ABSTRACT
Introduction: The gross Motor Measure (GMFM), and two validated scales that measure gross motor developmental changes in children with cerebral palsy (CP), and are utilized to quantify therapeutic intervention results. Objetive: To analyze gross motor function scores (GMFM-66), across condiction severity levels (GMFCS), considering age, type, and CP topographical distribution, at Santiago's Teletón Institute, from 2006 to 2010. Comparison of results with that obtained in the literature, and evaluate the eventual prognostic value of both scales. Methodology: A descriptive retrospective review study of a GMFM database was conducted. A total of 397 children with CP met the inclusion criteria. GMFM-66 scores and GMFCS levels were associated according to the selected variables, and were compared to the literature. Results: 63.2 percent of the children are under 6 years of age. Hemiplegic and diplegic patients concentrate 76.6 percent of cases. According to type, 84.9 percent of patients with spastic CP. Independently from the analyzed variable, the mediam values of GMFM-66 decrease two to four time as the severity of the pathology increases from level I to level IV. Quadriplegic children are distributed among levels III and IV. No hemiplegic patients were found at level V. Compared to other studies, no significant differences were found in GMFM-66 scores across GMFCS levels. Visually, the score diagram by GMFCS levels and age is similar to Rosenbaum's. Conclusion: GMFM-66 score distribution across GMFCS levels turns out as described in the literature, however caution is suggested if used as a predictive value for individual cases.
Introducción: Gross Motor Function Measure (GMFM) Y Gross Motor Function Classification System (GMFCS) son escalas validadas que miden cambios en el desarrollo motor grueso del niño con parálisis cerebral (PC); se utilizan para cuantificar resultados de las intervenciones terapéuticas. Objetivo: Analizar puntajes de función motora gruesa (GMFM-66) por nivel de severidad de la patología (GMFCS), según edad, tipo y distribución topográfica de PC del Instituto Teletón Santiago 2006-2010, comparar con resultados obtenidos en literatura y evaluar el eventual pronóstico de ambas escalas. Metodología: Estudio descriptivo, retrospectivo de revisión a base de datos del GMFM. Un total de 397 niños con PC cumplen con requisitos de inclusión. Se relacionan puntajes de GMFM-66 por niveles de GMFCS según variables consideradas y se compara con bibliografía. Resultados: 63,2 por ciento de los niños son menores de 6 años. Pacientes con diplejía y hemiplejía concentran 76,6 por ciento. Según tipo, el 84,9 por ciento son espásticos. Independientemente de la variable analizada, los valores de las medianas de GMFM-66 disminuye entre 2 y 4 veces a medida que aumenta la severidad de la patología de nivel I a V. Los niños con cuadriplejía se distribuyen de nivel III a V; no se observan diferencias significativas de puntajes GMFM-66 por niveles de GMFCS; visualmente, el diagrama de puntajes por nivel de GMFCS y edad sería similar a Rosenbaum. Conclusión: La distribución de puntajes obtenidos en GMFM-66 por nivel de GMFCS guarda relación con estudios descritos en la literatura, pero se sugiere cautela al usarlos como valor predictivo para los casos individuales.
Subject(s)
Humans , Male , Adolescent , Female , Infant , Child, Preschool , Child , Disability Evaluation , Motor Skills/physiology , Cerebral Palsy/physiopathology , Age Distribution , Cross-Sectional Studies , Predictive Value of Tests , Cerebral Palsy/classification , Reproducibility of Results , Retrospective Studies , Severity of Illness IndexABSTRACT
The aim of the present study was to identify the enzymes in human liver catalyzing hydroxylations of bile acids. Fourteen recombinant expressed cytochrome P450 (CYP) enzymes, human liver microsomes from different donors, and selective cytochrome P450 inhibitors were used to study the hydroxylation of taurochenodeoxycholic acid and lithocholic acid. Recombinant expressed CYP3A4 was the only enzyme that was active towards these bile acids and the enzyme catalyzed an efficient 6alpha-hydroxylation of both taurochenodeoxycholic acid and lithocholic acid. The Vmax for 6alpha-hydroxylation of taurochenodeoxycholic acid by CYP3A4 was 18.2 nmol/nmol P450/min and the apparent Km was 90 microM. Cytochrome b5 was required for maximal activity. Human liver microsomes from 10 different donors, in which different P450 marker activities had been determined, were separately incubated with taurochenodeoxycholic acid and lithocholic acid. A strong correlation was found between 6alpha-hydroxylation of taurochenodeoxycholic acid, CYP3A levels (r2=0.97) and testosterone 6beta-hydroxylation (r2=0.9). There was also a strong correlation between 6alpha-hydroxylation of lithocholic acid, CYP3A levels and testosterone 6beta-hydroxylation (r2=0.7). Troleandomycin, a selective inhibitor of CYP3A enzymes, inhibited 6alpha-hydroxylation of taurochenodeoxycholic acid almost completely at a 10 microM concentration. Other inhibitors, such as alpha-naphthoflavone, sulfaphenazole and tranylcypromine had very little or no effect on the activity. The apparent Km for 6alpha-hydroxylation of taurochenodeoxycholic by human liver microsomes was high (716 microM). This might give an explanation for the limited formation of 6alpha-hydroxylated bile acids in healthy humans. From the present results, it can be concluded that CYP3A4 is active in the 6alpha-hydroxylation of both taurochenodeoxycholic acid and lithocholic acid in human liver.
Subject(s)
Cytochrome P-450 Enzyme System/metabolism , Lithocholic Acid/metabolism , Microsomes, Liver/metabolism , Mixed Function Oxygenases/metabolism , Taurochenodeoxycholic Acid/metabolism , Cytochrome P-450 CYP3A , Cytochrome P-450 Enzyme System/biosynthesis , Cytochrome P-450 Enzyme System/genetics , Gene Expression , Humans , Hydroxylation , Kinetics , Mixed Function Oxygenases/biosynthesis , Mixed Function Oxygenases/genetics , Taurodeoxycholic Acid/metabolismABSTRACT
A mitochondrial cytochrome P450 fraction catalyzing 1 alpha- and 27-hydroxylation but not 24-hydroxylation of 25-hydroxyvitamin D3 was purified from pig kidney. The ratio between the 1 alpha- and 27-hydroxylase activities was the same in all purification steps including a side fraction. Attempts to separate the 1 alpha- and 27-hydroxylase activities were unsuccessful. A monoclonal antibody directed against purified pig liver CYP27 recognized a protein of the same apparent M(r) and immunoprecipitated both the 1 alpha- and 27-hydroxylase activities towards 25-hydroxyvitamin D3 in the purified kidney enzyme fraction as well as in a solubilized, crude cytochrome P450 extract considered to represent the major part of the 25-hydroxyvitamin D3 hydroxylases in kidney mitochondria. Taken together, the results from the purification and the experiments with CYP27 antibody, substrate inhibition, and recombinant expressed human liver CYP27 strongly indicate that CYP27 is able to catalyze 1 alpha-hydroxylation but not 24-hydroxylation of 25-hydroxyvitamin D3 in kidney. In conclusion, the results provide evidence for a role for CYP27 as a major renal mitochondrial 25-hydroxyvitamin D3 1 alpha-hydroxylase.
Subject(s)
25-Hydroxyvitamin D3 1-alpha-Hydroxylase/metabolism , Cytochrome P-450 Enzyme System/metabolism , Kidney Cortex/enzymology , Mitochondria, Liver/enzymology , Mitochondria/enzymology , Steroid Hydroxylases/metabolism , 25-Hydroxyvitamin D3 1-alpha-Hydroxylase/immunology , 25-Hydroxyvitamin D3 1-alpha-Hydroxylase/isolation & purification , Animals , Antibodies, Monoclonal , Blotting, Western , Cholestanetriol 26-Monooxygenase , Chromatography, Ion Exchange , Cloning, Molecular , Cytochrome P-450 Enzyme System/immunology , Cytochrome P-450 Enzyme System/isolation & purification , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Humans , Kinetics , Molecular Weight , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Steroid Hydroxylases/immunology , Steroid Hydroxylases/isolation & purification , SwineABSTRACT
It was recently reported that both cholesterol 7 alpha-hydroxylase CYP7 and sterol 27-hydroxylase CYP27 are subject to negative feedback control by bile acids in isolated rat hepatocytes and rat liver (Twisk et al., (1995) Biochem. J. 305, 505-511; Vlahcevic et al. XIII International Bile Acid Meeting, September 30-October 2, 1994, Abstract 17). In the present work a series of experiments was performed to study whether there is a coordinate regulation of CYP7 and CYP27 also in rabbit liver. Treatments of rabbits with cholic acid or cholestyramine resulted in marked suppression and induction, respectively, of CYP7 mRNA. In contrast, there were no significant effects on the CYP27 mRNA expression, amount of CYP27 protein or mitochondrial 27-hydroxylase activity. Thus, there is an apparent difference between rat and rabbit with respect to regulation of CYP27 by bile acids. It is concluded that, in rabbit, there is little or no coordinate regulation of CYP7 and CYP27 at a transcriptional level, and that CYP27 is not subject to a negative feedback control by bile acids neither at a transcriptional nor at a posttranscriptional level.
Subject(s)
Cholesterol 7-alpha-Hydroxylase/genetics , Cholestyramine Resin/pharmacology , Cholic Acids/pharmacology , Cytochrome P-450 Enzyme System/genetics , Gene Expression Regulation, Enzymologic/drug effects , Liver/enzymology , Steroid Hydroxylases/genetics , Animals , Blotting, Northern , Blotting, Western , Cholestanetriol 26-Monooxygenase , Cholic Acid , DNA Probes , Male , RNA, Messenger/metabolism , Rabbits , RatsABSTRACT
A fraction of cytochrome P-450 catalysing an efficient 6 alpha-hydroxylation of taurine-conjugated 3 alpha,7 alpha-dihydroxy-5 beta- cholanoic acid (taurochenodeoxycholic acid) was partially purified from pig liver microsomes. The specific content of cytochrome P-450 was 6 nmol/mg protein and the preparation showed two major protein bands upon SDS/PAGE. These two bands were isolated after SDS/PAGE and protein blotting. The protein band with a molecular mass of 53 kDa had an N-terminal amino acid sequence and internal sequences resembling that of the cytochrome P-450 4A subfamily (CYP 4A). Polyclonal antibodies raised against this protein were able to, after SDS/PAGE and immunoblotting, detect the protein in microsomal fractions as well as in the purified cytochrome P-450 fraction. Furthermore, addition of these antibodies to a reconstituted system containing the cytochrome P-450 fraction, inhibited 6 alpha-hydroxylation of taurochenodeoxycholic acid by up to 90%. Experiments with irrelevant antibodies did not show inhibition of 6 alpha-hydroxylation. The purified cytochrome P-450 fraction catalysed in addition omega- and omega-1 hydroxylation of lauric acid and 6 alpha-hydroxylation of 3 alpha-hydroxy-5 beta-cholanoic acid (lithocholic acid). However, these hydroxylase activities were rather low compared to 6 beta-hydroxylation of taurochenodexycholic acid. The enzyme fraction did not show hydroxylase activities towards cholesterol and 5 beta-cholestane-3 alpha,7 alpha-diol. These results indicate that 6 alpha-hydroxylation of taurochenodeoxycholic acid is catalysed by a specific species of cytochrome P-450 that, according to N-terminal amino acid sequence as well as catalytic properties, could be a member of the CYP 4A subfamily.
Subject(s)
Cytochrome P-450 Enzyme System/metabolism , Microsomes, Liver/enzymology , Steroid Hydroxylases/metabolism , Amino Acid Sequence , Animals , Blotting, Western , Chromatography, High Pressure Liquid , Cytochrome P-450 Enzyme System/chemistry , Cytochrome P-450 Enzyme System/isolation & purification , Electrophoresis, Polyacrylamide Gel , Molecular Sequence Data , Protein Conformation , Steroid Hydroxylases/chemistry , Steroid Hydroxylases/isolation & purification , SwineABSTRACT
Para calcular la duración media de la protección brindada por diversos métodos de planificación familiar, es necesario usar estimaciones fiables de la frecuencia del coito en la población en cuestión. Antes no se contaba con esa información en México, América Central, Panamá y el Caribe. En un esfuerzo conjunto, los autores de este estudio realizaron encuestas entre usuarias de clínicas privadas de planificación familiar con el fin de determinar la frecuencia del coito y factores relacionados con ella. Mediante procedimientos de muestreo aleatorio, se reunieron datos correspondientes a 4.332 mujeres no esterilizadas que concurrían a servicios de planificación familiar en México, países de América Central, Panamá y el Caribe. Esos datos indicaron que la media del número de relaciones sexuales al mes era de 7,8. Se encontró que se producía una disminución de la frecuencia del coito entre las entrevistadas de más de 35 años de edad y también cuando sus compañeros sobrepasaban los 45 años. Las mujeres que vivían en unión estable con un compañero (cualquiera fuera la condición jurídica de esa unión) tenían relaciones más frecuentes. No se comprobó que el nivel de educación y el tipo de método contraceptivo usado fueran factores relacionados con loa frecuencia del coito
Subject(s)
Adult , Middle Aged , Humans , Male , Female , Coitus , Family Development Planning , Family Planning Services , Central America , Costa Rica , Dominican Republic , El Salvador , Guatemala , Honduras , Mexico , Nicaragua , PanamaABSTRACT
Calculating the average duration of the protection afforded by different family planning methods requires the use of reliable estimates of the frequency of coitus in the subject population. In the past, such information was unavailable in Mexico, Central America, Panama, and the Caribbean. The authors of this study together conducted surveys of women patients of private family plannig clinics to determine the frequency of coitus and associated factors. Random sampling procedures were used to assemble data on 4 332 nonsterile women presenting a family planning units in Mexico, countries of Central America, Panama, and the Caribbean. These data yielded a mean of 7.8 sexual acts a month. It was fond that this frequency declined among interviewees over 35 years of age and when their partners were over 45. Women living permanently with one companion (regardless of the legal status of the union) had coitus more frequently. No relationship was found between frequency of coitus and educational level or the contraceptive method employed