AMP hydrolysis reduction in blood plasma of breast cancer elderly patients after different treatments.

Adenine nucleotides are important signaling molecules that mediate biological functions in many conditions, including cancer. The enzymes CD39 and CD73 produce adenosine in the extracellular milieu that has a very important role in tumor development. This study aimed to evaluate nucleotide hydrolysis in the plasma blood of breast cancer elderly patients. In this prospective cohort study, we investigated the ectonucleotidases activity in breast cancer elderly patients, at the moment of diagnosis and after treatment. Control group consisted of elderly women without cancer diagnostic. The nucleotide hydrolysis assay was performed by the malachite green method and used ATP, ADP, or AMP as substrates. Paired t test or Wilcoxon rank-sum test was used. Our data showed that breast cancer patients presented high levels of ATP and AMP hydrolyses when compared to control group at the moment of diagnosis. When analyzing the differences between the samples at the time of diagnostic and 6 months after treatment, we observed a significant reduction on CD73 activity after all treatments used: surgery, chemotherapy, radiotherapy, or hormone therapy. The results with APCP, a specific CD73 inhibitor, showed that the AMP hydrolysis was inhibited in all conditions evaluated. We observed a diminished ADPase activity in the patients without metastasis when compared to metastatic breast cancer patients. The results showed that AMP hydrolysis was reduced in the blood plasma of breast cancer elderly patients after different treatments. This study strengthens the potential role of CD73 enzyme as a biomarker for breast cancer treatment response.

Malformações congênitas: principais etiologias conhecidas, impacto populacional e necessidade de monitoramento / Birth defects: main known etiologies, population impact and the need for monitoring

OBJETIVOS: Este artigo tem por objetivo revisar as causas, o impacto dos defeitos congênitos na morbimortalidade infantil e a importância da sua vigilância, bem como apresentar como nossa comunidade acadêmica está inserida neste monitoramento. MÉTODOS: Foi realizada uma revisão da literatura nas bases de dados Medline/PubMed e Scielo, incluindo publicações de janeiro de 2010 até maio de 2018, em português e inglês. As palavras-chave utilizadas foram: "birth defects", "etiology", "infant mortality", "teratogens", "congenital abnormalities", "defeitos congênitos", "ECLAMC", "genética". Foram incluídos também livros-textos e artigos relevantes na área, independente da data de publicação, assim como sites eletrônicos de relevância. Adicionalmente, descrevemos o funcionamento do programa de monitoramento de defeitos congênitos do Hospital São Lucas da PUCRS, vinculado ao Estudo Colaborativo Latino Americano de Malformações Congênitas (ECLAMC). RESULTADOS: 971 artigos foram localizados com as combinações das palavras-chave, destes 21 foram utilizados na revisão. No conteúdo da revisão incluímos as etiologias conhecidas dos defeitos congênitos, dentre elas, estão as genéticas, multifatoriais e teratógenos físicos, químicos e biológicos. Esses dados estão resumidos das tabelas 1 a 5 deste artigo. Com relação ao monitoramento de defeitos congênitos, o ECLAMC- PUCRS avaliou 3981 recém-nascidos no período de agosto de 2016 a dezembro de 2017, e a taxa observada de malformações foi de 3,77%. CONCLUSÃO: Os defeitos congênitos são uma causa importante de morbimortalidade em todos os países, inclusive no Brasil. A compreensão de suas etiologias contribui para o aconselhamento genético das famílias, para o estabelecimento do prognóstico e intervenções terapêuticas, assim como para sua prevenção.

OBJECTIVE: This article aims to review the impact of birth defects on infant morbidity and mortality and the importance of their surveillance, to review their causes, as well as to show how our academic community is included in this monitoring. METHODS: a literature review was conducted in the Medline / PubMed and Scielo databases, including publications from January 2010 to May 2018, in Portuguese and English. The keywords used were: "birth defects", "etiology", "infant mortality", "teratogens", "congenital abnormalities", "congenital defects", "ECLAMC", "genetics". Also included were textbooks and relevant articles in the area, regardless the date of publication, as well as relevant electronic sites. Additionally, we describe the operation of the congenital defect monitoring program of the São Lucas Hospital of PUCRS, linked to the Latin American Collaborative Study of Congenital Malformations (ECLAMC). RESULTS: 971 articles were found using the keywords, from those 21 were used in the review. In the content of the review we include the known etiologies of the congenital defects, among which are the genetic, multifactorial, and physical, chemical and biological teratogens. These data are summarized in Tables 1-5 of this article. With regard to the monitoring of congenital defects, ECLAMC-PUCRS evaluated 3981 newborns from August 2016 to December 2017, and the observed malformation rate was 3.77%. CONCLUSION: Congenital defects are an important cause of morbidity and mortality in all countries, including Brazil. Understanding their etiologies contributes to the genetic counseling of families, to the establishment of prognosis and therapeutic interventions, as well as to their prevention.

Pharmacological perspectives from Brazilian Salvia officinalis (Lamiaceae): antioxidant, and antitumor in mammalian cells.

Salvia officinalis (Lamiaceae) has been used in south of Brazil as a diary homemade, in food condiment and tea-beverage used for the treatment of several disorders. The objective of this study was to characterize chemical compounds in the hydroalcoholic (ExtHS) and aqueous (ExtAS) extract from Salvia officinalis (L.) by gas chromatography-mass spectrometry (GC-MS) and by high-resolution electrospray ionization mass spectrometry (ESI-QTOF MS/MS), evaluate in vitro ability to scavenge the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), catalase (CAT-like) and superoxide dismutase (SOD-like) activity, moreover cytotoxic by MTT assay, alterations on cell morphology by giemsa and apoptotic-induced mechanism for annexin V/propidium iodide. Chemical identification sage extracts revealed the presence of acids and phenolic compounds. In vitro antioxidant analysis for both extracts indicated promising activities. The cytotoxic assays using tumor (Hep-2, HeLa, A-549, HT-29 and A-375) and in non-tumor (HEK-293 and MRC-5), showed selectivity for tumor cell lines. Immunocytochemistry presenting a majority of tumor cells at late stages of the apoptotic process and necrosis. Given the results presented here, Brazilian Salvia officinalis (L.) used as condiment and tea, may protect the body against some disease, in particularly those where oxidative stress is involved, like neurodegenerative disorders, inflammation and cancer.

Stem-cell treatment in disc degeneration: What is the evidence? / Células tronco no tratamento de doença degenerativa discal: Qual é a evidência? / Células madre en el tratamiento de degeneración de disco: ¿Cual es la evidencia?

To review the potential role of stem cells in treating degenerative disc disease of the intervertebral disc (IVD). A review was performed of articles from the Medline database concerning stem cells and degenerative disc disease (DDD). To discuss the data, the papers were classified as: review, in vitro, experimental, and clinical. The currently available treatments were basically for symptom reduction, not to revert the IVD degenerative process. The use of mesenchymal stem cells (MSC) is being proposed as an option of treatment for DDD. In vitro studies have shown that the MSC are able to differentiate into NP cells and that the MSC also reduce the inflammatory levels of the degenerated IVD. Besides, experimental studies demonstrated that the MSC remained viable when injected into the IVD, and that they were able to regenerate partially from the degenerated IVD and its structure. The few clinical studies found in the literature presented diverging results. The use of MSC is being widely studied and shows promising results for the treatment of DDD. Although many advances are being achieved in studies in vitro and experimental, there is a lack of clinical studies to prove the role of MSC in DDD management.

Revisar o potencial papel das células-tronco para o tratamento de doença degenerativa do disco intervertebral (IVD). Foi realizada uma revisão dos trabalhos da base de dados Medline em relação a células-tronco e doença degenerativa discal (DDD). Para fins de discussão dos dados, os trabalhos foram divididos em: revisão, in vitro, experimentais e clínicos. Os tratamentos disponíveis atualmente focam basicamente na redução dos sintomas, e não na reversão do processo degenerativo do DIV. O uso de células-tronco mesenquimais (CTM) vem sendo proposto como uma opção de tratamento para DDD. Estudos in vitro demonstraram que as CTM são capazes de se diferenciarem em células do NP e que as CTM também diminuem os níveis inflamatórios do DIV degenerado. Além disso, estudos experimentais demonstraram que as CTM permaneciam viáveis quando injetadas no DIV e que eram capazes de regenerar parcialmente do DIV degenerado e sua estrutura. Os poucos estudos clínicos presentes na literatura apresentam resultados divergentes. O uso de CTM esta sendo amplamente estudado e mostra resultados promissores para o tratamento da DDD. Embora muitos avanços venham sendo alcançados em estudos in vitro e experimentais, ainda faltam estudos clínicos para comprovar o papel das CTM no manejo da DDD.

Revisar el papel potencial de las células madre en el tratamiento de la enfermedad degenerativa del disco intervertebral (IVD). Se realizó una revisión de los artículos de la base de datos Medline sobre células madre y la enfermedad degenerativa del disco (DDD). Para fines de discusión de los datos, los trabajos se dividieron en: revisión, in vitro, experimentales y clínicos. Los tratamientos disponibles actualmente enfocan básicamente la reducción de los síntomas, y no la reversión del proceso degenerativo del IVD. El uso de células madre mesenquimales (MSC) se propone como una opción de tratamiento para DDD. Estudios in vitro demostraron que las MSC son capaces de diferenciarse en células NP y que las MSC también reducen los niveles inflamatorios de la IVD degenerado. Además, estudios experimentales demostraron que las MSC se mantuvieron viables cuando inyectadas en el IVD y que eran capaces de regenerarse parcialmente del IVD degenerado y su estructura. Los pocos estudios clínicos presentes en la literatura presentan resultados divergentes. El uso de MSC se está estudiando ampliamente y muestra resultados prometedores en el tratamiento de la DDD. Aunque se hayan logrado muchos avances en estudios in vitro y experimentales, aún faltan estudios clínicos para comprobar el papel de las MSC en el manejo de la DDD.

Avaliação in vitro do potencial biológico da Salvia officinalis L. em células tumorais / In vitro evaluation of the biological potential of Salvia officinalis L. in tumor cells

Objetivos: Determinar a viabilidade celular frente ao efeito biológico do extrato hidroalcoólico de Salvia officinalis L. em culturas de células tumorais de laringe (Hep-2) e células de hepatoma humano (HepG2).Métodos: Células tumorais Hep-2 e HepG2 foram cultivadas em meio DMEM (Dulbecco's Modified Eagle Médium)suplementado com 10% soro fetal bovino inativado e 1% de antibiótico (Penicilina/Estreptomicina) em estufa a 37°Ce atmosfera umidificada com 5% de CO2. Na segunda etapa foram semeadas (5×104 células/mL) em placas de 96 poçosdurante o período de 24 horas até obtenção de 60-70% de confluência e realizou-se o tratamento das células com extratohidroalcoólico de Salvia officinalis L., controle negativo de etanol 80% (v/v) e controles positivos de peróxido de hidrogênio(H2O2), tert-butil hidroperóxido (t-BOOH), doxorrubicina e cisplatina. A viabilidade celular foi determinada pela reduçãodo MTT (brometo de 3-(4,5 dimetiltiazol-2-il)-2,5-difeniltetrazolio). Os resultados foram analisados pelo teste de Tukeyno programa SPSS v.19.Resultados: O extrato hidroalcoólico de Salvia officinalis L. mostrou atividade citotóxica para células tumorais Hep-2 IC500,38±0,02 mg/mL e para HepG2 IC50 0,54±0,03 mg/mL em comparação ao controle negativo, ficando acima do IC50 obtidopara seus controles positivos. A cisplatina apresentou IC50 abaixo do extrato de sálvia, porém para a obtenção do seu IC50aumentou-se o tempo de exposição em seis horas.Conclusões: Sugere-se que o extrato de Salvia officinalis L. tem atividade biológica em células tumorais, podendo serobjetivo de mais estudos com a finalidade de comprovar sua eficácia como possível agente para o tratamento do câncer.

Aims: To determine cell viability compared to the biological effect of Salvia officinalis L. extract on cultured tumor cells of the larynx (Hep-2) and in human hepatoma cells (HepG2). Methods: Tumor cells Hep-2 and HepG2 were grown in DMEM (Dulbecco’s Modified Eagle Medium) supplemented with 10% inactivated fetal bovine serum and 1% antibiotics (Penicillin/Streptomycin), incubated at 37°C and 5% CO2. In the second step they were plated (5×104 cells/mL) in 96 well plates during 24 hours to obtain 60-70% confluency, and treated with the hydroalcoholic extract of Salvia officinalis L., negative control with 80% ethanol (v/v) and positive control of hydrogen peroxide (H2O2), tert-butyl hydroperoxide (t-BOOH), doxorubicin and cisplatin. Cell viability was determined by MTT reduction (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). Results were analyzed by Tukey test using the SPSS v.19. Results: The hydroalcoholic extract of Salvia officinalis L. showed cytotoxic activity to tumor cells Hep-2 IC50 0,38±0.02 mg/mL for HepG2 IC50 0,54±0.03 mg/mL compared to negative control, staying above the IC50 obtained for their positive controls. Cisplatin showed IC50 below the extract of sage, but to obtain the IC50 exposure time was increased in six hours. Conclusions: The results of this study suggest that the extract of Salvia officinalis L. has biological activity against tumor cell; further studies are needed to confirm its efficacy and its potential role in cancer treatment.

Perfil da resposta terapêutica a medicamentos antirretrovirais envolvendo crianças e adolescentes infectados pelo HIV1 no Rio Grande do Sul / Profile of therapeutic response to antiretroviral drug treatment among HIV1?infected children and adolescents in Rio Grande do Sul

No fim da década de 80, muitas mulheres passaram a contrair o vírus HIV e este fato começou a delinear um novo perfil da epidemia da AIDS no Brasil. No entanto, muitos estudos sobre HIV/AIDS são realizados somente com a população adulta e dados sobre a infecção pediátrica ainda são bastante escassos. Este é um estudo transversal que objetiva traçar o perfil da resposta à terapia antirretroviral em uma população formada por crianças e adolescentes. Este trabalho foi desenvolvido por meio da análise dos resultados dos testes de genotipagem - disponíveis no banco de dados do Setor de Biologia Molecular do Laboratório Central do Estado do Rio Grande do Sul (LACEN-RS). Por meio dos testes de genotipagem foi possível observar algumas mutações de resistência aos antirretrovirais. O subtipo B apresentou maior prevalência, seguido pelo subtipo C. As mutações que ocorreram com maior frequência foram D67N e V118I, pertencentes à classe de medicamentos Nucleosídeos Inibidores da Transcriptase Reversa (NRTI). Foi possível observar que alguns medicamentos apresentaram maiores níveis de resistência simples: Zidovudina e Lamivudina. A importância da identificação de mutações de resistência se faz relevante na escolha dos medicamentos que serão utilizados, aumentando assim, as chances de sucesso da terapia.

In the late 80s, many women began to contract the HIV virus; this fact began to change the profile of AIDS epidemic in Brazil. However, many studies of HIV/AIDS are performed only on the adult population and data on pediatric HIV infection are still quite scarce. The aim of this cross-sectional study was to outline the response of the HIV infection to antiretroviral therapy in a population composed of children and adolescents. The work involved analyzing the results of virus genotyping assays, available in the database of the Section of Molecular Biology of the Central State Laboratory of Rio Grande do Sul (LACEN-RS). Subtype B was the most prevalent, followed by subtype C. Through genotyping, it was possible to observe some antiretroviral resistance mutations. The mutations that occurred most frequently were D67N and V118I, which confer resistance to medications of the class of Nucleoside Reverse Transcriptase Inhibitors (NRTI). The highest observed frequencies of resistance were to Zidovudine and Lamivudine. The identification of resistance mutations is of great relevance in the choice of drugs that will be used to treat a particular patient, to maximize the chance of a successful therapy.

Differentiation of human adipose-derived adult stem cells into neuronal tissue: does it work?

Adipose tissue contains many cells and proteins that are of value not only for their potential therapeutic applications, but also for the low cost of their harvest and delivery. Mesenchymal stem cells (MSC) were originally isolated from the bone marrow, although similar populations have been isolated from adipose and other tissues. At one time, neural tissues were not regarded as regenerative populations of cells. Therefore, the identification of cell populations capable of neuronal differentiation has generated immense interest. Adipose tissue may represent an alternative source of cells that are capable of neuronal differentiation, potentially enhancing its use in the treatment of neurological disease. The aim of this review is to cover the current state of knowledge of the differentiation potential of human adipose-derived stem (ADAS) cells, specifically their ability to give rise to neuronal cells in vitro. This review presents and discusses different protocols used for inducing human ADAS cells to differentiate in vitro, and the neuronal markers utilized in each system.

Transition of cervical carcinoma in situ to invasive cancer: role of p16 INK4a expression in progression and in recurrence.

To investigate the expression of p16(INK4a) in cervical carcinoma and its relation to the transition of carcinoma in situ to invasive carcinoma, and its role in recurrence of cervical lesions as well, a series of 90 patients with cervical carcinoma (49 with in situ lesion and 41 with invasive lesion) were selected from July 2001 and September 2002. Groups with in situ and invasive lesions were paired for a series of risk variables for cervical cancer and followed up for 60 months. The follow-up visits occurred every 6 months in the first three years and annually up to the fifth year. It was observed that 87.9% of the patients with invasive lesion showed overexpression of p16(INK4a), in comparison with 37.6% of those with in situ lesion (X(2): 13.68; 2 df; p=0.0002; OR: 12.08), demonstrating overexpression of p16(INK4a) as a risk of invasion of the basal layer by dysplastic cells. We also observed an association between overexpression of p16(INK4a) and staging of cancer (X(2): 18.38; 6 df; p=0.0003). A prospective analysis, when controlled for interaction with cervical lesion groups (by Cox regression), demonstrated a risk of recurrence of 4.83 times attributed to overexpression of p16(INK4a), albeit not statistically significant (p=0.14).

p16INK4A expression in cervical premalignant and malignant lesions.

p16INK4a is a cyclin-dependent kinase (CDK) inhibitor which decelerates cell cycle by inactivating CDKs that phosphorylate pRb. Human Papillomavirus persistent infection plays an important role on cervical carcinogenesis, mainly by the action of two viral oncoproteins, E6 and E7, which interact with p53 and pRb, respectively. Increasing expression of E6 and E7 in dysplastic cervical cells might thus be reflected by increased expression of p16INK4a. Recent studies revealed that p16INK4a expression could be a marker for dysplastic and neoplastic cervical cells. The aim of this study was to analyze p16INK4a expression in cervical preneoplastic and neoplastic lesions and correlate with lesion grade. Expression of p16INK4a was analyzed by immunohistochemistry. A total of 6 low-grade squamous intraepithelial lesion (LSIL), 21 high-grade squamous intraepithelial lesions (HSIL) and 27 cancer samples were studied. In HPV-positive cervical samples (n=48), p16INK4a expression was observed in 1 of 3 LSIL, in 18 of 19 HSIL and in all 26 cancer cases. These results are in accordance with the hypothesis that functional inactivation of pRb by HPV-E7 protein induces p16INK4a expression in cervical lesions. In our study, a statistically significant association was observed between cervical lesion grade and p16INK4a expression (P<0.001).