RESUMO
The evolution of larger mammals resulted in a corresponding increase in peripheral nerve length. To ensure optimal nervous system functionality and survival, nerve conduction velocities were likely to have increased to maintain the rate of signal propagation. Increases of conduction velocities may have required alterations in one of the two predominant properties that affect the speed of neuronal transmission: myelination or axonal diameter. A plausible mechanism to explain faster conduction velocities was a concomitant increase in axonal diameter with evolving axonal length. The carboxy terminal tail domain of the neurofilament medium subunit is a determinant of axonal diameter in large caliber myelinated axons. Sequence analysis of mammalian orthologs indicates that the neurofilament medium carboxy terminal tail contains a variable lysine-serine-proline (KSP) repeat sub-domain flanked by two highly conserved sub-domains. The number of KSP repeats within this region of neurofilament medium varies among species. Interestingly, the number of repeats does not change within a species, suggesting that selective pressure conserved the number of repeats within a species. Mapping KSP repeat numbers onto consensus phylogenetic trees reveals independent KSP expansion events across several mammalian clades. Linear regression analyses identified three subsets of mammals, one of which shows a positive correlation in the number of repeats with head-body length. For this subset of mammals, we hypothesize that variations in the number of KSP repeats within neurofilament medium carboxy terminal tail may have contributed to an increase in axonal caliber, increasing nerve conduction velocity as larger mammals evolved.
Assuntos
Axônios/ultraestrutura , Proteínas de Neurofilamentos/análise , Proteínas de Neurofilamentos/genética , Sequência de Aminoácidos , Animais , Humanos , Camundongos , Dados de Sequência Molecular , Filogenia , Ratos , Sequências Repetitivas de Aminoácidos , Alinhamento de SequênciaRESUMO
The ability of EB virus-transformed lymphoblasts with undetectable galactose-1-phosphate uridyltransferase (GALT) from 15 galactosaemic patients to oxidize [1-(14)C]galactose to 14CO2 was compared to that of cells from 7 normal subjects. The oxidation of galactose but not of glucose was markedly diminished by cells from Q188R homozygous galactosaemic patients but was not absent. After 2.5 h these cells liberated 14CO2 at nearly 3% and at 5 h up to 9% of normal. Cells from patients homozygous for the S135L mutation produced much larger amounts of 14CO2 (15-17% of normal) and were distinguishable from the Q188R homozygous cells. A cell line with a homozygous deletion of the GALT gene oxidized galactose at 7% of the normal rate, suggesting that pathways(s) other than GALT exist in these cells as well as Q188R homozygous cells for oxidation of galactose to CO2. Concentration dependence studies are consistent with the presence of a pathway that is unsaturable or has a very high Km The ability of 10(7) lymphoblasts with the S135L genotype to oxidize more than 7% of the sugar to 14CO2 in 5 h suggests the presence of residual GALT despite the inability to detect the activity by enzymatic analysis.
Assuntos
Galactose/metabolismo , Linfócitos/enzimologia , UTP-Hexose-1-Fosfato Uridililtransferase/deficiência , Adolescente , Adulto , Dióxido de Carbono/metabolismo , Radioisótopos de Carbono , Linhagem Celular Transformada , Criança , Pré-Escolar , Galactosemias/sangue , Galactosemias/genética , Glucose/metabolismo , Herpesvirus Humano 4 , Homozigoto , Humanos , Cinética , Mutação , Oxirredução , UTP-Hexose-1-Fosfato Uridililtransferase/genéticaRESUMO
The development of tools to follow and quantitate the fate of galactose in mammalian cells is crucial to the study and understanding of the inherited disorders of galactose metabolism. In this study we incubated normal human lymphoblasts with 1- or 2-(13)C galactose for 2.5 or 5 h and prepared TCA extracts of the cells. The various galactose metabolites were identified and quantified using a combination of proton, carbon and phosphorus NMR spectra. Galactose-1-phosphate (gal-1P), uridine diphosphogalactose, uridine diphosphoglucose and galactitol were present in the extracts. Average levels for gal-1P were around 10 nmol/mg protein and for uridine diphosphoglucose, uridine diphosphogalactose and galactitol in the range of 0.5-2 nmol/mg protein. Galactonate was never found in any conditions. Percentage labeling could be estimated for gal-1P and for the ribose carbons of AMP. The labeling agrees with a conversion of galactose to glucose through the Leloir pathway.
Assuntos
Galactose/metabolismo , Linfócitos/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Isótopos de Carbono , Extratos Celulares/química , Células Cultivadas , Galactitol/análise , Galactitol/metabolismo , Galactose/análise , Galactosefosfatos/análise , Galactosefosfatos/metabolismo , Humanos , Hidrogênio , Isótopos de Fósforo , Uridina Difosfato Galactose/análise , Uridina Difosfato Galactose/metabolismo , Uridina Difosfato Glucose/análise , Uridina Difosfato Glucose/metabolismoRESUMO
Mice deficient in galactose-1-phosphate uridyltransferase (GALT) demonstrate abnormal galactose metabolism but no obvious clinical phenotype. To further dissect the pathways of galactose metabolism in these animals, galactose oxidation and metabolite levels were studied in 16-day-old sucklings and the effect of a 4 week prior exposure to a 40% glucose or 40% galactose diet was determined in 7-week-old mice. Suckling GALT-deficient (G/G) mice slowly oxidized [1-14C]galactose to 14CO2, 4.0% of the dose when fed and 7.9% when fasted compared to normal animals 38.3 and 36.4% in 4 h, respectively. Plasma of G/G sucklings contained 11.1 mM galactose and erythrocyte galactose 1-phosphate levels were 28.2 and 31.9 mg/dl packed cells. Galactose, galactitol, galactonate, and galactose 1-phosphate were found in G/G suckling mouse tissues. The tissue galactose concentrations were 10% or less of that in plasma, suggesting that there was limited cellular entry of galactose. In 7-week-old fasted mice with 4 weeks prior exposure to glucose or galactose-containing diet, 4-h oxidation was 12.9 and 15.0% of the administered radiolabeled galactose, respectively. Normal animals oxidized 33.9 and 37.9% of the dose when fed the same diets, respectively. The ability of G/G mice to oxidize galactose in the absence of GALT activity suggests the presence of alternate metabolic pathways for galactose disposition. G/G mice fed the galactose-free 40% glucose diet had erythrocyte galactose 1-phosphate levels ranging from 6.4 to 17.7 mg/dl packed cells and detectable galactose and galactose metabolites in tissues, suggesting that these animals endogenously produced galactose. The plasma of 40% galactose-fed G/G mice contained 9.1 mM galactose with red blood cell galactose 1-phosphate averaging 43.6 mg/dl. Tissues of these animals also contained high levels of galactose and galactose 1-phosphate. Liver contained over 4 micromol/g galactonate but little galactitol. Despite the elevated galactose and galactose 1-phosphate, the animals tolerated the high-galactose diet and were indistinguishable from normal animals, exhibiting no manifestations of galactose toxicity seen in human GALT-deficient galactosemia. The data suggest that high galactose 1-phosphate levels do not cause galactose toxicity and that high galactitol in combination with galactose 1-phosphate may be a prerequisite. Absence of GALT appears necessary but insufficient to produce human galactosemic phenotype.
Assuntos
Galactosemias/genética , Galactosemias/metabolismo , Galactosefosfatos/metabolismo , UTP-Hexose-1-Fosfato Uridililtransferase/deficiência , Animais , Animais Lactentes , Encéfalo/metabolismo , Dióxido de Carbono/análise , Radioisótopos de Carbono , Dieta , Eritrócitos/metabolismo , Galactosefosfatos/administração & dosagem , Cromatografia Gasosa-Espectrometria de Massas , Heterozigoto , Homozigoto , Rim/metabolismo , Fígado/metabolismo , Camundongos , OxirreduçãoRESUMO
The persistent, dietary-independent elevation of galactose metabolites in patients with galactose-1-phosphate uridyltransferase (GALT) deficiency is probably secondary to de novo synthesis of galactose. Relatively constant steady-state levels of galactose metabolites in patients also suggest that non-GALT metabolic pathways must function to dispose of the galactose synthesized each day. The discovery of a patient with a rare deletion of the GALT gene provided a unique opportunity to examine the availability of any alternate galactose oxidative capacity both in vivo and in vitro. Utilizing genomic DNA from the patient, Southern blot data demonstrated that 10 of the 11 GALT exons were homozygously deleted. By measurement of 13CO2 in expired air for up to 24 h after an oral bolus of [1-13C]galactose, it was demonstrated that 17% of the galactose was metabolized, a value comparable to the 3-h elimination rate in a control subject. Furthermore, lymphoblasts prepared from the patient could also convert [1-14C]galactose to 14CO2. This unique study provides the first unambiguous evidence that another pathway exists in man that can be responsible for galactose disposal. Further knowledge of this alternate galactose oxidative route and its regulation may aid in formulating new strategies for the treatment of galactosemia.
Assuntos
Galactose/metabolismo , Galactosemias/enzimologia , Deleção de Genes , Mutação , UTP-Hexose-1-Fosfato Uridililtransferase/genética , Southern Blotting , Testes Respiratórios , Pré-Escolar , Primers do DNA/química , Feminino , Galactosemias/genética , Glucose/biossíntese , Humanos , Ativação Linfocitária , Espectrometria de Massas , Oxirredução , Reação em Cadeia da Polimerase , Reprodutibilidade dos TestesRESUMO
The ability of mice deficient in galactose-1-phosphate uridyltransferase (GALT) to metabolize galactose was determined in animals weaned to a mouse chow diet for a 4-wk period. When given [14C]galactose intraperitoneally, these animals slowly oxidized the sugar, excreting only 5.5% of the dose as 14CO2 in 4 h, whereas normal animals excreted 39.9%. These results mimic those seen in human galactosemic patients given isotopic galactose. When given 10 micromol of [1-13C]galactose, normal animals excrete small amounts of labeled galactose and galactonate but no galactitol in urine whereas GALT-deficient mice excrete significant amounts of all of these as labeled compounds in urine. When challenged with galactose, only about 20% of the dose is excreted in urine, and even on the chow diet, significant amounts of galactose, galactonate, and galactitol are excreted in urine. These compounds are also found to be present in liver, kidney, and brain, except that galactonate is not found in brain. Galactose-1-phosphate accumulates in red blood cells to levels found in humans exposed to large amounts of galactose, and galactose-1-phosphate is found in increased amounts in liver, kidney, and brain of GALT-deficient animals. There was no difference in the hepatic concentration of uridine diphosphate galactose and uridine diphosphate glucose between normal and GALT-deficient mice. The explanation for the presence of galactose and its conversion products in tissues and urine of affected mice appears to be related to the presence of approximately 1.75% of galactose-containing carbohydrates in the chow, which becomes bioavailable to mice. Despite the presence of galactose and its metabolites in tissues and urine and impaired ability to oxidize the sugar, the GALT-deficient animals are indistinguishable from normal animals and do not exhibit the phenotype of humans with GALT-deficiency galactosemia.
Assuntos
Galactose/metabolismo , Galactosemias/genética , UDPglucose-Hexose-1-Fosfato Uridiltransferase/deficiência , UDPglucose-Hexose-1-Fosfato Uridiltransferase/genética , Animais , Encéfalo/metabolismo , Dióxido de Carbono/análise , Isótopos de Carbono , Radioisótopos de Carbono , Eritrócitos/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Heterozigoto , Homozigoto , Humanos , Rim/metabolismo , Cinética , Fígado/metabolismo , CamundongosRESUMO
Urinary excretion of galactitol was determined in 95 normals (N/N), 67 galactosemic (G/G), and 39 compound heterozygotes for the Duarte and galactosemia genotype (D/G). Galactitol excretion is age-dependent in both normal individuals and patients with classic galactosemia on lactose-restricted diets. In galactosemic patients who are homozygous for the Q188R mutation, urinary galactitol levels were fivefold to 10-fold higher than those of normal subjects of comparable age. All but a few patients with classic galactosemia with the Q188R mutation and another mutant G allele had urinary excretion comparable to the Q188R homozygous patients. African-American galactosemic patients with the S135L mutation of the galactose-1-phosphate uridyltransferase (GALT) gene also excreted abnormal quantities of galactitol. Most subjects with a Duarte allele and a G allele excrete normal amounts of the sugar alcohol. There is a correlation between galactitol excretion and red blood cell (RBC) galactose-1-phosphate (gal-1-P). Plasma galactitol was also elevated in galactosemic patients (3.4 to 23.2 micromol/L; undetectable in normal individuals). In contrast to the decrease in urinary galactitol with age, plasma levels remain in a narrow concentration range with no significant difference with age. Urine and plasma galactitol distinguish galactosemic patients from normals. In addition, urinary galactitol excretion may be an important parameter for the assessment of steady-state galactose metabolism in galactosemia.
Assuntos
Galactitol/sangue , Galactosemias/genética , Mutação Puntual , Deleção de Sequência , UTP-Hexose-1-Fosfato Uridililtransferase/genética , Adolescente , Fatores Etários , Substituição de Aminoácidos , População Negra , Criança , Pré-Escolar , Códon de Terminação , Galactitol/urina , Galactosemias/sangue , Galactosemias/urina , Genótipo , Humanos , Lactente , Recém-Nascido , Valores de Referência , UTP-Hexose-1-Fosfato Uridililtransferase/deficiência , Estados Unidos , População BrancaRESUMO
Spanish word recognition scores were determined from 10 native Spanish-speaking listeners' written and oral responses to four Auditec lists (A, B, C, and D) of two-syllable Spanish words. An analysis of variance revealed that the word recognition scores derived from the oral responses were the same when scored by 15 native English-speaking judges with a knowledge of Spanish and when scored by 15 native English-speaking judges without a knowledge of Spanish (p > 0.01). To evaluate the accuracy with which these two groups of judges scored the oral responses, the absolute values of the numeric differences between the word recognition scores derived from the oral and written responses were determined. An analysis of variance of these absolute difference scores revealed a significant main effect of judges' knowledge of Spanish (p < .01). Although the difference between the groups of judges was statistically significant, this difference is of little clinical significance. The word recognition scores for written and oral responses differed on average by two percentage points when oral responses were scored by judges with a knowledge of Spanish and by three percentage points when oral responses were scored by the group without a knowledge of Spanish. These data indicate that English-speaking audiologists are competent to judge the accuracy of Spanish-speaking listeners' oral responses to Spanish word recognition measures.
Assuntos
Idioma , Testes de Discriminação da Fala , Adulto , Análise de Variância , Comparação Transcultural , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Leitura , Reprodutibilidade dos Testes , VocabulárioRESUMO
Since minoxidil was previously shown to raise plasma renin activity (PRA) but not the plasma aldosterone concentration (PAC), the effect of minoxidil on the rate of aldosterone metabolic clearance was studied. After the addition of minoxidil to the antihypertensive regimen of 7 hypertensive patients, the aldosterone metabolic clearance rate increased from 1,110 +/- 91 to 1,570 +/- 180 liters/day (p less than 0.01), an overall increase of 41%. Hepatic blood flow, estimated from the clearance from plasma of indocyanine green, was increased in a group of minoxidil-treated patients compared to a comparable group not treated with minoxidil (p less than 0.01). Thus, although minoxidil induces an increase in PRA, PAC does not change, since aldosterone metabolic clearance increases. Minoxidil increased hepatic blood flow, and it was probably by this mechanism that the drug accelerated the rate of aldosterone clearance.