RESUMO
The effects of blue light emitter diode (LED) light exposure on retinal pigment epithelial cells (RPE cells) were examined to detect cellular damage or change and to clarify its mechanisms. The RPE cells were cultured and exposed by blue (470 nm) LED at 4.8 mW/cm(2). The cellular viability was determined by XTT assay and cellular injury was determined by the lactate dehydrogenase activity in medium. Intracellular reactive oxygen species (ROS) generation was determined by confocal laser microscope image analysis using dihydrorhodamine 123 and lipid peroxidation was determined by 4-hydroxy-2-nonenal protein-adducts immunofluorescent staining (HNE). At 24 h after 50 J/cm(2) exposures, cellular viability was significantly decreased to 74% and cellular injury was significantly increased to 365% of control. Immediately after the light exposure, ROS generation was significantly increased to 154%, 177%, and 395% of control and HNE intensity was increased to 211%, 359%, and 746% of control by 1, 10, and 50 J/cm(2), respectively. These results suggest, at least in part, that oxidative stress is an early step leading to cellular damage by blue LED exposure and cellular oxidative damage would be caused by the blue light exposure at even lower dose (1, 10 J/cm(2)).
Assuntos
Células Epiteliais/metabolismo , Células Epiteliais/efeitos da radiação , Peroxidação de Lipídeos/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Espécies Reativas de Oxigênio/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Epitélio Pigmentado da Retina/efeitos da radiação , Animais , Bovinos , Dano ao DNA , Células Epiteliais/citologia , Luz , Oxirredução , FototerapiaRESUMO
BACKGROUND: Although the morbidity of bowel ischemic events after glue embolization has been suggested, a causal relationship between glue and ischemia has not been clearly established. PURPOSE: To evaluate the efficiency and safety of transcatheter arterial embolization with n-butyl cyanoacrylate (NBCA-TAE) for upper gastrointestinal hemorrhage (GIH). MATERIAL AND METHODS: Between October 2006 and October 2012, 21 patients with upper GIH underwent NBCA-TAE, and endoscopic data were obtained within 30 days of follow-up. Shock index prior to and immediately after NBCA-TAE were compared to determine changes in hemodynamics. Days to Forrest type III, as assessed by follow-up endoscopy, was used as an indicator of the healing process. Other clinical outcomes included days for starting ingestion and for hospital discharge. RESULTS: Sixteen gastric and five duodenal ulcers, classified into Forrest type I, were treated. Immediate hemostasis was achieved in all the patients, and no re-bleeding occurred within the follow-up period. Shock index significantly (P < 0.001) improved from before (0.99 ± 0.076) to immediately after NBCA-TAE (0.67 ± 0.038). Sequential mucosal healing processes were observed in all the patients, and the number of days to Forrest type III was 9.6 ± 7.1. The number of days for starting ingestion and hospital discharge was 9.0 ± 4.5 and 15 ± 7.7 days, respectively. CONCLUSION: NBCA-TAE is an effective and safe method for the control of nonvariceal upper GIH, in terms of contribution to hemodynamics and healing process of the gastroduodenal mucosa.
Assuntos
Pressão Sanguínea , Embolização Terapêutica/métodos , Embucrilato/uso terapêutico , Frequência Cardíaca , Úlcera Péptica Hemorrágica/terapia , Úlcera Péptica/complicações , Adulto , Idoso , Idoso de 80 Anos ou mais , Meios de Contraste/administração & dosagem , Feminino , Seguimentos , Humanos , Óleo Iodado/administração & dosagem , Tempo de Internação/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Úlcera Péptica Hemorrágica/etiologia , Resultado do Tratamento , CicatrizaçãoRESUMO
The effects of DNA repair and transcription gene abnormalities in human pre-natal life have never been studied. Trichothiodystrophy (TTD) is a rare (affected frequency of 10(-6)) recessive disorder caused by mutations in genes involved in nucleotide excision repair (NER) pathway and in transcription. Based on our novel clinical observations, we conducted a genetic epidemiologic study to investigate gestational outcomes associated with TTD. We compared pregnancies resulting in TTD-affected offspring (n = 24) with respect to abnormalities during their antenatal and neonatal periods to pregnancies resulting in their unaffected siblings (n = 18), accounting for correlation, and to population reference values. Significantly higher incidence of several severe gestational complications was noted in TTD-affected pregnancies. Small for gestational age (SGA) <10th percentile [Relative risk (RR ) = 9.3, 95% CI = 1.4-60.5, p = 0.02], SGA <3rd percentile (RR = 7.2, 95% CI = 1.1-48.1, p = 0.04), and neonatal intensive care unit (NICU) hospitalization (RR = 6.4, 95% CI = 1.4-29.5, p = 0.02) occurred more frequently among TTD-affected neonates compared with their unaffected siblings. Compared with reference values from general obstetrical population, pregnancies that resulted in TTD-affected infants were significantly more likely to be complicated by hemolysis, elevated liver enzymes and low platelets (HELLP) syndrome (RR = 35.7, 95% CI = 7.6-92.5, p = 0.0002), elevated mid-trimester maternal serum human chorionic gonadotropin (hCG) levels (RR = 14.3, 95% CI = 7.0-16.6, p < 0.0001), SGA <3rd percentile (RR = 13.9, 95% CI = 7.4-21.1, p < 0.0001), pre-term delivery (<32 weeks) (RR = 12.0, 95% CI = 4.9-21.6, p < 0.0001), pre-eclampsia (RR = 4.0, 95% CI = 1.6-7.4, p = 0.006), and decreased fetal movement (RR = 3.3, 95% CI = 1.6-5.2, p = 0.0018). Abnormal placental development is an underlying mechanism that may explain the constellation of observed complications in our study. Thus, we hypothesize that TTD DNA repair and transcription genes play an important role in normal human placental development.
Assuntos
Reparo do DNA/genética , Desenvolvimento Fetal/genética , Transcrição Gênica , Síndromes de Tricotiodistrofia/embriologia , Síndromes de Tricotiodistrofia/genética , Adulto , Demografia , Família , Feminino , Humanos , Nascido Vivo , Pessoa de Meia-Idade , Gravidez , Resultado da Gravidez , Valores de Referência , Adulto JovemRESUMO
It is very rare that endometriotic lesions in the rectovaginal septum cause ileus. We report a case of bowel obstruction due to endometriotic lesions in the rectovaginal septum in a 22-year-old woman whose barium enema presented with apple-core-like findings. Diagnostic and treatment modalities were discussed. Preoperative and postoperative gonadotropin-releasing hormone analog and aromatase inhibitor therapy promote relief of clinical symptoms, a reduction of tumor volume and a better approach to radical surgery.
Assuntos
Endometriose/complicações , Íleus/etiologia , Doenças Retais/complicações , Endometriose/diagnóstico por imagem , Feminino , Humanos , Íleus/diagnóstico por imagem , Radiografia , Doenças Retais/diagnóstico por imagem , Adulto JovemRESUMO
We have developed a protein-synthesizing system reconstituted from recombinant tagged protein factors purified to homogeneity. The system was able to produce protein at a rate of about 160 microg/ml/h in a batch mode without the need for any supplementary apparatus. The protein products were easily purified within 1 h using affinity chromatography to remove the tagged protein factors. Moreover, omission of a release factor allowed efficient incorporation of an unnatural amino acid using suppressor transfer RNA (tRNA).
Assuntos
Sistema Livre de Células , Proteínas Recombinantes/metabolismo , Aminoácidos/química , Cromatografia de Afinidade , Eletroforese em Gel de Poliacrilamida , Escherichia coli/metabolismo , Histidina/química , Plasmídeos/metabolismo , Biossíntese de Proteínas , RNA de Transferência/metabolismo , Aminoacil-RNA de Transferência/metabolismo , Ribossomos/metabolismo , Fatores de TempoRESUMO
OBJECTIVES: To prospectively assess the efficacy of transurethral holmium (Ho):YAG laser prostatectomy using a side-firing fiber in patients with bladder outlet obstruction due to benign prostatic enlargement (BPE) from the standpoint of urodynamics. METHODS: 32 male patients with BPE aged 53-83 (mean 69.4) years were operated on. All patients, excluding 3 with urinary retention, were evaluated with the International Prostatic Symptom Score (IPSS), Quality of Life (QOL) score and uroflowmetry up to 12 months postoperatively, and a pressure/flow study was performed before and 3 months after the operation. RESULTS: The total IPSS score, QOL score, average and maximum flow rates improved significantly (p<0.0001) at 12 months postoperatively. In the pressure/flow study, detrusor opening pressure, maximum detrusor pressure, detrusor pressure at maximum flow, minimum urethral opening pressure, and Abrams-Griffiths number decreased significantly (p<0.0001, p = 0.0001, p<0.0001, p = 0.0019 and p<0.0001, respectively) 3 months postoperatively. Detrusor instability disappeared in 12 of 17 patients and remained in 2. CONCLUSIONS: Transurethral Ho:YAG laser prostatectomy was found to be effective for the treatment of bladder outlet obstruction due to BPE.
Assuntos
Terapia a Laser/métodos , Hiperplasia Prostática/cirurgia , Ressecção Transuretral da Próstata/métodos , Obstrução do Colo da Bexiga Urinária/cirurgia , Idoso , Idoso de 80 Anos ou mais , Hólmio , Humanos , Terapia a Laser/instrumentação , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Hiperplasia Prostática/complicações , Resultado do Tratamento , Obstrução do Colo da Bexiga Urinária/etiologia , Obstrução do Colo da Bexiga Urinária/fisiopatologia , Urodinâmica/efeitos da radiaçãoRESUMO
alpha-Glucosidase (AGH) inhibitory study by natural anthocyanin extracts was done. As the result of a free AGH assay system, 12 anthocyanin extracts were found to have a potent AGH inhibitory activity; in particular, Pharbitis nil (SOA) extract showed the strongest maltase inhibitory activity, with an IC(50) value of 0.35 mg/mL, as great as that of Ipomoea batatas (YGM) extract (IC(50) = 0.36 mg/mL). Interestingly, neither extract inhibited the sucrase activity at all. For the immobilized assay system, which may reflect the pharmacokinetics of AGH at the small intestine, SOA and YGM extracts gave more potent maltase inhibitory activities than those of the free AGH assay, with IC(50) values of 0.17 and 0.26 mg/mL, respectively. Both extracts also inhibited alpha-amylase action, indicating that anthocyanins would have a potential function to suppress the increase in postprandial glucose level from starch.
Assuntos
Antocianinas/farmacologia , Inibidores Enzimáticos/farmacologia , Inibidores de Glicosídeo Hidrolases , Acilação , Diabetes Mellitus Tipo 2/terapia , Humanos , Cinética , Pigmentos Biológicos , Extratos Vegetais/farmacologiaRESUMO
Four diacylated pelargonidin (Pg: SOA-4 and SOA-6), cyanidin (Cy: YGM-3), and peonidin (Pn: YGM-6) 3-sophoroside-5-glucosides isolated from the red flowers of the morning glory, Pharbitis nil cv. Scarlett O'Hara (SOA), and the storage roots of purple sweet potato, Ipomoea batatas cv. Ayamurasaki (YGM), were subjected to an alpha-glucosidase (AGH) inhibitory assay, in which the assay was performed with the immobilized AGH (iAGH) system to mimic the membrane-bound AGH at the small intestine. As a result, the acylated anthocyanins showed strong maltase inhibitory activities with IC(50) values of <200 microM, whereas no sucrase inhibition was observed. Of these, SOA-4 [Pg 3-O-(2-O-(6-O-(E-3-O-(beta-D-glucopyranosyl)caffeyl)-beta-D-glucopyranosyl)-6-O-E-caffeyl-beta-D-glucopyranoside)-5-O-beta-D-glucopyranoside] possessed the most potent maltase inhibitory activity (IC(50) = 60 microM). As a result of a marked reduction of iAGH inhibitory activity by deacylating the anthocyanins, that is, Pg (or Cy or Pn) sophoroside-5-glucoside, acylation of anthocyanin with caffeic (Caf) or ferulic (Fer) acid was found to be important in the expression of iAGH (maltase) inhibition. In addition, the result that Pg-based anthocyanins showed the most potent maltase inhibition, with an IC(50) value of 4.6 mM, and the effect being in the descending order of potency of Pg > Pn/Cy strongly suggested that no replacement at the 3'(5')-position of the aglycon B-ring may be essential for inhibiting iAGH action.
Assuntos
Antocianinas/farmacologia , Inibidores Enzimáticos/farmacologia , Inibidores de Glicosídeo Hidrolases , Acilação , Diabetes Mellitus Tipo 2/terapia , Humanos , Extratos Vegetais/farmacologiaRESUMO
Aurones are plant flavonoids that provide yellow color to the flowers of some popular ornamental plants, such as snapdragon and cosmos. In this study, we have identified an enzyme responsible for the synthesis of aurone from chalcones in the yellow snapdragon flower. The enzyme (aureusidin synthase) is a 39-kilodalton, copper-containing glycoprotein catalyzing the hydroxylation and/or oxidative cyclization of the precursor chalcones, 2',4',6',4-tetrahydroxychalcone and 2',4',6',3,4-pentahydroxychalcone. The complementary DNA encoding aureusidin synthase is expressed in the petals of aurone-containing varieties. DNA sequence analysis revealed that aureusidin synthase belongs to the plant polyphenol oxidase family, providing an unequivocal example of the function of the polyphenol oxidase homolog in plants, i.e., flower coloration.
Assuntos
Benzofuranos/metabolismo , Magnoliopsida/enzimologia , Sequência de Aminoácidos , Catálise , Catecol Oxidase/química , Catecol Oxidase/metabolismo , Ciclização , DNA Complementar , Precursores Enzimáticos/química , Precursores Enzimáticos/genética , Precursores Enzimáticos/isolamento & purificação , Precursores Enzimáticos/metabolismo , Genes de Plantas , Hidroxilação , Magnoliopsida/genética , Oxigenases de Função Mista/química , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/isolamento & purificação , Oxigenases de Função Mista/metabolismo , Dados de Sequência Molecular , Peso Molecular , Pigmentação , Estruturas Vegetais/enzimologia , Plantas/enzimologia , Homologia de Sequência de AminoácidosRESUMO
Four multiparous late-lactation Holstein cows were fed a basal ration designed to be co-limiting in intestinally absorbable supplies of methionine and lysine. Cows were supplemented with no amino acids, lysine by abomasal infusion to 140% of the calculated intestinally absorbable requirement, methionine by abomasal infusion to 140% of requirement, or both amino acids in a 4 x 4 Latin square design with 28-d periods. Unsupplemented cows consumed 23.8 kg/d of dry matter and produced 36.9 kg/d of milk containing 3.70% fat, 3.22% protein, and 4.82% lactose. Cows ate less dry matter and produced less milk and milk lactose, and tended (P = .06 or .08) to produce less milk protein when abomasally infused with methionine alone or together with lysine. Infusion of lysine alone resulted in production values numerically between those of unsupplemented cows and those cows supplemented with methionine alone or together with lysine. Evaluation of the results with two metabolic models of dairy cows indicated that performance of unsupplemented cows may have been limited by delivery of metabolizable or digestible protein, or intestinally absorbable lysine, isoleucine, or histidine, depending on the metabolic model used to evaluate animal performance. Regardless, results are consistent with those using nonruminant species, which have shown that imbalanced profiles of intestinally absorbable amino acids are associated with reduced dry matter intake and animal performance. Results also show that negative effects on performance of lactating dairy cows can occur if methionine is supplied at levels substantially in excess of calculated intestinally absorbable requirements, either alone or together with lysine.
Assuntos
Abomaso/efeitos dos fármacos , Bovinos/crescimento & desenvolvimento , Comportamento Alimentar/efeitos dos fármacos , Lisina/farmacologia , Metionina/farmacologia , Rúmen/metabolismo , Abomaso/metabolismo , Ração Animal , Animais , Indústria de Laticínios , Proteínas Alimentares/metabolismo , Ingestão de Energia/efeitos dos fármacos , Metabolismo Energético , Feminino , Fermentação , Lactação , Leite/química , Paridade , Distribuição AleatóriaRESUMO
A cDNA corresponding to 1-Cys peroxiredoxin, an evolutionarily conserved thiol-specific antioxidant enzyme, was isolated from buckwheat (Fagopyrum esculentum Moench), a dicotyledonous plant species belonging to the Polygonaceae family. The cDNA, which we have designated as FePer1, contains a major open reading frame capable of encoding a polypeptide of 219 residues with a predicted molecular mass of 24.3kDa. The deduced primary structure of FePer1 polypeptide shows a high level (about 70%) of sequence homology to other recently identified plant 1-Cys peroxiredoxins. FePer1 also exhibits a significant level of sequence similarity to non-plant 1-Cys peroxiredoxins, sharing 52 and 42% identities with mammalian and fungal 1-Cys peroxiredoxins, respectively. As for all 1-Cys peroxiredoxins identified from various organisms, the amino acid sequence proposed to constitute the active site of the enzyme is highly conserved in FePer1 polypeptide. The gene corresponding to FePer1 cDNA is a single-copy gene in the buckwheat genome. Its expression is regulated in a seed-specific and temporal manner during seed development. FePer1 gene is induced transiently for a short period immediately after seed imbibition.
Assuntos
Fagopyrum/genética , Germinação/genética , Peroxidases/genética , Sementes/genética , Sequência de Aminoácidos , Sequência de Bases , Northern Blotting , Clonagem Molecular , Sequência Conservada , DNA Complementar/química , DNA Complementar/genética , DNA de Plantas/genética , Evolução Molecular , Fagopyrum/enzimologia , Fagopyrum/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Peroxirredoxinas , Filogenia , Proteínas de Plantas/genética , RNA de Plantas/genética , RNA de Plantas/metabolismo , Sementes/enzimologia , Sementes/crescimento & desenvolvimento , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosRESUMO
OBJECTIVE: Allogenic blood transfusion can transmit viral infection or cause immunological side effects. Recently, improved operative techniques have required less frequent transfusions in thoracic aortic aneurysm surgery. This study examined the efficacy of using autologous platelet-rich plasma in thoracic aortic aneurysm surgery. METHOD: Eight patients underwent nine operations using an autologous platelet-rich plasma program. The control group consisted of 15 historic patients matched for operative procedure and age. All operations were performed by the same surgeon. The platelet-rich plasma program required the collection of platelet-rich plasma prior to the infusion of heparin; platelet-rich plasma transfusions were administered following neutralization by heparin. RESULTS: The volume of platelet-rich plasma averaged 252 +/- 14.3 ml; total platelets in the platelet-rich plasma were 2.27 +/- 0.20 x 10(11) cells. The median number of homologous red blood cells transfused during the operative day was 0 units (range 0 to 12) in the platelet-rich plasma group and 3 units (range 0 to 25) in the controls. The median number of homologous fresh frozen plasma was 0 units (range 0 to 20) in the platelet-rich plasma group, and 5 units (range 0 to 30) in the controls. The platelet-rich plasma group received significantly fewer transfusions. CONCLUSION: Autologous platelet-rich plasma transfusion was an effective way to reduce homologous blood transfusions in thoracic aortic aneurysm surgery.
Assuntos
Aneurisma da Aorta Torácica/cirurgia , Transfusão de Sangue Autóloga/normas , Adulto , Feminino , Humanos , Pessoa de Meia-Idade , Plasmaferese , Contagem de PlaquetasRESUMO
Ca2+/calmodulin-dependent protein kinases (CaMKs) are believed to play important roles in the development and function of the nervous system. We report here the identification and expression of mouse CaMKIbeta (mCaMKIbeta), in particular mCaMKIbeta2, an isoform of mCaMKIbeta. During embryogenesis, the mCaMKIbeta2 gene is expressed mainly in the nervous system, including brain, spinal cord, trigeminal ganglion, and retina. Within the CNS, the expression of mCaMKIbeta2 is detected in the mantle zone, but not in the ventricular zone, suggesting its possible involvement in the differentiation of neurons. In the adult brain, mCaMKIbeta2 transcripts are detected at high levels in the anterior olfactory nuclei, piriform cortex, septal nuclei, bed nuclei of the stria terminalis, hippocampal pyramidal cells, dentate granule cells, amygdala, hypothalamic nuclei, parabrachial nucleus, and nucleus of the solitary tract. The distinct gene expression pattern suggests that mCaMKIbeta2 may also be involved in different mature neuronal functions from other CaMKs. In addition, mCaMKI/beta2 proteins are localized to the cytoplasm and nuclei, but not to nucleoli, suggesting that mCaMKIbeta2 proteins might be involved in the cytoplasmic and nuclear signal transduction of the nervous system.
Assuntos
Proteínas Quinases Dependentes de Cálcio-Calmodulina/análise , Isoenzimas/análise , Sistema Nervoso/enzimologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Encéfalo/enzimologia , Proteína Quinase Tipo 1 Dependente de Cálcio-Calmodulina , Proteínas Quinases Dependentes de Cálcio-Calmodulina/química , Proteínas Quinases Dependentes de Cálcio-Calmodulina/genética , Núcleo Celular/enzimologia , Clonagem Molecular , Citoplasma/enzimologia , DNA Complementar/química , DNA Complementar/genética , Isoenzimas/química , Isoenzimas/genética , Camundongos , Dados de Sequência Molecular , Sistema Nervoso/crescimento & desenvolvimento , Células PC12 , RNA Mensageiro/análise , Ratos , Retina/enzimologia , Medula Espinal/enzimologia , Distribuição Tecidual , Transfecção , Gânglio Trigeminal/enzimologiaRESUMO
Amino acid assignments of metazoan mitochondrial codons AGA/AGG are known to vary among animal species; arginine in Cnidaria, serine in invertebrates and stop in vertebrates. We recently found that in the mitochondria of the ascidian Halocynthia roretzi these codons are exceptionally used for glycine, and postulated that they are probably decoded by a tRNA(UCU). In order to verify this notion unambig-uously, we determined the complete RNA sequence of the mitochondrial tRNA(UCU) presumed to decode codons AGA/AGG in the ascidian mitochondria, and found it to have an unidentified U derivative at the anticodon first position. We then identified the amino acids attached to the tRNA(U*CU), as well as to the conventional tRNAGly(UCC) with an unmodified U34, in vivo. The results clearly demonstrated that glycine was attached to both tRNAs. Since no other tRNA capable of decoding codons AGA/AGG has been found in the mitochondrial genome, it is most probable that this tRNA(U*CU) does actually translate codons AGA/AGG as glycine in vivo. Sequencing of tRNASer(GCU), which is thought to recognize only codons AGU/AGC, revealed that it has an unmodified guanosine at position 34, as is the case with vertebrate mitochondrial tRNASer(GCU) for codons AGA/AGG. It was thus concluded that in the ascidian, codons AGU/AGC are read as serine by tRNASer(GCU), whereas AGA/AGG are read as glycine by an extra tRNAGly(U*CU). The possible origin of this unorthodox genetic code is discussed.
Assuntos
Códon , Glicina/metabolismo , Mitocôndrias/genética , RNA de Transferência de Glicina/metabolismo , Animais , Sequência de Bases , Dados de Sequência Molecular , Conformação de Ácido Nucleico , RNA de Transferência de Glicina/química , RNA de Transferência de Serina/química , RNA de Transferência de Serina/genética , Serina/metabolismo , UrocordadosRESUMO
BACKGROUND: Drosophila neurospecific receptor tyrosine kinases (RTKs), Dror and Dnrk, as well as Ror1 and Ror2 RTKs, isolated from human neuroblastoma, have been identified as a structurally related novel family of RTKs (Ror-family RTKs). Thus far, little is known about the expression and function of mammalian Ror-family RTKs. RESULTS: We have identified murine Ror-family RTKs, mRor1 and mRor2. Both mRor1 and mRor2 genes are induced upon neuronal differentiation of P19EC cells. During neuronal differentiation in vitro, the expression of mRor2 is transiently induced, although that of mRor1 increases continuously. During embryogenesis, the mRor1 gene is expressed in the developing nervous system within restricted regions and in the developing lens epithelium. The expression of mRor1 is sustained in the nervous system and is also detected in non-neuronal tissues after birth. In contrast, the expression of mRor2 is detected mainly in the developing nervous system within broader regions and declines after birth. Possible relationships of mRor1 and mRor2 genes with previously identified mutants have also been examined. CONCLUSIONS: The developmental expressions of mRor1 and mRor2, in particular in the nervous system, are differentially regulated, reflecting their expression patterns in vitro. mRor1 and mRor2 may thus play differential roles during the development of the nervous system.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Sistema Nervoso/embriologia , Receptores de Superfície Celular/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Células COS , Divisão Celular , Células Cultivadas , Mapeamento Cromossômico , Clonagem Molecular , DNA Complementar/metabolismo , Embrião de Mamíferos/anatomia & histologia , Embrião de Mamíferos/metabolismo , Ligação Genética , Humanos , Immunoblotting , Hibridização in Situ Fluorescente , Camundongos , Modelos Genéticos , Dados de Sequência Molecular , Neuroblastoma/metabolismo , Fosforilação , Ratos , Receptores Proteína Tirosina Quinases , Receptores Órfãos Semelhantes a Receptor Tirosina Quinase , Homologia de Sequência de Aminoácidos , Fatores de Tempo , Distribuição Tecidual , Tirosina/metabolismoRESUMO
BACKGROUND: Multiple organ failure (MOF) is the most serious complication in severe acute pancreatitis, contributing to its high mortality. It has been suggested that changes of high-energy phosphates, intracellular pH, and intracellular cation homeostasis are closely related to hepatocellular injury associated with MOF. METHODS: Phosphorus metabolites, intracellular pH (pHi), and intracellular Na+ concentration ([Na+]i) were measured in rat livers in vivo using 31P and 23Na NMR spectroscopy after deoxycholic acid (DCA)-induced pancreatitis or intraperitoneal injection (ip) of pancreatitis-induced ascitic fluid (PAF). RESULTS: Two hours after induction of DCA-pancreatitis, the liver experienced significant intracellular acidosis (pHi = 6.99 +/- 0.16) and sodium loading (75 +/- 9 mM) and a reduction in its energy state (beta-ATP/Pi = 0.2 +/- 0.03 and Pi = 164 +/- 12). Although ip injection of PAF into healthy rats did not induce systemic hypotension, the livers under these conditions also developed severe disturbances in hepatocellular ion homeostasis and depletion of its bioenergetics. The longer the abdomen was exposed to the PAF, the worse the changes were. At 3 h after ip injection of PAF, hepatic [Na+]i significantly increased (42 +/- 3 mM) along with a significant decrease in pHi (7.30 +/- 0. 03). At 6 h after ip injection of PAF, the hepatic beta-ATP/Pi ratio decreased to 0.34 +/- 0.05 and Pi increased to 97 +/- 27. CONCLUSIONS: PAF induced severe hepatocellular acidosis, rapid accumulation of hepatic intracellular sodium, impaired hepatic cytosolic phosphorylation potential, and increased hepatic utilization of ATP. These effects may account for the eventual development of liver dysfunction associated with necrotizing pancreatitis.
Assuntos
Líquido Ascítico/metabolismo , Fígado/fisiopatologia , Pancreatite/fisiopatologia , Doença Aguda , Animais , Líquido Ascítico/fisiopatologia , Ácido Desoxicólico , Metabolismo Energético/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Membranas Intracelulares/metabolismo , Fígado/metabolismo , Espectroscopia de Ressonância Magnética , Masculino , Pancreatite/induzido quimicamente , Pancreatite/metabolismo , Fósforo/metabolismo , Ratos , Ratos Sprague-Dawley , Sódio/metabolismoRESUMO
Following the characterization of dihydroisocoumarin constituents, two secoiridoid glucoside complexes, called hydramacrosides A and B, were isolated from the leaves of Hydrangea macrophylla Seringe var. thunbergii Makino. The absolute stereostructures of hydramacrosides A and B were elucidated on the basis of chemical and physicochemical evidence, which included the application of the 13C-NMR glycosylation shift rule of 1,1'-disaccharides and the modified Mosher's method. Hydramacrosides A and B exhibited an inhibitory effect on histamine release from rat mast cells induced by an antigen-antibody reaction.
Assuntos
Glucosídeos/farmacologia , Antagonistas dos Receptores Histamínicos/farmacologia , Plantas Medicinais/química , Piranos/farmacologia , Acetilação , Animais , Reações Antígeno-Anticorpo/efeitos dos fármacos , Exsudatos e Transudatos/citologia , Exsudatos e Transudatos/efeitos dos fármacos , Exsudatos e Transudatos/metabolismo , Glucosídeos/química , Antagonistas dos Receptores Histamínicos/química , Hidrólise , Técnicas In Vitro , Espectroscopia de Ressonância Magnética , Mastócitos/efeitos dos fármacos , Mastócitos/metabolismo , Conformação Molecular , Folhas de Planta/química , Piranos/química , RatosRESUMO
BACKGROUND: Liver dysfunction may be an early event or the end result of multiple organ dysfunction (MOD) in necrotizing pancreatitis. This study measured the early changes in hepatocellular ions and energetics associated with such conditions. METHODS: Twenty-five rats, prepared with a 23Na and 31P double-tuned nuclear magnetic resonance surface coil secured over the dome of the liver, were randomized into 5 groups: control, 10 and 20 minutes of total inflow ischemia, pancreatitis induced by deoxycholic acid (DCA), and sham-DCA (saline injection). Dysprosium-TTHA3- solution was used to separate the intracellular and extracellular sodium peaks. RESULTS: In rat liver, 20 minutes of total inflow occlusion caused irreversible depletion of high-energy phosphates. Changes at 2 hours after the onset of DCA-pancreatitis are compared with changes after 20 minutes of ischemia (mean +/- SEM). Although the DCA-pancreatitis animals did not become hypotensive until 1 hour after the induction of pancreatitis, the changes in hepatic intracellular ions and energetics began soon after such an insult. At 2 hours after the onset of pancreatitis, hepatocellular pHi and [NA+]i were 6.99 +/- 0.16 and 78.4 +/- mmol/L, respectively (P < .01, compared with sham animals). A similar pattern of changes in hepatic bioenergetics also occurred. After the onset of pancreatitis, the hepatic cytostolic phosphorylation potential decreased with time (y = 0.654 - 0.004t, where t is time in minutes and r2 = 0.967 and the rate of hepatic hydrolysis of adenosine triphosphate increased progressively (y = 0.702t + 91.363, where t is time in minutes and r2 = 0.969. These changes correlated well with the accumulated [Na]i. CONCLUSIONS: Unresuscitated necrotizing pancreatitis caused severe hepatocellular acidosis, profound sodium accumulation, and bioenergy depletion early in its course. These effects were as severe as those induced by total liver ischemia. Liver dysfunction may be an early, not terminal, event of MOD in necrotizing pancreatitis.