RESUMO
BACKGROUND: Necrotizing enterocolitis (NEC) and periventricular hemorrhagic infarction (PVHI) are complications of prematurity associated with poor neurodevelopmental outcomes. OBJECTIVES: We characterized temporal and causal associations between NEC and type of PVHI as well as associations with outcomes. METHODS: This was a multicenter retrospective study of infants with birth weight <1,500 g and diagnosis of PVHI by a pediatric radiologist at 3 neonatal intensive care units in North Carolina, USA, between January 1998 and December 2004. NEC was confirmed using radiological and surgical pathology findings. Infants were assessed by 3 years using the Bayley Scales of Infant Development, second edition. RESULTS: 35 of 112 (31%) of study patients developed NEC. NEC was diagnosed subsequent to PVHI by a median of 16.6 days (95% CI [9.5, 24.9], p < 0.0001). Indomethacin use and the presence of bilateral PVHI were associated with an increased risk of subsequent NEC (OR 2.8, 95% CI [1.1, 7.2] and OR 2.4, 95% CI [1.1, 5.7], respectively). Having bilateral versus unilateral PVHI was associated with a 2.34-fold increased risk of death (95% CI [1.27, 4.33], p = 0.007). NEC was not associated with worse motor outcomes in this population. Overall, the probability of a mental development index >70 was greatest for infants with unilateral PVHI versus bilateral PVHI, although the presence of NEC was associated with worse cognitive outcomes in both groups. CONCLUSIONS: Premature infants with PVHI often subsequently develop NEC, especially if they have bilateral PVHI and are exposed to indomethacin. While NEC results in worse neurodevelopmental outcomes, PVHI severity appears more important to the outcome of these infants.
Assuntos
Deficiências do Desenvolvimento/etiologia , Enterocolite Necrosante/complicações , Hemorragias Intracranianas/complicações , Distribuição de Qui-Quadrado , Enterocolite Necrosante/fisiopatologia , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Recém-Nascido de muito Baixo Peso , Hemorragias Intracranianas/fisiopatologia , North Carolina , Estudos RetrospectivosRESUMO
PURPOSE: Mammalian cells at mitosis contain chromatin in compacted form and are hypersensitive to ionizing radiation. Previous research had shown some chemicals that induce chromatin compaction within interphase cells act as radiosensitizers. Of these agents, cantharidin (LS-1), which is an inhibitor of protein phosphatases 1 (PP1) and 2A (PP2A), showed good radiosensitizing activity at non-toxic doses. Cantharidin and 13 additional structural analogues (LS-2-14) were tested for their radiosensitizing activity on tumour cells in vitro. MATERIALS AND METHODS: Twelve of the 14 cantharidin analogues were synthesized in the authors' laboratory. Various concentrations of the drugs were screened for toxicity and radiosensitizing effectiveness with asynchronous DU-145 (human prostate carcinoma) cells. More detailed radiobiological studies of the more potent agents were performed with HT-29 (human colon carcinoma) cells since they could be readily synchronized. The radiosensitization of G1 phase HT-29 cells was measured after a 2-h exposure to the more potent drugs and reductions of the surviving fraction after an acute dose of 2 Gy (SF2Gy) served to estimate their relative effectiveness. The increase in phosphorylation of histone 1 (H1) and histone 3 (H3) induced by these drug exposures was measured by Western blotting of protein extracts. Drug-induced change in chromatin morphology was visualized by electron microscopy, and the alkaline comet assay (which measures DNA single-strand breaks) was employed to measure the radiation sensitivity of cellular chromatin in the drug-treated cells. RESULTS: Of the 14 cantharidin analogues tested, LS-1, LS-2 and LS-5 at concentrations of 3-20 microM showed little or no toxicity, produced elevated levels of H1 and H3 phosphorylation, and effected significant radiosensitization at low radiation dose. The chromatin in tumour cells treated with LS-5 became visibly compacted and its DNA was about 1.6 times more sensitive to radiation-induced strand breakage relative to that of control cells. CONCLUSIONS: The results confirm the authors' earlier studies that showed an increase in tumour cell intrinsic radiosensitivity by exposure to agents that promote chromatin compaction. LS-5 was identified as the optimal radiosensitizing agent of this class of compounds. Radiosensitization was correlated with chromatin compaction and elevated phosphorylation of H1 and H3. The DNA in drug-treated cells exhibited an enhanced sensitivity to radiation-induced single-strand breakage.
Assuntos
Cantaridina/análogos & derivados , Cantaridina/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Neoplasias do Colo/patologia , Neoplasias da Próstata/patologia , Tolerância a Radiação/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos da radiação , Relação Dose-Resposta a Droga , Relação Dose-Resposta à Radiação , Raios gama , Humanos , Masculino , Radiossensibilizantes/farmacologiaRESUMO
BACKGROUND: The usefulness of serum insulinlike growth factor (IGF)-system-peptide measurement to assess the adequacy of nutritional intake in premature infants with chronic lung disease bronchopulmonary dysplasia (BPD) was assessed. METHODS: Twenty-nine premature infants had serial measurements taken of their serum IGF-1, insulinlike growth factor binding protein (IGFBP)-2, and IGFBP-3 concentrations between 2 and 6 weeks of age. Regression analyses were used to examine the relation between nutritional parameters and IGF-1, IGFBP-2, and IGFBP-3 concentrations in premature infants with and without BPD. RESULTS: The group of infants with BPD (n = 12) did not differ from infants without BPD (n = 17) in gestational age or weight at entry, but gained less weight during the study period. In infants without BPD, IGF-1 correlated positively with protein intake (r = 0.50) and caloric intake (r = 0.41) over the 3 days before sample collection and with weight change over the previous week (r = 0.46). In contrast, infants with BPD showed a significant correlation between IGF-1 and weight change (r = 0.54) only. There was a significant negative correlation between IGFBP-2 and protein intake in infants without BPD (r = -0.50) and in infants with BPD (r = -0.41). Negative correlations between IGFBP-2 and both weight change (r = -0.64) and caloric intake (r = -0.43) over the previous week were found only in the group of infants without BPD. IGFBP-3 correlated positively with weight changes and protein intake in both groups but correlated with caloric intake only in the group without BPD. Multiple regression analyses were used to determine significant independent variables associated with IGF-1, IGFBP-2, and IGFBP-3. In infants without BPD, significant independent predictors of IGFBP-2 were 7-day weight change and 2-day protein intake; 3-day caloric intake was the only significant independent predictor for IGFBP-3. For infants with BPD, 3-day weight gain was the only independent variable associated with serum IGF-1. Protein intake in the week before sample collection was an independent predictor of IGFBP-2 and 3-day weight change and 2-day protein intake were independent predictors of IGFBP-3. CONCLUSIONS: These results confirm that changes in serum IGF-1, IGFBP-2, and IGFBP-3 reflect the nutritional status of premature infants and demonstrate that the relation between these proteins and nutritional intake differs in premature infants with and without BPD. Refinement of these observations by future studies may permit a more accurate determination of the protein and caloric intake sufficient for growth and repair after injury in premature infants with lung disease.
Assuntos
Displasia Broncopulmonar/fisiopatologia , Ingestão de Energia/fisiologia , Recém-Nascido Prematuro/sangue , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/sangue , Fator de Crescimento Insulin-Like I/análise , Biomarcadores , Displasia Broncopulmonar/sangue , Estudos de Casos e Controles , Estudos de Coortes , Proteínas Alimentares/administração & dosagem , Proteínas Alimentares/sangue , Humanos , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/sangue , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/sangue , Estado Nutricional , Radioimunoensaio , Análise de Regressão , Aumento de PesoRESUMO
Focal small bowel perforation (FSBP) is a life-threatening event that predominantly affects extremely low birth weight (ELBW) infants. Histopathology from surgical specimens of ileum with FSBP shows a healthy mucosa overlying a thinned muscularis with segmental degeneration. Clinical data strongly support an association between early postnatal administration of dexamethasone (EPD) and FSBP. Additional risk factors, including gestational age, administration of prophylactic indomethacin, and severity of illness, may be synergistic with EPD for the pathogenesis of perforations. Animal models of dexamethasone administration show morphologic changes in the ileum, similar to those seen in ELBW infants, including increased mucosal maturation and thinning of the muscularis. These tissue-specific differences may be mediated by a perturbation in growth factor expression or accumulation. In support of this hypothesis, dexamethasone has been associated with increased IGF-I immunolocalization in the mucosa and decreased immunolocalization in the muscularis. The known growth-promoting functions of IGF-I are consistent with the observed dexamethasone-associated changes within both the mucosa and the muscularis. Ongoing studies in this animal model are exploring the potential mechanisms by which dexamethasone might affect IGF-I availability.
Assuntos
Dexametasona/farmacologia , Íleo/anormalidades , Íleo/efeitos dos fármacos , Íleo/embriologia , Animais , Modelos Animais de Doenças , Mucosa Gástrica/metabolismo , Idade Gestacional , Humanos , Indometacina/uso terapêutico , Recém-Nascido , Recém-Nascido de muito Baixo Peso , Fator de Crescimento Insulin-Like I/biossíntese , Camundongos , Modelos Biológicos , Cloreto de Sódio/farmacologiaRESUMO
Glucocorticoid exposure accelerates the maturation of small bowel mucosa. We hypothesized that IGF-I, a mitogen and differentiating peptide expressed in small bowel, mediates steroid-induced change within the developing ileum. To investigate this possibility, we intraperitoneally administered 1 microg/gm/d of dexamethasone (DEX) or equal volumes of saline to litter-mate newborn mice. The animals were killed on d 1-3 of life and their ilea were harvested and prepared for microscopy. Tissue sections of ileum were examined for morphologic analyses, mucin staining, immunolocalization of IGF-I and -II, proliferating cell nuclear antigen (PCNA), terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), and in situ hybridization for IGF-I transcripts. Morphologic comparisons showed increases in goblet cell number, total cell number, and TUNEL-positive cells within the mucosa of DEX-treated animals. In contrast, the number of smooth muscle nuclei per cross-section was unchanged with DEX treatment despite a reduction in the number of PCNA-positive nuclei and an increased bowel circumference. These findings suggest the muscularis stretches to accommodate increasing bowel diameter. IGF-I peptide was localized to the mesenchyme of all control animals. After 48 h of DEX treatment, IGF-I was detected in the epithelia whereas mesenchymal IGF-I localization appeared diminished. In situ hybridization analyses for IGF-I transcripts showed no differences in localization between the groups. We conclude that DEX administration differentially affects adjacent tissues in the newborn ileum and that the associated changes in IGF-I localization are consistent with its participation in this process.
Assuntos
Animais Recém-Nascidos , Dexametasona/administração & dosagem , Íleo/efeitos dos fármacos , Fator de Crescimento Insulin-Like I/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Animais , Dexametasona/farmacologia , Íleo/anatomia & histologia , Íleo/metabolismo , Imuno-Histoquímica , Hibridização In Situ , Marcação In Situ das Extremidades Cortadas , Fator de Crescimento Insulin-Like I/genética , Mucosa Intestinal/anatomia & histologia , Mucosa Intestinal/metabolismo , Camundongos , Antígeno Nuclear de Célula em Proliferação/metabolismo , RNA Mensageiro/genéticaRESUMO
Insulin-like growth factor (IGF)-stimulated lung fibroblast proliferation may be regulated by locally produced IGF-binding proteins (IGFBPs) during lung development. Recent evidence has shown that many growth factors participate in the regulation of cell proliferation by regulating IGFBPs. Because platelet-derived growth factor-BB (PDGF-BB) is highly expressed during lung development and is known to regulate IGFBP-4 production by lung cells, we examined the mechanisms by which PDGF-BB regulates ICFBP-4 production using primary cultures of 19-day gestation rat lung fibroblasts. Exposure of fetal rat lung fibroblasts to PDGF-BB increased IGFBP-4 mRNA transcript abundance by 3.6- and 2.4-fold at 18 and 40 hours, respectively. Addition of Rp-adenosine-3'-5'-cyclic monophosphothioate triethylamine (rp-cAMPS), a competitive inhibitor of protein kinase A, blunted the PDGF-BB-stimulated increase in conditioned medium (CM) IGFBP-4 and the increase in IGFBP-4 mRNA. Proteolysis of IGFBP-4 was detected in aliquots of cell-free CM from cells exposed to SFM for 48 hours. IGFBP-4 proteolysis was inhibited by EDTA and 1,10-phenanthroline and was accentuated by the addition of IGF-I and IGF-II and, to a lesser extent, by des(1-3)IGF-I. Exposure of cells to PDGF-BB for 48 hours resulted in an inhibition of IGFBP-4 proteolysis that was associated with a decrease in the concentration of IGF-I in CM. These studies demonstrate that PDGF-BB increases the accumulation of ICFBP-4 in fetal rat lung fibroblasts CM through increased production and by inhibiting IGF-mediated IGFBP-4 proteolysis.
Assuntos
AMP Cíclico/análogos & derivados , Fibroblastos/efeitos dos fármacos , Proteína 4 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Fator de Crescimento Derivado de Plaquetas/farmacologia , Animais , Becaplermina , Células Cultivadas , Meios de Cultivo Condicionados/química , AMP Cíclico/farmacologia , Ácido Edético/farmacologia , Fibroblastos/citologia , Fibroblastos/metabolismo , Proteína 4 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Fator de Crescimento Insulin-Like I/farmacologia , Fator de Crescimento Insulin-Like II/farmacologia , Pulmão/citologia , Pulmão/efeitos dos fármacos , Pulmão/embriologia , Fragmentos de Peptídeos/farmacologia , Fenantrolinas/farmacologia , Proteínas Proto-Oncogênicas c-sis , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Tionucleotídeos/farmacologiaRESUMO
Focal small bowel perforation (FSBP) occurs most commonly in the ileum of extremely low-birth-weight (ELBW) infants. Early postnatal dexamethasone (EPD) administration results in an increased risk for FSBP in this patient population, but the mechanism by which this occurs is unknown. Infants with FSBP have healthy mucosa but thinned smooth muscle, suggesting a mechanism involving the muscularis propria for these perforations. One explanation for these findings would be that dexamethasone alters the tissue availability of pertinent growth factors to the smooth muscle. To explore this possibility, we administered dexamethasone or saline by intraperitoneal injection to newborn mice for 3 days (dosed at 1 microg/g of body weight/day) to simulate EPD protocols. The animals were sacrificed after 72 h of treatment and their ileums harvested and prepared for microscopy. Immunolocalization was performed for three related growth factors (epidermal growth factor [EGF], heparin-binding EGF [h-EGF], and transforming growth factor alpha [TGF-alpha]) and their common receptor. We found TGF-alpha to be abundant and discretely localized in the muscularis propria in control animals but to be diminished in dexamethasone-treated animals. EGF-receptor immunostaining was also decreased with dexamethasone but there was minimal to no detection of EGF or h-EGF in either treatment condition. Surgical and autopsy specimens of the ileum were obtained from seven ELBW infants who either received EPD or not. These tissues were used for immunolocalization of the same growth factors and similar distributions for TGF-alpha were observed in several of these cases. These findings are consistent with an autocrine role for TGF-alpha in ileal smooth muscle proliferation and suggest a mechanism by which EPD might mediate smooth muscle thinning.
Assuntos
Animais Recém-Nascidos/metabolismo , Dexametasona/farmacologia , Íleo/efeitos dos fármacos , Recém-Nascido de muito Baixo Peso/metabolismo , Músculo Liso/efeitos dos fármacos , Fator de Crescimento Transformador alfa/metabolismo , Animais , Dexametasona/administração & dosagem , Fator de Crescimento Epidérmico/metabolismo , Receptores ErbB/metabolismo , Fator de Crescimento Semelhante a EGF de Ligação à Heparina , Humanos , Íleo/metabolismo , Técnicas Imunoenzimáticas , Recém-Nascido , Injeções Intraperitoneais , Injeções Intravenosas , Peptídeos e Proteínas de Sinalização Intercelular , Camundongos , Camundongos Endogâmicos C57BL , Músculo Liso/metabolismoRESUMO
Insulin-like growth factor (IGF) binding proteins (IGFBPs) are expressed in fetal lung and may provide important post-translational regulation of IGF-induced mitogenesis during lung organogenesis. Because of the observation that growth factors can control cell growth through regulation of IGFBPs, we examined IGFBP production by fetal lung fibroblasts following stimulation by peptide growth factors important for fetal lung growth and development. Fetal lung fibroblasts were cultured in serum-free medium supplemented with various growth factors for up to 48 h, and IGFBPs in conditioned medium (CM) were analyzed by ligand blot and immunoblot techniques. Accumulation of CM IGFBP-3 was increased and IGFBP-2 decreased by incubation with either keratinocyte growth factor (KGF) or epidermal growth factor (EGF). The effect of these factors on IGFBP-3 accumulation increased with time but the effects of KGF on CM IGFBP-2 decreased over 48 h of incubation. CM IGFBP-4 was increased by 24 and 48 h incubation with basic fibroblast growth factor (bFGF; 2.1- and 2.7-fold increases at 24 and 48 h, respectively) and platelet-derived growth factor-BB (PDGF-BB; 4.2- and 14.9-fold increases at 24 and 48 h, respectively), and 48 h incubation with EGF (6.3-fold increase). In 48-h coincubation experiments, EGF in combination with PDGF-BB or with bFGF, and bFGF in combination with PDGF-BB, resulted in IGFBP-4 accumulations twice that expected from a summation of the effects of either growth factor alone (IGFBP-4 increased 9.8-, 4.0-, and 1.8-fold by PDGF-BB, EGF, and bFGF, respectively; and 27.1-, 37.3-, and 13.0-fold by PDGF-BB plus EGF, PDGF-BB plus bFGF, and EGF plus bFGF, respectively). These results suggest synergistic effects of these growth factors on IGFBP-4 accumulation in fetal lung fibroblast CM. Because IGFBPs are known to regulate DNA synthesis, we speculate that peptide growth factors may alter cell proliferation in fetal lung, in part through their effect on IGFBPs.
Assuntos
Feto/metabolismo , Substâncias de Crescimento/metabolismo , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/biossíntese , Pulmão/metabolismo , Animais , Células Cultivadas , Feto/citologia , Fibroblastos/metabolismo , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/metabolismo , Pulmão/citologia , Pulmão/embriologia , Peptídeos/metabolismo , RatosRESUMO
The insulin-like growth factors (IGF-I and IGF-II) may play an important role in postpneumonectomy compensatory lung growth by translating hormonal inputs and mechanical forces into cellular proliferation signals. We examined the mRNA abundance of IGF-I, IGF-II, and IGF binding proteins (IGFBPs) in lungs of rats on postoperative days 1, 2, 3, 5, and 7 following left pneumonectomy (PNX) or shamoperation (SC) and in normal animals (CON). There was no difference in the abundance of lung IGF-I mRNA (measured by Northern analysis) or serum IGF-I (measured by radioimmunoassay (RIA)) between SC and PNX animals. IGF-II mRNA abundance was initially decreased following PNX (73% decrease compared to SC animals on day 1, p < .05) and then rose to approach SC group values on subsequent days. Transcripts for IGFBP-2, -3, -4, -5, and -6 were decreased in both the SC and PNX groups compared to CON animals on the day following pneumonectomy, then rose back to baseline by postoperative day 2-3. Tissue IGFBPs, measured by ligand blot analyses, were not different in either the SC or PNX groups. In contrast, all serum IGFBP bands were increased on postoperative day 1 following either sham or PNX surgery. In addition, serum IGFBP-4 was increased in PNX animals compared to the SC group on days 1 and 2 (increase of 38% and 78%, respectively, p < .05). We conclude that the changes observed in lung IGF and IGFBP expression following pneumonectomy do not represent major.
Assuntos
Fator de Crescimento Insulin-Like II/biossíntese , Fator de Crescimento Insulin-Like I/biossíntese , Pulmão/metabolismo , Pulmão/fisiologia , Regeneração/fisiologia , Animais , Northern Blotting , Fator de Crescimento Insulin-Like I/metabolismo , Fator de Crescimento Insulin-Like II/metabolismo , Pulmão/anatomia & histologia , Masculino , Tamanho do Órgão/fisiologia , Pneumonectomia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Chronic injury to the developing lung results in cell proliferation and characteristic architectural changes. It is likely that growth factors produced and acting locally are important to these processes. Insulin-like growth factors I and II (IGF-I and IGF-II) are peptide growth factors expressed by lung cells. Roles for IGF-I and IGF-II in lung injury are suggested by their expression during lung development and by studies showing changes in IGF-I expression by activated alveolar macrophages, and increases in IGF-II peptide in oxidant arrested alveolar epithelial cells. To investigate whether the expression of IGF-I and IGF-II are changed with hyperoxic exposure, newborn rats were exposed to 80-90% oxygen for up to 6 wk and Northern hybridization analyses, in situ hybridization histochemistry, immunohistochemical staining, and reverse transcription-polymerase chain reaction (RT-PCR) studies were performed. Northern hybridization analyses of RNA extracted from whole lung showed increases in IGF-I and IGF-II mRNAs with prolonged hyperoxia. In situ hybridization histochemistry and immunohistochemical staining demonstrated spatial patterns of IGF-I and IGF-II expression similar to those seen during fetal lung development. In addition, alveolar macrophages express IGF-I and type II epithelial cells express IGF-II in control and oxygen-injured lung. These results suggest that in lung injury resident lung cells may re-express IGFs in a manner reminiscent of fetal development, and activated inflammatory cells may contribute to the proliferative response through autocrine and paracrine mechanisms.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Fator de Crescimento Insulin-Like II/genética , Fator de Crescimento Insulin-Like I/genética , Pulmão/crescimento & desenvolvimento , Oxigênio/farmacologia , Animais , Animais Recém-Nascidos , Líquido da Lavagem Broncoalveolar/citologia , Hibridização In Situ/métodos , Fator de Crescimento Insulin-Like I/análise , Fator de Crescimento Insulin-Like II/análise , Pulmão/citologia , Pulmão/efeitos dos fármacos , Macrófagos Alveolares/química , Músculo Liso/química , RNA Mensageiro/análise , Ratos , Ratos Sprague-DawleyRESUMO
To examine the changes in coenzyme A profile and the possible corrective effects of carnitine supplementation in the genetic disorders of mitochondrial beta-oxidation, we carried out experiments using an inhibitor of multiple acyl-CoA dehydrogenase enzymes, methylenecyclopropaneacetic acid (MCPA), in rat hepatocytes. MCPA irreversibly inhibited ketone synthesis from straight-chain fatty acids (butyrate, octanoate, palmitate) and branched-chain fatty acids (alpha-ketoisocaproate) with a parallel 70-90% reduction of hepatocyte acetyl-CoA levels. Alone, MCPA or substrates halved free CoA levels to 15% of total CoA and doubled short- and medium-chain acyl-CoA levels to 30% of total CoA. With MCPA plus substrates combined, free CoA levels were 10% of total CoA, and short- and medium-chain acyl-CoA levels were 45% of total CoA. Comparable changes in CoA profiles were found in a patient with a severe genetic defect in beta-oxidation. Neither the suppression of ketogenesis nor the alterations in CoA profiles induced by MCPA inhibition could be corrected by carnitine supplementation.
Assuntos
Acil-CoA Desidrogenases/antagonistas & inibidores , Carnitina/farmacologia , Coenzima A/metabolismo , Ciclopropanos/farmacologia , Hipoglicinas/farmacologia , Fígado/metabolismo , Acil-CoA Desidrogenases/metabolismo , Animais , Inibidores Enzimáticos/farmacologia , Cetonas/metabolismo , Erros Inatos do Metabolismo Lipídico/metabolismo , Mitocôndrias Hepáticas/metabolismo , Ratos , Ratos Sprague-Dawley , Compostos de Espiro/farmacologiaRESUMO
Lung development requires a complex series of developmentally controlled interactions that involve mechanical forces, genetic and endocrine influences, and cell-cell communication. At each step, cell-matrix or cell-cell signaling mediated by peptide growth factors and extracellular matrix components is crucial in directing cell proliferation, differentiation, and migration. Although a comprehensive understanding of how these components interact to guide lung organogenesis has been elusive, the action and control of peptide growth factors in autocrine and paracrine signaling, mesenchymal-epithelial interactions in controlling branching morphogenesis, cell-cell communication in the regulation of cellular differentiation, and factors regulating pattern formation are being clarified.
Assuntos
Pulmão/embriologia , Animais , Comunicação Celular/fisiologia , Matriz Extracelular/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Substâncias de Crescimento/genética , Substâncias de Crescimento/fisiologia , Humanos , Mesoderma/fisiologia , Morfogênese/fisiologiaRESUMO
DMP 811 exhibited high binding affinity for the angiotensin II subtype receptor AT1 in rat adrenal tissues with an IC50 of 6 nM, but not for the subtype receptor AT2. In the isolated rabbit aorta, DMP 811 inhibited the contractile response to angiotensin II selectively and noncompetitively with a KB value of 0.1 nM. In conscious renal hypertensive rats, DMP 811 decreased blood pressure with i.v. and p.o. ED30s of 0.005 and 0.03 mg/kg, respectively (p.o. ED30 for losartan = 0.59 mg/kg). In conscious furosemide-treated dogs, DMP 811 given either at 0.3 or 1 mg/kg p.o. decreased blood pressure. DMP 811 has oral bioavailabilities of 7 and 29% in rats and dogs, respectively, after a solution dose and 8 and 13%, respectively, after a suspension or capsule dosing. Our study indicates that DMP 811 is a selective and insurmountable AT1 receptor antagonist and is a 20-fold more potent orally-active antihypertensive agent than losartan.
Assuntos
Antagonistas de Receptores de Angiotensina , Imidazóis/farmacocinética , Tetrazóis/farmacocinética , Animais , Bioensaio , Pressão Sanguínea/efeitos dos fármacos , Cães , Cobaias , Hipertensão Renovascular , Imidazóis/sangue , Imidazóis/farmacologia , Masculino , Coelhos , Ensaio Radioligante , Ratos , Ratos Sprague-Dawley , Tetrazóis/sangue , Tetrazóis/farmacologiaRESUMO
The insulin-like growth factors (IGF-I and IGF-II) participate in the control of cell proliferation in normal and neoplastic lung cells. To examine the role of IGF binding proteins (IGFBPs) in modulating IGF actions in lung, we examined the production and regulation of IGFBPs from A549 cells, a human adenocarcinoma-derived lung cell line. Ligand blot and immunoblot analysis of conditioned media (CM) from A549 cells demonstrated IGFBP bands of relative molecular mass (M(r)) approximately 39-43,000 (IGFBP-3), 34,000 (IGFBP-2), 30,000 (IGFBP-1), and 24,000 (IGFBP-4). IGFBP-3 abundance in A549 cell CM increased following exposure to IGF-I and IGF-II (3.0- and 1.8-fold, respectively) without a change in IGFBP-3 transcript abundance, suggesting IGFBP-3 is post-transcriptionally regulated. Cycloheximide almost completely abrogated the IGF-I-stimulated increase in CM IGFBP-3, suggesting that ongoing protein synthesis is necessary for the IGF-I-stimulated increase in IGFBP-3 abundance. Increases in IGFBP-3 occurred by at least two mechanisms, through activation of the type 1 IGF receptor and by a type 1 IGF receptor independent mechanism. The increase in IGFBP-3 was due, in part, to activation of the type 1 IGF receptor because blocking type 1 IGF receptor activation with an antibody (alpha IR3) diminished the IGF-I-induced increase in IGFBP-3 and insulin, at doses that stimulate the type 1 IGF receptor, increased IGFBP-3 abundance. The increase in IGFBP-3 was partially independent of type 1 IGF receptor activation because [QAYL]-IGF-I, an analog of IGF-I that binds the type 1 IGF receptor but not IGFBP-3, was less potent than IGF-I in stimulating IGFBP-3 abundance, and IGF-II, which binds IGFBP-3 normally, but binds the type 1 IGF receptor with lower affinity than IGF-I, was nearly equipotent to IGF-I in its stimulation of IGFBP-3 accumulation at low concentrations. These results suggest that ligand binding decreases IGFBP-3 clearance or increases IGFBP-3 accumulation in CM. IGF-I decreased IGFBP-4 abundance in A549 cell CM without decreasing IGFBP-4 mRNA transcripts and without increasing the amount of cell-associated IGFBP-4. To determine whether the decrease in IGFBP-4 was due to increased degradation, cell-free CM was incubated with and without IGF-I, and IGFBP-4 abundance measured by ligand and immunoblot analyses.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/biossíntese , Proteína 4 de Ligação a Fator de Crescimento Semelhante à Insulina/biossíntese , Fator de Crescimento Insulin-Like I/farmacologia , Neoplasias Pulmonares/metabolismo , Pulmão/metabolismo , Adenocarcinoma/metabolismo , Sequência de Bases , Contagem de Células , Divisão Celular , Meios de Cultivo Condicionados/química , Cicloeximida/farmacologia , DNA/biossíntese , Humanos , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Proteína 4 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/análise , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/biossíntese , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/genética , Fator de Crescimento Insulin-Like II/farmacologia , Pulmão/citologia , Pulmão/efeitos dos fármacos , Neoplasias Pulmonares/patologia , Dados de Sequência Molecular , Inibidores da Síntese de Proteínas/farmacologia , RNA Mensageiro/biossíntese , Receptor IGF Tipo 1/fisiologia , Células Tumorais CultivadasAssuntos
Antipsicóticos/efeitos adversos , Isoxazóis/efeitos adversos , Carbonato de Lítio/efeitos adversos , Piperidinas/efeitos adversos , Antipsicóticos/uso terapêutico , Confusão/induzido quimicamente , Creatina Quinase/sangue , Quimioterapia Combinada , Febre/induzido quimicamente , Humanos , Isoxazóis/uso terapêutico , Carbonato de Lítio/uso terapêutico , Síndrome Maligna Neuroléptica/etiologia , Piperidinas/uso terapêutico , Risperidona , Esquizofrenia/tratamento farmacológicoRESUMO
Nine low birth weight infants with neutropenia born to mothers with preeclampsia were treated with granulocyte-colony stimulating factor, 10 micrograms/kg intravenously, within 24 hours of birth and at 24-hour intervals for a maximum of three doses if neutropenia persisted. The absolute neutrophil count increased significantly in eight of the nine infants within 6 hours, and neutrophilia was sustained for at least 72 hours after administration of a single dose of granulocyte-colony stimulating factor.
Assuntos
Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Recém-Nascido de Baixo Peso , Neutropenia/terapia , Pré-Eclâmpsia , Feminino , Humanos , Recém-Nascido de Baixo Peso/sangue , Recém-Nascido , Contagem de Leucócitos , Neutropenia/etiologia , Neutrófilos , GravidezRESUMO
Insulin-like growth factor (IGF)-I and IGF-II are small peptide growth factors that interact with a specific membrane receptor, the type 1 IGF receptor, to stimulate cellular proliferation and/or differentiation. The actions of these growth factors and their availability to their receptors are modulated by specific binding proteins, IGF binding protein (IGFBP)-1 through -6, which together with the IGFs and IGF receptors form the IGF system. We have analyzed RNA extracted from fetal (gestation day 16 [E16] through 22 [E22]) and adult (60-day-old) rat lung for expression of each component of the IGF system. IGF-I and -II RNAs are expressed throughout fetal development. IGF-I mRNA remained relatively constant in fetal and adult lung, whereas IGF-II RNA decreased in later gestation to levels below detection by Northern analyses in adult lung. Type 1 IGF receptor expression varied little through all ages studied, whereas the type 2 IGF receptor RNA displayed developmental regulation with a decline in expression with advancing age. IGFBP-1 transcripts were not detected in fetal or adult lung. IGFBP-2 RNA was expressed from E16 to E22, although its abundance decreased in late gestation and in adult lung, with the lowest levels of expression on day E22. IGFBP-3, -4, and -5 had similar profiles of RNA abundance, with fetuses at ages E21 and E22 displaying higher levels of transcript abundance as compared with those aged E17 to E20; the lowest RNA abundance was seen at E20.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Proteínas de Transporte/biossíntese , Regulação da Expressão Gênica no Desenvolvimento , Pulmão/crescimento & desenvolvimento , Receptores de Somatomedina/biossíntese , Somatomedinas/biossíntese , Animais , Sequência de Bases , Desenvolvimento Embrionário e Fetal , Feminino , Idade Gestacional , Proteína 1 de Ligação a Fator de Crescimento Semelhante à Insulina , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina , Fator de Crescimento Insulin-Like I/biossíntese , Fator de Crescimento Insulin-Like II/biossíntese , Pulmão/embriologia , Pulmão/metabolismo , Dados de Sequência Molecular , Gravidez , RNA Mensageiro/biossíntese , Ratos , Receptor IGF Tipo 1/biossíntese , Receptor IGF Tipo 2/biossínteseRESUMO
Cellular proliferation and differentiation in the fetus and newborn are influenced by many factors. Increasingly, peptide growth factors are thought to play an important role in cell-to-cell communication during fetal life, promoting cellular proliferation, differentiation, and migration within tissues. Each growth factor may play various roles depending on the time in gestation, the type of receptor expressed by the target tissue, and the presence or absence of other permissive or inhibitory growth factors. In addition, growth factor production is regulated by nutritional factors, encouraging or inhibiting growth depending on the energy and oxygen supply. Although the consequences of fetal exposure to tobacco, caffeine, and other drugs on growth have been well documented, the effect of low-grade maternal immune system activation is less well understood. Postnatally, adequate standards for the growth of premature infants are lacking. Each of these areas is discussed in light of recent publications.
Assuntos
Desenvolvimento Embrionário e Fetal/fisiologia , Substâncias de Crescimento/fisiologia , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido/crescimento & desenvolvimento , Recém-Nascido Prematuro/crescimento & desenvolvimento , Cafeína/efeitos adversos , Diferenciação Celular , Divisão Celular , Metabolismo Energético , Feminino , Humanos , Troca Materno-Fetal , Consumo de Oxigênio , Gravidez , Fumar/efeitos adversosRESUMO
We have employed a method of enrichment that allows us to significantly increase the rate of reductive polychlorinated biphenyl (PCB) dechlorination. This method shortens the time required to investigate the effects that culture conditions have on dechlorination and provides an estimate of the potential activity of the PCB-dechlorinating anaerobes. The periodic supplementation of sterile sediment and PCB produced an enhanced, measurable, and sustained rate of dechlorination. We observed volumetric rates of the dechlorination of 2,3,6-trichlorobiphenyl (2,3,6-CB) to 2,6-dichlorobiphenyl (2,6-CB) of more than 300 mumol liter day when the cultures were supplemented daily. A calculation of this activity that is based on an estimate of the number of dechlorinating anaerobes present indicates that 1.13 pmol of 2,3,6-CB was dechlorinated to 2,6-CB day bacterial cell. This rate is similar to that of the reductive dechlorination of 3-chlorobenzoate by Desulfomonile tiedjei. Methanogenesis declined from 585.3 to 125.9 mumol of CH(4) liter day, while dechlorination increased from 8.2 to 346.0 mumol of 2,3,6-CB dechlorinated to 2,6-CB liter day.