RESUMO Contexto: Helicobacter pylori (H. pylori) é uma bactéria gram-negativa associada à etiologia de várias patologias do trato gastrointestinal, e cepas positivas para cagA (cagA+) são encontradas em populações com úlceras gástricas e lesões pré-cancerígenas, induzindo respostas pró-inflamatórias. O desenvolvimento de neoplasias está relacionado à desregulação do microRNA (miRNA), indicando miRNA-629 altamente expresso. O artigo tem como objetivo correlacionar o nível de expressão do miRNA-629 com a presença de H. pylori e o marcador de patogenicidade cagA. Métodos: Foram avaliadas 203 amostras de biópsia gástrica de indivíduos com tecido gástrico normal (n=60), gastrite (n=96) e câncer gástrico (n=47) de ambos os sexos e com mais de 18 anos. As amostras foram subdivididas de acordo com a presença ou ausência de H. pylori, detectado por reação em cadeia da polimerase (PCR). O RNA foi extraído usando um kit comercial e quantificado. O DNA complementar (cDNA) foi sintetizado usando kits comerciais, e a expressão relativa foi calculada usando o método 2-ΔΔCt. Resultados: Indivíduos infectados com H. pylori têm nove vezes mais chances de desenvolver câncer gástrico. Pacientes com câncer parecem ter diminuição da expressão do miRNA-629; no entanto, a presença da bactéria não influenciaria essa redução. Indivíduos no grupo do câncer apresentaram menor expressão do miRNA-629 quando cagA+; no entanto, no grupo controle, a expressão foi maior quando cagA+. Conclusão: H. pylori é um fator envolvido na etiologia e progressão das doenças gástricas. A redução na expressão do miRNA-629 em pacientes com câncer ocorre independentemente da presença da bactéria, mas quando o marcador de patogenicidade cagA está presente, induz mudanças na expressão gênica do respectivo miRNA. Contexto H (H gramnegativa gram negativa gastrointestinal cagA+ (cagA+ précancerígenas, précancerígenas pré cancerígenas, cancerígenas pré-cancerígenas próinflamatórias. próinflamatórias pró inflamatórias. inflamatórias pró-inflamatórias miRNA, miRNA , (miRNA) miRNA629 629 miRNA-62 expresso Métodos 20 n=60, n60 n n=60 60 (n=60) n=96 n96 96 (n=96 n=47 n47 47 (n=47 1 anos PCR. PCR . (PCR) quantificado cDNA (cDNA comerciais 2ΔΔCt. 2ΔΔCt ΔΔCt 2 ΔΔCt. 2-ΔΔCt Resultados entanto controle Conclusão presente (cagA (miRNA miRNA62 62 miRNA-6 n6 n=6 6 (n=60 n=9 n9 9 (n=9 n=4 n4 4 (n=4 (PCR miRNA6 miRNA- n= (n=6 (n= (n

ABSTRACT Background: Helicobacter pylori (H. pylori) is a gram-negative bacterium associated with the etiology of several gastrointestinal tract pathologies, and cagA-positive (cagA+) strains are found in populations with gastric ulcers and precancerous lesions, inducing pro-inflammatory responses. The development of neoplasms is related to microRNA (miRNA) dysregulation, indicating highly expressed miRNA-629. The article aims to correlate the expression level of miRNA-629 with the presence of H. pylori and the pathogenicity marker cagA. Methods: 203 gastric biopsy samples were evaluated from individuals with normal gastric tissue (n=60), gastritis (n=96), and gastric cancer (n=47) of both genders and over 18 years old. The samples were subdivided according to the presence or absence of H. pylori, detected by polymerase chain reaction (PCR). RNA was extracted using a commercial kit and quantified. Complementary DNA (cDNA) was synthesized using commercial kits, and the relative expression was calculated using the 2-ΔΔCt method. Results: Individuals infected with H. pylori are nine times more likely to develop gastric cancer. Cancer patients appeared to have decreased expression of miRNA-629; however, the presence of the bacterium would not influence this reduction. Individuals in the cancer group showed lower miRNA-629 expression when cagA+; however, in the control group, the expression was higher when cagA+. Conclusion: H. pylori is a factor involved in the etiology and progression of gastric diseases. Reduction in miRNA-629 expression in cancer patients occurs independent of the presence of the bacterium, but when the cagA pathogenicity marker is present, it induces changes in the gene expression of the respective miRNA. Background H (H gramnegative gram negative pathologies cagApositive positive cagA+ (cagA+ lesions proinflammatory pro inflammatory responses miRNA (miRNA dysregulation miRNA629. miRNA629 629. 629 miRNA-62 Methods 20 n=60, n60 n n=60 , 60 (n=60) n=96, n96 n=96 96 (n=96) n=47 n47 47 (n=47 1 old PCR. PCR . (PCR) quantified cDNA (cDNA kits 2ΔΔCt ΔΔCt 2 method Results however reduction Conclusion diseases present (cagA miRNA62 62 miRNA-6 n6 n=6 6 (n=60 n9 n=9 9 (n=96 n=4 n4 4 (n=4 (PCR miRNA6 miRNA- n= (n=6 (n=9 (n= (n

BACKGROUND: Recently, the importance of cytogenetics has grown in the diagnosis, prognosis and treatment of leukemias and myelodysplastic syndromes. 5-azacytidine is a drug that has well-known cytogenetical effects and is approved in the treatment of myelodysplastic syndromes. To date, no studies have been performed to evaluate the impact of 5-azacytidine on the chromosomes of patients with hematological neoplasias. This study aimed to investigate the effects of 5-azacytidine on chromosomes of patients with different hematological malignancies using G-band analyses to identify possible cytogenetical alterations. METHODS: The peripheral blood of 18 patients with hematological malignancies and 18 controls was collected in heparinized tubes. 5-azacytidine was added, at a final concentration of 10-5M, to cultures 7 hours prior to harvest. RESULTS: Uncoiled centromeric/pericentromeric heterochromatin of chromosomes-1, 9 and 16 occurred more frequently in the patients than in controls. This higher frequency of uncoiled heterochromatin was statistically significant (p-value = 0.004) for chromosome-9. Conversely, we observed that the fragile site at 19q13 was more frequent in controls (p-value = 0.0468). CONCLUSIONS: The results of this study suggest that satellite sequences, located in the heterochromatin of chromosome-9, are hypomethylated in hematological malignancies. This hypomethylation may contribute to the disease, activating transposable elements and/or promoting genomic instability, enabling the loss of heterozygosity of important tumor suppressor genes. An investigation of the 19q13 region may help to understand whether or not the predominant occurrence of the fragile site at 19q13 in controls is due to hypermethylation of this region.

An inflammatory process has been involved in numerous neurodegenerative disorders such as Parkinson's disease, stroke and Alzheimer's disease (AD). In AD, the inflammatory response is mainly located in the vicinity of amyloid plaques. Cytokines, such as interleukin-8 (IL-8) and interleukin-1α (IL-1α), have been clearly involved in this inflammatory process. Polymorphisms of several interleukin genes have been correlated to the risk of developing AD. The present study investigated the association of AD with polymorphisms IL-8 -251T > A (rs4073) and IL-1α-889C > T (rs1800587) and the interactive effect of both, adjusted by the Apolipoprotein E genotype. 199 blood samples from patients with AD, 146 healthy elderly controls and 95 healthy young controls were obtained. DNA samples were isolated from blood cells, and the PCR-RFLP method was used for genotyping. The genotype distributions of polymorphisms IL-8, IL-1α and APOE were as expected under Hardy-Weinberg equilibrium. The allele frequencies did not differ significantly among the three groups tested. As expected, the APOE4 allele was strongly associated with AD (p < 0.001). No association of AD with either the IL-1α or the IL-8 polymorphism was observed, nor was any interactive effect between both polymorphisms. These results confirm previous studies in other populations, in which polymorphisms IL-8 -251T > A and IL-1α-889C > T were not found to be risk factors for AD.

Helicobacter pylori is an important human pathogen that causes chronic gastritis and is associated with the development of peptic ulcer disease and gastric malignancies. The oral cavity has been implicated as a potential H. pylori reservoir and may therefore be involved in the reinfection of the stomach, which can sometimes occur following treatment of an H. pylori infection. The objectives of this paper were (i) to determine the presence of H. pylori in the oral cavity and (ii) to examine the relationship between oral H. pylori and subsequent gastritis. Gastric biopsies, saliva samples and dental plaques were obtained from 78 dyspeptic adults. DNA was extracted and evaluated for the presence of H. pylori using polymerase chain reaction and Southern blotting methods. Persons with gastritis were frequently positive for H. pylori in their stomachs (p < 0.0001) and there was a statistically significant correlation between the presence of H. pylori in gastric biopsies and the oral cavity (p < 0.0001). Our results suggest a relationship between gastric infection and the presence of this bacterium in the oral cavity. Despite this, H. pylori were present in the oral cavity with variable distribution between saliva and dental plaques, suggesting the existence of a reservoir for the species and a potential association with gastric reinfection.

The presence of human papillomavirus (HPV) was evaluated in 65 samples of prostate tumours and six samples of prostates with benign prostatic hyperplasia from individuals from Northern Brazil. We used a highly sensitive test, the Linear Array HPV Genotyping Test, to detect 37 high and low-risk HPV types. In this study, only 3% of tumour samples showed HPV infection. Our findings support the conclusion that, despite the high incidence of HPV infection in the geographic regions studied, HPV was not associated with a higher risk of prostate cancer. To our knowledge, this is the first study evaluating the frequency of HPV detection in prostatic tissue of individuals from Brazil.

Helicobacter pylori is a spiral-shaped Gram-negative bacterium. It colonizes the gastric mucosa of humans and persists for decades if not treated. Helicobacter pylori infection affects more than half of the world's population and invariably results in chronic gastritis. The cagA gene is present in about 60 to 70% of H. pylori strains; it encodes a high-molecular-weight protein (120 to 140 kDa) and several investigators have noted a correlation between strains that possess cagA and the severity of gastric mucosal inflammation. We examined the relation between cagA status in H. pylori strains and chronic gastritis with inflammatory processes in children from Marília, São Paulo, Brazil. One-hundred-twenty-one children were analyzed histopathologically and by polymerase chain reaction (PCR) to detect H. pylori and cagA. We then looked for an association between cagA presence and inflammatory infiltration. Using histology and PCR, we found 47% H. pylori positive infection; 29 children were diagnosed with chronic gastritis, while 28 showed normal mucosa by histopathological analysis. CagA presence was genotyped in both groups, and an inflammatory infiltrate was studied in all infected children with chronic gastritis. We found cagA strains in 20 of 29 (69%) children with chronic gastritis and 18 of 28 (64%) with normal mucosa, demonstrating a strong relationship between the strains and the inflammatory process. We found a positive association between an inflammatory process associated with H. pylori of cagA+ strains and chronic gastritis development.

A linhagem celular Hep-2 é formada por células de carcinoma da laringe e é muito utilizada em modelos de carcinogênese e mutagenêse. Para avaliar o potencial proliferativo desta linhagem, desenvolvemos uma metodologia citogenética (técnica do sobrenadante) para obtenção de metáfases a partir de células que, ao entrarem em mitose, perdem adesão celular, ficando em suspensão no meio de cultura. Através deste procedimento, foram contadas 2000 células, correspondendo a um índice mitótico (IM) de 22.2% . Apesar de o IM obtido por esta técnica não ter sido estatisticamente diferente do IM obtido por outras metodologias citogenéticas clássicas, a técnica do sobrenadante é vantajosa porque elimina o uso de alguns reagentes utilizados na obtenção de metáfases e também diminui o consumo de reagentes de manutenção desta linhagem.

The Hep-2 cell line is derived from laryngeal carcinoma cells and is often utilized as a model in carcinogenesis and mutagenesis tests. To evaluate the proliferative potential of this line, we developed a cytogenetic methodology (drifter technique) to obtain metaphases from cells that loose cellular adhesion when they underwent mitosis in culture. By this procedure, 2000 cells were counted, resulting in a mitotic index (MI) of 22.2%. Although this MI was not statistically different from the one obtained using either a classical cytogenetic method or a cell synchronization technique, the drifter technique has the advantage of not requiring the use of some reagents for the obtention of metaphases and also of diminishing the consumption of maintenance reagents for this cell line.

O artigo apresenta uma revisão sucinta dos aspectos genéticos da doença de Alzheimer e da metodologia empregada. Três genes distintos foram responsabilizados pela afecção até o momento: o da APP<FONT FACE="Symbol">-</FONT> responsável pela substância precursora da b-amilóide, a qual se deposita intensamente no cérebro dos afetados e está associada ao quadro demencial <FONT FACE="Symbol">-</FONT>, o gene da presenilina 1 (PS1) e o da presenilina 2 (PS2), proteínas de membrana celular. O gene da PS1 é responsável por cerca de 40% dos casos familiares e de acometimento precoce da DA . Os genes da ApoE4, da a-2-macroglobulina e da catepsina D, envolvidos no metabolismo da b-amilóide, foram caracterizados como fatores de risco para a DA. O gene da ApoE4 é fator de risco em cerca de 50% dos casos de DA esporádicos e de acometimento tardio. Muitos outros genes foram ainda associados à DA e são apresentados brevemente. São discutidas a conduta <FONT FACE="Symbol">-</FONT> quanto ao aconselhamento genético para familiares de afetados <FONT FACE="Symbol">-</FONT> e a utilização de diagnóstico molecular na predisposição genética à afecção. É apresentado como mecanismo comum às síndromes progeróides genéticas, como a DA, a alteração da atividade dos genes ribossômicos.

This paper concisely reviews the genetics of Alzheimer Disease (AD). So far, three different genes have been implicated in AD. Specifically, the gene that codes for APP <FONT FACE="Symbol">-</FONT> the amyloid beta precursor protein, which is associated to dementia through intense brain deposition <FONT FACE="Symbol">-</FONT> the presenilin 1 gene (PS1) and the presenilin 2 gene (PS2). Both presenilin genes code for cellular membrane proteins and PS1 are responsible for approximately 40% of the early-onset familial cases. Genes that code for ApoE4, alpha-2 macroglobulin and cathepsin D, which are all involved in APP metabolism, have been considered as risk factors for AD. The ApoE4 gene is a risk factor in about 50% of the AD sporadic and late-onset cases. Several other genes that have been associated to AD are briefly discussed. Different strategies for genetic counseling of relatives are considered, and finally, ribosomal gene alterations are discussed as possible mechanisms underlying genetic progeroid syndromes such as AD.