Wound-healing gene family expression differences between fetal and foreskin cells used for bioengineered skin substitutes.

For tissue engineering, several cell types and tissues have been proposed as starting material. Allogenic skin products available for therapeutic usage are mostly developed with cell culture and with foreskin tissue of young individuals. Fetal skin cells offer a valuable solution for effective and safe tissue engineering for wounds due to their rapid growth and simple cell culture. By selecting families of genes that have been reported to be implicated in wound repair and particularly for scarless fetal wound healing including transforming growth factor-beta (TGF-beta) superfamily, extracellular matrix, and nerve/angiogenesis growth factors, we have analyzed differences in their expression between fetal skin and foreskin cells, and the same passages. Of the five TGF-beta superfamily genes analyzed by real-time reverse transcription-polymerase chain reaction, three were found to be significantly different with sixfold up-regulated for TGF-beta2, and 3.8-fold for BMP-6 in fetal cells, whereas GDF-10 was 11.8-fold down-regulated. For nerve growth factors, midkine was 36-fold down-regulated in fetal cells, and pleiotrophin was 4.76-fold up-regulated. We propose that fetal cells present technical and therapeutic advantages compared to foreskin cells for effective cell-based therapy for wound management, and overall differences in gene expression could contribute to the degree of efficiency seen in clinical use with these cells.

Covalent cell surface functionalization of human fetal osteoblasts for tissue engineering.

The chemical functionalization of cell-surface proteins of human primary fetal bone cells with hydrophilic bioorthogonal intermediates was investigated. Toward this goal, chemical pathways were developed for click reaction-mediated coupling of alkyne derivatives with cellular azido-expressing proteins. The incorporation via a tetraethylene glycol linker of a dipeptide and a reporter biotin allowed the proof of concept for the introduction of cell-specific peptide ligands and to follow the reaction in living cells. Tuning the conditions of the click reaction resulted in chemical functionalization of living human fetal osteoblasts with excellent cell survival.

Hipertensión y edema pulmonar de altura. Rol de la disfunción endotelial y de la programación fetal [Pulmonary hypertension and lung edema at high altitude. Role of endothelial dysfunction and fetal programming].

High altitude constitutes an exciting natural laboratory for medical research. While initially, the aim of high-altitude research was to understand the adaptation of the organism to hypoxia and find treatments for altitude-related diseases, over the past decade or so, the scope of this research has broadened considerably. Two important observations led to the foundation for the broadening of the scientific scope of high-altitude research. First, high-altitude pulmonary edema (HAPE) represents a unique model which allows studying fundamental mechanisms of pulmonary hypertension and lung edema in humans. Secondly, the ambient hypoxia associated with high-altitude exposure facilitates the detection of pulmonary and systemic vascular dysfunction at an early stage. Here, we review studies that, by capitalizing on these observations, have led to the description of novel mechanisms underpinning lung edema and pulmonary hypertension and to the first direct demonstration of fetal programming of vascular dysfunction in humans.

Surface modification of biomaterials for conjugation with human fetal osteoblasts.

Surface functionalization of hydroxyapatite (HA) and beta-tricalcium phosphate (TCP) bioceramics with chemical ligands containing a pyrrogallol moiety was developed to improve the adhesion of bone cell precursors to the biomaterials. Fast and biocompatible copper-free click reaction with azido-modified human fetal osteoblasts resulted in improved cell binding to both HA and TCP bioceramics, opening the way for using this methodology in the preparation of cell-engineered bone implants.

Phenotypic and functional analysis of human fetal liver hematopoietic stem cells in culture.

Steady-state hematopoiesis and hematopoietic transplantation rely on the unique potential of stem cells to undergo both self-renewal and multilineage differentiation. Fetal liver (FL) represents a promising alternative source of hematopoietic stem cells (HSCs), but limited by the total cell number obtained in a typical harvest. We reported that human FL nonobese diabetic/severe combined immunodeficient (NOD/SCID) repopulating cells (SRCs) could be expanded under simple stroma-free culture conditions. Here, we sought to further characterize FL HSC/SRCs phenotypically and functionally before and following culture. Unexpanded or cultured FL cell suspensions were separated into various subpopulations. These were tested for long-term culture potential and for in vivo repopulating function following transplantation into NOD/SCID mice. We found that upon culture of human FL cells, a tight association between classical stem cell phenotypes, such as CD34(+) /CD38(-) and/or side population, and NOD/SCID repopulating function was lost, as observed with other sources. Although SRC activity before and following culture consistently correlated with the presence of a CD34(+) cell population, we provide evidence that, contrary to umbilical cord blood and adult sources, stem cells present in both CD34(+) and CD34(-) FL populations can sustain long-term hematopoietic cultures. Furthermore, upon additional culture, CD34-depleted cell suspensions, devoid of SRCs, regenerated a population of CD34(+) cells possessing SRC function. Our studies suggest that compared to neonatal and adult sources, the phenotypical characteristics of putative human FL HSCs may be less strictly defined, and reinforce the accumulated evidence that human FL represents a unique, valuable alternative and highly proliferative source of HSCs for clinical applications.

Fetal testosterone (2D:4D) as predictor of cognitive reflection

The Cognitive Reflection Test (CRT) is a test introduced by S. Frederick (2005) Cognitive reflection and decision making, J Econ Perspect 19(4): 25-42. The task is designed to measure the tendency to override an intuitive response that is incorrect and to engage in further reflection that leads to the correct response. The consistent sex differences in CRT performance may suggest a role for gonadal hormones, particularly testosterone. A now widely studied putative marker for fetal testosterone is the second-to-fourth digit ratio (2D:4D). This paper tests to what extent 2D:4D, as a proxy for prenatal exposure to testosterone, can predict CRT scores in a sample of 623 students. After controlling for sex, we observe that a lower 2D:4D (reflecting a higher exposure to testosterone) is significantly associated with a higher number of correct answers. The result holds for both hands? 2D:4Ds. In addition, the effect appears to be sharper for females than for males. We also control for patience and math proficiency, which are significantly related to performance in the CRT. But the effect of 2D:4D on performance in CRT is not reduced with these controls, implying that these variables are not mediating the relationship between digit ratio and CRT.

Produção e qualidade da videira 'Superior Seedless' sob restrição hídrica na fase de maturação

O objetivo deste trabalho foi avaliar o efeito das condições de deficit hídrico, na fase de maturação da uva, sobre a produção e qualidade da uva 'Superior Seedless' entre julho e novembro de 2007. O experimento foi realizado em delineamento de blocos ao acaso, com quatro repetições, em arranjo fatorial (3x3) + 1: três épocas de alteração da aplicação das lâminas de irrigação (21, 13 e 5 dias antes da colheita); três lâminas de irrigação (100, 50 e 0% da evapotranspiração da cultura); e um tratamento controle (manejo de irrigação adotado pelo produtor). As épocas de irrigação e as lâminas de irrigação utilizadas influenciaram a firmeza das bagas e a acidez titulável. A interrupção da irrigação, aos 13 ou 21 dias antes da colheita, resultou em produtividade, qualidade de frutos e eficiência do uso da água semelhante às obtidas pelo produtor, assim, pode ser adotada para economia da água de irrigação na Região do Submédio do Vale do São Francisco.

Hyperammonemia: regulation of argininosuccinate synthetase and argininosuccinate lyase genes in aggregating cell cultures of fetal rat brain.

Hyperammonemia in the brain leads to poorly understood alterations of nitric oxide (NO) synthesis. Arginine, the substrate of nitric oxide synthases, might be recycled from the citrulline produced with NO by argininosuccinate synthetase (AS) and argininosuccinate lyase (AL). The regulation of AS and AL genes during hyperammonemia is unknown in the brain. We used brain cell aggregates cultured from dissociated telencephalic cortex of rat embryos to analyze the regulation of AS and AL genes in hyperammonemia. Using RNase protection assay and non-radioactive in situ hybridization on aggregate cryosections, we show that both AS and AL genes are induced in astrocytes but not in neurons of aggregates exposed to 5 mM NH4Cl. Our work suggests that the hyperammonemic brain might increase its recycling of citrulline to arginine.

Is screening for fetal anomalies reliable in HIV-infected pregnant women? A multicentre study.

OBJECTIVE: To assess the impact of HIV infection on the reliability of the first-trimester screening for Down syndrome, using free beta-human chorionic gonadotrophin, pregnancy-associated plasma protein-A and fetal nuchal translucency, and of the second-trimester screening for neural tube defects, using alpha-fetoprotein. PATIENTS AND METHODS: Multicentre study comparing the multiples of the median of markers for Down syndrome and neural tube defect screening among 214 HIV-infected pregnant women and 856 HIV-negative controls undergoing a first-trimester Down syndrome screening test, and 209 HIV-positive women and 836 HIV-negative controls with a risk evaluation for neural tube defect. The influence of treatment, chronic hepatitis and HIV disease characteristics were also evaluated. RESULTS: Multiples of the median medians for pregnancy-associated plasma protein-A and beta-human chorionic gonadotrophin were lower in HIV-positive women than controls (0.88 vs. 1.05 and 0.84 vs. 1.09, respectively; P < 0.005), but these differences had no impact on risk estimation; no differences were observed for the other markers. No association was found between HIV disease characteristics, antiretroviral treatment use at the time of screening or chronic hepatitis and marker levels. CONCLUSION: Screening for Down syndrome during the first trimester and for neural tube defect during the second trimester is accurate for HIV-infected women and should be offered, similar to HIV-negative women.

Proliferative and Osteogenic Differentiation Potentials of Human Fetal Bone Cells

BACKGROUND. Human primary fetal bone cells (hFBC) are being characterized for use in bone tissue regeneration. Unlike human mesenchymal stem cells (hMSC), hFBC are partially differentiated with high expansion and regeneration potential. To date, proliferative and osteoblastic differentiation capacities of fetal bone cells remain poorly examined. The goal of this study was to define an environmental culture conditions for optimal proliferation and production of extracellular bone matrix leading to efficient bone repair. METHODS. Human primary FBC derived from our dedicated, consistent banks of bone cells comprising several fetal donors. For proliferation study, monolayer cultures of both cell types were expanded in DMEM or α- MEM media. Osteoblastic differentiation potentials of both hFBC and hMSC were evaluated through RT-PCR. Regulation of osteogenic differentiation by protein ligands Wnt3a and Wnt5a was studied by ALP enzymatic activity measurement. RESULTS. Evaluation of the proliferation rate demonstrated that hFBC proliferated more rapidly in α-MEM medium. Regarding growth factors that could stimulate cell proliferation rate, we observed that PDGF, FGF2 and Wnt3a had positive effects on proliferation of hFBC. Gene expression analysis demonstrated a higher expression of runx2 in hFBC cultured in basal conditions, which was was similar than that was observed in hMSC in osteoinductive culture conditions. Expression of sox9 was very low in hBFC and hMSC, compared to expression observed in fetal cartilage cells. Looking at osteogenic differentiation capacity, ALP activity was positively regulated byWnt5awhen hFBCwere cultured inα-MEM, but not in DMEM. Conversely, Wnt3a was shown to block the effect of osteogenic inductors on differentiation of both cell types. CONCLUSION. Data presented in this study indicate that the proliferation and differentiation of fetal and mesenchymal stem cells is optimal in α- MEM. Evidence for a pre-differentiated state of hBFC was given by extracellular matrix spontaneous mineralization as well as by higher ALP activity levels observed for these cells in baseline culture conditions, in comparison with hMSC. As we showed that, in vitro, hFBC express a higher capacity to differentiate in osteoblasts, they represent an attractive and promising prospect for fundamental research, and specifically for a new generation of skeletal tissue engineering.

Maturação e qualidade de uvas para suco em condições tropicais, nos primeiros ciclos de produção

O objetivo deste trabalho foi caracterizar a maturação e a qualidade das uvas das cultivares Isabel Precoce e BRS Cora, enxertadas sobre 'IAC 572', para a determinação do ponto de colheita. O experimento foi realizado no Vale do Submédio São Francisco, nos ciclos de produção de novembro de 2009 a março de 2010 e de junho a setembro de 2010, em delineamento experimental de blocos ao acaso, com quatro repetições constituídas por cinco cachos. Os cachos foram coletados, periodicamente, a partir do início da maturação, que correspondeu, na 'Isabel Precoce', aos 54, 61, 68, 71, 74 e 77 dias após a frutificação (DAF), no primeiro ciclo, e aos 49, 56, 63, 67, 71, 74 e 77 DAF, no segundo. Na 'BRS Cora', os cachos foram coletados aos 61, 68, 71, 74, 77 e 82 DAF, no primeiro ciclo, e aos 53, 60, 65, 70, 74 e 78 DAF, no segundo. As uvas de 'BRS Cora' são mais ácidas do que as de 'Isabel Precoce'; porém, apresentam maiores teores de sólidos solúveis e de açúcares solúveis. O ponto de colheita da 'Isabel Precoce' ocorreu aos 77 DAF, em ambos os períodos de produção; na 'BRS Cora', ocorreu aos 82 DAF, no primeiro semestre, e foi antecipado em quatro dias no segundo.

Seleção de cultivares de laranja doce de maturação precoce por índices de desempenho

O objetivo deste trabalho foi selecionar cultivares de laranja doce de maturação precoce, adequadas para o mercado de frutas in natura e para o processamento industrial, por meio de índices de desempenho. Índices de desempenho para citros foram estabelecidos com base em dados coletados em experimento conduzido na região sudoeste do Estado de São Paulo, envolvendo 12 cultivares de laranja doce de maturação precoce. Resultados pioneiros foram obtidos na identificação de cultivares superiores. Em comparação com a laranja 'Hamlin', cultivar padrão de maturação precoce, identificaram-se as laranjas 'Valência 2' e 'Salustiana' com potencial para o mercado de frutas in natura, e a laranja 'Westin', para o processamento industrial.

Ponto de colheita e maturação de frutos de camu-camu colhidos em diferentes estádios

O objetivo deste trabalho foi determinar o ponto de colheita e caracterizar a pós-colheita de frutos de camu-camu (Myrciaria dubia) colhidos em diferentes estádios de maturação. A colheita dos frutos foi realizada em quatro estádios de maturação, definidos pela cor da casca: verde, verde-avermelhada, vermelho-esverdeada e roxa. O armazenamento foi feito em câmaras de refrigeração a 22±1°C e 85±5% UR. Utilizou-se delineamento experimental inteiramente casualizado, em parcelas subdivididas no tempo, com cinco períodos de armazenamento: 0, 3, 6, 9 e 12 dias. Foram avaliados: atividade respiratória; produção de etileno; coloração da casca verificada pelo ângulo de cor e coordenadas de cromaticidade a* e b*; firmeza; perda de massa de matéria fresca; teores de clorofilas, antocianinas, sólidos solúveis e ácido ascórbico; acidez titulável; e incidência de podridão. Os picos de produção de CO2 e etileno ocorreram após a colheita. Os teores de clorofilas e antocianinas variaram com a mudança na coloração da casca de verde para roxa, o que confirmou a maturação dos frutos. Os teores de ácido ascórbico variaram de 759,02 mg por 100 g, no estádio verde, a 1.071,12 mg por 100 g, no roxo. Os frutos colhidos totalmente roxos têm reduzida vida pós-colheita. A maior qualidade pós-colheita do camu-camu é obtida quando os frutos são colhidos com coloração vermelho-esverdeada.