Towards liver-directed gene therapy: Retrovirus-mediated gene transfer into human hepatocytes

Liver-directed gene therapy is being considered in the treatment of inherited metabolic diseases. One approach we are considering is the transplantation of autologous hepatocytes that have been genetically modified with recombinant retroviruses ex vivo. We describe, in this report, techniques for isolating human hepatocytes and efficiently transducing recombinant genes into primary cultures. Hepatocytes were isolated from tissue of four different donors, plated in primary culture, and exposed to recombinant retroviruses expressing either the LacZ reporter gene or the cDNA for rabbit LDL receptor. The efficiency of gene transfer under optimal conditions, as determined by Southern blot analysis, varied from a maximum of one proviral copy per cell to a minimum of 0.1 proviral copy per cell. Cytochemical assays were used to detect expression of the recombinant derived proteins, E. coli β-galactosidase and rabbit LDL receptor. Hepatocytes transduced with the LDL receptor gene expressed levels of receptor protein that exceeded the normal endogenous levels. The ability to isolate and genetically modify human hepatocytes, as described in this report, is an important step towards the development of liver-directed gene therapies in humans.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/45540/1/11188_2005_Article_BF01233625.pd

Consumo de combustível na operação de subsolagem realizada antes e depois de diferentes sistemas de preparo periódico do solo Fuel consumption in the subsoiling operation accomplished before and after different preparation systems of periodic soil tillage

A subsolagem tem aumentado nos últimos anos de forma indiscriminada, faltando estudos que norteiem os melhores procedimentos para que novos problemas não sejam acrescentados devido a subsolagens inadequadas ou mesmo em solos onde a operação é desnecessária, e principalmente buscar redução no consumo de combustível. Dessa forma, este trabalho teve o objetivo de avaliar o consumo de combustível na operação de subsolagem efetuada antes e depois de diferentes sistemas de preparo periódico do solo, classificado como Nitossolo Vermelho distroférrico. Os preparos foram realizados com arado de discos, arado de discos mais uma gradagem de nivelamento, grade pesada, grade pesada mais gradagem de nivelamento e escarificador. A realização da subsolagem depois dos sistemas de preparo periódico requereu 15% menos de potência na barra de tração. A subsolagem depois dos diferentes sistemas de preparo economizou 16,5% de combustível por área. O deslizamento das rodas motrizes e a velocidade média operacional obtiveram melhor desempenho quando se realizou a subsolagem depois do preparo do solo.<br>The subsoiling has had a great increase in the last few years, but there is still a lack of studies that guide the best procedures so that new problems do not appear because of inadequate subsoiling or even in soil where the same operation is unnecessary, and primarily look for a reduction in the consumption of fuel. Thus, this work had the objective of evaluating the fuel consumption in the operation of subsoiling performed before and after different systems of preparation of a regular land classified as Nitossolo Red Distroférrico. The periodic tillage systems were: plowing with disks, plowing with disks followed by one leveling, plowing harrow, plowing harrow followed by leveling and chiseling. The accomplishment of the subsoiling after the preparation of periodic systems required 15% less power in the bar of traction. The subsoiling after the different systems of preparation saved 16.5% of fuel per area. The slip of the driving wheels and average operational speed obtained better performance when it took the subsoiling after preparing the soil

Demanda energética na subsolagem realizada antes e depois de diferentes sistemas de preparo periódico do solo Energy demand in the subsoiling performed before and after different systems of periodic soil tillage

A subsolagem é uma das operações mecanizadas de elevado custo e demanda energética por área, tradicionalmente utilizada pelos agricultores antes do preparo do solo na descompactação de camadas adensadas. Este trabalho teve como objetivo avaliar a demanda energética na operação de subsolagem realizada antes e depois de diferentes sistemas de preparo periódico num solo classificado como Nitossolo Vermelho Distroférrico. Os sistemas de preparo periódico foram: aração com discos; aração com discos mais uma gradagem de nivelamento; grade aradora; grade aradora mais gradagem de nivelamento e escarificação. O trator utilizado como fonte de potência no experimento foi um Valmet 128 (4x2tda). A demanda energética por área foi menor na subsolagem realizada depois do preparo periódico do solo, proporcionando uma economia de 21,9%. A realização da subsolagem depois do preparo periódico do solo resultou numa diminuição da exigência de força de tração em 21,1% e da potência disponível na barra de tração em 15%.<br>Subsoiling is one of the mechanized operations of high cost and energy demand per area, traditionally utilized by farmers before soil tillage in the decompactation of hardened layers. This research was intended to evaluate the energy demand in the subsoiling operation performed before and after different systems of periodic tillage in a soil classified as Distroferric Red Nitossol. The periodic tillage systems were: plowing with disks; plowing with disks plus one leveling; plowing harrow; plowing harrow plus leveling and chiseling. The tractor utilized as a power source in the experiment was a Valmet 128 (4x2tda). The demand for energy area was lower in subsoiling held after the regular preparation of the soil, providing a saving of 21.9%. The completion of subsoiling after the preparation of the soil resulted in a decrease in demand for power to pull in 21% and 15% of the power available in the bar of traction

The role of puberty on physical and brain development: A longitudinal study in male Rhesus Macaques

This study examined the role of male pubertal maturation on physical growth and development of neurocircuits that regulate stress, emotional and cognitive control using a translational nonhuman primate model. We collected longitudinal data from male macaques between pre- and peri-puberty, including measures of physical growth, pubertal maturation (testicular volume, blood testosterone -T- concentrations) and brain structural and resting-state functional MRI scans to examine developmental changes in amygdala (AMY), hippocampus (HIPPO), prefrontal cortex (PFC), as well as functional connectivity (FC) between those regions.Physical growth and pubertal measures increased from pre- to peri-puberty. The indexes of pubertal maturation -testicular size and T- were correlated at peri-puberty, but not at pre-puberty (23 months). Our findings also showed ICV, AMY, HIPPO and total PFC volumetric growth, but with region-specific changes in PFC. Surprisingly, FC in these neural circuits only showed developmental changes from pre- to peri-puberty for HIPPO-orbitofrontal FC. Finally, testicular size was a better predictor of brain structural maturation than T levels -suggesting gonadal hormones-independent mechanisms-, whereas T was a strong predictor of functional connectivity development. We expect that these neural circuits will show more drastic pubertal-dependent maturation, including stronger associations with pubertal measures later, during and after male puberty

Hastes instrumentadas para a mensuração da resistência mecânica do solo Instrumented shanks for soil mechanical resistance measurements

A presença de camada de solo compactada pode significar sérios problemas ao desenvolvimento do sistema radicular das culturas em geral. A correção pode ser feita por métodos biológicos ou mecânicos. Uma das formas de diagnóstico da sua presença é com o uso de penetrômetros que exigem tempo considerável para a obtenção de dados. Com o advento dos recursos de espacialização das informações na forma de mapas, a necessidade de amostragem passou a ser ainda maior. Este trabalho reporta a busca de uma solução alternativa ao uso do penetrômetro. Envolve a geração de mapas de resistência mecânica do solo ao deslocamento de uma haste em três profundidades, baseado em três hastes instrumentadas, um receptor de GPS e um sistema de aquisição de dados. O sistema é acoplado a uma barra porta-ferramenta montada no engate de três pontos do trator. A configuração mostrou ser mecanicamente bastante simples e confiável. Dados de um teste inicial em campo demonstraram que o sistema pode produzir mapas que denotam a variabilidade espacial da referida resistência nas áreas amostradas, com alta resolução, densidade de amostragem elevada e com capacidade operacional ainda mais elevada se comparada com levantamentos feitos com métodos convencionais.<br>The presence of soil compaction may be considered a serious problem for the development of crop root systems. It can be alleviated by biological or mechanical methods (such as tillage). One of the most popular diagnostic methods is based on standardized cone penetrometer measurements, which requires a considerable amount of time to obtain the data. With the advent of site-specific field management, high-density measurements have become a necessity. This work reports on an alternative approach to use a cone penetromenter. It involves the generation of maps of soil mechanical resistance at three depths using three instrumented blades, a GPS receiver and a data logger. The system is attached to a toolbar that is installed to the tractor's three-point hitch. It has proven to be mechanically simple and reliable. Maps indicating the variability of soil mechanical resistance with relatively high spatial resolution (as compared to traditional methods) were generated in a reasonable amount of time based on data collected during initial field tests