Hyaluronidases and hyaluronan synthases expression is inversely correlated with malignancy in lung/bronchial pre-neoplastic and neoplastic lesions, affecting prognosis

We collected a series of 136 lung/bronchial and 56 matched lung parenchyma tissue samples from patients who underwent lung/bronchial biopsies and presented invasive carcinoma after lung surgery. The lung/bronchial samples included basal cell hyperplasia, squamous metaplasia, moderate dysplasia, adenomatous hyperplasia, severe dysplasia, squamous cell carcinoma and adenocarcinoma. Matched lung parenchyma tissue samples included 25 squamous cell carcinomas and 31 adenocarcinomas. Immunohistochemistry was performed to analyze for the distribution of hyaluronidase (Hyal)-1 and −3, and hyaluronan synthases (HAS)-1, −2, and −3. Hyal-1 showed significantly higher expression in basal cell hyperplasia than in moderate dysplasia (P=0.01), atypical adenomatous hyperplasia (P=0.0001), or severe dysplasia (P=0.03). Lower expression of Hyal-3 was found in atypical adenomatous hyperplasia than in basal cell hyperplasia (P=0.01) or moderate dysplasia (P=0.02). HAS-2 was significantly higher in severe dysplasia (P=0.002) and in squamous metaplasia (P=0.04) compared with basal cell hyperplasia. HAS-3 was significantly expressed in basal cell hyperplasia compared with atypical adenomatous hyperplasia (P=0.05) and severe dysplasia (P=0.02). Lower expression of HAS-3 was found in severe dysplasia compared with squamous metaplasia (P=0.01) and moderate dysplasia (P=0.01). Epithelial Hyal-1 and −3 and HAS-1, −2, and −3 expressions were significantly higher in pre-neoplastic lesions than in neoplastic lesions. Comparative Cox multivariate analysis controlled by N stage and histologic tumor type showed that patients with high HAS-3 expression in pre-neoplastic cells obtained by lung/bronchial biopsy presented a significantly higher risk of death (HR=1.19; P=0.04). We concluded that localization of Hyal and HAS in lung/bronchial pre-neoplastic and neoplastic lesions was inversely related to malignancy, which implied that visualizing these factors could be a useful diagnostic procedure for suspected lung cancer. Finalizing this conclusion will require a wider study in a randomized and prospective trial.

Violence against adolescents: an analysis based on the categories gender and generation / La violencia contra adolescentes: un análisis a la luz de las categorías género y generación / Violência contra adolescentes: uma análise à luz das categorias gênero e geração

Exploratory and descriptive study based on quantitative and qualitative methods that analyze the phenomenon of violence against adolescents based on gender and generational categories. The data source was reports of violence from the Curitiba Protection Network from 2010 to 2012 and semi-structured interviews with 16 sheltered adolescents. Quantitative data were analyzed using SPSS software version 20.0 and the qualitative data were subjected to content analysis. The adolescents were victims of violence in the household and outside of the family environment, as victims or viewers of violence. The violence was experienced at home, mostly toward girls, with marked overtones of gender violence. More than indicating the magnitude of the issue, this study can give information to help qualify the assistance given to victimized people and address how to face this issue.

Objetivo Analizar la violencia contra los adolescentes a la luz de las categorías de género y generación. Método Estudio exploratorio, descriptivo, de abordaje cuantitativo y cualitativo que. Las fuentes de datos fueron las denuncias de violencia mantenidos por la Red de Protección en Curitiba entre los años 2010-2012 y entrevistas semi-estructuradas con 16 adolescentes alojados. Las variables cuantitativas se analizaron mediante el programa SPSS y los cualitativos por la análisis de contenido. Resultados Los adolescentes fueron sometidos a la violencia en el hogar y en el exterior, como víctimas o espectadores. La violencia fue más frecuente en el hogar, centrándose principalmente en las chicas con matices marcados de violencia de género. Conclusión Más que encontrar la magnitud del problema, el estudio puede servir de base para calificar la asistencia a las personas víctimas de este fenómeno.

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Objetivo Analisar a violência contra o adolescente à luz das categorias gênero e geração. Método Estudo exploratório, descritivo, de abordagem quantitativa e qualitativa. As fontes de dados foram as notificações de violência da Rede de Proteção do município de Curitiba, de 2010 a 2012, e entrevistas semiestruturadas com 16 adolescentes abrigados. As variáveis quantitativas foram analisadas pelo software SPSS e os dados qualitativos através da análise de conteúdo. Resultados Os adolescentes foram submetidos à violência no ambiente doméstico e fora dele, como vítimas ou como espectadores. Prevaleceu no domicílio, incidindo principalmente sobre as meninas, com marcada conotação de violência de gênero. Conclusão Mais que constatar a magnitude do problema, o estudo pode fornecer subsídios para qualificar a assistência prestada aos sujeitos vitimizados e subsidiar o enfrentamento do fenômeno. .

Immunohistochemical detection of human telomerase reverse transcriptase in oral cancer and pre-cancer.

Purpose: Telomerase is a specialized ribonucleoprotein complex that stabilizes telomeres by adding "TAG" repeats to the end of chromosomes. The catalytic subunit of telomerase is human telomerase reverse transcriptase (hTERT), whose expression is the critical determinant of telomerase activity. Telomeres and telomerases play an important role in the longevity of cell and are known to conform "immortalization" on neoplastic cells. Although there exists a lot of information on telomerase in oral cancer, very little is known about their expression in leukoplakia and oral submucous fibrosis (OSF). This study addresses this lacuna. Materials and Methods: In this preliminary study, immunohistochemistry (IHC) was used to detect the expression of hTERT protein in oral squamous cell carcinoma (OSCC) (n=30), leukoplakia (n=15), OSF (n=15) and normal oral mucosa (n=10). The cellular localization of immunostain, intensity of stain, mean nuclear labeling index (LI) and mean nuclear labeling score (LS) of hTERT protein were studied. A total number of 1000 cells were counted in each slide. All the data were analyzed using SPSS software version 10.0.2. The cellular localization of cytoplasmic/nuclear/both of hTERT stain, staining intensity and LI were compared across the groups using Pearson's χ2 test. The mean LI and LS for OSF, leukoplakia, OSCC and normal were compared using analysis of variance (ANOVA). A P-value <0.05 was considered to be statistically significant. Results: The mean nuclear LI increased from OSF (22.46±4.53), through normal (28.3±12.3) to OSCC (47.56±21.30) (P=0.002) and from normal (28.3±12.3), through leukoplakia (44.06±14.6), to OSCC (47.56±21.30) (P=0.00). The mean nuclear labeling score was observed to increase from OSF (37.8±15), through normal (64.9±30.7), to OSCC samples (106.9±29.77) (P=0.00) and from normal (64.9±30.7), through leukoplakia (85.6±25.1) to OSCC samples (106.9±29.77) (P=0.00). Conclusion: There was increased expression of hTERT protein in OSCC and leukoplakia samples when compared to normal oral mucosa. The cellular localization, LI and LS in OSF were significantly different from OSCC and leukoplakia.

Water extracts of cabbage and kale inhibit ex vivo H2O2-induced DNA damage but not rat hepatocarcinogenesis

The chemopreventive potential of water extracts of the Brassica vegetables cabbage and kale was evaluated by administering their aqueous extracts in drinking water ad libitum to Wistar rats submitted to Ito’s hepatocarcinogenesis model (CB group and K group, respectively - 14 rats per group). Animals submitted to this same model and treated with water were used as controls (W group - 15 rats). Treatment with the vegetable extracts did not inhibit (P > 0.05) placental glutathione S-transferase-positive preneoplastic lesions (PNL). The number of apoptotic bodies did not differ (P > 0.05) among the experimental groups. Ex vivo hydrogen peroxide treatment of rat livers resulted in lower (P < 0.05) DNA strand breakage in cabbage- (107.6 ± 7.8 µm) and kale- (110.8 ± 10.0 µm) treated animals compared with control (120.9 ± 12.7 µm), as evaluated by the single cell gel (comet) assay. Treatment with cabbage (2 ± 0.3 µg/g) or kale (4 ± 0.2 µg/g) resulted in increased (P < 0.05) hepatic lutein concentration compared with control (0.5 ± 0.07 µg/g). Despite the absence of inhibitory effects of cabbage and kale aqueous extracts on PNL, these Brassica vegetables presented protection against DNA damage, an effect possibly related to increased hepatic lutein concentrations. However, it must be pointed out that the cause-effect relationship between lutein levels and protection is hypothetical and remains to be demonstrated.

Immunohistochemical Analysis of Nuclear Factor, p38, and Cyclin D1 Proteins in Premalignant Lesions and Carcinomas of the Colorectal Mucosa / 대한소화기학회지

BACKGROUND/AIMS: Nuclear factor-kappa B p65 (NF-kappa B p65), nuclear factor-kappa B1 p50 (NF-kappa B p50) have been shown to play a role in cell proliferation, apoptosis, cytokine production, and oncogenesis. Recently, p38 mitogen-activated protein kinase (MAPK)/ NF-kappa B/ cyclin D1 signaling pathway has been shown to play an important part in the pathogenesis of human cancers. This study was designed to investigate the expression of NF-kappa B p65, NF-kappa B p50, p38 MAPK alpha, and cyclin D1 proteins in premalignant lesions of colon and colorectal adenocarcinoma. METHODS: Paraffin sections of 20 normal mucosa, 20 low-grade tubular adenoma, 20 high-grade tubular adenoma and 64 adenocarcinoma tissues were analysed immunohistochemically for the expression of NF-kappa B p65, NF-kappa B p50, p38 MAPK alpha, and cyclin D1 proteins. RESULTS: The expression of NF-kappa B p65, NF-kappa B p50, and p38 MAPK alpha proteins were significantly higher in adenocarcinoma tissue in comparison with that in normal mucosa, low-grade tubular adenoma, and high-grade tubular adenoma tissues. Expression of NF-kappa B p50 was more frequent in poorly differentiated histologic grade, presence of nodal metastasis, and advanced stage. Expression of p38 MAPK alpha protein was higher in advanced tumor stage, presence of nodal metastasis and advanced stage. Synchronous expression of NF-kappa B p65, NF-kappa B p50, p38 MAPK alpha, and cyclin D1 proteins were significantly higher in adenocarcinoma tissue. CONCULSIONS: With the increased expression of NF-kappa B p65, NF-kappa B p50, and p38 MAPK alpha proteins, p38 MAPK/ NF-kappa B/ cyclin D1 signaling pathway may play a role in the pathogenesis of colorectal carcinoma.

A relação do uso crônico de fenobarbital com áreas potencialmente pré-neoplásicas em fígado de ratos / Potentially pre-neoplasics areas in rat's liver associated to chronic use of phenobarbital

RACIONAL: O fenobarbital é utilizado em modelos experimentais não só por ser um importante agente promotor da carcinogênese em fígado de ratos, como também por ser não-genotóxico, órgão-específico e dose-dependente. OBJETIVOS: Avaliar o efeito da administração diária de fenobarbital em ratos, desde o nascimento até os 24 meses de idade, na ausência concomitante de administração de agentes químicos iniciadores da carcinogênese. MATERIAL E MÉTODOS: Um grupo controle de ratos machos Wistar recebeu dieta básica e a esta, do outro grupo, foi adicionado diariamente, fenobarbital a 0,05 por cento, durante 24 meses. Cortes dos lobos médio e direito do fígado foram submetidos ao processamento histológico e corados pela hematoxilina-eosina e coloração imunoistoquímica para a glutationa S-transferase forma placentária. RESULTADOS: Detectaram-se áreas glutationa S-transferase forma placentária positivas em ambos os grupos e as imagens foram analisadas quanto ao número e à extensão da superfície, mediante análise de imagem por histomorfometria. CONCLUSÃO: O uso crônico de fenobarbital não alterou o número de áreas glutationa S-transferase forma placentária positivas, havendo, no entanto, aumento no tamanho médio de áreas glutationa S-transferase forma placentária positivas, com conseqüente aumento da superfície glutationa S-transferase forma placentária positiva, sendo este aumento provavelmente relacionado a maior capacidade evolutiva dessas lesões e possível irreversibilidade das mesmas.

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 percent 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.

Altered expression of thioredoxin reductase-1 in dysplastic bile ducts and cholangiocarcinoma in a hamster model

Thioredoxin reductase 1 (TrxR) is a homodimeric selenoenzyme catalyzing thioredoxin (Trx) in an NADPHdependent manner. With regard to carcinogenesis, these redox proteins have been implicated in cell proliferation, transformation and anti-apoptosis. In the present study, using a hamster cholangiocarcinoma (ChC) model, we evaluated the immunohistochemical expression pattern of TrxR in precancerous lesions and ChCs as well as in normal bile ducts. The goal of this study was to determine the potential role and importance of TrxR in cholangiocarcinogenesis. For the ChC model, we obtained liver tissue specimens with dysplastic bile ducts prior to the development of ChC 8 weeks after initiation of the experiment and ChC samples at 27 weeks. The immunohistochemical analysis showed diffuse cytoplasmic overexpression of TrxR in the dysplastic bile duct epithelial cells as well as in cholangiocarcinoma; this was comparable to the negative or weakly positive in normal and type 1 hyperplastic bile ducts. However, TrxR appeared to be considerably down-regulated in the ChCs when compared to the higher expression observed in the dysplastic bile ducts. Therefore, these results suggest that TrxR overexpression followed by down-regulation might be an important event in cholangiocarcinogenesis, especially at early stages including the cellular transformation of candidate bile ducts. Further studies are however required to determine whether TrxR may be a potential target molecule for chemoprevention against cholangiocarcinogenesis. In addition, the molecular mechanism as well as the importance of the loss of TrxR in the development of cholangiocarcinoma, following dysplastic transformation of bile duct cells, also remains to be clarified.

Lipid peroxidation and antioxidant enzyme status in oral carcinoma patients.

OBJECTIVE: To measure the lipid peroxidation and endogenous antioxidant enzyme status in oral carcinoma and the protective role of exogenous antioxidants. MATERIAL AND METHODS: 20 new cases of histologically proven oral squamous cell carcinoma, 20 of leukoplakia and 20 age and sex matched healthy conrols were included. Intra oral pH of patients and controlled were measured by quantitative litmus paper test and serum was analysed for malonialdehyde (MDA), super oxide bismutase (SOD), catalase and glutathione peroxidase (GP). Patients with leukoplakia were treated with exogenous antioxidants for 3 months and the same were reassessed. RESULTS: Oral pH of oral cancer patients was neutral (PH-7) but that of leukoplakia and controls were mildly acidic (6.64 and 6.58 respectively). Serum malonialdehyde levels were highest in oral cancer group. With antioxidant enzymes super oxide bismutase, catalase and glutathione peroxidase different pattern was noticed. Antioxidant enzymes remained almost the same (P > 0.005 each) in patients with leukoplakia after 3 months of vitamin A,C and E. but there was marginal increase in catalase level (P<0.05). CONCLUSION: This study shows the positive benefit of vitamin (A,C,E) and nutrition supplementation on the antioxidant enzyme defense system hence prevention of oral carcinogenesis in patients with leukoplakia.

Helicobacter pylori and Telomerase Activity in Intestinal Metaplasia of the Stomach

BACKGROUND: Helicobacter pylori (H. pylori) has been considered a definitive carcinogen in gastric cancer. Telomerase is activated in gastric cancer and some premalignant gastric lesions, including intestinal metaplasia (IM). In this study, we evaluated the relationships of both H. pylori infection and telomerase activity with endoscopic and histologic features in IM. The effects of H. pylori eradication on endoscopic, histologic and biochemical changes were evaluated. METHODS: Endoscopic biopsies were obtained from 43 patients with IM for rapid urease, histologic and telomerase tests. The endoscopic and histologic features, H. pylori infection and telomerase were assessed. After H. pylori eradication, 15 patients were re-evaluated and compared after 4 months. RESULTS: Thirty-four (79.1%) patients were infected with H. pylori. The incidence of H. pylori infection was borderline correlated to the severity of IM (p=0.076). Telomerase was elevated in eight (18.6%) patients. Telomerase tends to be high in subtype III and endoscopic grade III of IM. After H. pylori eradication, endoscopic extent (p=0.039) and histologic severity (p=0.074) showed improvements, and telomerase decreased significantly (p=0.0001). CONCLUSION: Our data suggest that telomerase is associated with the severity and extent of IM and that H. pylori eradication improves the endoscopic and histologic features in IM, and decreases telomerase activity. H. pylori eradication can be considered one of the methods to prevent gastric cancer in patients with H. pylori-infected IM. Further long-term and large-scaled study will be needed.

Telomerase is strongly activated in hepatocellular carcinoma but not in chronic hepatitis and cirrhosis

Telomerase is highly activated in human immortal cell lines and tumor tissues, whereas it is not activated in primary cell strains and many tumor-adjacent tissues. It is suggested that telomerase activation is one of the critical steps in malignant transformation. In the present study, the telomerase activity was investigated in hepatocellular carcinoma tissues and non-tumor liver tissues from Korean patients with chronic hepatitis and cirrhosis. Eighty two liver tissues (24 chronic hepatitis specimens, 34 cirrhosis specimens, and 24 hepatocellular carcinomas) were obtained from 23 chronic viral hepatitis patients, 19 cirrhosis patients (including 7 liver transplants), and 24 patients with hepatocellular carcinoma, of which the surrounding non-tumor liver tissues were available in 16 patients (1 chronic hepatitis and 15 cirrhosis). As negative controls, 3 normal liver tissues were included. Protein from liver specimens was purified by a detergent lysis method as described elsewhere, and telomerase activity was measured in 2 diluents of each sample (1:1 and 1:100) by a telomeric repeat amplification protocol (TRAP). Telomerase was strongly activated in 79% (19/24) of the hepatocellular carcinomas, while weakly in 8% (2/24) of the chronic hepatitis tissues and in 24% (8/34) of the cirrhosis tissues. All of 3 normal control livers showed no telomerase activation. No relationship could be observed between the enhancement of telomerase activity and tumor nature. None of the chronic heaptitis or cirrhosis patients with mild telomerase activation in the liver have developed hepatocellular carcinoma for at least 2 years of follow-up period. These results suggest that the strong enhancement of telomerase activity may be a critical part of hepatocarcinogenesis, although the exact mechanism of such high activation in hepatocellular carcinoma is not clear. In addition, further study will be necessary to clarify the reason why no telomerase activity detectable by a conventional TRAP can be seen in some hepatocellular carcinoma.