RESUMO
Records collected between 1997 and 2010 were used to determine the effect of calving period on heifer (n = 1,019) and steer (n = 771) progeny from the Gudmundsen Sandhills Laboratory, Whitman, NE. Progeny were classified as being born in the first, second, or third 21-d period of the spring calving season within year. Heifer birth BW was lightest (P < 0.01) for heifers born in the first period. Birth to weaning ADG tended (P = 0.10) to be least for heifers born in the first calving period; however, weaning BW decreased (P = 0.03) with advancing calving period. Weaning to prebreeding ADG tended (P = 0.07) to be least for heifers born in the first period; however, prebreeding BW was greatest (P < 0.01) for calves born in the first period. Heifer ADG from the beginning of the breeding season to pregnancy diagnosis was greater (P = 0.03) for heifers born in the third vs. first calving period. Heifers cycling at the beginning of the breeding season decreased (P < 0.01) with advancing calving date (70, 58, and 39%, respectively) and 45 d pregnancy rates were lowest (P = 0.02) for heifers born in the third calving period (90, 86, and 78%, respectively). Birth date of the first calf of the heifer and birth BW decreased (P < 0.01) if the heifer was born in the first calving period. First calf progeny had the greatest (P ≤ 0.10) weaning BW if born to a heifer born in the first calving period. As steer calving period advanced, weaning BW decreased (P < 0.01). Calving period did not affect (P = 0.81) feedlot ADG. As calving period advanced, HCW, marbling score, and yield grade decreased (P < 0.01). The percentage of steers grading USDA small marbling was not affected (P = 0.13) by calving period; however, the percentage of steers grading USDA modest marbling or greater and the total carcass value declined (P ≤ 0.01) as calving period advanced. Heifer calves born during the first 21 d of the spring calving season had greater weaning, prebreeding, and precalving BW; greater percent cycling before breeding; and greater pregnancy rates compared with heifers born in the third period. First calf progeny also had an earlier birth date and greater weaning BW. Steer calves born during the first 21 d of the calving season had greater weaning, final, and carcass weights; greater marbling scores; a greater percentage grading USDA modest or greater; and greater carcass value. Calving period of progeny significantly impacts performance.
Assuntos
Bovinos/crescimento & desenvolvimento , Desmame , Aumento de Peso , Criação de Animais Domésticos , Animais , Bovinos/fisiologia , Feminino , Masculino , Nebraska , Parto , Gravidez , Taxa de Gravidez , Estações do Ano , Fatores de TempoRESUMO
Studies in numerous species provide evidence that diet during development can mediate physiological changes necessary for puberty. In cattle, several studies have reported inverse correlations between postweaning growth rate and age at puberty and heifer pregnancy rates. Thus, postweaning growth rate was determined to be an important factor affecting age of puberty, which in turn influences pregnancy rates. This and other research conducted during the late 1960s through the early 1980s indicated puberty occurs at a genetically predetermined size, and only when heifers reach their target BW can increased pregnancy rates be obtained. Guidelines were established indicating replacement heifers should achieve 60 to 65% of their expected mature BW by breeding. Traditional approaches for postweaning development of replacement heifers used during the last several decades have primarily focused on feeding heifers to achieve or exceed an appropriate target BW and thereby maximize heifer pregnancy rates. Intensive heifer development systems may maximize pregnancy rates, but not necessarily optimize profit or sustainability. Since inception of target BW guidelines, subsequent research demonstrated that the growth pattern heifers experience before achieving a critical target BW could be varied. Altering rate and timing of BW gain can result in compensatory growth periods, providing an opportunity to decrease feed costs. Recent research has demonstrated that feeding replacement heifers to traditional target BW increased development costs without improving reproduction or subsequent calf production relative to development systems in which heifers were developed to lighter target BW ranging from 50 to 57% of mature BW.
Assuntos
Fenômenos Fisiológicos da Nutrição Animal/fisiologia , Bovinos/fisiologia , Ração Animal , Animais , Bovinos/crescimento & desenvolvimento , Dieta/veterinária , Feminino , Gravidez , Maturidade Sexual/fisiologiaRESUMO
Two hundred ninety-nine Angus-based, nulliparous heifers (253 ± 2 kg initial BW) from 3 production years were utilized to compare traditional postweaning dry lot (DL) development with a more extensive winter grazing system utilizing a combination of corn residue and winter range (EXT). Heifers developed in the DL were offered a common diet after the weaning period for 208 d in yr 1, 194 d in yr 2, and 150 d in yr 3 until breeding. Heifers developed in EXT grazed corn residue for 135 d in yr 1, 106 d in yr 2, and 91 d in yr 3, and then fed in the DL until breeding (yr 1) or grazed dormant winter grass for approximately 60 d before being fed in the DL (yr 2 and 3). All 3 years, heifers were estrus synchronized, with timed AI performed in yr 1. In yr 2 and 3, estrus was detected and those detected in estrus were artificially inseminated approximately 12 h later. Heifers were exposed to bulls 10 d after the last AI for 60 d while grazing summer pasture. During the winter grazing period, EXT heifers gained less (P = 0.01) BW than DL heifers and EXT heifers had lighter (P = 0.02) BW at breeding. Fewer (P < 0.01) EXT heifers reached puberty before breeding. Conception to AI was not different (P = 0.23); however, AI pregnancy rate tended (P = 0.08) to be less in EXT heifers. Final pregnancy rates were not different (P = 0.38) between treatment groups. Although EXT heifers had lighter (P = 0.02) BW at pregnancy diagnosis; however, they did compensate with greater (P = 0.05) ADG after breeding, resulting in similar (P = 0.22) precalving BW. Winter development system did not influence (P > 0.10) percentage of calving in the first 21 d, calf birth date, and calf birth BW, or dystocia score. Pregnancy rate after the second breeding season was not different (P = 0.56) between treatments. Heifer development using extended winter grazing reduced (P < 0.01) the cost of producing a pregnant heifer by $45 compared with DL.
Assuntos
Criação de Animais Domésticos/métodos , Bovinos/fisiologia , Dieta/veterinária , Reprodução/fisiologia , Ração Animal , Animais , Animais Recém-Nascidos , Peso ao Nascer/fisiologia , Peso Corporal/fisiologia , Dieta/economia , Feminino , Masculino , Gravidez , Estações do AnoRESUMO
Two experiments at 2 Nebraska locations evaluated effects of heifer development system on growth and pregnancy rate. In Exp. 1, heifers (n=270, BW=225 ± 2 kg) grazed winter Sandhills range (WR) or west central Nebraska corn residue (CR) with a supplement (0.45 kg/animal; 31% CP; 80 mg·animal(-1)·d(-1) of monensin). In Exp. 2, heifers (n=180, BW = 262 ± 3 kg) grazed eastern Nebraska WR or CR with a supplement (0.45 to 0.90 kg/d; 31% CP; 80 to 160 mg·animal(-1)·d(-1) of monensin). The CR heifers tended to have less (P=0.10) ADG compared with WR heifers before breeding in Exp. 1; however, prebreeding ADG was similar (P=0.77) in Exp. 2. Prebreeding BW, percentage of mature BW at breeding, and pregnancy determination BW were similar (P ≥ 0.14) for CR and WR in both experiments. Percentage of heifers pubertal at breeding, AI conception, and AI pregnancy rate (Exp. 2) and final pregnancy rate in both experiments were also similar (P ≥ 0.27) for CR and WR heifers. Precalving BW, percentage of calves born in the first 21 d, calf birth date, calf birth BW, and dystocia score were all similar (P ≥ 0.21) for CR and WR heifers in both experiments. Cow BW at weaning, calf weaning BW, adjusted 205-d calf BW, and second season pregnancy rates were not affected (P ≥ 0.16) by treatment. Heifer development system did not affect (P ≥ 0.56) the cost of producing 1 pregnant heifer in Exp. 1 or 2. Development on CR may reduce ADG before breeding, but did not affect pregnancy rate. Heifer development using CR or WR postweaning resulted in similar reproductive performance and development cost.
Assuntos
Ração Animal , Criação de Animais Domésticos/métodos , Bovinos/crescimento & desenvolvimento , Dieta/veterinária , Criação de Animais Domésticos/economia , Fenômenos Fisiológicos da Nutrição Animal , Animais , Feminino , Gravidez , Taxa de Gravidez , Maturidade Sexual/fisiologia , Aumento de PesoRESUMO
A 2 × 2 factorial arrangement of treatments was used to evaluate effects of dam winter grazing system and supplementation in the last third of gestation on subsequent BW gain, feed efficiency, and reproduction in heifer progeny. Crossbred cows (yr 1, n = 109; yr 2, n = 114; yr 3, n = 116) grazed range (WR) or corn residue (CR) during winter and within grazing treatment received 0.40 kg/d of 31% CP (DM basis) cubes (PS) or no supplement (NS). Heifer calves (yr 1, n = 56; yr 2, n = 56; yr 3, n = 54) grazed dormant pasture for 114 d postweaning and were individually fed for 87 d before a 45-d natural service breeding. Dam PS reduced (P = 0.04) heifer birth date and CR tended to increase (P = 0.07) heifer birth BW. Both PS and CR increased (P ≤ 0.05) heifer weaning BW; however, adjusted 205-d weaning BW was only lighter (P = 0.03) if the dam grazed WR with NS. Heifers from PS dams tended to be younger (P = 0.09) at puberty than NS, and there was a trend (P = 0.11) for more heifers to be pubertal by breeding if the dam grazed WR with PS compared with other treatment groups. Heifers from WR-NS dams tended to weigh less (P ≤ 0.09) at breeding and at pregnancy diagnosis than WR PS. There was a trend (P = 0.13) for pregnancy rate to be greater for heifers born to PS dams. Individually fed heifer DMI was not affected (P = 0.25) by treatment; however, heifers from dams that grazed CR with PS gained the least BW (P = 0.04) during individual feeding and had the smallest (P = 0.03) G:F. In contrast, there were no differences (P > 0.15) in feed efficiency when expressed as residual feed intake. The first calf birth and weaning BW of the heifer was unaffected (P > 0.15) by dam treatment. Heifers from dams that grazed WR with NS tended to have lighter (P = 0.09) BW before the second breeding season but similar (P = 0.97) pregnancy rates. Cows grazing CR with NS produced the most valuable heifer calf at weaning; however, heifers from cows that grazed WR with NS cost the least to develop per pregnant heifer. Winter grazing system and late gestation supplementation affected heifer progeny BW, feed efficiency, and fertility.
Assuntos
Criação de Animais Domésticos/métodos , Fenômenos Fisiológicos da Nutrição Animal , Bovinos/fisiologia , Dieta/veterinária , Fenômenos Fisiológicos da Nutrição Materna , Prenhez , Ração Animal/análise , Criação de Animais Domésticos/economia , Animais , Feminino , Gravidez , Efeitos Tardios da Exposição Pré-Natal , Estações do AnoRESUMO
Developmental programming is the concept that a maternal stimulus or insult at a critical period in fetal development has long-term effects on the offspring. Historically, considerable effort has been made to understand how nutrition influences health and productivity during the postnatal period. Whereas maternal nutrition during pregnancy plays an essential role in proper fetal and placental development, less is known about how maternal nutrition affects the health and productivity of the offspring. Conceptus growth is sensitive to direct and indirect effects of maternal dietary intake. Even from the earliest stages of embryonic life, when nutrient requirements for conceptus growth are negligible, alterations in tissue composition can occur, influencing future growth of the compromised organ system. Not only is neonatal health compromised, but subsequent health may also be programmed because offspring from undernourished dams have exhibited poor growth and productivity and have developed significant diseases later in life. Although the literature is now evolving, with increasing evidence of how maternal nutrient restriction impairs several prenatal physiological variables, few studies have evaluated postnatal growth and development in livestock species, and fewer have evaluated it in beef cattle. In addition, very few studies have evaluated restriction of specific components of the diet during pregnancy (such as protein) on offspring growth and performance. This review focuses on how maternal nutrition affects conceptus growth and postnatal responses in beef cattle.
Assuntos
Bovinos/crescimento & desenvolvimento , Desenvolvimento Fetal/fisiologia , Fenômenos Fisiológicos da Nutrição Materna/fisiologia , Animais , Animais Recém-Nascidos/crescimento & desenvolvimento , Peso ao Nascer/fisiologia , Bovinos/fisiologia , Proteínas Alimentares/farmacologia , Suplementos Nutricionais , Feminino , Desenvolvimento Fetal/efeitos dos fármacos , Fenômenos Fisiológicos da Nutrição Materna/efeitos dos fármacos , Músculo Esquelético/embriologia , Músculo Esquelético/crescimento & desenvolvimento , Organogênese/fisiologia , Placentação , GravidezRESUMO
Ground, raw soybeans (SB), or dried distillers grain plus solubles (DDGS) were utilized in heifer development diets to determine the effect of dietary fat and protein source on hormone and follicle characteristics and ADG. The experiment was conducted over 2 yr with 100 June-born heifers (199 +/- 2 kg initial BW, n = 50 per yr). The experimental periods were 157 and 207 d in yr 1 and 2, respectively. Heifers were provided a dietary supplement (DM basis) of 1.23 kg of SB and 0.40 kg of corn or 1.65 kg of DDGS between weaning and breeding. Estrus was synchronized with 2 injections of PGF(2alpha) 14 d apart. Dominant follicles were measured and aspirated via transvaginal ultrasonography 60 h after the second PGF(2alpha) injection. Heifers were exposed to bulls beginning 14 d after aspiration for 45 d. Heifer ADG was greater (P = 0.02) for DDGS heifers in yr 1, but was similar (P = 0.47) in yr 2. However, there was no difference (P = 0.35) in final BW in either year. There was no difference (P >or= 0.67) in follicle size, follicle hormone concentrations, or pregnancy rate (88%) between yr 1 and 2. Serum estrogen at 48 or 60 h after PGF(2alpha) injection were similar (P >or= 0.91); however, LH at 60 h in yr 2 tended to be greater (P = 0.07) for DDGS heifers. The percentage of heifers experiencing an LH surge 48 and 60 h after PGF(2alpha) injection was not affected (P >/= 0.40) by treatment. Calf production was not affected (P >or= 0.20) by developmental diet. In summary, DDGS and SB have similar effects on hormone and follicle characteristics at the inclusion rates used in these studies.
Assuntos
Ração Animal , Bovinos/fisiologia , Proteínas Alimentares/farmacologia , Folículo Ovariano/fisiologia , Animais , Peso ao Nascer/fisiologia , Bovinos/crescimento & desenvolvimento , Suplementos Nutricionais , Estrogênios/sangue , Feminino , Fertilidade/fisiologia , Hormônio Foliculoestimulante/sangue , Hormônio Luteinizante/sangue , Gravidez , Taxa de Gravidez , Glycine maxRESUMO
Three experiments were conducted to determine the effect of increasing field pea level in high-concentrate finishing cattle diets on ADG, DMI, G:F, and carcass traits, and to estimate the NE of field pea. In Exp. 1, 118 yearling heifers (417.9 +/- 2.4 kg initial BW) were blocked by initial BW and assigned randomly to 1 of 4 treatments (0, 10, 20, or 30% dry-rolled field pea, DM basis; 4 pens/treatment). In Exp. 2, 143 beef steers (433 +/- 19 kg initial BW) were blocked by BW and assigned randomly to 1 of 4 treatments (0, 10, 20, or 30% dry-rolled field pea, DM basis; 6 pens/treatment). In Exp. 3, 80 beef steers (372.4 +/- 0.4 kg initial BW) were assigned randomly to 1 of 4 treatments (0, 18, 27, or 36% cracked field pea, DM basis; 4 pens/treatment). Field pea replaced a portion of the grain (dry-rolled and high moisture corn, dry-rolled corn, and barley and barley sprouts; Exp. 1, 2, and 3, respectively) and protein supplement. In Exp. 1, field pea inclusion decreased DMI linearly (P = 0.03), whereas ADG and G:F were not affected by treatment (P > or = 0.17); however, dietary NE(g) increased quadratically with increasing field pea level (P = 0.04). Fat thickness responded quadratically (P = 0.008) where heifers fed 20% field pea had greatest fat thickness and 30% field pea inclusion the least. Marbling tended (P < or = 0.09) to respond quadratically as field pea increased. No differences (P > or = 0.17) were observed for HCW, LM area, or KPH. In Exp. 2, DMI, ADG, G:F, dietary NE(g), HCW, marbling, LM area, 12th-rib fat, and USDA yield grade (YG) were unaffected by dietary field pea inclusion (P > or = 0.12). In Exp. 3, marbling score increased linearly (P = 0.05), fat thickness increased quadratically (P = 0.01), and YG tended to increase (P = 0.07) quadratically as field pea increased. Field pea inclusion did not affect (P > or = 0.38) DMI, ADG, G:F, dietary NE(g), HCW, or LM area. These results indicate that field pea can be included successfully into rations at levels up to 36% of DM without negatively affecting growth performance and most carcass characteristics of finishing beef cattle; however, effects on marbling score were variable. These data also indicate the energy content of field peas is similar to cereal grains, such as corn and barley, when included in high-concentrate finishing diets.
Assuntos
Ração Animal , Bovinos/crescimento & desenvolvimento , Carne , Pisum sativum/metabolismo , Tecido Adiposo/fisiologia , Animais , Peso Corporal/fisiologia , Bovinos/fisiologia , Feminino , Análise dos Mínimos Quadrados , Masculino , Distribuição AleatóriaRESUMO
A 2 x 2 factorial study evaluated effects of cow wintering system and last trimester CP supplementation on performance of beef cows and steer progeny over a 3-yr period. Pregnant composite cows (Red Angus x Simmental) grazed winter range (WR; n = 4/yr) or corn residue (CR; n = 4/yr) during winter and within grazing treatment received 0.45 kg/d (DM) 28% CP cubes (PS; n = 4/yr) or no supplement (NS; n = 4/yr). Offspring steer calves entered the feedlot 14 d postweaning and were slaughtered 222 d later. Precalving BW was greater (P = 0.02) for PS than NS cows grazing WR, whereas precalving BCS was greater (P < 0.001) for cows grazing CR compared with WR. Calf birth BW was greater (P = 0.02) for CR than WR and tended to be greater (P = 0.11) for PS than NS cows. Prebreeding BW and BCS were greater (P
Assuntos
Criação de Animais Domésticos/métodos , Bovinos/fisiologia , Proteínas Alimentares/administração & dosagem , Suplementos Nutricionais , Métodos de Alimentação/veterinária , Estações do Ano , Criação de Animais Domésticos/economia , Fenômenos Fisiológicos da Nutrição Animal , Animais , Métodos de Alimentação/economia , Feminino , Masculino , Gravidez , Distribuição Aleatória , Reprodução/fisiologiaRESUMO
We have been investigating the function and gene expression of connexins by vascular wall cells, especially Connexin43 (Cx43) in bovine aortic endothelial cells (BAEC). In this study, we tested the effects of carbenoxolone (CBN), a gap junction communication (GJC) blocker on the junctional transfer of Lucifer yellow in BAEC. CBN is a water-soluble derivative of the liquorice-root extract 18-alpha-glycyrrhetinic acid. CBN rapidly abolished dye-transfer in the scrape-load transfer assay (a measure of GJC) in a reversible and dose-dependent fashion. We then asked whether the BAEC might somehow compensate for the loss of junctional communication by altering the expression of connexins. Thus, we treated BAEC with 100 microM CBN in serum free medium and determined the total Cx43 cellular distribution (immunostaining) and protein content (immunoblotting). Besides changes in distribution, by 6 h, Cx43 content levels increased to 166% +/- 22% (P < 0.0001) of controls. RNA blot data showed two-three fold increases in Cx43 message in BAEC after 6 h of CBN treatment, suggesting transcriptional control. Since CBN has structural similarities to corticosteroids, we tested both aldosterone and prednisolone but neither drug increased Cx43 levels, suggesting that the CBN response was not due to a generalized steroid effect. Staurosporine inhibited the CBN-induced increase in Cx43 content, suggesting a role for kinases in the signaling pathway. Further studies with inhibitors indicated that PKA but not PKC was implicated. In summary, CBN blocks junctional communication and modulates Cx43 expression in BAEC. These results suggest a feedback mechanism for control of connexin expression based on junctional patency.
Assuntos
Carbenoxolona/farmacologia , Conexina 43/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Junções Comunicantes/metabolismo , Aldosterona/farmacologia , Animais , Bovinos , Células Cultivadas , Proteínas Quinases Dependentes de AMP Cíclico/fisiologia , Relação Dose-Resposta a Droga , Células Endoteliais/metabolismo , Regulação da Expressão Gênica , Prednisolona/farmacologia , Transdução de SinaisRESUMO
Numerous apolipoproteins associate with amyloid plaques. A minor high-density lipoprotein-associated protein, glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD), has recently been described by the authors and others. Since GPI-PLD is synthesized by, and secreted from, pancreatic islet beta cells, the present study examined the hypothesis that GPI-PLD associates with islet amyloid. GPI-PLD immunoreactivity was examined in pancreatic tissues from type 2 diabetic and non-diabetic humans. GPI-PLD binding to heparan sulphate proteoglycan was determined in the absence or presence of heparan sulphate or heparin. Fibril formation from human islet amyloid polypeptide was determined in the absence or presence of GPI-PLD. In non-diabetics, GPI-PLD immunoreactivity was present and co-localized with insulin, as opposed to co-localizing with amyloid in diabetics. No immunoreactivity for apolipoprotein A-I was present in islet cells or islet amyloid. Heparan sulphate proteoglycan, which is commonly present in most amyloid, bound GPI-PLD in vitro. GPI-PLD inhibited the formation of amyloid fibrils from synthetic islet amyloid polypeptide in vitro. GPI-PLD is therefore present in islet amyloid and appears to derive from local production from islets. This localization likely derives from interaction between GPI-PLD and heparan sulphate proteoglycan. Since GPI-PLD also inhibited islet amyloid polypeptide fibril formation in vitro, it is concluded that GPI-PLD may play a role in islet amyloid formation in type 2 diabetes.
Assuntos
Amiloide/metabolismo , Diabetes Mellitus Tipo 2/enzimologia , Ilhotas Pancreáticas/enzimologia , Fosfolipase D/metabolismo , Amiloide/biossíntese , Amiloide/efeitos dos fármacos , Proteoglicanas de Heparan Sulfato/metabolismo , Humanos , Polipeptídeo Amiloide das Ilhotas Pancreáticas , Microscopia Confocal , Fosfolipase D/farmacologiaRESUMO
Avian leukosis type A virus-derived retroviral vectors have been used to introduce genes into cells expressing the corresponding avian receptor tv-a. This includes the use of Replication-Competent Avian sarcoma-leukosis virus (ASLV) long terminal repeat (LTR) with Splice acceptor (RCAS) vectors in the analysis of avian development, human and murine cell cultures, murine cell lineage studies and cancer biology. Previously, cloning of genes into this virus was difficult due to the large size of the vector and sparse cloning sites. To overcome some of the disadvantages of traditional cloning using the RCASBP-Y vector, we have modified the RCASBP-Y to incorporate "Gateway" site-specific recombination cloning of genes into the construct, either with or without HA epitope tags. We have found the repetitive "att" sequences, which are the targets for site-specific recombination, do not impair the production of infectious viral particles or the expression of the gene of interest. This is the first instance of site-specific recombination being used to generate retroviral gene constructs. These viral constructs will allow for the efficient transfer and expression of cDNAs needed for functional genomic analyses.
Assuntos
Vírus do Sarcoma Aviário/genética , Clonagem Molecular/métodos , Vetores Genéticos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Regulação da Expressão Gênica , Proteínas de Fluorescência Verde , Humanos , Proteínas Luminescentes , Camundongos , Dados de Sequência MolecularRESUMO
We have been studying the relationships between cell growth and the expression of the gap junction protein Connexin43 (Cx43) in cultured bovine aortic endothelial cells (BAEC). As part of these studies, we examined the effect of the growth inhibitory cytokine TGF-beta1 on Cx43 expression. We have shown recently that TGF-beta treatment increases Cx43 mRNA and synthesis, content, and half-life of the protein within 24 h, which leads, over the course of days, to an accumulation of Cx43 in large, intensely immunostaining vesicles, filling much of the perinuclear cytoplasmic space. In the current study, based on their distribution and markers, we identified these vesicles as lysosomes/autophagosomes. Cx43 immunostaining and staining with a fluorescent probe for acidic compartments are coincident, as retention of a fluorescent-labeled low-density lipoprotein occurs in a similar pattern and the same staining pattern can be detected in the treated cells using other markers for lysosomal compartments. TEM revealed prominent lysosomal figures with considerable heterogeneous material. After withdrawal of TGF-beta, the accumulated Cx43 was cleared only slowly, with some brightly immunoreactive cells remaining even after 72 h. The prolonged appearance (based on immunoreactivity in situ and in immunoblots) of intact vesicular Cx43 in the treated cells suggests decreased degradation, resulting from impaired lysosomal activity. These data not only emphasize the importance of the lysosome in connexin degradation, but also show that TGF-beta can cause an alteration in lysosomal functioning, with implications for cellular metabolism.
Assuntos
Conexina 43/metabolismo , Endotélio Vascular/efeitos dos fármacos , Lisossomos/efeitos dos fármacos , Fator de Crescimento Transformador beta/farmacologia , Animais , Aorta , Bovinos , Conexina 43/efeitos dos fármacos , Endotélio Vascular/citologia , Endotélio Vascular/ultraestrutura , Humanos , Lisossomos/metabolismo , Lisossomos/patologia , Microscopia Eletrônica , Fatores de Tempo , Fator de Crescimento Transformador beta1RESUMO
The major purpose of this paper is to explore the potential value of benefit-cost evaluation for stormwater quality management decisions at a local level. A preliminary benefit-cost analysis (BCA) screening method is used for maximum extent practicable (MEP) analysis, identifying promising management practices, and identifying societal and economic tradeoffs for local stormwater problems. Ballona Creek, a major urban storm drain in Los Angeles, California, USA, is used to illustrate the practicality of the benefit-cost evaluation. The Ballona Creek example demonstrates the economic limits of stormwater management in an urban region and attests to the value of coordinated basinwide management compared to uncoordinated management by individual landowners. Evaluation results suggest that in urban areas, the benefit of stormwater quality improvements might be far greater if accompanied by comprehensive redesign of drainage networks and neighboring land uses. In this case, benefit-cost analysis is found to be useful for evaluating and understanding stormwater management alternatives despite the uncertainties in characterizing stormwater quality and the effects of stormwater management on improving receiving water quality.
RESUMO
Intercellular communication may be regulated by the differential expression of subunit gap junction proteins (connexins) which form channels with differing gating and permeability properties. Endothelial cells express three different connexins (connexin37, connexin40, and connexin43) in vivo. To study the differential regulation of expression and synthesis of connexin37 and connexin43, we used cultured bovine aortic endothelial cells which contain these two connexins in vitro. RNA blots demonstrated discordant expression of these two connexins during growth to confluency. RNA blots and immunoblots showed that levels of these connexins were modulated by treatment of cultures with transforming growth factor-ss1. To examine the potential ability of these connexins to form heteromeric channels (containing different connexins within the same hemi-channel), we stably transfected connexin43-containing normal rat kidney (NRK) cells with connexin37 or connexin40. In the transfected cells, both connexin proteins were abundantly produced and localized in identical distributions as detected by immunofluorescence. Double whole-cell patch-clamp studies showed that co-expressing cells exhibited unitary channel conductances and gating characteristics that could not be explained by hemi-channels formed of either connexin alone. These observations suggest that these connexins can readily mix with connexin43 to form heteromeric channels and that the intercellular communication between cells is determined not only by the properties of individual connexins, but also by the interactions of those connexins to form heteromeric channels with novel properties. Furthermore, modulation of levels of the co-expressed connexins during cell proliferation or by cytokines may alter the relative abundance of different heteromeric combinations.
Assuntos
Comunicação Celular/fisiologia , Conexina 43/fisiologia , Conexinas/fisiologia , Endotélio Vascular/fisiologia , Animais , Aorta , Bovinos , Células Cultivadas , Immunoblotting , Técnicas de Patch-Clamp , RNA Mensageiro/análise , Ratos , Proteína alfa-5 de Junções Comunicantes , Proteína alfa-4 de Junções ComunicantesRESUMO
To study the gap junction protein connexin37 (Cx37), we stably transfected cell lines with constructs of human Cx37 containing the epitope tag FLAG (DYKDDDDK). A Cx37 construct containing the FLAG moiety at the carboxyl terminus (Cx37F) was expressed in BWEM cells, and did not substantially alter the levels of endogenous Cx43 in these cells. Immunostaining showed that Cx37F colocalized with Cx43 at cell-cell contacts. Pulse-chase metabolic labeling and immunoprecipitation with anti-FLAG antibodies indicated that Cx37F was synthesized as a protein that ran at 35.9 +/- 0.9 kDa on reducing SDS-PAGE but chased into a slower migrating band at 38.0 +/- 1.0 kDa. This shift in mobility was due to phosphorylation on serine residues, based on [(32)P]-metabolic labeling, immunoprecipitation, and phosphoamino acid analyses. The transition to the phosphoCx37F correlated with a loss of solubility in 1% Triton X-100. Based on the [(35)S]-methionine pulse-chase experiments, the half-life of the labeled Cx37F was approximately 3 h, which is within the range reported for other connexins. Analysis of dye injection experiments indicated that dye transfer was reduced in Cx37-transfected cells in comparison to parental BWEM cells, suggesting that formation of heteromeric Cx37-Cx43 channels reduced the molecular permeability of communication between these cells. Moreover, the similarities of previously demonstrated kinetic details and modification of Cx43 to our new data regarding Cx37 provide evidence for a commonality in processing and assembly steps of these two connexins.
Assuntos
Conexinas/metabolismo , Epitopos/metabolismo , Animais , Western Blotting , Linhagem Celular , Conexinas/biossíntese , Conexinas/genética , Humanos , Cinética , Oligopeptídeos , Peptídeos/química , Processamento de Proteína Pós-Traducional , Ratos , Transfecção , Proteína alfa-4 de Junções ComunicantesRESUMO
OBJECTIVE: The aim of this study was to analyze the prognostic significance of DNA ploidy in patients with endometrial cancer. METHODS: Between October 1988 and January 1997, DNA ploidy was determined prospectively in 208 women who were staged surgically by a standard protocol that included pelvic and para-aortic lymphadenectomy. Median follow-up was 48 months. RESULTS: Diploid tumors were identified in 154 (74%) patients and aneuploid tumors in 54 (26%). Patients with aneuploid tumors had a significantly higher prevalence of metastases to the cervix, adnexa, and omentum, malignant pelvic cytology, and advanced surgical stage. Patients with aneuploid tumors had a 4.5 times higher prevalence of pelvic lymph node metastases and a 5.8 times higher prevalence of para-aortic lymph node metastases. A significantly higher proportion of patients with aneuploid tumors was diagnosed with recurrent or progressive endometrial cancer (22.2 versus 6.5%, P = 0.002). Patients with aneuploid tumors had a significantly lower rate of survival from cancer death (P = 0.038) with 83% versus 94% surviving 5 years. CONCLUSION: Patients with aneuploid tumors are at high risk for lymph node metastases and should be surgically staged, including pelvic and para-aortic lymphadenectomy. Aneuploidy confers a risk for endometrial cancer death and these patients should be candidates for clinical trials evaluating treatment following surgery.
Assuntos
Neoplasias do Endométrio/genética , Ploidias , Idoso , Neoplasias do Endométrio/mortalidade , Neoplasias do Endométrio/patologia , Feminino , Seguimentos , Humanos , Metástase Linfática , Pessoa de Meia-Idade , Prognóstico , Estudos Prospectivos , Taxa de SobrevidaRESUMO
BACKGROUND: Medical management for invasive pulmonary aspergillosis (IPA) is often unsatisfactory. Antifungal therapy may be unable to eradicate IPA in the immunocompromised or neutropenic patient. METHODS: We retrospectively reviewed the surgical management of IPA in 13 immunocompromised patients at our institution. Twelve patients underwent perioperative bone marrow transplantation (4 autologous, 8 allogenic). All 13 patients received antifungal therapy. Eleven patients were neutropenic at the time of operation. RESULTS: The mean interval from diagnosis of aspergillosis to operation was 42 days (range, 3 to 135 days). Eighteen operations were performed on the 13 patients. Seven patients had resections from multiple pulmonary sites, whereas 6 had a single lesion resected. The average lesion resected was 3.7 cm in greatest diameter (range, 1 to 9 cm). After a mean follow-up of 21 months (range, 0 to 9 years), 3 patients (23%) are alive with no evidence of aspergillosis, 6 patients (46%) died without evidence of aspergillosis, and 4 patients (31%) died secondary to aspergillus infection. All 4 patients who died of aspergillus infection received an allogenic bone marrow transplantation. Two patients with direct extrapulmonic extension of IPA at time of operation died of recurrent aspergillus infections. Three of 4 patients who died of aspergillus infection had an absolute neutrophil count less than 1,300 cells/microL at time of operation. The mean absolute neutrophil count of the patients who cleared the aspergillus infection was 5,538 cells/microL. The mean survival of allogenic bone marrow transplant recipients was 5.2 months, and for recipients of autografts was 51.4 months. CONCLUSIONS: In this series, surgical resection of IPA cleared the aspergillus infection in 69% of the patients. Neutropenia, extrapulmonic extension of IPA, and allogenic bone marrow transplantation may predict a worse prognosis. Surgical resection of IPA in immunocompromised patients is an effective form of therapy in a properly selected patient population.
Assuntos
Aspergilose/cirurgia , Hospedeiro Imunocomprometido , Pneumopatias Fúngicas/cirurgia , Adolescente , Adulto , Antifúngicos/uso terapêutico , Aspergilose/tratamento farmacológico , Transplante de Medula Óssea/imunologia , Criança , Pré-Escolar , Intervalo Livre de Doença , Feminino , Seguimentos , Humanos , Contagem de Leucócitos , Pneumopatias Fúngicas/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Neutropenia/imunologia , Neutrófilos/patologia , Seleção de Pacientes , Pneumonectomia/métodos , Prognóstico , Recidiva , Estudos Retrospectivos , Taxa de Sobrevida , Fatores de Tempo , Transplante Autólogo , Transplante HomólogoRESUMO
Renal medullary carcinoma is an aggressive neoplasm of the renal pelvis arising in patients with a history of sickle-cell trait. The authors report a case of renal medullary carcinoma with positive urinary cytology. Due to the location of the tumor in the renal pelvis and the loosely cohesive nature of poorly differentiated neoplasms, the presence of renal medullary carcinoma in a urinary cytology specimen is not surprising. The cytologic characteristics as well as the ultrastructural features are described.
Assuntos
Carcinoma Medular/secundário , Neoplasias Renais/patologia , Neoplasias Renais/urina , Adolescente , Carcinoma Medular/urina , Feminino , HumanosRESUMO
OBJECTIVE: To analyze the role of surgery alone, including pelvic and para-aortic lymphadenectomy, in patients with endometrial cancer who did not receive radiotherapy. METHODS: Between August 1987 and January 1997, 225 women with disease clinically confined to the uterus were staged surgically by a standard protocol that included pelvic and para-aortic lymphadenectomy in women with high risk factors. No radiation was administered before or after surgery. RESULTS: The combination of preoperative endometrial biopsy grade and gross depth of myometrial invasion identified 123 (55%) high-risk patients who had lymphadenectomy and 102 (45%) low-risk patients who did not. Eighteen (15%) high-risk patients had lymph node metastases and received postoperative systemic therapy. Three low-risk, eight high-risk-node-negative, and no high-risk-node-positive patients were diagnosed with recurrent cancer, corresponding to 5-year recurrence-free proportions of 0.95, 0.89, and 1.00, respectively. Although sample sizes and limited follow-up limit conclusions, the experience to date suggests a high rate of survival in all three groups. CONCLUSION: Our preliminary experience indicates that even high-risk patients have an excellent prognosis when treated with surgery, including pelvic and para-aortic lymphadenectomy, without radiotherapy.