Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Toxoplasma gondii in experimentally infected Bos taurus and Bos indicus semen and tissues

Dezoito bovinos foram inoculados com Toxoplasma gondii e distribuídos aleatoriamente em três grupos de seis bovinos cada: GI (2,5x10(5) oocistos da cepa P), GII (5,0x10(6) taquizoítos da cepa RH) e GIII (controle). Exames clínicos, sorológicos e parasitêmicos foram realizados. Pesquisas do parasito, por meio da bioprova e pela técnica de Reação em Cadeia pela Polimerase (PCR), foram realizadas no sêmen e em fragmentos de musculatura esquelética, linfonodos, cérebro, retina, baço, fígado, pulmão, testículo, epidídimo e vesícula seminal. Amostras de sangue e sêmen foram colhidas nos dias -2, -1, 1, 3, 5, 7, 14 e, semanalmente, até o 84º dia pós-infecção (DPI). Os bovinos inoculados (GI e GII) apresentaram hipertermia do 3º ao 16º DPI. Anticorpos contra T. gondii foram detectados (IFI) no 5º DPI (1:16), em ambos grupos inoculados (oocistos e taquizoítos), atingindo picos de 1:4096 no 7º DPI. Surtos parasitêmicos ocorreram em todos os bovinos infectados, principalmente do 7º ao 28º DPI, independente da cepa e inóculo utilizados. O bioensaio revelou a presença do parasito em amostras seminais dos bovinos infectados com oocistos (GI) e taquizoítos (GII), em diversas datas experimentais, entre o 7º e 84º DPI. Parasitismo tissular por T. gondii foi diagnosticado por meio da bioprova e pela técnica da PCR, em vários fragmentos de tecidos e/ou órgãos. Os achados sugerem a possibilidade da ocorrência da transmissão sexual do T. gondii na espécie bovina.Eighteen young steers were inoculated with Toxoplasma gondii and randomly distributed into three groups of six animals each: GI, 2.5x10(5) P strain oocysts, GII, 5.0x10(6) RH strain tachyzoites, and GIII (Control). Clinical, serological and parasitemia exams were realized. Parasite investigation by bioassay and PCR was realized on semen and fragments of skeletal musculature, lymph nodes, brain, retina, spleen, liver, lung, testicle, epididymis and seminal vesicle. Blood and semen samples were collected on days -2, -1, 1, 3, 5, 7, 14 and weekly thereafter, up to postinfection day (PID) 84. The inoculated steers (GI and GII) presented hyperthermia from PID 3 to 16. Antibodies against T. gondii were detected through the indirect fluorescence antibody test (IFAT) on PID 5 (1:16) in both inoculated groups (oocysts and tachyzoites), reaching peaks of 1:4096 on PID 7. Parasitemia outbursts occurred in all infected bovines, principally from PID 7 to 28, independent of the strain and inoculate used. Bioassays revealed the presence of parasites in semen samples of animals infected with oocysts (GI) and tachyzoites (GII) on several experimental days between PID 7 and 84. Tissue parasitism by T. gondii was diagnosed by bioassay and the PCR technique in several organ and tissue fragments. These findings suggest the possibility of sexual transmission of T. gondii in the bovine species

Rectal temperatures in postpartum cows

O trabalho teve como objetivo correlacionar dados da parturição com o acompanhamento diário da temperatura retal no pós-parto imediato em fêmeas bovinas. Foram selecionadas aleatoriamente 180 vacas leiteiras provenientes de sete propriedades, em Araçatuba (SP) e região, sendo monitoradas durante o puerpério precoce, no período de setembro de 1999 a julho de 2000. Do primeiro ao décimo dia pós-parto a temperatura retal (TR) de todos os animais foi aferida no intervalo das 05:00 às 08:00 horas da manhã, utilizando-se termômetro eletrônico (M525 - GLA Agricultural Electronics, San Luis Obispo, CA 93401-7500). Foi considerada acima da normalidade TR superior a 39,5°C, sendo colhidas informações sobre eventuais distocias, partos gemelares, retenção de membranas fetais, alteração do estado geral, ingestão de alimentos, produção de leite e descarga vaginal. Houve associação significativa entre descarga vaginal purulenta ou sanguinolenta e estado febril, sendo que dos 180 animais observados, 26 (14,4%) apresentaram corrimento vaginal patológico, destes dez (38,5%) tiveram febre, enquanto que os bovinos sem presença de secreção alterada tiveram elevação na temperatura corpórea em 17,5% dos casos. Também ocorreu associação significativa entre parto distócico e placenta retida, 37 vacas (20,6%) mostraram parturição difícil e, destas, oito (21,6%) retiveram as membranas fetais, enquanto as fêmeas que pariram normalmente tiveram 4,2% (seis animais) de retenção de secundinas. Constatou-se que vacas apenas com retenção placentária não apresentaram sinais clínicos, exceto presença de uma transitória redução no apetite e da produção láctea. Este monitoramento diário, através da aferição da temperatura retal, consiste em importante instrumento adicional, podendo indicar precocemente alterações reprodutivas, como possíveis infecções uterinas, maximizando a produção leiteira e a performance reprodutiva futura.The purpose of this study was to evaluate parturition data with the rectal temperature in the early postpartum period of dairy cows. One hundred and eighty cows were randomly selected between September 1999 and July 2000, in seven dairy farms located in the Northwest region of São Paulo, Brazil. For the first ten days postpartum, rectal temperature (RT) was taken between 5:00 and 8:00 a.m. using an electronic thermometer (M525 - GLA Agricultural Electronics, San Luis Obispo, CA 93401-7500). Cows with RT>39,5°C were enrolled and other signs like calving difficulties, dystocia due to twins, fetal membrane retention, ill appearance, off-feed, milk production and vaginal discharge were recorded. Significant association was observed between vaginal discharge and fever. Considering 180 animals, 26 (14,4%) showed vaginal discharge, among them ten (38,5%) were pyretic, while in cows without vaginal discharge only 27 (17,5%) showed higher rectal temperatures. Difficult parturition and retained membranes also were statistically different, 37 cows (20,6%) presented dystocia, among them eight (21,6%) had retention of the afterbirth, while in the cows with normal parturition only six (4,2%) showed retention of the fetal membranes. It was observed that cows with or without retained fetal membranes did not show clinical signs less a transitory reduction of appetite and milk production. This daily monitoring of rectal temperature represents an aditional tool indicating precociously reprodutive alterations, like uterine infections, maximizing milk production and future uterine performance.

Mechanical stretch promotes fetal type II epithelial cell differentiation via shedding of HB-EGF and TGF-α

The mechanisms by which mechanical forces promote fetal lung development are not fully understood. Here, we investigated differentiation of fetal type II epithelial cells via the epidermal growth factor receptor (EGFR) in response to mechanical strain. First, we showed that incubation of embryonic day (E) 19 fetal type II cells with recombinant heparin-binding EGF-like growth factor (HB-EGF) or transforming growth factor (TGF)-α, but not with amphiregulin (AR), betacellulin (BTC) or epiregulin (EPR), increased fetal type II cell differentiation, as measured by surfactant protein B/C mRNA and protein levels. Next, we demonstrated that 5% cyclic stretch of E19 monolayers transfected with plasmid encoding alkaline phosphatase (AP)-tagged ligands shed mature HB-EGF and TGF-α into the supernatant and promoted type II cell differentiation. Release of these ligands was also observed in E19 cells subjected to higher degrees of cyclic strain, but not in cells exposed to continuous stretch. Interestingly, the addition of fibroblasts to type II cell cultures did not enhance release of HB-EGF. Whereas HB-EGF shedding was also detected in E18 cells exposed to 5% cyclic stretch, release of this ligand after 2.5% sustained stretch was restricted to cells isolated on E18 of gestation. In addition, mechanical stretch released EGF, AR and BTC. We conclude that mechanical stretch promotes fetal type II cell differentiation via ectodomain shedding of HB-EGF and TGF-α. The magnitude of shedding varied depending on gestational age, ligand, and strain protocol. These studies provide novel mechanistic information potentially relevant to fetal lung development and to mechanical ventilation-induced lung injury