RESUMO
Two experiments were conducted to determine the influence of supplemental nonprotein N (NPN) provided daily (D) or every other day (2D) on ruminant performance and N efficiency. Treatments included an unsupplemented control (CON) and a urea (28.7% CP) or biuret (28.6% CP) supplement provided D or 2D at 0700. In Exp. 1, five wethers (39 +/- 1 kg BW) were used in an incomplete 5 x 4 Latin square with four 24-d periods to determine the influence of supplemental NPN source and supplementation frequency (SF) on the efficiency of N use in lambs consuming low-quality grass straw (4% CP). The amount of CP supplied by each supplement was approximately 0.10% of BW/d (averaged over a 2-d period). In Exp. 2, 80 Angus x Hereford cows (540 +/- 8 kg BW) in the last third of gestation were used to determine the effect of NPN source and SF on cow performance. The NPN treatments were formulated to provide 90% of the estimated degradable intake protein requirement. The supplemented treatments received the same amount of supplemental N over a 2-d period; therefore, the 2D treatments received double the quantity of supplemental N on their respective supplementation day than the D treatments. In Exp. 1, total DM, OM, and N intake; DM, OM, and N digestibility; N balance; and digested N retained were greater (P < 0.03) for supplemented than for CON wethers, with no difference (P > 0.05) between NPN sources or SF. Plasma urea-N (PUN) was increased with N supplementation compared with CON (P < 0.01), and urea treatments had greater PUN than biuret (P < 0.01). In addition, PUN was greater (P = 0.02) for D than for 2D treatments. In Exp. 2, pre- and postcalving (within 14 d and 24 h after calving, respectively) cow weight and body condition score change were more positive (P < 0.05) for supplemented groups than for CON. These results suggest that supplements containing urea or biuret as the primary source of supplemental N can be effectively used by lambs and cows consuming low-quality forage, even when provided every other day.
Assuntos
Biureto/administração & dosagem , Bovinos/fisiologia , Proteínas Alimentares/metabolismo , Nitrogênio/metabolismo , Ovinos/metabolismo , Ureia/administração & dosagem , Ração Animal , Animais , Biureto/efeitos adversos , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Esquema de Medicação/veterinária , Feminino , Masculino , Necessidades Nutricionais , Gravidez , Distribuição Aleatória , Rúmen/metabolismo , Ureia/efeitos adversosRESUMO
Five steers (491 +/- 21 kg BW) were used in an incomplete 5 x 4 Latin square with four 24-d periods to determine the influence of supplemental non-protein N (NPN) source and supplementation frequency (SF) on nutrient intake and site of digestion in steers consuming low-quality grass straw (4% CP). Treatments (TRT) included an unsupplemented control and a urea- or biuret-containing supplement placed directly into the rumen daily (D) or every other day (2D) at 0700. The NPN treatments were formulated to provide 90% of the estimated degradable intake protein requirement. Daily TRT were supplemented CP at 0.04% of BW/d, whereas the 2D TRT were supplemented at 0.08% of BW every other day. Therefore, all supplemented TRT received the same quantity of supplemental CP over a 2-d period. Forage OM intake was not affected (P > 0.05) by NPN supplementation, NPN source, or SF; however, total OM and N intake were increased (P < 0.01) with CP supplementation. Duodenal flow of N was greater (P = 0.04) with CP supplementation compared with the control. In addition, duodenal bacterial N flow was increased with CP supplementation (P = 0.04) and for biuret compared with urea (P < 0.01). Bacterial efficiency (g bacterial N/kg OM truly digested in the rumen) was greater (P = 0.05) for biuret than for urea. Apparent total-tract N digestibility was increased with NPN supplementation (P < 0.01) but not affected by NPN source or SF. These results suggest that urea or biuret can be used effectively as a supplemental N source by steers consuming low-quality forage.
Assuntos
Bactérias/metabolismo , Biureto/administração & dosagem , Bovinos/metabolismo , Digestão , Rúmen/metabolismo , Ureia/administração & dosagem , Ração Animal/normas , Animais , Suplementos Nutricionais , Esquema de Medicação/veterinária , Masculino , Nitrogênio/metabolismo , Necessidades Nutricionais , Distribuição Aleatória , Rúmen/microbiologiaRESUMO
Five ruminally and duodenally cannulated steers (491 +/- 21 kg BW) were used in an incomplete 5 x 4 Latin square with four 24-d periods to determine the influence of supplemental nonprotein N (NPN) source and supplementation frequency (SF) on the dynamics of ruminal fermentation in steers consuming low-quality grass straw (4% CP). Treatments (TRT) included an unsupplemented control (CON) and a urea or biuret supplement that were placed directly into the rumen at 0700 daily (D) or every other day (2D). The NPN treatments were formulated to provide 90% of the estimated degradable intake protein requirement; therefore, the urea and biuret treatments received the same amount of supplemental N over a 2-d period. Daily TRT were supplemented with CP at 0.04% of BW/d, whereas the 2D TRT were supplemented at 0.08% of BW every other day. Forage was provided at 120% of the previous 5-d average intake in two equal portions at 0715 and 1900. Ruminal fluid was collected 0, 3, 6, 9, 12, and 24 h after supplementation on a day of and a day before supplementation for all TRT. Ruminal NH3-N increased (P < 0.04) with CP supplementation on the day all supplements were provided and on the day on which only daily supplements were provided compared with the CON. However, an NPN source x SF interaction (P = 0.03) on the day all supplements were provided indicated that NH3-N increased at a greater rate for urea as SF decreased compared with biuret. Ruminal NH3-N on the day only daily supplements were provided was greater (P = 0.02) for D compared with 2D. On the day all supplements were provided, D increased (P = 0.05) ruminal indigestible acid detergent fiber passage rate and ruminal fluid volume compared with 2D. These results suggest that urea or biuret can be used effectively as a supplemental N source by steers consuming low-quality forage without adversely affecting ruminal fermentation, even when provided every other day.
Assuntos
Ração Animal/normas , Biureto/administração & dosagem , Bovinos/metabolismo , Duodeno/metabolismo , Rúmen/metabolismo , Ureia/administração & dosagem , Animais , Bactérias/metabolismo , Biureto/efeitos adversos , Suplementos Nutricionais , Digestão , Esquema de Medicação/veterinária , Duodeno/microbiologia , Fermentação/efeitos dos fármacos , Cinética , Masculino , Necessidades Nutricionais , Distribuição Aleatória , Rúmen/microbiologia , Ureia/efeitos adversosRESUMO
Developmental dyslexia is a language-based learning disability, and a number of candidate dyslexia susceptibility genes have been identified, including DYX1C1, KIAA0319, and DCDC2. Knockdown of function by embryonic transfection of small hairpin RNA (shRNA) of rat homologues of these genes dramatically disrupts neuronal migration to the cerebral cortex by both cell autonomous and non-cell autonomous effects. Here we sought to investigate the extent of non-cell autonomous effects following in utero disruption of the candidate dyslexia susceptibility gene homolog Dyx1c1 by assessing the effects of this disruption on GABAergic neurons. We transfected the ventricular zone of embryonic day (E) 15.5 rat pups with either Dyx1c1 shRNA, DYX1C1 expression construct, both Dyx1c1 shRNA and DYX1C1 expression construct, or a scrambled version of Dyx1c1 shRNA, and sacrificed them at postnatal day 21. The mothers of these rats were injected with BrdU at either E13.5, E15.5, or E17.5. Neurons transfected with Dyx1c1 shRNA were bi-modally distributed in the cerebral cortex with one population in heterotopic locations at the white matter border and another migrating beyond their expected location in the cerebral cortex. In contrast, there was no disruption of migration following transfection with the DYX1C1 expression construct. We found untransfected GABAergic neurons (parvalbumin, calretinin, and neuropeptide Y) in the heterotopic collections of neurons in Dyx1c1 shRNA treated animals, supporting the hypothesis of non-cell autonomous effects. In contrast, we found no evidence that the position of the GABAergic neurons that made it to the cerebral cortex was disrupted by the embryonic transfection with any of the constructs. Taken together, these results support the notion that neurons within heterotopias caused by transfection with Dyx1c1 shRNA result from both cell autonomous and non-cell autonomous effects, but there is no evidence to support non-cell autonomous disruption of neuronal position in the cerebral cortex itself.
Assuntos
Proteínas de Transporte/genética , Córtex Cerebral/anormalidades , Córtex Cerebral/metabolismo , Neurônios/metabolismo , Ácido gama-Aminobutírico/metabolismo , Animais , Animais Recém-Nascidos , Proteínas de Transporte/antagonistas & inibidores , Córtex Cerebral/crescimento & desenvolvimento , Regulação para Baixo/genética , Neurônios/citologia , Interferência de RNA/fisiologia , RNA Interferente Pequeno/genética , Ratos , Ratos Wistar , Transfecção/métodosRESUMO
Transcription of the surfactant protein-C (SP-C) gene is restricted to Type II epithelial cells in the adult lung. We have shown previously that the 0.32-kilobase pair (kb) mouse SP-C promoter is functional in transient transfection assays of the lung epithelial cell-derived cell line, MLE-15, and that thyroid transcription factor 1 (TTF-1) transactivates promoter activity. The 0.32-kb SP-C promoter can be separated into a proximal promoter region (-230 to +18) and an enhancer region (-318 to -230). Three DNase I footprints were mapped in the promoter region (C1 through C3) and two in the enhancer region (C4 and C5). We now show that nuclear factor I (NFI) family members bind to both individual NFI half-sites in footprints C1, C3, and C5, and to a composite site in footprint C4 by competition gel retardation and antibody supershift analyses. Mutational analysis of the 0.32-kb mouse SP-C promoter and transient transfection of MLE-15 cells demonstrated that the NFI binding sites are required for promoter activity in this cell type. Site-specific mutation of the proximal or distal NFI sites drastically reduced transactivation by a co-transfected NFI-A expression vector in HeLa cells. These data indicate that NFI family member(s), binding to sites in both the promoter and enhancer regions, regulate SP-C gene expression in a process independent of TTF-1.
Assuntos
Proteínas Estimuladoras de Ligação a CCAAT , Proteínas de Ligação a DNA/fisiologia , Proteolipídeos/genética , Surfactantes Pulmonares/genética , Fatores de Transcrição/fisiologia , Transcrição Gênica , Animais , Sequência de Bases , Sítios de Ligação , Desoxirribonuclease I/metabolismo , Células HeLa , Humanos , Camundongos , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Fatores de Transcrição NFI , Proteínas Nucleares , Regiões Promotoras Genéticas , Transfecção , Proteína 1 de Ligação a Y-BoxRESUMO
Linear bone growth was studied in male mice possessing a controlled ovine metallothionein 1a promoter-ovine growth hormone (oMT1a-oGH) transgene. Transgene expression was activated at weaning by the addition of 25 mM zinc sulfate to drinking water; transgenic and control mice received the zinc supplementation. The ulna, humerus, and tibia were excised at 10-day intervals until 130 days from control and from mice hemizygous for oMT1a-oGH. Bones from mice overexpressing growth hormone (GH) were 11-20% longer than those from controls (P less than 0.01) at 130 days. Transgenic mice exhibited both an enhanced rate of bone growth and a growth period of greater duration, i.e., the ulna, tibia, and humerus from oMT1a-oGH mice grew at an accelerated rate for an additional 20-40 days relative to the same bones from control mice. The bones from both groups were characterized by isometric growth patterns. Genetic size scaling revealed that the observed differences in bone growth were directly related to the mature size of the bone, suggesting that the bones possess an inherent growth pattern that is followed even in the presence of elevated GH.