BMI-1 expression increases in oral leukoplakias and correlates with cell proliferation

Abstract Oral leukoplakia (OL) is a white lesion of an indeterminate risk not related to any excluded (other) known diseases or disorders that carry no increased risk for cancer. Many biological markers have been used in an attempt to predict malignant transformation; however, no reliable markers have been established so far. Objective To evaluate cell proliferation and immortalization in OL, comparing non-dysplastic (Non-dys OL) and dysplastic OL (Dys OL). Methodology This is a cross-sectional observational study. Paraffin-embedded tissue blocks of 28 specimens of Non-dys OL, 33 of Dys OL, 9 of normal oral mucosa (NOM), 17 of inflammatory hyperplasia (IH), and 19 of oral squamous cell carcinomas (OSCC) were stained for Ki-67 and BMI-1 using immunohistochemistry. Results A gradual increase in BMI-1 and K-i67 expression was found in oral carcinogenesis. The immunolabeling for those markers was higher in OSCC when compared with the other groups (Kruskal-Wallis, p<0.05). Ki-67 expression percentage was higher in OL and in IH when compared with NOM (Kruskal-Wallis/Dunn, p<0.05). Increased expression of BMI-1 was also observed in OL when compared with NOM (Kruskal-Wallis/Dunn, p<0.05). No differences were observed in expression of both markers when non-dysplastic and dysplastic leukoplakias were compared. A significant positive correlation between Ki-67 and BMI-1 was found (Spearman correlation coefficient, R=0.26, p=0.01). High-grade epithelial dysplasia was associated with malignant transformation (Chi-squared, p=0.03). Conclusions These findings indicate that BMI-1 expression increases in early oral carcinogenesis and is possibly associated with the occurrence of dysplastic changes. Furthermore, our findings indicate that both Ki-67 and BMI-1 are directly correlated and play a role in initiation and progression of OSCC.

TGF-ß1 and its association with clinicopathological features, proliferative activity and prognosis in oral squamous cell carcinoma: an immunohistochemical study

Introduction: The prognostic value of transforming growth factor beta-1 (TGF-ß1) in oral cancer remains unclear. Therefore, the aim of this study was to evaluate TGF-ß1 expression in oral squamous cell carcinoma (OSCC) samples and its association with clinicopathological data, tumor proliferative activity, and patients' prognosis. Methods: A total of 68 patients with histopathological diagnosis of OSCC were included, as well as 9 cases of normal oral mucosa for comparison purposes. The OSCC sample was categorized according to patients' outcomes in favorable prognosis (n=30) or unfavorable prognosis (n=38). Immunohistochemical staining for TGF-ß1 and Ki-67 were performed. The slides were semi-quantitatively and quantitatively evaluated for TGF-ß1 and Ki-67, respectively. Results: TGF-ß1 was significantly increased in OSCC compared to normal oral mucosa (<0.01). An inverse correlation was found between TGF-ß1 and Ki67 staining in OSCC (p=0.01). No association was found between TGF-ß1 expression and OSCC clinicopathological features, prognosis or survival. Conclusions: TGF-ß1 had no prognostic value and appears to maintain its suppressive role concerning cell proliferation. (AU)

Collagen I and III in women with diastasis recti

OBJECTIVES: Interest in elucidating the etiology of hernias has encouraged countless studies of musculoaponeurotic structures in individuals with and without hernias. Studies of hernia patients have firmly demonstrated a correlation between hernias and collagen alterations in their fascia. Diastasis recti is an increased width of the abdominal midline that is exclusively composed of interlacing aponeurotic expansions of the anterolateral abdominal muscles. The condition is common among women undergoing abdominoplasty, and many factors, not only mechanical, play a role. The goal of this study is to evaluate and compare collagen type I and III levels in the midline fascia of women with and without diastasis recti to report their possible influence on this condition. METHODS: This is a case-control study nested within a surgical cohort of 18 women with diastasis recti and 18 women without the condition (cases and controls, respectively). Fascia from the midline of the abdominal wall was collected and analyzed through immunohistochemistry using polyclonal antibodies to collagen type I and III. RESULTS: Both type I and type III collagen were less abundant in women with diastasis recti than in those without the condition, and the difference was statistically significant (p<0.001). CONCLUSION: Low collagen type I and type III levels in the midline of the abdominal wall may play a key role in the development of diastasis recti.

Peritoneal adhesions type i, iii and total collagen on polypropylene and coated polypropylene meshes: experimental study in rats / Aderências peritoneais tipo i, iii e colágeno total em telas de polipropileno e revestidas: estudo experimental em ratos

ABSTRACT Background: Hernia correction is a routinely performed treatment in surgical practice. The improvement of the operative technique and available materials certainly has been a great benefit to the quality of surgical results. The insertion of prostheses for hernia correction is well-founded in the literature, and has become the standard of treatment when this type of disease is discussed. Aim: To evaluate two available prostheses: the polypropylene and polypropylene coated ones in an experimental model. Methods: Seven prostheses of each kind were inserted into Wistar rats (Ratus norvegicus albinus) in the anterior abdominal wall of the animal in direct contact with the viscera. After 90 days follow-up were analyzed the intra-abdominal adhesions, and also performed immunohistochemical evaluation and videomorphometry of the total, type I and type III collagen. Histological analysis was also performed with hematoxylin-eosin to evaluate cell types present in each mesh. Results: At 90 days the adhesions were not different among the groups (p=0.335). Total collagen likewise was not statistically different (p=0.810). Statistically there was more type III collagen in the coated polypropylene group (p=0.039) while type I was not different among the prostheses (p=0.050). The lymphocytes were statistically more present in the polypropylene group (p=0.041). Conclusion: The coated prosthesis was not different from the polypropylene one regarding the adhesion. Total and type I collagen were not different among the groups, while type III collagen was more present on the coated mesh. There was a greater number of lymphocytes on the polypropylene mesh.

RESUMO Racional: A correção herniária é tratamento realizado rotineiramente na prática cirúrgica. O aprimoramento da técnica operatória e dos materiais disponíveis trouxe grande benefício na qualidade dos resultados cirúrgicos. A inserção de próteses para correção herniária é bem embasada na literatura e tornou-se o padrão de tratamento. Objetivo: Avaliar em modelo experimental dois tipos de próteses diferentes, de polipropileno e polipropileno revestido. Métodos: Foram inseridas sete próteses de cada tipo em ratos Wistar (Ratus norvegicus albinus) na parede abdominal anterior do animal em contato direto com as vísceras. Após o seguimento de 90 dias analisaram-se as aderências intra-abdominais, bem como avaliação por imunoistoquímica e videomorfometria do colágeno total, tipo I e tipo III. Também, fez-se análise histológica com hematoxylina-eosina para avaliação dos tipos celulares presentes em cada tela. Resultados: Aos 90 dias as aderências não foram diferentes entre os grupos (p=0,335). O colágeno total igualmente não foi estatisticamente diferente (p=0,810). O colágeno tipo III foi estatisticamente maior no grupo polipropileno revestido (p=0,039) enquanto o tipo I não diferiu entre as próteses (p=0,050). Os linfócitos foram estatisticamente mais presentes no grupo polipropileno (p=0,041). Conclusão: A prótese revestida não foi diferente da de polipropileno na variável aderência. O colágeno total e tipo I não foram diferentes entre os grupos enquanto que o colágeno tipo III foi mais presente na tela revestida. O número de linfócitos foi maior na tela de polipropileno.

Impact of highly active antiretroviral therapy in the development and remission of oral plasmablastic lymphoma.

Plasmablastic lymphoma (PBL) represents a rare type of non-Hodgkin lymphoma associated with human immunodeficiency virus (HIV) infection. The impact of highly active antiretroviral therapy (HAART) in this tumor is poorly known due to its small incidence. This study reports a case of a 33-year-old HIV-positive woman who was referred to the Stomatology Department complaining about a painful gingival growth and cervical nodule both with 20 days of evolution. The lesions appeared 7 months after the patient stopped HAART. The final diagnosis was PBL. After resuming HAART for 45 days, the gingival lesion presented complete remission. The patient continued with HAART alongside chemotherapy. At 24 months follow-up, the patient was stable. The dental surgeon plays an essential role in orientation and retention in care of HIV patients once the adherence of HAART seems to play an important role in PBL development and response to treatment.

Clinicopathological analysis of salivary gland tumors over a 15-year period

Abstract Salivary gland tumors (SGT) are rare neoplasms that generate interest due to their histopathological diversity and clinical behavior. The aims of the present study were to investigate clinicopathological aspects of SGTs diagnosed at a tertiary health center and compare the findings with epidemiological data from different geographic locations. Cases of tumor in the head and neck region at a single health center in the period between 1995 and 2010 were reviewed. Patient gender, age and ethnic group as well as anatomic location, histological type and clinical behavior of the tumor were recorded. Availability of complete information about these aspects was considered the inclusion criteria. Descriptive statistical analysis of the data was performed using the frequencies of categorical variables. Among the 2168 cases of tumors in the head and neck region, 243 (11.20%) cases were diagnosed in the salivary glands, 109 of which met the inclusion criteria: 85 (78%) benign tumors and 24 (22%) malignant tumors. Mean patient age was 46.47 years. The female gender accounted for 56 cases (51.4%) and the male gender accounted for 53 (48.3%). The major salivary glands were affected more (75.2%) than the minor glands. The most frequent benign and malignant SGTs were pleomorphic adenoma (81.2%) and adenoid cystic carcinoma (58.3%), respectively. In conclusion, pleomorphic adenoma and adenoid cystic carcinoma are the most frequent benign and malignant lesions, respectively. Comparing the present data with previous studies on SGTs, one may infer that some demographic characteristics and the predominance of malignant tumors vary in different geographic regions.

Prevalence of iffection with caga-positive heliobacter pylori strains among chhildren and adolescents in southern Brazil / Prevalência da infecção por cepas de Helicobacter pylori cagA-positivo em crianças e adolescentes do Sul do Brasil

Context Helicobacter pylori (H. pylori) has a worldwide distribution, but the prevalence of infection, virulence factors, and clinical presentation vary widely according to the studied population. In Brazil, a continental country composed of several ethnicities and cultural habits, the behavior of infection also appears to vary, as many other studies have shown. Objectives Describe the prevalence of infection with cagA-positive H. pylori strains in a group of children and adolescents who underwent esophagogastroduodenoscopy in Porto Alegre, Rio Grande do Sul. Methods Fifty-four gastric biopsy specimens of children and adolescents with H. pylori infection demonstrated by histology, urease test and molecular analysis were tested for the presence of cagA positive H. pylori strains by the polymerase chain reaction method. Results he prevalence of cagA-positive H. pylori was 29.6% (95% confidence interval, 18 to 43.6%). There were no statistically significant differences in clinical or demographic characteristics or in the endoscopic and histological features of patients infected with cagA-positive strains as compared with those infected by cagA-negative strains. Conclusions he study showed a low prevalence of infection with cagA-positive H. pylori strains among children and adolescents who underwent EGD in southern Brazil, in comparison to studies conducted with children from other regions of Brazil. There was no association between the presence of cagA-positive strains and more severe clinical presentations in the studied sample. .

Contexto Helicobacter pylori (H. pylori) tem distribuição geográfica universal, embora a prevalência da infecção, os fatores de virulência, bem como a apresentação clínica, variem de acordo com a população estudada. No Brasil, um país continental composto por várias etnias e hábitos culturais diversos, o comportamento da infecção também parece variar, como muitos estudos têm demonstrado. Objetivos Descrever a prevalência da infecção por cepas de H. pylori cagA-positivo em um grupo de crianças e adolescentes submetidos a esofagogastroduodenoscopia em Porto Alegre, Rio Grande do Sul. Métodos Cinquenta e quatro (54) fragmentos de biópsia gástrica com presença de H. pylori demonstrada pela análise histológica, teste da urease e análise molecular foram testados para a presença de cepas de H. pylori cagA-positivo pelo método da reação em cadeia da polimerase. Resultados prevalência de cepas de H. pylori cagA-positivo foi de 29,6% (intervalo de confiança de 95%, 18% a 43,6%). Não houve diferenças estatisticamente significativas nas características clínicas e demográficas e nos achados endoscópicos e histológicos entre os pacientes infectados por cepas de H. pylori cagA-positivo em comparação com os cagA-negativo. Conclusões O estudo demonstrou uma baixa prevalência de infecção por cepas de H. pylori cagA-positivo nas crianças e adolescentes submetidas a esofagogastroduodenoscopia no Sul do Brasil em comparação com os estudos realizados com crianças de outras regiões do Brasil. Não houve associação entre a presença de cepas cagA-positivo e desfechos clínicos desfavoráveis na amostra estudada. .

Prevalence of thyroid-related abnormalities in adults with celiac disease: a cross-sectional study / Prevalência de alterações da tireoide em adultos com doença celíaca

An increased prevalence of autoimmune disorders, including thyroid diseases, has been reported in patients with celiac disease (CD). This study aimed to identify the prevalence of thyroid-related abnormalities in adults with CD.METHODS: Thirty-nine subjects with biopsy-proven CD, three men (7.7%) and 36 women (92.3%), answered a questionnaire. All patients were followed at the outpatient clinic of Hospital de Clínicas de Porto Alegre. Thyroid-related abnormalities were evaluated by serum thyroid stimulating hormone (TSH), free thyroxine (FT4)and anti-thyroperoxidase antibodies (anti-TPO) levels.RESULTS: Three subjects had grade 1 lesions; one had grade 2; 13 had grade 3a; eight had grade 3b; and 14 had grade 3c, according to the modified Marsh classification. Mean age was 50.8 ± 12.9 years. Twenty-nine (74.4%) subjects were euthyroid. Ten subjects (25.6%) had hypothyroidism: eight of them (20.5%) werealready taking levothyroxine, and subclinical hypothyroidism was identified during the study in the other two patients (5.1%). No patients were hyperthyroid. Mean serum FT4 levels were 1.15 ± 0.25 ng/dl. Median serum TSH levels were 2.39 (0.06-6.64)IU/ml. Serum anti-TPO levels were positive in six patients (15.4%): one with hypothyroidism, one with subclinical hypothyroidism, and four with normal thyroid function. No factors associated with thyroid-related abnormalities were identified. CONCLUSION: This study showed a high prevalence of thyroid-related abnormalities, as did other studies. Since anti-thyroid antibodies were positive in some patientswith normal thyroid function, periodic evaluation of thyroid function in CD patients is recommended.

INTRODUÇÃO: Tem-se observado um aumento na prevalência de doenças autoimunes, incluindo doenças da tireoide, em pacientes com doença celíaca (DC).O objetivo deste estudo foi identificar a prevalência de alterações da tireoide em adultos com DC.MÉTODOS: Trinta e nove indivíduos com diagnóstico comprovado de DC, três homens (7,7%) e 36 mulheres (92,3%), responderam a um questionário. Todos os pacientes foram acompanhados no ambulatório do Hospital de Clínicas de Porto Alegre, alterações relacionadas à tireoide foram detectadas por meio da medição dos níveis de hormônio estimulador da tireoide (TSH), tiroxina livre (T4 livre) e intraanticorpos antitireoperoxidase (anti-TPO), no soro.RESULTADOS: Três pacientes apresentavam lesões tipo 1; um paciente apresentava tipo 2; 13 pacientes, tipo 3a; oito pacientes, tipo 3b; e 14 tipo 3c, de acordo com a classificação modificada de Marsh. A média de idade foi de 50,8 ± 12,9 anos. Vinte e nove (74,4%) pacientes apresentaram tireoide nomal. Dez indivíduos (25,6%) apresentaram hipotireoidismo: oito pacientes (20,5%) já usavam levotiroxina e dois pacientes (5,1%) com hipotireoidismo subclínico foram identificados durante a avaliação. Nenhum paciente apresentou hipertireoidismo. Os níveis médios séricos de T4L foram 1,15 ± 0,25 ng/dl. A mediana do TSH sérico foi 2,39 (0,06-6,64) UI/ml. Níveis séricos de anti-TPO foram positivos em seis pacientes (15,4%), um com hipotireoidismo, um com hipotireoidismo subclínico e quatro com função tireoidiana normal. Não se identificou fator associado a anormalidades relacionadas à tireoide. CONCLUSÃO: Este estudo demonstrou uma alta prevalência de anormalidades tireoidianas, como observado em outros estudos. Uma vez que os anticorpos antitireoide estavam presentes em alguns pacientes com função tireoidiana normal, recomenda-se a avaliação periódica da função tireoidiana em pacientes com DC.

Avaliação da expressão imunoistoquímica da proteína p16INK4a no adenocarcinoma de esôfago / Protein p16INK4a immunohistochemical expression in adenocarcinoma of the esophagus

INTRODUÇÃO: O adenocarcinoma de esôfago apresenta aumento de frequência nas últimas décadas, particularmente em países desenvolvidos. O esôfago de Barrett é reconhecido como a principal lesão precursora e o estudo da sequência metaplasia-displasia-adenocarcinoma mostra a ocorrência de alterações genéticas desde suas fases mais incipientes. As alterações no p16INK4a são relatadas como frequentes no esôfago de Barrett e no carcinoma de esôfago. OBJETIVO: Verificar a prevalência da expressão imunoistoquímica da proteína p16INK4a em exames anatomopatológicos de pacientes com adenocarcinoma de esôfago. MÉTODO: A população do estudo foi constituída de 37 pacientes com adenocarcinoma de esôfago. A expressão da proteína p16 foi detectada por meio de análise imunoistoquímica, com anticorpo primário p16INK4aAb-7, clone 16P07, NeoMarkers e avaliada de acordo com o Sistema de Escore de Imunorreatividade (Immunoreactive scoring system - IRS) modificado. RESULTADOS: No grupo houve predominância de pacientes do sexo masculino (86,5 por cento) e a maioria dos casos correspondia a estádios avançados (III e IV = 67,5 por cento). Em 12 casos (32,4 por cento) foi identificada expressão imunoistoquímica da proteína p16INK4a. Não foi observada relação significativa entre a perda da expressão da proteína p16INK4a e o grau de diferenciação histológica (p=0,81) nem com o estadiamento da doença (p=0,485). CONCLUSÃO: Ocorre perda da expressão imunoistoquímica da proteína p16INK4a, corroborando as informações de que a inativação do gene p16 é um evento frequente e que pode exercer papel importante na carcinogênese do adenocarcinoma de esôfago.

BACKGROUND: The esophageal adenocarcinoma shows an increasing frequence in the last decades, specially in the developed countries. The Barrett´s esophagus is accepted as the major premalignant lesion and the metaplasia-dysplasia-adenocarcinoma sequence presents a lot of genetic changes since its early events. The alterations in p16INK4a are frequent in Barrett´s esophagus and esophageal carcinoma. AIM: To verify the prevalence of the immunohistochemical expression of the p16INK4a protein in patients with esophageal adenocarcinoma. METHODS: The study population consisted of 37 patients with resected esophageal adenocarcinoma. The p16INK4a protein expression was determined by immunohistochemistry using primary antibody p16INK4aAb-7, clone 16P07 NeoMarkers and assessed according to the Immunoreactive scoring system (IRS). RESULTS: Of 37 analyzed patients, the most were male (86,5 percent) and the advanced disease was predominant (stages III and IV = 67,5 percent). In 12 (32,4 percent) the immunohistochemistry was positive for p16INK4a.There was no significative relation between the protein expression and the degrees of histological differentiation of the biopsies and surgical especimens (p=0,81) neither with the staging (p=0,485). CONCLUSION: The lost of the immunohistochemical expression of the p16INK4a protein in this study suggests that p16 is enroled in the carcinogenesis of the adenocarcinoma of esophagus.

Chlamydia trachomatis and human papillomavirus coinfection: association with p16INK4a and Ki67 expression in biopsies of patients with pre-neoplastic and neoplastic lesions

The objective of this study was to identify the frequency of coinfection by human papillomavirus (HPV) and Chlamydia trachomatis (CT) in cervical lesions and relate it with immunohistochemical expression of p16INK4a and Ki67, both oncogenicity markers. A cross-sectional study with 86 women from primary care units in southern Brazil was conducted. Cervical swabs were collected for HPV-DNA and CT-DNA detection, through the polymerase chain reaction technique (PCR). The immunohistochemical analysis was performed on biopsy cervical tissue material to identify the expression of p16INK4a and Ki67 cell cycle markers. About 83 percent were positive for HPV-DNA and 19 percent had coinfection with CT-DNA. Among coinfected women, 56 percent expressed p16INK4a. There was a statistically significant association between the histological grade of the lesion and Ki67 expression. All high-grade lesions, 50 percent of low-grade lesions and 31 percent of negative biopsies expressed Ki67 (p = 0.004). A total of 37 percent of coinfected women expressed both markers. In conclusion, although more than half of the coinfected patients have expressed p16INK4a and more than one third have expressed both markers, these results suggest no association between those variables. However, other studies involving larger samples are necessary to corroborate such findings.

O modelo experimental de carcinogênese gástrica induzido por n-methyl-n-nitrosourea em ratos F344 e camundongos C3H é válido para os ratos Wistar? / Experimental model of gastric carcinogenesis with N-methyl-N-nitrosourea for F344 rats and C3H mices is valid for Wistar rats?

INTRODUÇÃO: O N-metil-N-nitrosourea (MNU) tem ação cancerígena direta, induzindo tumores em várias espécies em uma variedade de órgãos, incluindo o estômago de ratos. Tratamento do MNU na água de beber por 25-42 semanas, seletivamente, induz carcinoma gástrico glandular de ratos F344 e camundongos C3H. OBJETIVO: Estabelecer um modelo experimental para indução seletiva de câncer no estômago glandular de ratos Wistar com MNU. MÉTODOS: Um total de 48 ratos Wistar machos com oito semanas, foram utilizados no presente estudo. MNU (Sigma-Aldrich) foi dissolvido em DMSO e liberada água potável ad libitum por um período variando de 16 a 70 semanas. Após 16 semanas, quatro ratos foram selecionados aleatoriamente e mortos. Depois, de seis em seis semanas, quatro animais também foram mortos até 70 semanas. RESULTADOS: A taxa de sobrevivência foi superior a 90 por cento. Ocorreu a indução de dois adenocarcinomas, um carcinoma espinocelular e um sarcoma. A incidência de adenocarcinoma gástrico foi de 4,5 por cento (0,5 a 15). CONCLUSÕES: O modelo experimental de carcinogênese gástrica em ratos Wistar, utilizando MNU dissolvido na água, não mostrou viabilidade prática neste estudo, devido à baixa taxa de adenocarcinoma gástrico que ocorreu.

BACKGROUND: The N-methyl-N-nitrosourea (MNU) is a direct acting carcinogen, inducing tumors in several species in a variety of organs, including stomach of rats. Treatment of MNU in the drinking water for 25-42 weeks selectively induced glandular gastric carcinoma in F344 rats and C3H mice. AIM: To establish an experimental model for selective MNU induction of glandular stomach cancer in Wistar rats. METHODS: A total of 48 males eight-week-old Wistar rats were used in the present study. MNU (Sigma-Aldrich) was dissolved in DMSO and provided as the drinking water ad libitum for a period ranging from 16 to 70 weeks. After 16 weeks, four rats were randomly selected and killed. After every six weeks four animals were killed until 70 weeks. RESULTS: Survival rate was higher than 90 percent. It had the induction of two adenocarcinomas, one squamous cell carcinoma and one sarcoma. The incidence of gastric adenocarcinoma was 4.5 percent (0.5 to 15). CONCLUSIONS: The experimental model of gastric carcinogenesis in Wistar rats, using MNU dissolved in water, showed not practice viability in this study due to the low rate of gastric adenocarcinoma.

Adipocyte morphometric evaluation and angiogenesis in the omentum transposed to the breast: a preliminary study

OBJECTIVE: The purpose of this study was to describe the probable mechanism of the volume increase of laparoscopically harvested omentum flaps used to treat breast deformities. METHODS: A histological analysis of omentum samples was performed to study the volume increase of laparoscopically harvested omentum flaps. Samples were harvested immediately after the transposition of the omentum from the abdominal cavity to the breast region and during the second surgical procedure for breast symmetrization of eight patients submitted to the transposition of the omentum flap. Changes in the morphometric measurements of the adipocytes (perimeter, diameter, and area), microvascular density (as measured by the CD31 endothelial marker), and immunohistochemical expression of VEGF were documented. RESULTS: The increases in adipocyte size and microvascular density were statistically significant (P < 0.012). The expression levels of VEGF were lower in the second set of samples when compared to the first set, but the differences were not statistically significant (P < 0.093). CONCLUSION: These results demonstrate an increase in cellular volume as measured by adipocyte perimeter, diameter, and area. Moreover, the increase in the number of vessels in the second set of samples suggests that neoangiogenesis was stimulated by the initial increase in VEGF expression levels observed in the first set of samples. The increase in VEGF expression in the flap may have been caused by adipocyte hypertrophy resulting from neoangiogenesis.

Expressão imunohistoquímica de P53 e Ki-67 na carcinogênese esofágica induzida pela dietilnitrosamina: modelo experimental / Immunohistochemical expression of P53 and Ki-67 in diethylnitrosamine-induced carcinomas esophageal: experimental model

OBJETIVO: Avaliar a expressão imunohistoquímica de p53 e ki-67 na carcinogênese esofágica induzida quimicamente através do uso de dietilnitrosamina, em um grupo de 100 camundongos fêmeas. MÉTODOS: O estudo experimental foi realizado com quatro grupos de animais, onde os grupos I e II foram considerados controles, sendo diferenciados por gavagem esofágica, uma vez semana, com água fria (temperatura ambiente) ou quente (60º-70ºC). E os grupos III e IV foram considerados estudos, os quais receberam dietilnitrosamina por três dias consecutivos semanalmente, também sendo diferenciados por gavagem, uma vez por semana, com água fria ou quente. O estudo apresentou datas progressivas de sacrifícios com coleta de peças esofágicas, que iniciava aos 30 dias de experimento e terminava aos 150 dias. Demonstrou-se que não houve diferença na incidência tumoral quando foi acrescida a variável temperatura da água; provavelmente devido ao episódio único semanal que era adicionado ao animal em experimentação. RESULTADOS: A análise imunohistoquímica do p53 não evidenciou diferença estatística durante a evolução da carcinogênese até 150 dias, porém quando analisado a relação com alterações patológicas demonstra-se que apresenta significância em relação à patologia baixo grau de displasia, alto grau e carcinoma. CONCLUSÃO: A análise imunohistoquímica do ki-67 demonstrou diferença estatística durante a evolução da carcinogênese a partir do dia 120 de experimento e quando analisada a relação com alterações patológicas demonstrou-se que apresenta significância também em relação à lesão intraepitelial de alto grau e carcinoma.

OBJECTIVE:To evaluate the expression of P53 and Ki-67 during esophageal diethylnitrosamine (DEN)-induced carcinogenesis in 100 mice by immunohistochemistry. METHODS: The animals were assigned to 4 groups, receiving water and food ad libitum. Control groups I and II received weekly esophageal gavage with cold (room temperature) or hot (60-70ºC) water, respectively. Experimental groups III and IV were treated with DEN for 3 consecutive days during the week, and one weekly gavage as above. The mice were sacrificed in different periods from day 30 to day 150 after the beginning of the experiment, for collection of esophageal samples which were then submitted to microscopic and immunohistochemical analyses. The temperature of the water administered by gavage was not related to the frequency of esophageal tumors. RESULTS:The expression of Ki-67 was significantly higher in high-grade intraepithelial lesion (I.L.), and the expression of P53 was also higher in low-grade I.L. CONCLUSION:The results emphasize the direct relationship of the carcinogenic process with early cell alterations detected by immunohistochemistry.

Laparoscopic treatment of Poland's syndrome using the omentum flap technique

OBJECTIVE: For patients with Poland's syndrome, a transverse skin fold in the anterior axillary pillar, infra-clavicular depression and an anomalous breast contour are the most uncomfortable disfigurements. This study aims to demonstrate that superior aesthetic results can be achieved by using a laparoscopically harvested omentum flap to treat this condition. METHODS: From a prospectively maintained clinical database of patients undergoing a laparoscopic omentum flap procedure for breast reconstruction, all of the patients with Poland's syndrome were identified and their outcomes were studied. RESULTS: Thirteen consecutive patients with Poland's syndrome were treated and evaluated regarding breast contour, reconstruction of the anterior axillary pillar and filling of the infra-clavicular depression. Implants were employed beneath the flap in 76 percent of cases to improve symmetry. In 23 percent of cases, a contra-lateral mastopexy was performed, and in 15 percent of cases, a breast implant was used. The consistency of the flap is similar to natural breast tissue and only a small incision in the breast fold is needed. The majority of patients (85 percent) were female, with a mean age of 26 (18-53). The flap is extremely malleable, adapts to irregular surfaces, and has a long vascular pedicle. Additionally, its removal does not leave a scar at the donor site as the removal of muscular flaps does. For example, the removal of the latissimus dorsi flap causes a deformity in the dorsal contour. The mean operative time was 201 minutes (80-350) and the mean hospital stay was 2.3 days (1-5). CONCLUSIONS: The outcomes of these patients revealed that the omentum flap technique provided superior amelioration of the deformities caused by Poland's syndrome when compared with other reconstructive options.

A normativa do Hospital de Clínicas de Porto Alegre para o armazenamento e utilização de materiais biológicos humanos e informações associadas em pesquisa: uma proposta interdisciplinar / The Hospital de Clínicas de Porto Alegre normative for the storage and use of human biological materials and their associated information in research: an interdisciplinary approach

Este artigo apresenta a justificativa e o processo de elaboração do marco normativo do Hospital de Clínicas de Porto Alegre (HCPA) para o armazenamento e utilização de materiais biológicos humanos e suas informações associadas em atividades de pesquisa. Um grupo de trabalho multiprofissional se reuniu e discutiu todos os aspectos referentes a esta questão que envolve as atividades de biobanco e de biorrepositórios. Como produto deste trabalho é apresentada a Normativa Institucional.

This article presents the rationale and framework for developing the Hospital de Clinicas de Porto Alegre (HCPA) normative for storage and use of human biological material and their associated information on research activities. A multidisciplinary task force was established and all issues related to biobanks and biorepositories activities in periodic meetings were discussed; the final result is presented as an Institutional Normative.

Immunoreactivity of cytokeratins 7 and 20 in goblet cells and columnar blue cells in patients with endoscopic evidence of Barrett's esophagus / Imunoreatividade das citoqueratinas 7 e 20 nas células caliciformes e células colunares azuis em pacientes com evidência endoscópica de esôfago de Barrett

CONTEXT: Barrett's esophagus is characterized by the presence of goblet cells. However, when alcian-blue is utilized, another type of cells, called columnar blue cells, is frequently present in the distal esophagus of patients with endoscopic evidence of Barrett's esophagus. Cytokeratin 7 and 20 immunoreactivity has been previously studied in areas of intestinal metaplasia at the esophagogastric junction. However, the expression of these cytokeratins in columnar blue cells has not been characterized. OBJECTIVE: To compare the expression of cytokeratin 7 and 20 in goblet cells and columnar blue cells in patients with endoscopic evidence of Barrett's esophagus. METHODS: Biopsies from 86 patients with endoscopic evidence of Barrett's esophagus were evaluated. The biopsies were stained for cytokeratin 7 and 20. RESULTS: Goblet cells were present in 75 cases and columnar blue cells in 50 cases. Overall, cytokeratin 7 expression was similar in goblet cells and columnar blue cells (P = 0.25), while cytokeratin 20 was more common in goblet cells (P <0.001). In individuals with both cell types, however, cytokeratin 7 staining was the same in goblet and columnar blue cells in 95 percent of the cases, and cytokeratin 20 staining was the same in 77 percent. CONCLUSION: Goblet cells and columnar blue cells have similar immunohistochemical staining patterns for cytokeratins 7 and 20 in patients with endoscopic evidence of Barrett's esophagus.

CONTEXTO: Esôfago de Barrett é caracterizado pela presença de células caliciformes. Entretanto, quando "alcian blue" é utilizado, outro tipo de células, chamadas células colunares azuis, estão frequentemente presentes no esôfago distal de pacientes com evidência endoscópica de esôfago de Barrett. A imunoreatividade das citoqueratinas 7 e 20 tem sido estudada previamente em áreas de metaplasia intestinal na junção esôfago-gástrica. Entretanto, a expressão destas citoqueratinas nas células colunares azuis não foi caracterizada. OBJETIVO: Comparar a expressão das citoqueratinas 7 e 20 nas células caliciformes e células colunares azuis em pacientes com evidência endoscópica de esôfago de Barrett. MÉTODOS: Biopsias de 86 pacientes com evidência endoscópica de esôfago de Barrett foram avaliadas. Estas foram coradas com citoqueratinas 7 e 20. RESULTADOS: Células caliciformes estavam presentes em 75 casos e células colunares azuis em 50 casos. Ao todo, a expressão da citoqueratina 7 foi similar nas células caliciformes e células colunares azuis (P = 0,25), enquanto que a da citoqueratina 20 foi mais comum nas células caliciformes (P<0,001). Por outro lado, em indivíduos apresentando ambos os tipos de células, a coloração da citoqueratina 7 foi a mesma nas células caliciformes e células colunares azuis em 95 por cento dos casos, e a coloração da citoqueratina 20 foi a mesma em 77 por cento. CONCLUSÃO: As células caliciformes e as células colunares azuis têm padrões similares de coloração imunoistoquímica para citoqueratina 7 e 20 em pacientes com evidência endoscópica de esôfago de Barrett.

Immunohistochemical evaluation for P53 and VEGF (Vascular Endothelial Growth Factor) is not prognostic for long term survival in end stage esophageal adenocarcinoma / Avaliação imunoistoquímica do P53 e VEGF (Fator de Crescimetno Endotelial Vascular) não representa fator prognóstico para sobrevida a longo prazo na fase terminal do adenocarcinoma do esôfago

OBJECTIVES: To correlate the expression of p53 protein and VEGF with the prognosis of patients submitted to curative resection to treat esophageal adenocarcinoma. METHODS: Forty-six patients with esophageal adenocarcinoma, submitted to curative resection, were studied. The expressions of p53 protein and VEGF were assessed by immunohistochemistry in 52.2 percent and 47.8 percent of tumors, respectively. RESULTS: P53 protein and VEGF expressions coincided in 26 percent of the cases, and no correlation between these expressions was observed. None of the clinicopathological factors showed a significant correlation with p53 protein or VEGF expressions. There was no significant association between p53 protein and VEGF expressions and long-term survival. CONCLUSION: The expression of p53 protein and VEGF did not correlate with prognosis in esophageal adenocarcinoma patients submitted to curative resection.

OBJETIVO: Correlacionar a expressão do p53 e VEGF com o prognóstico de pacientes submetidos à operação curativa para tratar adenocarcinoma do esôfago. MÉTODO: Foram estudados 46 pacientes com adenocarcinoma de esôfago, submetidos à ressecções curativas. As expressões do p53 e VEGF foram assessadas por imunoistoquímica em 52.2 por cento e 47.8 por cento dos tumors, respectivamente . RESULTADOS: As expressões de ambos coincidiram em 26 por cento dos casos sem correlação entre elas. Os fatores clinicopatológicos estudados não mostraram correlação significante. Não houve associação significante entre as expresses do p53 e VEGF na sobrevida a longo prazo. CONCLUSÃO: As expressões do p53 e VEGF não se correlacionaram com o prognóstico do adenocarcinoma do esôfago nos pacientes operados com ressecções curativas.

Osso liofilizado bovino não-desmineralizado com células-tronco mesenquimais para engenharia tecidual: estudo experimental em sítio heterotópico / Bovine non-demineralized lyophilized bone with mesenchymal stem cells for tissue engineering: experimental study in heterotopic site

Objetivo: Avaliar o uso de células da medula óssea, com potencial osteogênico, agregadas a estrutura tridimensional de osso liofilizado bovino não-desmineralizado para engenharia tecidual óssea. Método: Os animais doadores de células da medula óssea, assim como os animais receptores dos construtos ósseos, foram camundongos de linhagem isogênica C57Bl/6. Utilizou-se modelo experimental heterotópico, com a implantação de construtos de osso liofilizado bovino não-desmineralizado (OL) no plano subcutâneo no dorso dos animais. Foram organizados 4 grupos de comparação (n=10 animais em cada grupo): 1) OL isoladamente (grupo controle); 2) OL + células mononucleares da medula (CMM); 3) OL + células-tronco mesenquimais (CTM); 4) OL + células-tronco mesenquimais diferenciadas em meio osteoindutor (CTMdif). A aferição foi realizada após 5 semanas, com avaliação histológica e determinação da atividade de fosfatase alcalina. Resultados: A avaliação histológica não mostrou diferença entre os grupos de comparação, com a observação em todas as amostras de tecido conjuntivo fibroso rico em neovasos estendendo-se por entre as trabéculas ósseas, sem osteoblastos ou osteócitos viáveis e sem neoformação óssea. Os resultados da atividade de fosfatase alcalina também não mostraram diferença entre os grupos de comparação, com análise de variância entre os grupos mostrando p=0,867. Conclusões: Os resultados mostraram que, no modelo estudado e com os métodos utilizados, a adição de células da medula óssea com potencial osteogênico sobre estrutura de osso liofilizado bovino não-desmineralizado não agregou propriedades osteogênicas ao material. Este estudo não confirmou a perspectiva inicial de utilizá-lo como estrutura tridimensional e carreadora celular na engenharia tecidual óssea, sendo necessários estudos subseqüentes que o avaliem em outros modelos experimentais, e que explorem separadamente cada etapa metodológica que possa influir no sucesso da engenharia tecidual óssea.

Objective: To evaluate the use of bone marrow cells with osteogenic potential seeded on bovine nondemineralized lyophilized bone scaffolds for bone tissue engineering. Method: Bone marrow cells donors, as well as the receptors of the bone constructs were C57BI/6 isogenic line mice. A heterotopic experimental model was used, with implantation of the constructs into subcutaneous pouches on the backs of the animals. Four comparison groups were set (n=10 animals each group): 1) LB alone (control group); 2) LB + marrow mononuclear cells (MMC); 3) LB + mesenchymal stem cells (MST); 4) LB + mesenchymal stem cells differentiated in osteoinductive medium (MSTdif). The constructs were harvested 5 weeks after implantation for histological analysis and alkaline phosphatase activity test. Results: The histological analysis did not show differences among the comparison groups. In all samples fibrous connective tissue rich in neovessels was observed extending through bone trabeculae, without viable osteoblasts or osteocytes and without new bone formation. Likewise, results of the alkaline phosphatase activity have not shown any difference among comparison groups, with the analysis of variance between groups showing p value=0.867. Conclusions: In this experimental model and with the methods used, the addition of bone marrow cells with osteogenic potential to a bovine non-demineralized lyophilized bone structure did not add osteogenic properties to the material. The initial perspective of using it as a scaffold for bone tissue engineering could not be confirmed, and further studies are required to assess it in other experimental models, and to explore separately each methodological step that might influence the success of bone tissue engineering.

Utilização de células-tronco autólogas de medula óssea na regeneração do nervo tibial de coelhos mediante técnica de tubulização com prótese de silicone / Applyng of autologous stem cells from bone marrow in the regeneration of tibial nerve of rabbits using the tubulization technique with silicone tube

Neste estudo é apresentado um modelo experimental de defeito agudo em nervo periférico para avaliação da regeneração nervosa mediante técnica de tubulização associada à inoculação de células-tronco autólogas de medula óssea. Foram utilizados 12 coelhos Nova Zelândia albinos, submetidos à secção bilateral e ao afastamento de 5mm do nervo tibial e posterior reparo mediante utilização de câmara de silicone. Internamente à prótese de tubulização do nervo tibial esquerdo em todos os animais, foram inoculadas células-tronco autólogas de medula óssea, coletadas a partir do úmero. Como grupo controle (nervo tibial direito), mediante aplicação da mesma técnica de reparo, solução de NaCl 0,9 por cento foi administrada internamente à prótese. Após 30 dias de observação, os animais foram eutanasiados e foi realizada a avaliação histológica dos segmentos nervosos por meio das colorações de hematoxilina-eosina, luxol fast blue e azul de toluidina. Com os resultados, foi possível concluir que o transplante de células-tronco autólogas associado à técnica de tubulização apresenta vantagens no processo de regeneração nervosa periférica.

This study presents an experimental model of an acute deffect in a peripheral nerve to evaluate neural regeneration using a tubulization technique associated with the inoculation of autologous stem cells from bone marrow. A total of 12 New Zealand white rabbits underwent a bilateral dissection of the tibial nerve followed by repair with silicone tubulization. On the left tibial nerve of all animals, the tube was filled with autologous bone marrow-derived stem cells collected from the humerus. For control, using the same repair technique, the tubes were filled with a NaCl solution in the right tibial nerve. After 30 days of observation, the animals were euthanized and a histological evaluation of the collected nerve segments was performed by staining with hematoxylin-eosin, luxol fast blue, and toluidine blue. From the results it is possible to conclude that the transplanted autologous stem cells associated with the tubulization technique present an advantage in the peripheral nerve regeneration process.