Supplemented tissue culture medium 199 is a better medium for in vitro maturation of oocytes from women with polycystic ovary syndrome women than human tubal fluid

Objective: To compare oocyte maturation, fertilization and cleavage rates, and embryonic developmental quality after culture of human immature oocytes from polycystic ovary syndrome (PCOS) patients in human tubal fluid (HTF) or tissue culture medium (TCM) 199. Design: Prospective, randomized, controlled trial. Setting: University hospital. Patient(s): Thirteen women undergoing 23 in vitro maturation cycles, from whom 119 oocytes were retrieved. Intervention(s): Cumulus-enclosed germinal vesicle-stage oocytes matured in TCM-199-supplemented or HTF-supplemented media. Main Outcome Measure(s): Oocyte maturation and fertilization rates, embryonic developmental quality. Result(s): Significant differences were observed between TCM 199 and HTF regarding maturation rate (82% vs. 56.9%), fertilization rate (70% vs. 39.4%), and embryo quality (81.3% vs. 41.7%). Conclusion(S): Human tubal fluid medium, although widely used for embryo fertilization and maintenance in IVF techniques, is not,in appropriate medium for the maturation of oocytes obtained from PCOS patients in nonstimulated cycles. (Fertil Steril(R) 2009;91:509-13. (C)2009 by American Society for Reproductive Medicine.)

Bone quality associated with daily intake of coffee: a biochemical, radiographic and histometric study

Caffeine induces loss of calcium and influences the normal development of bone. This study investigated the effects of coffee on bone metabolism in rats by biochemical measurement of calcium, bone densitometry and histometry. Male rats, born of female treated daily with coffee and with coffee intake since born, were anesthetized, subjected to extraction of the upper right incisor, and sacrificed 7, 21 and 42 days after surgery. Blood and urine samples were taken, and their maxilla radiographed and processed to obtain 5-µm-thick semi-serial sections stained with hematoxylin and eosin. The volume and bone quality were estimated using an image-analysis software. The results showed significantly greater amount of calcium in the plasma (9.40 ± 1.73 versus 9.80 ± 2.05 mg%) and urine (1.00 ± 0.50 versus 1.25 ± 0.70 mg/24 h) and significantly less amount in bone (90.0 ± 1.94 versus 86.0 ± 2.12 mg/mg bone), reduced bone mineral density (1.05 ± 0.11 versus 0.65 ± 0.15 mmAL), and lower amount of bone (76.19 ± 1.6 versus 53.41 ± 2.1 %) (ANOVA; p≤0.01) in animals treated with coffee sacrificed after 42 days. It may be concluded that coffee/caffeine intake caused serious adverse effects on calcium metabolism in rats, including increased levels of calcium in the urine and plasma, decreased bone mineral density and lower volume of bone, thus delaying the bone repair process.

Melatonin in maturation media fails to improve oocyte maturation, embryo development rates and DNA damage of bovine embryos

Melatonin (MEL) acts as a powerful scavenger of free radicals and direct gonadal responses to melatonin have been reported in the literature. Few studies, however, have evaluated the effect of MEL during in vitro maturation (IVM) on bovine embryos. This study tested the addition of MEL to maturation medium (MM) with no gonadotropins on nuclear maturation and embryo development rates and the incidence of DNA damage in resulting embryos. Cumulus-oocyte complexes were aspirated from abattoir ovaries and cultured in MM (TCM-199 medium supplemented with 10% fetal calf serum - FCS) at 39ºC and 5% CO2 in air. After 24 hours of culture in MM with 0.5 µg mL-1 FSH and 5.0 µg mL-1 LH; 10-9 M MEL) or 10-9 M MEL, 0.5 µg mL-1 FSH and 5.0 µg mL-1 LH, the oocytes were stained with Hoechst 33342 to evaluate nuclear maturation rate. After in vitro fertilization and embryo culture, development rates were evaluated and the blastocysts were assessed for DNA damage by Comet assay. There was no effect of melatonin added to the MM, alone or in combination with gonadotropins, on nuclear maturation, cleavage and blastocyst rates. These rates ranged between 88% to 90%, 85% to 88% and 42% to 46%, respectively. The extent of DNA damage in embryos was also not affected by MEL supplementation during IVM. The addition of 10-9 M MEL to the MM failed to improve nuclear maturation and embryo development rates and the incidence of DNA damage in resulting embryos, but was able to properly substitute for gonadotropins during IVM.

Conteúdos de biologia em vestibulares seriados

A adoção de novos processos seletivos para o acesso de alunos às universidades só foi possível com a promulgação da Lei de Diretrizes e Bases da Educação Nacional n. 9.394/96. O vestibular seriado é uma dessas propostas alternativas, que prevê uma avaliação sistemática dos alunos ao término de cada ano do ensino médio. O propósito deste estudo foi caracterizar o vestibular seriado, hoje presente em 22 instituições públicas, quanto ao seu objetivo, estilos de avaliação e conteúdos da disciplina de Biologia distribuídos nos módulos referentes às três séries do ensino médio. Averiguamos que os diferentes programas analisados não apresentam uniformidade em diversos aspectos, tais como número de vagas, estrutura de avaliação e conteúdo programático. Verificamos, ainda, a ausência de consenso sobre a sequência a ser adotada para o ensino de Biologia e a resultante restrição para que o aluno possa preparar-se para as provas seriadas em mais de uma instituição.

Trypanocidal activity of Piper arboreum and Piper tuberculatum (Piperaceae)

In the scope of our ongoing research on bioactive agents from Brazilian flora, twenty-four extracts and fractions obtained from Piper arboreum Aub. and Piper tuberculatum Jacq. (Piperaceae) were screened for trypanocidal activity by using MTT colorimetric assay. The strongest activity was found in hexane fractions from the leaves of P. arboreum (IC50= 13.3 µg/ mL) and P. tuberculatum (IC50 = 17.2 µg/mL). Hexane fractions of the fruits of P. tuberculatum and P. arboreum showed potent toxic effects on epimastigote forms of Trypanosoma cruzi, with values of IC50 (µg/mL) of 32.2 and 31.3, respectively. Additionally, the phytochemical study of the hexane fraction of P. arboreum leaves furnished two pyrrolidine amides, piperyline (1) and 4,5-dihydropiperyline (2), which could be responsible, at least in part for the observed antiprotozoal activity.

Saúde gengival de adolescentes e a utilização de serviços odontológicos, Estado de São Paulo

OBJETIVO: Avaliar a associação de condições de saúde gengival com a utilização de serviço odontológico. MÉTODOS: Realizou-se levantamento epidemiológico de saúde bucal de 1.799 adolescentes, em 35 cidades do Estado de São Paulo, em 2002. A saúde gengival foi avaliada pela prevalência de sangramento na gengiva à sondagem e cálculo dentário (índice periodontal comunitário) e oclusão dentária (índice de estética dentária). A utilização de serviços odontológicos foi medida pelo índice de cuidado (O/CPO) para cada cidade. Análise multinível de regressão logística ajustou modelos explicativos para fatores associados aos desfechos de interesse. RESULTADOS: A prevalência de sangramento gengival à sondagem foi 21,5%; de cálculo dentário foi 19,4%. Os participantes do sexo masculino, negros e pardos, moradores em áreas rurais, residentes em domicílios aglomerados e com atraso escolar apresentaram chance significantemente mais elevada para os agravos que seus respectivos pares de comparação. Características de oclusão dentária também associaram com gengiva não-saudável: apinhamento dos segmentos incisais, mordida aberta vertical anterior, relação molar antero-posterior. Cidades com maior utilização de serviço odontológico tiveram menor proporção de adolescentes com sangramento gengival e cálculo. CONCLUSÕES: A utilização de serviços odontológicos foi significativamente associada a melhores condições de saúde gengival (sangramento e cálculo). Essa associação independeu das características sociodemográficas individuais e contextuais, e de oclusão dentária.

Insegurança alimentar, vínculo mãe-filho e desnutrição infantil em área de alta vulnerabilidade social

OBJETIVO: analisar a insegurança alimentar e o vínculo inadequado mãe-filho como dois potenciais determinantes da desnutrição em crianças de quatro a seis anos de idade. MÉTODOS: estudo de caso-controle desenvolvido em Escolas Municipais de Educação Infantil (EMEIs) no Jardim Jaqueline, área de alta vulnerabilidade social do município de São Paulo, Brasil. Foram aplicados a Escala Brasileira de Insegurança Alimentar e o Protocolo de Avaliação do Vínculo Mãe-filho, além de coletadas informações biológicas e socio-econômicas. Para verificação dos efeitos de cada variável independente e controle dos efeitos das demais variáveis incluídas no modelo, foi utilizado o modelo de regressão logística múltipla. RESULTADOS: verificou-se que tanto a insegurança alimentar familiar (OR=3,6) como o vínculo inadequado mãe-filho (OR=9,4) estiveram associados com a desnutrição infantil (p<0,05), mesmo após o controle para o peso ao nascimento da criança e idade, estado conjugal e trabalho maternos. CONCLUSÕES: tanto a insegurança alimentar familiar (OR=3,6) como o vínculo mãe-filho inadequado (OR=9,4) mostraram-se fatores determinantes da ocorrência da desnutrição na população estudada.

O Sistema Único de Saúde como observatório de direitos universais: uma reflexão a partir das Ciências Sociais

Há uma ampla bibliografia sobre a crise da modernidade. Uma certa vertente compreende que estão ameaçados vários pilares da vida social moderna, dentre eles as conquistas representadas pelos direitos universais. A literatura da Saúde Coletiva também aponta o Sistema Único de Saúde (SUS) como uma ocorrência contrária ao movimento mundial de flexibilização daqueles direitos. Os princípios de universalidade e integralidade, além de constituírem um desafio institucional, financeiro, político e social, expressam também a decisão da sociedade de implementar direitos universais. Utilizamos o instrumental teórico de Souza Santos para caracterizar o SUS como um observatório sobre a efetivação de tais direitos. Ressurge a importância do remodelamento institucional e das deliberações democráticas no estabelecimento do contrato social.

Effects of trans-10, cis-12 conjugated linoleic acid on gene expression and lipid metabolism of adipose tissue of growing pigs

The objective of the present study was to determine the effects of trans-10, cis-12 conjugated linoleic acid (CLA) in adipose tissue explant cultures of growing pigs on the following responses: lipogenesis (measured as rate of C-14-labeled glucose incorporation over a subsequent 2-h incubation in the presence or absence of insulin), lipolysis (release of non-esterified fatty acid over a 2-h incubation in the presence or absence of isoproterenol), activities of lipogenic enzymes, and mRNA abundance of fatty acid synthase (FAS). Adipose tissue explants from nine growing pigs (78 +/- 3 kg) were cultured in 199 medium with insulin, dexamethasone and antibiotics for 4, 12, 24, and 48 h. The treatments were 1) control: 100 mu M polyvinyl alcohol (PVA); 2) pGH: 100 ng/mL porcine growth hormone (pGH) plus 100 mu M PVA; 3) CLA200: 200 mu M trans-10, cis-12 CLA; 4) CLA50: 50 mu M trans-10, cis-12 CLA, and 5) LA: 200 mu M linoleic acid. Fatty acids were added along with PVA (2: 1), respectively, for 24 h. Explants were collected after each culture period and assayed for lipogenesis. Transcripts of FAS mRNA were quantified by real-time RT-PCR after 24 and 48 h. Lipolysis and activities of FAS, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and NADP-malate dehydrogenase were determined after 48 h. As expected, glucose incorporation was decreased (P < 0.05) in response to pGH treatment (positive control). LA had no effect on any parameter evaluated. Treatment with trans-10, cis-12 CLA decreased FAS activity (P < 0.05), but NADPH-generating enzymes were unaffected by treatments. Consistent with reduction in FAS activity, both lipid synthesis and FAS mRNA abundance were reduced with chronic CLA treatment, pGH increased baseline and stimulated lipolysis (P < 0.05) after 48 h of culture, while CLA treatment had no effect on non-esterified fatty acid release. Results of this study showed that trans-10, cis-12 CLA alters lipogenesis but has no effect on lipolysis in cultures of pig adipose tissue.

Adaptive Immunity against Leishmania Nucleoside Hydrolase Maps Its C-Terminal Domain as the Target of the CD4+ T Cell-Driven Protective Response

Nucleoside hydrolases (NHs) show homology among parasite protozoa, fungi and bacteria. They are vital protagonists in the establishment of early infection and, therefore, are excellent candidates for the pathogen recognition by adaptive immune responses. Immune protection against NHs would prevent disease at the early infection of several pathogens. We have identified the domain of the NH of L. donovani (NH36) responsible for its immunogenicity and protective efficacy against murine visceral leishmaniasis (VL). Using recombinant generated peptides covering the whole NH36 sequence and saponin we demonstrate that protection against L. chagasi is related to its C-terminal domain (amino-acids 199-314) and is mediated mainly by a CD4+ T cell driven response with a lower contribution of CD8+ T cells. Immunization with this peptide exceeds in 36.73 +/- 12.33% the protective response induced by the cognate NH36 protein. Increases in IgM, IgG2a, IgG1 and IgG2b antibodies, CD4+ T cell proportions, IFN-gamma secretion, ratios of IFN-gamma/IL-10 producing CD4+ and CD8+ T cells and percents of antibody binding inhibition by synthetic predicted epitopes were detected in F3 vaccinated mice. The increases in DTH and in ratios of TNF alpha/IL-10 CD4+ producing cells were however the strong correlates of protection which was confirmed by in vivo depletion with monoclonal antibodies, algorithm predicted CD4 and CD8 epitopes and a pronounced decrease in parasite load (90.5-88.23%; p = 0.011) that was long-lasting. No decrease in parasite load was detected after vaccination with the N-domain of NH36, in spite of the induction of IFN-gamma/IL-10 expression by CD4+ T cells after challenge. Both peptides reduced the size of footpad lesions, but only the C-domain reduced the parasite load of mice challenged with L. amazonensis. The identification of the target of the immune response to NH36 represents a basis for the rationale development of a bivalent vaccine against leishmaniasis and for multivalent vaccines against NHs-dependent pathogens.

Pimarane-type Diterpenes: Antimicrobial Activity against Oral Pathogens

Seven pimarane type-diterpenes re-isolated from Viguiera arenaria Baker and two semi-synthetic pimarane derivatives were evaluated in vitro against the following main microorganisms responsible for dental caries: Streptococcus salivarius, S. sobrinus, S. mutans, S. mitis, S. sanguinis and Lactobacillus casei. The compounds ent-pimara-8(14), 15-dien-19-oic acid (PA); ent-8(14), 15-pimaradien-3 beta-ol; ent-15-pimarene-8 beta, 19-diol; ent-8(14), 15-pimaradien-3 beta-acetoxy and the sodium salt derivative of PA were the most active compounds, displaying MIC values ranging from 2 to 8 mu g.mL(-1). Thus, this class of compounds seems promising as a class of new effective anticariogenic agents. Furthermore, our results also allow us to conclude that minor structural differences among these diterpenes significantly influence their antimicrobial activity, bringing new perspectives to the discovery of new natural compounds that could be employed in the development of oral care products.

Breast phantom with silicone implant for evaluation in conventional mammography

With the increased incidence of cancer and a similarly increased number of surgeries for insertion of silicone breast implants, it is necessary to assess the effect of such material within the breast tissue, particularly in mammography, because of the reduction in the power of breast cancer diagnosis. In this work, we introduce a breast phantom with silicone implants in order to evaluate the influence of the implant on the visibility of the main mammographic findings: fibers, microcalcifications and tumor masses. In this proposed phantom, the breast tissue was simulated using gel paraffin. In the optical density of phantom mammograms with implants, a reduction in breast tissue visibility was seen corresponding to 23% when compared to a phantom without silicone implants. This poor visibility was due to the X-ray beam scattering on silicone material; this effect produced a loss of visibility in the areas adjacent to the implant. It is expected that the proposed phantom model may be used as a device for the establishment of a technical standard for these types of procedures.

Concurrent sequence variation of TP53 and TP73 genes in anaplastic astrocytoma

Disruption or loss of tumor suppressor gene TP53 is implicated in the development or progression of almost all different types of human malignancies. Other members of the p53 family have been identified. One member, p73, not only shares a high degree of similarity with p53 in its primary sequence, but also has similar functions. Like p53, p73 can bind to DNA and activate transcription. Using PCR-SSCP and gene sequencing, we analyzed the TP53 and TP73 genes in a case of a grade III anaplastic astrocytoma that progressed to glioblastoma. We found a deletion of AAG at position 595-597 of TP53 (exon 6), resulting in the deletion of Glu 199 in the protein and a genomic polymorphism of TP73, identified as an A-to-G change, at position E8/+15 at intron 8 (IVS8-15A>G). The mutation found at exon 6 of the gene TP53 could be associated with the rapid tumoral progression found in this case, since the mutated p53 may inactivate the wild-type p53 and the p73 alpha protein, which was conserved here, leading to an increase in cellular instability.

H alpha spectropolarimetry of RY Tauri and PX Vulpeculae (Research Note)

Aims. To detect line effects using spectropolarimetry in order to find evidence of rotating disks and their respective symmetry axes in T Tauri stars. Methods. We used the IAGPOL imaging polarimeter along with the Eucalyptus-IFU to obtain spectropolarimetric measurements of the T Tauri stars RY Tau (two epochs) and PX Vul (one epoch). Evidence of line effects showing a loop in the Q-U diagram favors a compact rather than an extended source for the line photons in a rotating disk. In addition, the polarization position angle (PA) obtained using the line effect can constrain the symmetry axis of the disk. Results. RY Tau shows a variable H alpha double peak in 2004-2005 data. A polarization line effect is evident in the Q-U diagram for both epochs confirming a clockwise rotating disk. A single loop is evident in 2004 changing to a linear excursion plus a loop in 2005. Interestingly, the intrinsic PA calculated using the line effect is consistent between our two epochs (similar to 167 degrees). An alternative intrinsic PA computed from the interstellar polarization-corrected continuum and averaged between 2001-2005 yielded a PA similar to 137 degrees. This last value is closer to perpendicular to the observed disk direction (similar to 25 degrees), as expected from single scattering in an optically thin disk. For PX Vul, we detected spectral variability in H alpha along with non-variable continuum polarization when compared with previous data. The Q-U diagram shows a well-defined loop in H alpha associated with a counter-clockwise rotating disk. The symmetry axis inferred from the line effect has a PA similar to 91 degrees (with an ambiguity of 90 degrees). Our results confirm previous evidence that the emission line in T Tauri stars has its origin in a compact source scattered off a rotating accretion disk.