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OBJECTIVE: Sagittal suture synostosis (SSS) is the most common form of craniosynostosis. For older patients, the strategy for surgical correction needs to consider diminished growth dynamics of the skull and an active reconstruction cranioplasty aims to sustain stability for the active child. We describe our technique of biparietal meander expansion (BME) technique for SSS for patients older than 1 year and retrospectively reviewed the perioperative course as well as the subjective experience of patients and caregivers during follow-up. METHODS: The BME technique incorporates bilateral serpentine craniotomies and fixation of the consecutively expanded bone tongues with crossing sutures for patients with SSS older than 12 months of age at surgery. We reviewed patients undergoing this surgical technique for correction of SSS and collected data about the clinical course and performed a patients reported outcome measure (PROM) for patients or caregivers to evaluate subjective experience and outcome after surgical treatment. RESULTS: BME was performed in 31 patients (8 females; median age: 43 months; range 13-388). The mean length of operation was 172.7±43 minutes (range 115-294). Patients experienced no immediate complications or neurological morbidity after surgery. Considering a total of 21 completed PROM questionnaires, the head shape after surgery was evaluated as either "better" (57%) or "much better" (43%) compared to preoperatively. Eighty-one percent of patients or caregivers answered that the patient experiences no limitation in daily activities. Although 42.8% perceived the hospital as strenuous, 90.5% would choose to undergo this treatment again. CONCLUSION: BME is a feasible technique for older SSS patients resulting in immediate stability of the remodelled calvarium with a more normal head shape. The survey among caregivers or patients revealed a favourable subjectively experienced outcome after this type of surgical treatment of SSS in the more complex context of an older patient cohort.
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Craneosinostosis , Procedimientos de Cirugía Plástica , Niño , Craneosinostosis/diagnóstico por imagen , Craneosinostosis/cirugía , Craneotomía , Femenino , Humanos , Lactante , Estudios Retrospectivos , Cráneo/cirugía , SuturasRESUMEN
Objective: To investigate the safety and clinical efficacy of "zoning" style laminectomy by ultrasonic bone curette in patients with severe thoracic ossification of the ligamentum flavum(TOLF). Methods: The clinical data of 36 patients with severe TOLF treated by "zoning" style laminectomy at Department of Spinal Surgery,Zhengzhou Orthopaedic Hospital from October 2015 to October 2018 were respectively analyzed.There were 17 males and 19 females,aged(57.3±10.2)years(range:43 to 80 years).According to the anatomical characteristics of the thoracic ligamentum flavum and the pathological process of ossionization,each decompression segment was divided into the upper 1/3 area of the lamina,the bilateral area of the ossionum flavum,the transitional area,and the area of close contact between the ossionum flavum and the spinal cord.Different surgical strategies were used for decompression in turn.The modified Japanese Orthopedic Association (mJOA) was used to evaluate the neurological function status before and after surgery,to evaluate the surgical effect of patients,and to observe the surgical complications.Paired sample T test was used for data analysis. Results: All 36 patients successfully completed the operation,the operation time was (88.6±24.6) minutes(range:60 to 150 minutes).The intraoperative blood loss was (426.7±167.4) ml(range:250 to 800 ml).Follow-up time was (27.2±7.7) months(range:12 to 48 months).The mJOA score at the last follow-up was 9.0±1.5,which was statistically significant compared with the preoperative score 5.4±1.8 (t=13.59,P<0.01).The improvement rate of mJOA score was (65.7±22.1) %,of which 17 cases were excellent (47.2%),13 cases were good (36.1%),4 cases were normal (11.1%),2 cases were ineffective (5.6%).Ten patients had cerebrospinal fluid leakage during the separation or removal of dural ossification and were cured after a series of comprehensive conservative treatment.Two patients showed transient neurological deterioration,and the neurological function gradually recovered to the preoperative state after comprehensive treatment such as increasing the mean arterial pressure and using neurotrophic drugs.During the follow-up,no aggravation of neurological dysfunction and segmental kyphosis were found. Conclusions: The ultrasonic bone curette-assisted "zoning" style laminectomy for the treatment of severe TOLF can directly observed the position relationship between ossification of the ligamentum flavum and the spinal canal structure during the operation,and accurately guide the surgical decompression.It has the advantages of safe operation and complete decompression,which provides an important reference for the selection of clinical surgery.
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Ligamento Amarillo , Osificación Heterotópica , Femenino , Humanos , Laminectomía , Ligamento Amarillo/cirugía , Masculino , Osificación Heterotópica/cirugía , Osteogénesis , Estudios Retrospectivos , Vértebras Torácicas/cirugía , UltrasonidoRESUMEN
OBJECTIVE: To evaluate the clinical efficacy of anterior debridement combined with posterior atlantoaxial fusion for atlantoaxial Tuberculosis. METHODS: From February 2005 to February 2013, 7 patients, 3 males and 4 females, with atlantoaxial Tuberculosis underwent anterior debridement combined with posterior atlantoaxial fusion in Department of Orthopedics Zhengzhou University People's Hospital were selected.In the preoperative and final follow-up, Japanese Orthopaedic Association score (JOA), neck disability index (NDI) and Frankel Classification were used to evaluate neurological function and calculate improvement rate.At final follow-up, clinical efficacy was evaluated by Odom's grade.Situation of internal fixation, fusion of upper cervical were assessed by X-ray, CT scan and MRI scan. RESULTS: Bony fusion were achieved in 7 cases after operation in 12 months. Tuberculosis were reached clinical cure between 17 and 21 months. At follow The JOA score increased from (11.1±0.7) preoperatively to (15.3±0.5) in final follow-up(P<0.05), and the NDI decreased from (34.0±4.6) preoperatively to (10.1±1.3) in final follow-up (P<0.05). At last follow-up, according to Odom's standard, excellent were obtained in 5 cases, good 1 cases and ordinary 1 case. Frankel Classification of all cases improved from D class to E. CONCLUSIONS: The treatment of anterior retropharyngeal debridement combine with atlantoaxial fusion, and local anti-tuberculosis drug using intraoperative, not only could obtain reliable clinical efficacy, completly removal of lesions, but also obtain strong stability, which plays an important role in the treatment of atlantoaxial Tuberculosis.
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Articulación Atlantoaxoidea/anomalías , Anomalías Congénitas , Desbridamiento , Fusión Vertebral/métodos , Vértebras Torácicas/cirugía , Tuberculosis de la Columna Vertebral/cirugía , Artrodesis , Femenino , Fijación Interna de Fracturas , Humanos , Fijadores Internos , Imagen por Resonancia Magnética , Masculino , Periodo Posoperatorio , Procedimientos de Cirugía Plástica , Vértebras Torácicas/diagnóstico por imagen , Tomografía Computarizada por Rayos X , Resultado del Tratamiento , Tuberculosis de la Columna Vertebral/diagnóstico por imagenRESUMEN
The Young's modulus and fracture strength of Si(1-x)Ge(x) nanowires (NWs) as a function of Ge concentration were measured from tensile stress measurements. The Young's modulus of the NWs decreased linearly with increasing Ge content. No evidence was found for a linear relationship between the fracture strength of the NWs and Ge content, which is closely related to the quantity of interstitial Ge atoms contained in the wire. However, by removing some of the interstitial Ge atoms through rapid thermal annealing, a linear relationship could be produced. The discrepancy in the reported strength of Si and Ge NWs between calculated and experimented results could be related to SiO(2-x)/Si interfacial defects that are found in Si(1-x)Ge(x) NWs. It was also possible to significantly decrease the number of interfacial defects in the NWs by incorporating a surface passivated Al2O3 layer, which resulted in a substantial increase in fracture strength.
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UNLABELLED: Aerobic plate counts (APC), coliforms, Bacillus cereus, Escherichia coli and eight foodborne pathogens were tested in 1008 cheap and junk foods, including candies, dried cakes, chewing gum, chocolate, dried and seasoned seafood, ice cream, and sugary foods. APCs were positive for 342 samples (33·9%), and the majority of the counts were 2-3 log CFU g(-1) or ml(-1) (average: 1·10 log CFU g(-1) or ml(-1) ). Most samples (97·3%) contained no coliforms (average: 0·07 log CFU g(-1) or ml(-1) ). Bacillus cereus was detected in 68 samples (average: 0·14 log CFU g(-1) or ml(-1) ). Escherichia coli and Listeria monocytogenes were detected in 6 and 1 samples, respectively, whereas other foodborne pathogens were not isolated. The highest bacterial counts were associated with dried and seasoned seafood products and dried cakes, suggesting that appropriate regulations of these food types should be considered. Cheap and junk foods were produced mainly in developing countries, but there were no significant differences in the bacterial counts among different countries of origin. The presence of foodborne pathogens may pose a risk for children. These results suggest that there is cause for deeper concern about the safety of these foods and that effective countermeasures should be established to improve their microbiological safety. SIGNIFICANCE AND IMPACT OF THE STUDY: Food safety is especially important for children, but only limited information is available about the microbiological quality of cheap and junk foods that are consumed frequently by primary schoolchildren (e.g. dried cakes, candies and chocolates). The present study investigated the microbial quality of cheap and junk foods, and our results indicate that these foods are a potential health risk for children, therefore, deeper concern about the safety of these foods and effective countermeasures should be established to improve their microbiological safety. The present study may contribute to the development of an appropriate child food safety management system.
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Bacterias/clasificación , Bacterias/aislamiento & purificación , Microbiología de Alimentos , Inocuidad de los Alimentos , Niño , Recuento de Colonia Microbiana , Alimentos/economía , Humanos , República de CoreaRESUMEN
AIMS: To investigate roles of quorum-sensing (QS) system in Acinetobacter sp. strain DR1 and rifampicin-resistant variant (hereinafter DR1R). METHODS AND RESULTS: The DR1 strain generated three putative acyl homoserine lactones (AHLs), while the DR1R produced only one signal and QS signal production was abrogated in the aqsI (LuxI homolog) mutant. The hexadecane-degradation and biofilm-formation capabilities of DR1, DR1R, and aqsI mutants were compared, along with their proteomic data. Proteomics analysis revealed that the AHL lactonase responsible for degrading QS signal was highly upregulated in both DR1R and aqsI mutant, also showed that several proteins, including ppGpp synthase, histidine kinase sensors, might be under the control of QS signalling. Interestingly, biofilm-formation and hexadecane-biodegradation abilities were reduced more profoundly in the aqsI mutant. These altered phenotypes of the aqsI mutant were restored via the addition of free wild-type cell supernatant and exogenous C(12) -AHL. CONCLUSIONS: The QS system in strain DR1 contributes to hexadecane degradation and biofilm formation. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report to demonstrate that a specific QS signal appears to be a critical factor for hexadecane degradation and biofilm formation in Acinetobacter sp. strain DR1.
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Acinetobacter/crecimiento & desarrollo , Acinetobacter/metabolismo , Alcanos/metabolismo , Biopelículas , Percepción de Quorum/fisiología , Acinetobacter/efectos de los fármacos , Acinetobacter/genética , Acil-Butirolactonas/metabolismo , Alcanos/farmacología , Adhesión Celular , Genes Bacterianos/genética , Interacciones Hidrofóbicas e Hidrofílicas/efectos de los fármacos , Mutación , Fenotipo , Proteómica , Transducción de Señal , Contaminantes Químicos del Agua/farmacologíaRESUMEN
Choline is an essential nutrient for phospholipids and acetylcholine biosynthesis in normal development of fetus. In the present study, we investigated the functional characteristics of choline transport system and inhibitory effect of cationic drugs on choline transport in rat conditionally immortalized syncytiotrophoblast cell line (TR-TBT). Choline transport was weakly Na(+) dependent and significantly influenced by extracellular pH and by membrane depolarization. The transport process of choline is saturable with Michaelis-Menten constants (K(m)) of 68microM and 130microM in TR-TBT 18d-1 and TR-TBT 18d-2 respectively. Choline uptake in the cells was inhibited by unlabeled choline and hemicholinium-3 as well as various organic cations including guanidine, amiloride and acetylcholine. However, the prototypical organic cation tetraethylammonium and cimetidine showed very little inhibitory effect of choline uptake in TR-TBT cells. RT-PCR revealed that choline transporter-like protein 1 (CTL1) and organic cation transporter 2 (OCT2) are expressed in TR-TBT cells. The transport properties of choline in TR-TBT cells were similar or identical to that of CTL1 but not OCT2. CTL1 was also detected in human placenta. In addition, several cationic drugs such as diphenhydramine and verapamil competitively inhibited choline uptake in TR-TBT 18d-1 with K(i) of 115microM and 55microM, respectively. Our results suggest that choline transport system, which has intermediate affinity and weakly Na(+) dependent, in TR-TBT seems to occur through a CTL1 and this system may have relevance with the uptake of pharmacologically important organic cation drugs.
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Colina/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Trofoblastos/metabolismo , Acetilcolina/farmacología , Animales , Secuencia de Bases , Transporte Biológico Activo/efectos de los fármacos , Proteínas de Transporte de Catecolaminas en la Membrana Plasmática/genética , Proteínas de Transporte de Catecolaminas en la Membrana Plasmática/metabolismo , Proteínas de Transporte de Catión/genética , Proteínas de Transporte de Catión/metabolismo , Línea Celular , Colina/farmacología , Cartilla de ADN/genética , Femenino , Hemicolinio 3/farmacología , Humanos , Cinética , Potenciales de la Membrana , Proteínas de Transporte de Membrana/genética , Transportadores de Anión Orgánico Sodio-Independiente/genética , Transportadores de Anión Orgánico Sodio-Independiente/metabolismo , Proteínas de Transporte de Catión Orgánico/genética , Proteínas de Transporte de Catión Orgánico/metabolismo , Transportador 2 de Cátion Orgánico , Embarazo , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sodio/metabolismo , Trofoblastos/citología , Trofoblastos/efectos de los fármacosRESUMEN
Human aminoacyl-tRNA synthetases (ARSs) are normally located in cytoplasm and are involved in protein synthesis. In the present work, we found that human methionyl-tRNA synthetase (MRS) was translocated to nucleolus in proliferative cells, but disappeared in quiescent cells. The nucleolar localization of MRS was triggered by various growth factors such as insulin, PDGF, and EGF. The presence of MRS in nucleoli depended on the integrity of RNA and the activity of RNA polymerase I in the nucleolus. The ribosomal RNA synthesis was specifically decreased by the treatment of anti-MRS antibody as determined by nuclear run-on assay and immunostaining with anti-Br antibody after incorporating Br-UTP into nascent RNA. Thus, human MRS plays a role in the biogenesis of rRNA in nucleoli, while it is catalytically involved in protein synthesis in cytoplasm.
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Nucléolo Celular/enzimología , Metionina-ARNt Ligasa/metabolismo , ARN Ribosómico/biosíntesis , Anticuerpos/farmacología , Transporte Biológico/efectos de los fármacos , División Celular , Células Cultivadas , Nucleótidos de Desoxiuracil/inmunología , Nucleótidos de Desoxiuracil/metabolismo , Factor de Crecimiento Epidérmico/farmacología , Técnica del Anticuerpo Fluorescente , Humanos , Immunoblotting , Insulina/farmacología , Proteínas Nucleares/química , Factor de Crecimiento Derivado de Plaquetas/farmacología , ARN Polimerasa I/metabolismo , ARN Nuclear/químicaRESUMEN
The two Pacific anguilloid sishes Anguilla japonica and Astroconger myriaster, belonging to the different families, appear to have identical chromosome numbers (2n = 38) and karyotypes, including one pair of conspicuous heteromorphic chromosomes in females. Cytophotometric measurements, however, indicate a considerable difference in DNA content between the two species.
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Cromosomas , ADN/análisis , Anguilas , Animales , Femenino , Cariotipificación , Cromosomas Sexuales , Especificidad de la EspecieRESUMEN
The poly(methyl urethane) acrylate oligomer was obtained by the reaction of methyl acrylate oligomer and 2-isocyanatoethyl methacrylate. Synthesis of poly(methyl urethane) acrylate oligomer was done with 2-mercaptoethanol (2-MEOH), methyl acrylate, 2,2'-azobisisobutyronitrile (AIBN, initiator) and dibutyltin dilaurate as a catalyst. Then 2-MEOH was used for functional chain transfer agent. The structure and property of the synthesized oligomers were characterized by FT-IR, FT-NMR, rheometer, and DSC. In this study, by synthetic method including the addition of 2-isocyanatoethyl methacrylate, thermal behavior of synthesized material was improved more than that reported in the previous study. Poly(methyl urethane) oligomer can be used for UV curable coatings, inks and adhesives. UV curable coating have high resistance against weather, ozone, aging, frictional wear, and heat. Besides they can absorb the shock and resist rust according to the thickness of film. It is used as an adhesive, paint, optical fiber coating agent, and waterproof agent because of these advantages at the present time.
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BACKGROUND AND OBJECTIVE: CYP3A, the drug-metabolizing enzyme is an important factor in the pharmacokinetics of many drugs. Polymorphism of the CYP3A5 gene is known to influence the functionality of the CYP3A5 enzymes. The full extent of CYP3A5 genetic polymorphism was analysed in a Korean population. METHODS: Specific polymerase chain reaction-restriction fragment length polymorphism tests for CYP 3AP1 through CYP3A5*7 or direct sequencing were used to identify reported CYP3A5 variant alleles, using 194 unrelated samples. RESULTS AND DISCUSSION: The most frequent single nucleotide polymorphism (SNP) was 6986A>G (CYP3A5*3). The next most frequent SNP was 31611C>T. Haplotype analysis using detected SNPs revealed that the most frequent haplotype was *3A (frequency: 0.724), followed by *1E (frequency: 0.211), *3C (frequency: 0.034) and *1A (frequency: 0.023). We did not find CYP3AP1*3, CYP3A5*6, or *7 in this Korean sample. CONCLUSION: A large proportion of Koreans may have relatively low levels of metabolically active CYP3A5 protein and therefore may be at risk of high levels of drugs metabolized by this enzyme, after administration of conventional doses.
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Pueblo Asiatico/genética , Sistema Enzimático del Citocromo P-450/genética , Frecuencia de los Genes , Polimorfismo de Nucleótido Simple , Alelos , Citocromo P-450 CYP3A , Genotipo , Haplotipos , Humanos , Japón , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Análisis de Secuencia de ADNRESUMEN
BACKGROUND AND OBJECTIVE: A randomized and prospective study was performed to compare anaesthetic characteristics and stress hormone responses of two anaesthetic techniques. METHODS: Forty-two patients undergoing day case excisional biopsy of breast mass were randomly assigned to receive propofol-remifentanil or sevoflurane-N2O. Anaesthesia was induced and maintained either with sevoflurane and 50% N2O in oxygen or with target-controlled remifentanil and propofol in 50% oxygen and air. Anaesthetic depth was monitored by the bispectral index. RESULTS: The times for induction (2.9 vs. 1.7 min) and for laryngeal mask insertion (5.7 vs. 3.3 min) were longer in the sevoflurane-N2O group than in the propofol-remifentanil group. However, apnoea (57.1% vs. 9.5%) and bradycardia (23.8% vs. 0%) were more prevalent with propofol-remifentanil. In the sevoflurane-N2O group, the emergence times to a verbal response (10.6 vs. 3.7 min), to extubation (11.8 vs. 4.0 min) and to orientation (14.7 vs. 4.8 min) were longer than in the propofol-remifentanil group. There were significantly more nausea (38.1% vs. 4.8%) and vomiting (19.2% vs. 0%) in the sevoflurane-N2O group than in the propofol-remifentanil group. The time to discharge was similar although there was less postoperative pain in the sevoflurane-N2O group. There were no differences in the perioperative cortisol responses in the two groups. CONCLUSIONS: Smoother induction of anaesthesia was seen with sevoflurane-N2O. Propofol-remifentanil showed a quicker emergence with less nausea/vomiting. There were similar perioperative cortisol responses in the two anaesthetic techniques.
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Anestésicos Intravenosos , Mama/patología , Éteres Metílicos , Óxido Nitroso , Piperidinas , Propofol , Adulto , Procedimientos Quirúrgicos Ambulatorios , Periodo de Recuperación de la Anestesia , Anestesia General , Apnea/inducido químicamente , Biopsia/efectos adversos , Bradicardia/inducido químicamente , Bradicardia/tratamiento farmacológico , Electroencefalografía , Femenino , Humanos , Hidrocortisona/sangre , Estudios Prospectivos , Remifentanilo , Sevoflurano , Factores de TiempoRESUMEN
High doses of intravenous protamine cause generalized vascular permeability changes in brain and other organs, and concomitant hypoproteinemia. The present investigations test the hypothesis that protamine has a dual action of both binding serum proteins and of undergoing absorptive-mediated transcytosis through microvascular endothelial barriers. Binding of albumin to protamine was demonstrated using equilibrium dialysis, and protamine was shown to selectively augment the uptake of albumin, but not sucrose, in isolated bovine or human brain capillaries. In contrast, the anionic macromolecule, dextran sulfate, resulted in an increased capillary uptake of both albumin and sucrose in vitro. The selective effects of protamine on albumin transport were also documented in vivo using an external organ technique; the intravenous injection of 1.5 mg/kg protamine resulted in a marked and selective influx of albumin into brain, heart, kidney, lung, and liver, and the increased albumin transport exceeded the increased sucrose uptake in some organs by an order of magnitude. The transcytosis of protamine through the cerebral microvascular barrier was documented with an internal carotid artery perfusion technique. In summary, these studies provide evidence for protamine-mediated vectorial transport of albumin through microvascular barriers in brain and other organs.
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Encéfalo/metabolismo , Protaminas/metabolismo , Albúmina Sérica/metabolismo , Animales , Transporte Biológico , Encéfalo/irrigación sanguínea , Antígenos CD4/metabolismo , Capilares/metabolismo , Bovinos , Sulfato de Dextran/farmacología , Humanos , Ratas , Ratas Sprague-Dawley , Proteínas Recombinantes/metabolismo , Sacarosa/metabolismoRESUMEN
Scg1, the product of the Saccharomyces cerevisiae SCG1 (also called GPA1) gene, is homologous to the alpha subunits of G proteins involved in signal transduction in mammalian cells. Scg1 negatively controls the pheromone response pathway in haploid cells. Either pheromonal activation or an scg1 null mutation relieves the negative control and leads to an arrest of cell growth in the G1 phase of the cell cycle. Expression of rat G alpha s was previously shown to complement the growth defect of scg1 null mutants while not allowing mating. We have extended this analysis to examine the effects of the short form of G alpha s (which lacks 15 amino acids present in the long form), G alpha i2, G alpha o, and Scg1-mammalian G alpha hybrids. In addition, we have found that constructs able to complement scg1 are also able to inhibit the response to pheromone and mating when expressed in a wild-type SCG1 strain. Overexpression of Scg1 has a similar inhibitory effect. These results are consistent with a model proposed for the action of Scg1 as the alpha component of a heterotrimeric G protein in which the beta gamma component (Ste4/Ste18) activates the pheromone response after dissociation from Scg1. They suggest that the G alpha constructs able to complement scg1 can interact with beta gamma to prevent activation of the pathway but are unable to interact with pheromone receptors to activate the pathway.
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Proteínas de Unión al GTP/genética , Péptidos/fisiología , Feromonas/fisiología , Saccharomyces cerevisiae/genética , Transducción de Señal , Animales , Secuencia de Bases , Cruzamientos Genéticos , Proteínas de Unión al GTP/metabolismo , Expresión Génica , Prueba de Complementación Genética , Vectores Genéticos , Sustancias Macromoleculares , Factor de Apareamiento , Modelos Genéticos , Datos de Secuencia Molecular , Mutación , Sondas de Oligonucleótidos , Plásmidos , Multimerización de Proteína , Ratas , Proteínas Recombinantes de Fusión/metabolismo , Mapeo Restrictivo , Saccharomyces cerevisiae/fisiologíaRESUMEN
An IgA1 half-molecule, which is composed of a deleted alpha1 chain linked with a disulfide bond to an intact kappa chain, was detected in a patient (Cha). The molecular weights of the paraprotein and the isolated alpha1 chain were estimated to be 75 000 and 53 000, respectively. Identification of tyrosine as the C-terminal amino acid and the presence of idiotypic determinants in the abnormal alpha1 chain indicated that the molecule would have an intact N-terminal variable region and a C-terminal region. Furthermore, no cleavage of the abnormal protein into Fab and Fc by proteolytic enzyme isolated from Neisseria gonorrhoeae suggested the absence of a "hinge" region in the abnormal alpha1 chain.
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Inmunoglobulina A , Cadenas Pesadas de Inmunoglobulina , Cadenas alfa de Inmunoglobulina , Secuencia de Aminoácidos , Cromatografía en Gel , Deleción Cromosómica , Humanos , Inmunoglobulina A/genética , Alotipos de Inmunoglobulinas , Cadenas kappa de Inmunoglobulina , Peso Molecular , Neisseria gonorrhoeae/enzimología , Péptido Hidrolasas/metabolismoRESUMEN
Previously, tamoxifen (TAM) has been shown to induce apoptosis through elevation of intracellular Ca2+ in HepG2 human hepatoblastoma cells. In this study we investigated the role of reactive oxygen species (ROS) in the TAM-induced apoptosis, and interrelationship between intracellular Ca2+ and ROS. TAM induced a slow and sustained increase in intracellular ROS level. An antioxidant, N-acetylcysteine significantly inhibited both ROS production and apoptosis induced by TAM, suggesting that ROS may play an essential role in the TAM-induced apoptosis. In a time frame ROS generation followed intracellular Ca2+ increase, and the extracellular and intracellular Ca2+ chelation with EGTA and BAPTA/AM, respectively, completely inhibited the TAM-induced ROS production, indicating that intracellular Ca2+ may mediate the ROS generation. Inhibitors of NAD(P)H oxidase, diphenylene iodonium, phenylarsine oxide and neopterine, significantly blocked the TAM-induced ROS generation and apoptosis, implying that this oxidase may act as a source enzyme for the production of ROS. These results suggest that non-phagocytic NAD(P)H oxidase may play a novel role as a mediator of the apoptosis associated with intracellular Ca2+ in HepG2 cells.
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Antineoplásicos Hormonales/farmacología , Apoptosis/efectos de los fármacos , Hepatoblastoma , Neoplasias Hepáticas , NADPH Oxidasas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Tamoxifeno/farmacología , Apoptosis/fisiología , Calcio/metabolismo , Humanos , Células Tumorales Cultivadas/citología , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/enzimologíaRESUMEN
To describe quantitatively the in vivo distribution and elimination of insulin, high-performance liquid chromatography (HPLC) separation was applied to the pharmacokinetic study of human insulin labeled with 125I at tyrosine A14 (A14-125I-insulin) as a tracer. Intact A14-125I-insulin levels were determined by HPLC and trichloroacetic acid (TCA) precipitation in plasma and various tissues after its intravenous bolus injection into mice. TCA precipitation consistently overestimated the intactness of A14-125I-insulin compared with HPLC, possibly due to the presence of both a TCA-precipitable intermediate degradation product of labeled insulin found in HPLC elution profiles and reported high-molecular-weight forms of labeled insulin in plasma. Thus, TCA precipitation gave a considerably lower total plasma clearance (Cltot) value than HPLC. The half-life of A14-125I-insulin was prolonged by a simultaneous injection of 8 U/kg unlabeled insulin, and labeled insulin behaved similarly to [14C]inulin (an extracellular fluid marker). The concentration time profiles of HPLC-separated labeled insulin in plasma were analyzed by a noncompartmental moment method, and both Cltot and steady-state apparent volume distribution (VDss) of A14-125I-insulin were considerably decreased by unlabeled insulin coadministration. In particular, VDss of labeled insulin decreased by 79%, similar to that of inulin (181 ml/kg), suggesting that the nonspecific binding of labeled insulin to tissues was so small that VDss of labeled insulin was reduced to the extracellular fluid volume (approximately 20% of the body weight) when its receptor binding was blocked effectively by unlabeled insulin.(ABSTRACT TRUNCATED AT 250 WORDS)
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Insulina/análogos & derivados , Animales , Precipitación Química , Cromatografía Líquida de Alta Presión , Insulina/administración & dosificación , Insulina/farmacocinética , Inulina/farmacocinética , Masculino , Tasa de Depuración Metabólica , Ratones , Receptor de Insulina/fisiología , Distribución Tisular , Tirosina/análisisRESUMEN
An in vivo model of chronic hypoglycemia was used to investigate changes in blood-brain barrier (BBB) glucose transport activity and changes in the expression of GLUT1 mRNA and protein in brain microvasculature occurring as an adaptive response to low circulating glucose levels. Chronic hypoglycemia was induced in rats by constant infusion of insulin via osmotic minipumps; control animals received infusions of saline. The criterion for chronic hypoglycemia was an average blood glucose concentration of < 2.3 mmol/l (42 mg/dl) after 5 days. The average blood glucose concentration at the end of the experimental period in the rats selected for study was 2.0 +/- 0.1 mmol/l (36 +/- 1 mg/dl) vs. 4.9 +/- 0.1 mmol/l (88 +/- 1 mg/dl) in the controls. Internal carotid artery perfusion studies demonstrated an increase in the BBB permeability-surface area (PS) product of 40% (P < 0.0005) in the chronically hypoglycemic animals as compared with controls. Western blotting of solubilized isolated brain capillaries demonstrated a 51% increase (P < 0.05) in immunoreactive BBB GLUT1 in the chronically hypoglycemic rats, and Northern blotting of whole-brain poly(A+) mRNA revealed a 50% increase in the GLUT1-to-actin ratio in the insulin-treated group (P < 0.05). Northern blotting analysis of microvessel-depleted total brain poly(A+) showed that the increase in GLUT1 mRNA in the chronically hypoglycemic rats was restricted to the BBB. The present study demonstrates increased expression of GLUT1 mRNA and protein at the BBB in chronic hypoglycemia and suggests that this increase is responsible for the compensatory increase in BBB glucose transport activity that occurs with chronically low circulating blood glucose levels.
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Barrera Hematoencefálica , Regulación de la Expresión Génica , Hipoglucemia/metabolismo , Proteínas de Transporte de Monosacáridos/metabolismo , ARN Mensajero/metabolismo , Animales , Glucemia/metabolismo , Northern Blotting , Western Blotting , Encéfalo/irrigación sanguínea , Encéfalo/metabolismo , Capilares , Transportador de Glucosa de Tipo 1 , Insulina/administración & dosificación , Masculino , Proteínas de Transporte de Monosacáridos/genética , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND AND AIMS: Glomerular filtration rate (GFR) provides the most accurate estimation of renal function. This study investigated the clinical characteristics of patients with impaired renal function having a normal serum creatinine level. We also validated whether the new Modification of Diet in Renal Disease (MDRD) formula can be applied in a healthy general population. MATERIAL AND METHODS: A total 393 participants who had serum creatinine concentration below 132.6 micromol/L without underlying diseases were randomly selected on an address basis in Ansan City. According to the level of GFR, they were divided into 3 groups and we analyzed their clinical characteristics. In 75 subjects, who were randomly selected 25 cases in each group based on GFR estimated by Cockcroft-Gault (C-G) formula, true GFR was measured using the 99mTc-DTPA renal clearance method. RESULTS: A total 393 (male: 106, female: 287) participants were as follows: GFR < 60 ml/min/1.73 m2; 4% (n = 25); 60 < or = GFR < 90 ml/min/1.73 m2; 26.2% (n = 103); GFR > or = 90 ml/min/1.73 m2; 67.4% (n = 265). In the group of decreased GFR, the mean age was older (67.4+/-10.7 vs. 48.7+/-12.8 vs. 39.4+/-8.2 years, p < 0.001), the gender was male (90.33+/-28.77 vs. 110.55+/-31.64, p < 0.001), and amount of proteinuria more increased (0.61 (0.56) vs. 0.33 (0.34) vs. 0.38 (0.33) gm/day, p = 0.007). The accuracy and precision of each formula were assessed by the difference in GFR measured by the 99mTc-DTPA renal clearance method--estimated GFR by each formula (deltaGFR), and the coefficient of determination (r2) of different predictive equations. The results were as follows: deltaGFR = -14.78+/-46.03, r2 = 0.79 (24-hour urinary creatinine clearance), deltaGFR=-16.79+/-57.32, r2 = 0.66 (100/serum creatinine), deltaGFR = 9.54+/-39.18, r2 = 0.87 (C-G formula), deltaGFR = -12.30+/-54.31, r2 = 0.66 (AASK formula), deltaGFR = 8.70+/-37.62, r2 = 0.79 (MDRD formula). Multiple linear regression analysis and logistic regression analysis showed that age, serum creatinine, total cholesterol and 24-hour urinary protein excretion were independently related to GFR and associated with a significant increase in the risk of decrement of GFR. CONCLUSIONS: From these results, a more accurate assessment of renal function should be required in a population characterized by older age, male gender and more proteinuria. The MDRD study formula and Cockcroft-Gault formula have greater accuracy and precision with true GFR, and this equation can be applied in subjects with healthy general population.