

PURPOSE: Cofilin is a cytoskeletal protein whose overexpression has been associated with aggressiveness in several types of malignancies. Here, we established and optimized a simple semi-quantitative immunohistochemistry (SQ-IHC) method for cofilin quantification in tumor biopsies, and applied it in a retrospective cohort of NSCLC patients aiming at validating the use of cofilin-1 as a prognostic biomarker. METHODS: The SQ-IHC method for cofilin-1 quantification was established and applied in a NSCLC cohort. An archival collection of biopsies from 50 patients with clinicopathological information and 5 years follow-up was accessed. Association between cofilin-1 immunocontent and clinical outcome was assessed using standard Kaplan-Meier mortality curves and the log-rank test. To evaluate the robustness of our findings, three different partitional clustering strategies were used to stratify patients into two groups according to the biomarker expression level (hierarchical clustering, Kmeans and median cutoff). RESULTS: In all the three different partitional clustering we used, survival analysis showed that patient with high cofilin-1 immunocontent had a lower overall survival rate (P < 0.05), and could be used to discriminate between good and bad prognosis. No other correlation was found when the variables age, sex or histological type were tested in association with patients outcome or with cofilin immunocontent. CONCLUSIONS: Our method showed good sensitivity/specificity to indicate the outcome of patients according to their cofilin immunocontent in biological samples. Its application in a retrospective cohort and the results presented here are an important step toward the validation process of cofilin-1 as a prognostic biomarker.

[Effects of gadolinium chloride on sodium taurocholate-induced pancreatitis in rats]. / Efeitos do cloreto de gadolínio na pancreatite induzida por tauracolato de sódio em ratos.

OBJECTIVE: To evaluate the effects of the use of gadolinium chloride before and after induction of acute pancreatitis with sodium taurocholate 3% in rats. METHODS: Wistar rats were divided into five groups: SF--control with saline intra-ductal and IV; GD control with saline intra-ductal and gadolinium chloride IV; TS--with AP control induced by sodium taurocholate 3% and saline IV; GDTS--pre-treatment with GD (24 hours before the induction of AP) and TSGD--treatment with GD (1 hour after the induction of AP). Analysis was made in serum amylase, transaminases and TNF-α; determination of the MPO activity in lung tissue, lung and pancreatic histology. RESULTS: The number of dead animals before the end of the experiment was significantly higher in TSGD (P = 0.046). The scores of pancreatitis and lung damage were higher in the groups that used sodium taurocholate compared to groups with intra-ductal infusion of saline solution. There were no differences in other variables studied when comparing TS, GDTS and TSGD groups. CONCLUSION: The benefits with the use of gadolinium chloride as a prophylactic and therapeutic drug were not demonstrated.

Efeitos do cloreto de gadolínio na pancreatite induzida por tauracolato de sódio em ratos / Effects of gadolinium chloride on sodium taurocholate-induced pancreatitis in rats

OBJETIVO: Avaliar os efeitos do uso de cloreto de gadolínio como pré-tratamento e tratamento em um modelo experimental de pancreatite em ratos induzida por tauracolato de sódio a 3 por cento. MÉTODOS: Ratos Wistar foram divididos em cinco grupos: grupo SF - controle com solução fisiológica intra-ductal e IV; grupo TS - controle com PA induzida por tauracolato de sódio a 3 por cento e solução fisiológica a 0,9 por cento IV; grupo GD - controle com SF intra-ductal e cloreto de gadolínio IV; grupo GDTS - pré-tratamento com GD (24h antes da indução da PA) e grupo TSGD - tratamento com GD (1h após a indução da PA). Foi realizado dosagem sérica de amilase, transaminases e TNF-á; determinação da atividade da MPO no tecido pulmonar; histologia pancreática e pulmonar. RESULTADOS: O número de animais mortos antes do término previsto do experimento foi significativamente maior no grupo TSGD (p=0,046). Os escores de pancreatite e de dano pulmonar foram mais elevados nos grupos que utilizaram tauracolato em comparação aos grupos com infusão intra-ductal de solução salina. Não houve diferenças nas demais variáveis estudadas na comparação entre os grupos TS; GDTS e TSGD. CONCLUSÃO: Não foram demonstrados benefícios com o uso de cloreto de gadolínio de forma profilática e terapêutica.

OBJECTIVE: To evaluate the effects of the use of gadolinium chloride before and after induction of acute pancreatitis with sodium taurocholate 3 percent in rats. METHODS: Wistar rats were divided into five groups: SF - control with saline intra-ductal and IV; GD control with saline intra-ductal and gadolinium chloride IV; TS - with AP control induced by sodium taurocholate 3 percent and saline IV; GDTS - pre-treatment with GD (24 hours before the induction of AP) and TSGD - treatment with GD (1 hour after the induction of AP). Analysis was made in serum amylase, transaminases and TNF-á; determination of the MPO activity in lung tissue, lung and pancreatic histology. RESULTS: The number of dead animals before the end of the experiment was significantly higher in TSGD (P = 0.046). The scores of pancreatitis and lung damage were higher in the groups that used sodium taurocholate compared to groups with intra-ductal infusion of saline solution. There were no differences in other variables studied when comparing TS, GDTS and TSGD groups. CONCLUSION: The benefits with the use of gadolinium chloride as a prophylactic and therapeutic drug were not demonstrated.

Tumor necrosis factor alpha in experimental empyema thoracis.

PURPOSE: To determine serum and pleural concentrations of tumor necrosis factor alpha (TNF-α) in an experimental model of empyema induced by intrapleural inoculation of Staphylococcus aureus or Streptococcus pneumoniae. METHODS: Wistar rats were inoculated with S. aureus (SA group, 17 animals) or S. pneumoniae (SP group, 30 animals). The presence of free fluid or pus in the pleural space was investigated. TNF-α levels >150 pg/ml (minimum detection limit) were determined in pleural fluid and blood. Histopathological examination of pleural tissue was performed to determine the severity of infection. RESULTS: Serum TNF-α was >150 pg/ml in nine SA versus 10 SP rats. In pleural fluid, TNF-α was >150 pg/ml in 11 SA versus 19 SP rats. Pleural and serum TNF-α concentrations were significantly different in the SP group (P = 0.035), but not in the SA group (P = 0.727). Pleural TNF-α was similar in both groups (P = 0.92), but serum TNF-α was significantly higher in SA (P = 0.03). Out of 17 SA animals, 1 (5.8%) did not develop empyema, versus 4 (13.3%) out of 30 SP animals. A mild inflammatory response was predominant in both groups, but the inflammatory process was significantly more severe in SP (P = 0.012). However, TNF-α levels were not associated with severity of the inflammatory response. CONCLUSIONS: We describe a simple and effective rat model of empyema. TNF-α levels above 150 pg/ml in the pleural fluid are useful to confirm empyema, but cannot predict the severity of the inflammatory response. TNF-α levels below 150 pg/ml are useful to rule out empyema.

Immunohistochemical expression of HER-2/NEU-CERBB-2 in patients with adenocarcinoma of the stomach / Expressão imunoistoquímica do HER-2/NEU-CERBB-2 em pacientes com adenocarcinoma do estômago

Objectives: To determine the prevalence of Her-2/Neu-cerbb-2 in the gastric mucosa of patients with gastric adenocarcinoma in a brazilian patient group. Methods: The immunohistochemical expression of Her-2/Neu was studied in 37 formalin-fixed paraffin–embedded tissue sections. Results: The immunohistochemical reaction produced by the anti-HER- 2/Neu antibody was positive in two cases (5.4%). Conclusion: The low prevalence of Her-2/Neu observed in these southern brazilian cases is probably due to the great number of poorly differentiated cancers in this serie.

Objetivo: Determinar a prevalência do Her-2/Neu-CerbB-2 na mucosa de pacientes com adenocarcinoma de estômago em um grupo de doentes brasileiros. Métodos: A expressão imunoistoquímica do Her-2/Neu foi estudada em 37 amostras de tecidosfixados em formalina e embebidos em parafina. Resultados: A reação imunoistoquímica produzida pelo anticorpo HER-2/Neu foi positiva em dois pacientes (5.4%). Conclusão: A baixa prevalência Her-2/Neu observada neste grupo de pacientes é provavelmente devida ao grande número de tumores pouco diferenciados encontrados nesta série.

Immunohistochemical expression of HER-2/NEU-CERBB-2 in patients with adenocarcinoma of the stomach.

OBJECTIVES: To determine the prevalence of Her-2/Neu-cerbb-2 in the gastric mucosa of patients with gastric adenocarcinoma in a brazilian patient group. METHODS: The immunohistochemical expression of Her-2/Neu was studied in 37 formalin-fixed paraffin-embedded tissue sections. RESULTS: The immunohistochemical reaction produced by the anti-HER-2/Neu antibody was positive in two cases (5.4%). CONCLUSION: The low prevalence of Her-2/Neu observed in these southern brazilian cases is probably due to the great number of poorly differentiated cancers in this series.

Clubbed fingers: radiological evaluation of the nail bed thickness.

It is established that there is an increase in soft tissue of the clubbed digits as demonstrated on previous histopathological examinations. In the present study, the nail bed thickness was assessed and measured on plain radiographs of index fingers in two groups of patients: one group with lung disease and fingers clubbing and one group of normal controls. A vertical x-ray beam was used with a focus-film distance of 1.0 m, with the index finger placed in lateral view directly over the film, without anti-diffusion grid. Three investigators, blinded to prevent bias measured the thickness of soft tissues between the nail root and the terminal phalanx on the radiographs. This method was used to evaluate a group of 85 clinically clubbed (hyponychial angle > 192.0 degrees) adult patients with lung disease and a control group of a 100 normal adult individuals with no clubbing (hyponychial angle < 188.0 degrees). The mean nail bed thickness in the patients with clubbing (n = 85) was 3.88 +/- 0.55 mm (3.00-5.50 mm). In comparison, in the normal subjects (n = 100), the mean was 2.38 +/- 0.27 mm (1.75-3.10 mm), revealing a significant difference (P < 0.001). Only two normal individuals presented nail bed thickness >or=3.0 mm. A good interobserver agreement on the measurements was found (P > 0.900). The radiographic evaluation of the nail bed thickness was easily performed, with good interobserver concordance. It is possible to distinguish between clubbed from nonclubbed fingers, in vivo, using plain radiograph.

Electrocautery versus 23% NaOH infiltration to induce subglottic stenosis in a canine experimental model.

Subglottic stenosis (SGS) is defined as the narrowing of the lower larynx. Difficulties in the management of subglottic stenosis, especially in the pediatric population, justify the development of experimental models. The objective of this study was to compare the two methods of experimental subglottic stenosis induction. Twenty-three dogs were randomly selected and assigned by lottery to either one of the two groups: Gp I (n = 10) of electrocoagulation; and Gp II (n = 13) of 23% NaOH injection. In Gp I, self-interruption electrocoagulation was applied to one point in each of the four quadrants of the cricoid cartilage. In Gp II, 0.2 ml of 23% NaOH was injected in the submucosal layer in the anterior and posterior portions of the cricoid cartilage. Once a week, endoscopy was performed and the caliber of the subglottic region was measured using endotracheal tubes, and the injection was repeated if there were no signs of subglottic stenosis. The animals were killed on day 21; animals that developed respiratory distress were killed before day 21. One animal in Gp I died on day 14 after the injection and during transportation; two animals in Gp II died, one on day 7 due to a tracheoesophageal fistula, and the other of unknown causes on day 5. Significant subglottic stenosis (over 51% obstruction) was found in 67% of the animals in Gp I and in 64% of those in Gp II (P = 0.99). Median time to development of significant stenosis was 21 days in both groups, and required either two or three injections. Mean time for the performance of the procedures was significantly shorter (P < 0.01) in Gp I (mean: 6.36 min) than in Gp II (mean: 14.88 min). Electrocoagulation and 23% NaOH injection in the subglottic region were effective in the development of significant subglottic stenosis in dogs, both methods leading to stenosis in the same period of time and after the same number of procedures. However, electrocoagulation was the fastest method.

Hepatic outcomes after jejunoileal bypass: is there a publication bias?

BACKGROUND: One of the reasons why jejunoileal bypass (JIB) was abandoned were reports of liver failure. The aim of this study was to describe histological findings in the intraoperative and follow-up liver biopsies of a cohort of super-obese patients who had undergone JIB. METHODS: 50 consecutive patients underwent JIB. Samples of liver biopsies performed intraoperatively (41 patients) and in the follow-up (31 patients) were evaluated. Brunt's scale was used. RESULTS: Mean age at operation was 37.9 +/- 7.6 years, and 15 patients (30.6%) had diabetes type 2, 20 (40.8%) had dyslipidemia, 29 (59.2%) had high blood pressure, and one (0.5%) had hepatitis C. Mean BMI preoperatively was 52.8 +/- 7.5 kg/m(2). Mean follow-up time was 67.0 +/- 42.8 months. At the time of the latest liver biopsy, the mean BMI was 35.7 +/- 7.5 kg/m(2). The % excess weight loss (%EWL) was 62.4 +/- 20.0%. 8 deaths (16%) have occurred, none from liver-related complications. At liver biopsy during the JIB operation, NAFLD was confirmed in 36 patients (86.7%) and NASH in 13 (31.7%). In 25 patients with mean follow-up of 4.8 +/- 4.0 years, there was no statistically significant change in the liver histology regarding the extent of steatosis (P=0.20), steatohepatitis (P=0.74) and fibrosis (P=0.71). CONCLUSIONS: There was a significant metabolic improvement, maintenance of the %EWL, and no worsening of liver histology. There has possibly been a publication bias concerning liver outcomes, where the type of JIB and the concomitance of hepatitis C were not taken into account.

Lung oxidative response after acute coal dust exposure.

Coal dust exposure can induce an acute alveolar and interstitial inflammation that can lead to chronic pulmonary diseases. The objective of this study was to describe the acute and later effects of acute coal dust exposure in lung parenchyma and the involvement of reactive oxygen species in coal dust effects. Forty-eight male Wistar rats (200-250 mg) were separated into four groups: 48 h, 7 days, 30 days, and 60 days after coal dust instillation. Gross mineral coal dust (3 mg/0.5 mL saline) was administered directly in the lungs of the treatment group by intratracheal instillation. Control animals received only saline solution (0.5 mL). Lipid peroxidation was determined by the quantity of thiobarbituric acid-reactive species (TBARS), oxidative damage to protein was obtained by the determination of carbonyl groups, the total radical-trapping antioxidant parameter (TRAP) was estimated by luminol chemoluminescence emission, catalase activity was measured by the rate of decrease in hydrogen peroxide, and superoxide dismutase activity was assayed by the inhibition of adrenaline autooxidation. Histological evaluation of coal dust-treated rats demonstrated an inflammatory infiltration after 48 h of the exposure. Initially, this was a cellular infiltration suggestive of lymphocyte infiltration with lymphoid hyperplasia that remained until 7 days after induction. This initial response was followed by a chronic inflammatory infiltration characterized by aggregates of macrophages 30 days after induction. This inflammatory response tended to resolve 60 days after induction, being similar to that of control animals. During both the acute and chronic phases of lung inflammation we observed a decrease in the TRAP in the lung of coal dust-exposed animals compared to that in control animals. We also observed an activation of superoxide dismutase 60 days after coal dust exposition. TBARS were increased 60 days after coal dust exposure and protein carbonyl groups increased at all times after coal dust exposure (48 h, 7 days, 30 days, and 60 days). These data suggested a biphasic inflammatory response and the involvement of oxidative damage in coal dust-induced lung damage.

Induction of pleural empyema in rats by thoracentesis with intrapleural pressure monitoring.

Our objective was to evaluate experimental induction of empyema in rats by intrapleural inoculation of Staphylococcus aureus by means of thoracentesis with pleural pressure monitoring. Forty female albino Wistar rats, anesthetized with droperidol and fentanyl, underwent intrapleural inoculation of a 0.2-ml solution. Group I ( N=25) received 10(10) colony-forming units/ml of Staphylococcus aureus cultivated in brain-heart infusion agar (BHI); group II (N = 15), the control group, received sterile BHI agar. Rats were inoculated after right hemithorax thoracentesis with a needle connected to an oscilloscope for pressure monitoring to confirm that the needle was inside the pleural space. Animals were killed after 3 (groups Ia and IIa) to 5 (groups Ib and IIb) days with sodium pentobarbital. The macroscopic changes, amount of pleural fluid, and anatomopathological aspects of pleura and lungs were recorded, as well as death causes and bacteriological findings of pleural fluid for animals that died before the time established for euthanasia. In group I, three animals died of thoracentesis complications, and five others died in the first 24 h due to septicemia; blood and spleen cultures isolated the bacteria previously inoculated. In group II, there was one death of unknown cause. Of the 17 rats inoculated with bacteria, nine (group Ia) were killed on the 3rd day; all had bacteria in pleural fluid (volume 0.5-3.8 ml). The other eight rats (group Ib) were killed on the 5th day; three (39.5%) had pleural fluid with bacteria (0.5-1.5 ml), and five (60.5%) had no pleural fluid. Rats from group II killed on the 3rd (group IIa) and 5th (group IIb) days had no pleural fluid. Pathologic examination revealed inflammatory infiltrate (93.75%) and fibrin (18.75%) in pleura, and inflammatory interstitial infiltrate (12.5%) in the right lung in group I; no changes were observed in 93.3% of the lungs in group II. Macroscopic examination revealed only turbid and bloody pleural fluid (class I) without pleural adhesions. Pleural inflammatory infiltrate was found in rats that received the bacteria but had no fluid at necropsy (class 0). One control rat, although with no clinical signs of disease or pleural fluid, had signs of pleural and pulmonary infection at necropsy. We conclude that empyema may be induced in rats by the inoculation of Staphylococcus aureus by means of thoracentesis with pleural pressure monitoring. The highest amount of pleural fluid was observed 3 days after bacterial inoculation.