A novel multiplex PCR-RFLP method for simultaneous detection of the MTHFR 677 C > T, eNOS +894 G > T and - eNOS -786 T > C variants among Malaysian Malays

Background Hyperhomocysteinemia as a consequence of the MTHFR 677 C > T variant is associated with cardiovascular disease and stroke. Another factor that can potentially contribute to these disorders is a depleted nitric oxide level, which can be due to the presence of eNOS +894 G > T and eNOS −786 T > C variants that make an individual more susceptible to endothelial dysfunction. A number of genotyping methods have been developed to investigate these variants. However, simultaneous detection methods using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis are still lacking. In this study, a novel multiplex PCR-RFLP method for the simultaneous detection of MTHFR 677 C > T and eNOS +894 G > T and eNOS −786 T > C variants was developed. A total of 114 healthy Malay subjects were recruited. The MTHFR 677 C > T and eNOS +894 G > T and eNOS −786 T > C variants were genotyped using the novel multiplex PCR-RFLP and confirmed by DNA sequencing as well as snpBLAST. Allele frequencies of MTHFR 677 C > T and eNOS +894 G > T and eNOS −786 T > C were calculated using the Hardy Weinberg equation. Methods The 114 healthy volunteers were recruited for this study, and their DNA was extracted. Primer pair was designed using Primer 3 Software version 0.4.0 and validated against the BLAST database. The primer specificity, functionality and annealing temperature were tested using uniplex PCR methods that were later combined into a single multiplex PCR. Restriction Fragment Length Polymorphism (RFLP) was performed in three separate tubes followed by agarose gel electrophoresis. PCR product residual was purified and sent for DNA sequencing. Results The allele frequencies for MTHFR 677 C > T were 0.89 (C allele) and 0.11 (T allele); for eNOS +894 G > T, the allele frequencies were 0.58 (G allele) and 0.43 (T allele); and for eNOS −786 T > C, the allele frequencies were 0.87 (T allele) and 0.13 (C allele). Conclusions Our PCR-RFLP method is a simple, cost-effective and time-saving method. It can be used to successfully genotype subjects for the MTHFR 677 C > T and eNOS +894 G > T and eNOS −786 T > C variants simultaneously with 100% concordance from DNA sequencing data. This method can be routinely used for rapid investigation of the MTHFR 677 C > T and eNOS +894 G > T and eNOS −786 T > C variants.

The role of His-134, -147, and -150 residues in subunit assembly, cofactor binding, and catalysis of sheep liver cytosolic serine hydroxymethyltransferase

In an attempt to unravel the role of conserved histidine residues in the structure-function of sheep liver cytosolic serine hydroxymethyltransferase (SHMT), three site-specific mutants (H134N, H147N, and H150N) were constructed and expressed, H134N and H147N SHMTs had K-m values for L-serine, L-allo-threonine and beta-phenylserine similar to that of wild type enzyme, although the k(cat) values were markedly decreased, H134N SHMT was obtained in a dimeric form with only 6% of bound pyridoxal 5'-phosphate (PLP) compared with the wild type enzyme, Increasing concentrations of PLP (up to 500 mu M) enhanced the enzyme activity without changing its oligomeric structure, indicating that His-134 may be involved in dimer-dimer interactions, H147N SHMT was obtained in a tetrameric form but with very little PLP (3%) bound to it, suggesting that this residue was probably involved in cofactor binding, Unlike the wild type enzyme, the cofactor could be easily removed by dialysis from H147N SHMT, and the apoenzyme thus formed was present predominantly in the dimeric form, indicating that PLP binding is at the dimer-dimer interface, H150N SHMT was obtained in a tetrameric form with bound PLP, However, the mutant had very little enzyme activity (<2%). The k(cat)/K-m values for L-serine, L-allo-threonine and beta-phenylserine were 80-, 56-, and SS-fold less compared with wild type enzyme, Unlike the wild type enzyme, it failed to form the characteristic quinonoid intermediate and was unable to carry out the exchange of 2-S proton from glycine in the presence of H-4-folate. However, it could form an external aldimine with serine and glycine, The wild type and the mutant enzyme had similar K-d values for serine and glycine, These results suggest that His-150 may be the base that abstracts the alpha-proton of the substrate, leading to formation of the quinonoid intermediate in the reaction catalyzed by SHMT.

Passively harmonically mode-locked vertical-external-cavity surface-emitting laser emitting 1.1 ps pulses at 147 GHz repetition rate

Coupled-cavity passive harmonic mode-locking of a quantum well based vertical-external-cavity surface-emitting laser has been demonstrated, yielding an output pulse train of 1.5 ps pulses at a repetition rate of 80 GHz and with an average power of 80 mW. Harmonic mode-locking results from coupling between the main laser cavity and a cavity formed within the substrate of the saturable absorber structure. Mode-locking on the second harmonic of the substrate cavity allows a train of 1.1 ps pulses to be generated at a repetition rate of 147 GHz with 40 mW average power. © 2010 American Institute of Physics.

Sapientia, 1983, Vol. XXXVIII, nº 147 (número completo)

Contenido: El ser : su constitución, su expresión conceptual / Octavio N. Derisi – Las causas en Aristóteles y Santo Tomás / Gustavo E. Ponferrada – La esencia de Dios / Octavio N. Derisi – Educación y función docente en el pensamiento de Tomás de Aquino / William R. Darós – Notas y comentarios -- Bibliografía

80 C 147 U.S. Aeroplane carrier "Saratoga" and H.M.S. "Despatch" in lower chambers, Miraflores Locks - Panama Canal - 2/7/28

Leonard Carpenter Panama Canal Collection. Photographs: Views of Panama and the Canal. [Box 1] from the Special Collections & Area Studies Department, George A. Smathers Libraries, University of Florida.

Experimental study of the beta-delayed proton decays of Er-145,Er-147

The beta-delayed proton decays of Er-145,Er-147 have been studied experimentally using the Ni-58 + Mo-92 reaction at beam energy of 383 MeV. On the basis of a He-jet apparatus coupled with a tape transport system, the beta-delayed proton radioactivities both from the nu s(1/2) ground state and the nu h(11/2) isomer in Er-145,Er-147 were observed by proton-gamma coincidence measurements. By analyzing the time distributions of the 4(+) -> 2(gamma)(+) transitions in the granddaughter nuclei Dy-144,Dy-146, the half-lives of 1.0 +/- 0.3 s and 1.6 +/- 0.2 s have been deduced for the nu h(11/2) isomers in Er-145,Er-147, respectively.

Properties of the beta-Delayed Proton Decay of Er-147

The beta-delayed proton decay of Er-147 is studied experimentally using the Ni-58+Mo-92 reaction at a beam energy of 383 MeV. Based on a He-jet apparatus coupled with a tape transport system, the beta-delayed proton radioactivities both from the nu s(1/2) ground state and the nu h(11/2) isomer in Er-147 are identified by proton-gamma coincidence measurements. By analyzing the time distribution of the 4(+) -> 2(+) gamma transition in the grand-daughter nucleus Dy-146, a half-life of 1.6 +/- 0.2 s is determined for the nu h(11/2) isomer in Er-147. The half-life for the ground state of Er-147 is estimated to be 3.2 +/- 1.2 s.

The occurrence of intra-operative hypotension varies between hospitals: observational analysis of more than 147,000 anaesthesia.

BACKGROUND: Hypotension, a common intra-operative incident, bears an important potential for morbidity. It is most often manageable and sometimes preventable, which renders its study important. Therefore, we aimed at examining hospital variations in the occurrence of intra-operative hypotension and its predictors. As secondary endpoints, we determined to what extent hypotension relates to the risk of post-operative incidents and death. METHODS: We used the Anaesthesia Databank Switzerland, built on routinely and prospectively collected data on all anaesthesias in 21 hospitals. The three outcomes were assessed using multi-level logistic regression models. RESULTS: Among 147,573 anaesthesias, hypotension ranged from 0.6% to 5.2% in participating hospitals, and from 0.3% up to 12% in different surgical specialties. Most (73.4%) were minor single events. Age, ASA status, combined general and regional anaesthesia techniques, duration of surgery and hospitalization were significantly associated with hypotension. Although significantly associated, the emergency status of the surgery had a weaker effect. Hospitals' odds ratios for hypotension varied between 0.12 and 2.50 (P < or = 0.001), even after adjusting for patient and anaesthesia factors, and for type of surgery. At least one post-operative incident occurred in 9.7% of the procedures, including 0.03% deaths. Intra-operative hypotension was associated with a higher risk of post-operative incidents and death. CONCLUSION: Wide variations remain in the occurrence of hypotension among hospitals after adjustment for risk factors. Although differential reporting from hospitals may exist, variations in anaesthesia techniques and blood pressure maintenance may also have contributed. Intra-operative hypotension is associated with morbidities and sometimes death, and constant vigilance must thus be advocated.

Resolución N° 147 ¿Cuál es la fuente de consulta primaria cuando se verifica en las sociedades comerciales el requisito de la convocatoria?

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Decreto Rectoral No. 894 : Créase el progr ama de Especialización en Gestión del Mejoramiento y la Productividad

Generación de nuevos programas académicos que resulten adecuados para la realidad del país y se constituyan en fuentes de soluciones y desarrollos innovadores para el avance y crecimiento de la nación colombiana, programa de Especialización en Gestión del Mejoramiento y la Productividad