ABSTRACT
Uridine-cytidine kinase, an important catalyst in the compensation pathway of nucleotide metabolism, can catalyze the phosphorylation reaction of cytidine to 5'-cytidine monophosphate (CMP), but the reaction needs NTP as the phosphate donor. To increase the production efficiency of CMP, uridine-cytidine kinase gene from Thermus thermophilus HB8 and polyphosphate kinase gene from Rhodobacter sphaeroides were cloned and expressed in Escherichia coli BL21(DE3). Uridine-cytidine kinase was used for the generation of CMP from cytidine and ATP, and polyphosphate kinase was used for the regeneration of ATP. Then, the D403 metal chelate resin was used to adsorb Ni²⁺ to form an immobilized carrier, and the immobilized carrier was specifically combined with the recombinant enzymes to form the immobilized enzymes. Finally, single-factor optimization experiment was carried out to determine the reaction conditions of the immobilized enzyme. At 30 °C and pH 8.0, 60 mmol/L cytidine and 0.5 mmol/L ATP were used as substrates to achieve 5 batches of high-efficiency continuous catalytic reaction, and the average molar yield of CMP reached 91.2%. The above method has the advantages of low reaction cost, high product yield and high enzyme utilization rate, and has good applied value for industrial production.
Subject(s)
Cytidine Monophosphate , Metabolism , Escherichia coli , Genetics , Industrial Microbiology , Methods , Phosphotransferases (Phosphate Group Acceptor) , Metabolism , Uridine KinaseABSTRACT
O presente trabalho foi realizado com o objetivo de desenvolver um método para a determinação de CMP em leite UAT por meio da aplicação da espectroscopia de infravermelho próximo. Leites UAT de oito marcas diferentes foram utilizados para a construção dos modelos de calibração. Os resultados demonstram que, para o desenvolvimento de um modelo de calibração adequado para a determinação de CMP em leite UAT, deve-se utilizar, juntamente com o método de regressão PLS, o método de seleção de espectros máxima distância e os pré-tratamentos 2ª derivada e variável normal padronizada. Além disso, pôde-se determinar que as regiões do infravermelho próximo mais correlacionadas com os movimentos vibracionais dos aminoácidos presentes no CMP foram: 1100-1310; 1400-1430; 1490-1550; 1640-1680; 1780-1970; 2020-2100 e 2310-2350nm. Conclui-se que a espectroscopia de infravermelho próximo pode ser uma alternativa para a determinação de CMP em leite UAT, desde que haja um conjunto de calibração com amostras representativas da população a ser predita no futuro.(AU)
his work´s objective was to develop an UHT milk caseinomacropeptide determination method trough NIR spectroscopy application. Eight UHT milk trademarks are used to produce a mathematical calibration model. The results of NIR analysis suggested that to produce a suitable calibration model, partial least-square regression (PLSR) must be used, with maximum distance in wavelenght space to select spectra, pre - treatment with 2nd derivative and standard normal variant (SNV). Also, suitable near-infrared regions more correlated with CPM aminoacids vibrational movements: 1100-1310; 1400-1430; 1490-1550; 1640-1680; 1780-1970; 2020-2100; and 2310-2350nm. Therefore, NIR spectroscopy can be an alternative to caseinomacropeptide determination of UHT milk, since there was a representative calibration set with a large enough and representative sample of entire population to be predicted in the future.(AU)
Subject(s)
Cytidine Monophosphate , Milk , Spectrum Analysis/statistics & numerical dataABSTRACT
A presente comunicação objetivou avaliar a quantificação do caseínomacropeptídeo (CMP), bem como diferenciá-lo (devido à adulteração com soro) do pseudo-CMP (devido à proteólise bacteriana) em amostras de leite cru coletadas nos domicílios do sul do Brasil. Os resultados reforçam a necessidade de práticas higiênicas durante a ordenha e estocagem do leite. As amostras de leite estudadas não estavam adulteradas por adição de soro, mostrando que a análise por cromatografia de exclusão por tamanho deve ser complementada a fim de revelar a identidade do peptídeo (CMP ou pseudo-CMP). A contagem bacteriana total (TBC) também se mostrou útil como indicador da contaminação do leite por micro-organismos proteolíticos, uma vez que uma relação diretamente proporcional entre TBC e pseudo-CMP foi estabelecida.(AU)
Subject(s)
Cytidine Monophosphate/analysis , Mesophyll Cells/cytology , Milk/microbiologyABSTRACT
Nucleoside phosphorylases (NPases) were found to be induced in Enterobacter aerogenes DGO-04, and cytidine and cytidine 5'-monophosphate (CMP) were the best inducers. Five mmol/L to fifteen mmol/L cytidine or CMP could distinctly increase the activities of purine nucleoside phosphorylase (PNPase), uridine phosphorylase (UPase) and thymidine phosphorylase (TPase) when they were added into medium from 0 to 8 h. In the process of enzymatic synthesis of adenine arabinoside from adenine and uracil arabinoside with wet cells of Enterobacter aerogenes DGO-04 induced by cytidine or CMP, the reaction time could be shortened from 36 to 6 h. After enzymatic reaction the activity of NPase in the cells induced remained higher than that in the cells uninduced.
Subject(s)
Cytidine , Pharmacology , Cytidine Monophosphate , Pharmacology , Enterobacter aerogenes , Enzyme Induction , Pentosyltransferases , VidarabineABSTRACT
BACKGROUND: The aim of this study was to evaluate the effect of continuous infusion of ondansetron compared with bolus injection on the incidence of postoperative nausea and vomiting (PONV) in intravenous, patient-controlled analgesia (PCA). METHODS: Sixty three women undergoing laparoscopic-assisted vaginal hysterectomy were randomly allocated according to the method of ondansetron administration: bolus injection of ondansetron (8 mg) after the operation (Bolus group, n = 21); continuous infusion after ondansetron (8 mg) mixed to PCA (PCA 8 mix group, n = 22); and continuous infusion after ondansetron (16 mg) mixed to PCA (PCA 16 mix group, n = 20). The PONV were measured at 1 hr, 6 hr, 24 hr and 48 hr after operation and pain scores (visual analog scale, VAS) were checked. RESULTS: The incidence of PONV during 48 hr in the Bolus group (23.8%) and PCA 16 mix group (20.0%) were significantly lower than PCA 8 mix group (54.5%) (P < 0.05). The three groups showed similar VAS pain scores. CONCLUSIONS: Our results suggest that continuous infusion of ondansetron 16 mg is as effective as a bolus injection of ondansetron (8 mg) at preventing PONV in high-risk patients.
Subject(s)
Female , Humans , Analgesia, Patient-Controlled , Anesthesia , Arabinonucleotides , Cytidine Monophosphate , Hysterectomy, Vaginal , Incidence , Nausea , Ondansetron , Passive Cutaneous Anaphylaxis , Piperidines , Postoperative Nausea and Vomiting , Propofol , VomitingABSTRACT
With PVA as the carrier, the frozen beer yeast cells were immobilized for production of CTP from CMP. we explored the optimal condition of the immobilization from the aspects of the type, concentration of the PVA, and the immobilizing methods of cells In all 8 continuous batch of fermentation under the reactional condition of the immobilized cells, the conversion rate of CTP were maintained about 85% - 95%. Moreever, the storage stability of immobilized cells were investigated, and the products was also isolated and identifided by HPLC.
Subject(s)
Beer , Microbiology , Bioreactors , Microbiology , Cells, Immobilized , Cell Biology , Metabolism , Chromatography, High Pressure Liquid , Cytidine Monophosphate , Metabolism , Cytidine Triphosphate , Metabolism , Fermentation , Industrial Microbiology , Methods , Polyvinyl Alcohol , Saccharomyces cerevisiae , Cell Biology , MetabolismABSTRACT
Pregunta de investigación. ¿Es más rápida la respuesta clínica favorable en el tratamiento de la Parálisi Facial a Frigore si incluimos el fármaco Núcleo CMP?. Hipótesis: En pacientes consultantes de Fisioterapía- Hospital de Clínicas gestión 2001, con Parálisis Facial a Frigore emplear como parte del tratamiento el fármaco Núcleo CMP, además del tratamiento porFisioterapia y corticoesteroide: resulta favorable en cuanto a su recuperación más rápida; frente a pacientes que no recibieron el medicamaento y solo se les dio placebo. Objetivos. 1 Determinar la rapidez en la mejoría clínica en pacientes con parálisi facial a frigore tratados en la unidad de Fisioterapía que recibieron el fármaco Núcleo CMP y aquellos que no lo recibieron. Demostrar el tiempo en el cual se presenta la respuesta favorable en abos grupos de pacientes con parálisis facial que recibieron el fármaco y los queno lo recibieron. Diseño. Ensayo Clínico, simple ciego, randomizado. Lugar. Hospital de Clínicas- La Paz- Unidad Medicina Familiar y Rehabilitación. Material y métodos. 1. Etapa. Obtención de la muestra y randomización pormedio dela tabla de números aleatorios delpaquete de estadística STATS TM, con demostración de homogeneidad. 2. Etapa Examén clínico neurológico, cálculo de medidas de efecto. Resultados. Se aleatorizaron a Núcleo CMP (n=27) y placebo (n=30) un total de 57 pacientes. Concluyeron el estudio 24 asignados a placebo y 20 tratados con núcleo CMP. Para la variable de eficacia principal (tiempo transcurrido hasta la mejoría neurológica confirmada), el análisis de las observaciones durante un seguimiento de 8 semanas/pacientes, indico de manifiesto una diferencia significativa entre ambos grupos; favorables al núcleo CMP (p=0.0026). Confirmado por la prueba X2 con aceptación de la Hipótesis del estudio. Esta evolución favorable se observó en (38.9 porciento) de los 2o pacientes asigandos al núcleo CMP (p= 0.039), que desarrollaron una progresión clínica favorable confirmada desde el 10mo día de tratamiento y en (34.8 por ciento) de 24 pacientes tratados con placebo. La evolución primaria se confirmó mediante examén neurológico. Conclusión. Se cumple la hipótesis del estudio, por ser el fármaco efectivo en la recuperación temprana de la Parálisis facial a frigore, "según el estudi", su uso sin embargo dependerá del criterio médico y la comparación por metaanálisis, con resultados de estudios más grandes.
Subject(s)
Humans , Male , Female , Placebos , Cytidine Monophosphate , Facial Paralysis/complications , Facial Paralysis/diagnosis , Physical Therapy Department, HospitalABSTRACT
Angiogenin is a protein belonging to the superfamily of RNase A. The RNase activity of this protein is essential for its angiogenic activity. Although members of the RNase A family carry out RNase activity, they differ markedly in their strength and specificity. In this paper, we address the problem of higher specificity of angiogenin towards cytosine against uracil in the first base binding position. We have carried out extensive nano-second level molecular dynamics(MD) computer simulations on the native bovine angiogenin and on the CMP and UMP complexes of this protein in aqueous medium with explicit molecular solvent. The structures thus generated were subjected to a rigorous free energy component analysis to arrive at a plausible molecular thermodynamic explanation for the substrate specificity of angiogenin.
Subject(s)
Animals , Cattle , Cytidine Monophosphate/chemistry , Ligands , Models, Chemical , Models, Molecular , Protein Binding , Ribonuclease, Pancreatic/chemistry , Substrate Specificity , Thermodynamics , Uridine Monophosphate/chemistryABSTRACT
Idiopatic or HTLV-1 associated progressive spastic paraparesis does not have a clear etiology or treatment. To assess the effects of a medication containing cytidinmonophosphate, uridintriphosphate and vitamin B 12 in the treatment of progressive spastic. Patients with the disease were randomly assigned to receive the Nucleus CMP forte (containing dysodic cytidinmonophosphate 5 mg,trisodic uridintriphosphate 3 mg and hydroxicobalamin 2 mg) tid or placebo during 6 months. Gait, spasticity, degree of neurogenic bladder and somatosensitive evoked potentials were assessed during treatment. Forty six patients aged 25 to 79 years old were studied, 24 were female and 29 HTLV-1 positive. Twenty two were treated with the drug and the rest with placebo. Gait and spasticity improved in 7 of 22 patients receiving the drug and 1 of 24 receiving placebo (p<0.05). Neurogenic bladder improved in 10 of 22 receiving the drug and 4 patients treated with the drug and in two of seven treated with placebo. The medication caused a modest improvement in patients with progressive spastic paraparesis and was free of side effects
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Uridine Triphosphate/administration & dosage , Vitamin B 12/administration & dosage , Cytidine Monophosphate/administration & dosage , Paraparesis, Tropical Spastic/drug therapy , Urinary Bladder, Neurogenic/drug therapy , HTLV-I Infections/complications , HTLV-I Infections/drug therapy , Gait/drug effectsABSTRACT
Rats were perfused with glutaraldehyde. The lower incisors were dissected free. Mid-sagittal slice was cut through the entire incisor, deimneralized. The pigmentation zone was isolated and further sliced into cross sections. These sections were incubated for CMPase. CMPase reaction product were associated with the lateral cell memberane of subclass II and Subclass III. This localization confirm the role of extracellular space of SM as a route of enamel proteins during maturation