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1.
Equine Vet J ; 52(1): 41-45, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30895637

RESUMO

BACKGROUND: Infection and dehiscence of simple lacerations is common in horses, and consistently effective methods of prevention are yet to be found. Honey has been shown to promote wound healing when applied topically; however, intralesional application prior to wound closure has not been reported. OBJECTIVES: To examine whether intralesional application of medical grade honey (MGH) would reduce the incidence of infection and dehiscence following wound closure. STUDY DESIGN: Prospective, open-label randomised block design clinical study. METHODS: Lacerations, treated by field practitioners, were divided into treatment and control groups using block randomisation. Horses in the treatment group received a single intralesional treatment with l-mesitran gel (MGH). Data were collected at the time of wound closure and at suture removal. RESULTS: Data from 127 horses were included, 69 MGH-treated and 58 control cases. No adverse effects of the MGH were recorded. MGH-treated horses were more likely to completely heal (P = 0.006, odds ratio [OR] 3.40 95% confidence interval [CI] 1.41-8.20), to have no signs of infection (P = 0.007, OR 3.64, CI 1.42-9.26) and for the veterinarians to report some degree of satisfaction (P = 0.04, OR 2.72, CI 1.05-7.09) compared to control cases. Numbers needed to treat for complete healing was 5.1 (CI 2.8-40). MAIN LIMITATIONS: Clinical studies have inherent flaws compared to wound healing models, because of variability between wounds. There were more horses with limb injuries in the control group, although not statistically significant, this may have biased the results. Clinical satisfaction and signs of infection were subjective evaluations and evaluators were not blinded to the treatment group. CONCLUSIONS: Intralesional application of MGH to lacerations prior to wound closure may be beneficial in preventing infection and dehiscence. Larger, blinded studies focusing on wounds at a specific location with more objective assessment should be pursued.


Assuntos
Infecções Bacterianas/veterinária , Mel , Doenças dos Cavalos/prevenção & controle , Lacerações/veterinária , Cicatrização , Animais , Infecções Bacterianas/prevenção & controle , Feminino , Cavalos , Masculino
2.
Clin Genet ; 90(1): 84-9, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-26661508

RESUMO

Arthrogryposis multiplex congenita (AMC) is characterized by heterogeneous nonprogressive multiple joint contractures appearing at birth. We present a consanguineous Israeli-Druze family with several members presenting with AMC. A variable intra-familial phenotype and pected autosomal recessive inheritance prompted molecular diagnosis by whole-exome sequencing. Variant analysis focused on rare homozygous changes, revealed a missense variant in MYBPC1, NM_002465:c.556G>A (p.E286K), affecting the last nucleotide of Exon 8. This novel variant was not observed in the common variant databases and co-segregated as expected within the extended family. MYBPC1 encodes a slow skeletal muscle isoform, essential for muscle contraction. Heterozygous mutations in this gene are associated with distal arthrogryposis types 1b and 2, whereas a homozygous nonsense mutation is implicated in one family with lethal congenital contractural syndrome 4. We present a novel milder MYBPC1 homozygous phenotype.


Assuntos
Artrogripose/genética , Proteínas de Transporte/genética , Estudos de Associação Genética , Homozigoto , Mutação de Sentido Incorreto , Artrogripose/diagnóstico , Artrogripose/etnologia , Artrogripose/patologia , Sequência de Bases , Proteínas de Transporte/metabolismo , Pré-Escolar , Consanguinidade , Etnicidade , Exoma , Éxons , Feminino , Expressão Gênica , Genótipo , Humanos , Lactente , Israel , Masculino , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Linhagem , Fenótipo
3.
Bone Marrow Transplant ; 46(3): 330-337, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20436523

RESUMO

Allogeneic hematopoietic SCT (HSCT) has been proposed as a treatment for patients with mitochondrial neurogastrointestinal encephalomyopathy (MNGIE). HSCT has been performed in nine patients using different protocols with varying success. Based on this preliminary experience, participants of the first consensus conference propose a common approach to allogeneic HSCT in MNGIE. Standardization of the transplant protocol and the clinical and biochemical assessments will allow evaluation of the safety and efficacy of HSCT as well as optimization of therapy for patients with MNGIE.


Assuntos
Transplante de Células-Tronco/normas , Humanos , Pseudo-Obstrução Intestinal/genética , Pseudo-Obstrução Intestinal/cirurgia , Encefalomiopatias Mitocondriais/genética , Encefalomiopatias Mitocondriais/cirurgia , Distrofia Muscular Oculofaríngea , Oftalmoplegia/congênito
4.
Pharmacogenomics ; 10(12): 1979-86, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19958096

RESUMO

Pharmacogenomic biomarkers are becoming increasingly common in medicine and drug development. However, there is a genuine concern that the healthcare workforce will be ill-equipped to translate this information to clinical practice. As a result, a major effort is underway to educate future healthcare professionals on pharmacogenomics. This paper describes the development of a year-long course that aims to instill the fundamental concepts of this rapidly growing field into the minds of undergraduate students. This course offers the advantage of exposing students to the concepts of pharmacogenomics prior to their enrollment in PhD, PharmD or MD/DO graduate programs.


Assuntos
Educação de Graduação em Medicina/métodos , Educação em Farmácia/métodos , Farmacogenética/educação , Ensino/métodos , Currículo/tendências
6.
Mol Genet Metab ; 93(3): 275-81, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18078773

RESUMO

Pompe disease is an autosomal recessive disorder of glycogen metabolism caused by a deficiency of the lysosomal enzyme acid alpha-glucosidase (GAA). It presents at any age, with variable rates of progression ranging from a rapidly progressive course, often fatal by one-year of age, to a more slowly, but nevertheless relentlessly progressive course, resulting in significant morbidity and premature mortality. In infants, early initiation of enzyme replacement therapy is needed to gain the maximum therapeutic benefit, underscoring the need for early diagnosis. Several new methods for measuring GAA activity have been developed. The Pompe Disease Diagnostic Working Group met to review data generated using the new methods, and to establish a consensus regarding the application of the methods for the laboratory diagnosis of Pompe disease. Skin fibroblasts and muscle biopsy have traditionally been the samples of choice for measuring GAA activity. However, new methods using blood samples are rapidly becoming adopted because of their speed and convenience. Measuring GAA activity in blood samples should be performed under acidic conditions (pH 3.8-4.0), using up to 2 mM of the synthetic substrate 4-methylumbelliferyl-alpha-D-glucoside or glycogen (50 mg/mL), in the presence of acarbose (3-9 microM) to inhibit the isoenzyme maltase-glucoamylase. The activity of a reference enzyme should also be measured to confirm the quality of the sample. A second test should be done to support the diagnosis of Pompe disease until a program for external quality assurance and proficiency testing of the enzymatic diagnosis in blood is established.


Assuntos
Glucana 1,4-alfa-Glucosidase/sangue , Doença de Depósito de Glicogênio Tipo II/diagnóstico , Técnicas de Laboratório Clínico , Humanos , Lactente
7.
Am J Hum Genet ; 81(6): 1133-43, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17999356

RESUMO

Very-long-chain acyl-coenzyme A dehydrogenase (VLCAD) deficiency is an inborn mitochondrial fatty-acid beta-oxidation (FAO) defect associated with a broad mutational spectrum, with phenotypes ranging from fatal cardiopathy in infancy to adolescent-onset myopathy, and for which there is no established treatment. Recent data suggest that bezafibrate could improve the FAO capacities in beta-oxidation-deficient cells, by enhancing the residual level of mutant enzyme activity via gene-expression stimulation. Since VLCAD-deficient patients frequently harbor missense mutations with unpredictable effects on enzyme activity, we investigated the response to bezafibrate as a function of genotype in 33 VLCAD-deficient fibroblasts representing 45 different mutations. Treatment with bezafibrate (400 microM for 48 h) resulted in a marked increase in FAO capacities, often leading to restoration of normal values, for 21 genotypes that mainly corresponded to patients with the myopathic phenotype. In contrast, bezafibrate induced no changes in FAO for 11 genotypes corresponding to severe neonatal or infantile phenotypes. This pattern of response was not due to differential inductions of VLCAD messenger RNA, as shown by quantitative real-time polymerase chain reaction, but reflected variable increases in measured VLCAD residual enzyme activity in response to bezafibrate. Genotype cross-analysis allowed the identification of alleles carrying missense mutations, which could account for these different pharmacological profiles and, on this basis, led to the characterization of 9 mild and 11 severe missense mutations. Altogether, the responses to bezafibrate reflected the severity of the metabolic blockage in various genotypes, which appeared to be correlated with the phenotype, thus providing a new approach for analysis of genetic heterogeneity. Finally, this study emphasizes the potential of bezafibrate, a widely prescribed hypolipidemic drug, for the correction of VLCAD deficiency and exemplifies the integration of molecular information in a therapeutic strategy.


Assuntos
Acil-CoA Desidrogenase de Cadeia Longa/deficiência , Acil-CoA Desidrogenase de Cadeia Longa/genética , Bezafibrato/uso terapêutico , Hipolipemiantes/uso terapêutico , Erros Inatos do Metabolismo Lipídico/genética , Acil-CoA Desidrogenase de Cadeia Longa/química , Acil-CoA Desidrogenase de Cadeia Longa/metabolismo , Animais , Células Cultivadas , Ácidos Graxos/metabolismo , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/enzimologia , Fibroblastos/patologia , Terapia Genética/métodos , Genótipo , Humanos , Erros Inatos do Metabolismo Lipídico/enzimologia , Modelos Moleculares , Reação em Cadeia da Polimerase , RNA Mensageiro/genética , Ratos , Pele/citologia , Pele/enzimologia , Pele/patologia
8.
Br J Radiol ; 80(954): e115-6, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17684070

RESUMO

A 5-week-old baby with coagulopathy due to hepatic mitochondrial disorder developed a bladder haematoma secondary to suprapubic urine aspiration. Unlike previously reported cases, the haematoma manifested itself only as concentric thickening of the bladder wall without an intraluminal component. The ultrasound and CT features are described. Recognition of these findings is important when discussing the differential diagnosis of bladder wall thickening.


Assuntos
Hematoma , Doenças da Bexiga Urinária , Bexiga Urinária , Biópsia por Agulha/efeitos adversos , Biópsia por Agulha/métodos , Hematoma/diagnóstico por imagem , Hematoma/patologia , Humanos , Lactente , Hepatopatias/complicações , Hepatopatias/patologia , Masculino , Doenças Mitocondriais/complicações , Doenças Mitocondriais/patologia , Tomografia Computadorizada por Raios X/métodos , Ultrassonografia , Bexiga Urinária/diagnóstico por imagem , Bexiga Urinária/patologia , Doenças da Bexiga Urinária/diagnóstico por imagem , Doenças da Bexiga Urinária/patologia
9.
J Pediatr ; 150(6): 656-8, 658.e1, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17517257

RESUMO

Six families with prolidase deficiency (PD) and chronic lung disease are reported, a previously unrecognized association. In one family with a classic cystic fibrosis (CF) phenotype, no evidence for CF Transmembrane Conductance Regulator (CFTR)-related mutations could be found. Chronic lung disease and CFTR-mutation negative CF may be associated with PD.


Assuntos
Fibrose Cística/enzimologia , Dipeptidases/deficiência , Pneumopatias/enzimologia , Adulto , Fibrose Cística/genética , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Humanos , Lactente , Pneumopatias/genética , Masculino , Linhagem , Fenótipo
10.
Neurology ; 68(2): 99-109, 2007 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-17151339

RESUMO

BACKGROUND: Pompe disease is a progressive metabolic neuromuscular disorder resulting from deficiency of lysosomal acid alpha-glucosidase (GAA). Infantile-onset Pompe disease is characterized by cardiomyopathy, respiratory and skeletal muscle weakness, and early death. The safety and efficacy of recombinant human (rh) GAA were evaluated in 18 patients with rapidly progressing infantile-onset Pompe disease. METHODS: Patients were diagnosed at 6 months of age and younger and exhibited severe GAA deficiency and cardiomyopathy. Patients received IV infusions of rhGAA at 20 mg/kg (n = 9) or 40 mg/kg (n = 9) every other week. Analyses were performed 52 weeks after the last patient was randomized to treatment. RESULTS: All patients (100%) survived to 18 months of age. A Cox proportional hazards analysis demonstrated that treatment reduced the risk of death by 99%, reduced the risk of death or invasive ventilation by 92%, and reduced the risk of death or any type of ventilation by 88%, as compared to an untreated historical control group. There was no clear advantage of the 40-mg/kg dose with regard to efficacy. Eleven of the 18 patients experienced 164 infusion-associated reactions; all were mild or moderate in intensity. CONCLUSIONS: Recombinant human acid alpha-glucosidase is safe and effective for treatment of infantile-onset Pompe disease. Eleven patients experienced adverse events related to treatment, but none discontinued. The young age at which these patients initiated therapy may have contributed to their improved response compared to previous trials with recombinant human acid alpha-glucosidase in which patients were older.


Assuntos
Doença de Depósito de Glicogênio Tipo II/tratamento farmacológico , Doença de Depósito de Glicogênio Tipo II/mortalidade , Cuidados Paliativos/estatística & dados numéricos , Medição de Risco/métodos , Assistência Terminal/estatística & dados numéricos , alfa-Glucosidases/administração & dosagem , Relação Dose-Resposta a Droga , Europa (Continente)/epidemiologia , Humanos , Lactente , Recém-Nascido , Israel/epidemiologia , Análise de Sobrevida , Taxa de Sobrevida , Taiwan/epidemiologia , Resultado do Tratamento , Estados Unidos/epidemiologia
12.
Mutat Res ; 610(1-2): 85-92, 2006 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-16890479

RESUMO

Some hexavalent chromium [Cr(VI)]-containing compounds are lung carcinogens. Once within cells, Cr(VI) is reduced to trivalent chromium [Cr(III)] which displays an affinity for both DNA bases and the phosphate backbone. A diverse array of genetic lesions is produced by Cr including Cr-DNA monoadducts, DNA interstrand crosslinks (ICLs), DNA-Cr-protein crosslinks (DPCs), abasic sites, DNA strand breaks and oxidized bases. Despite the large amount of information available on the genotoxicity of Cr, little is known regarding the molecular mechanisms involved in the removal of these lesions from damaged DNA. Recent work indicates that nucleotide excision repair (NER) is involved in the processing of Cr-DNA adducts in human and rodent cells. In order to better understand this process at the molecular level and begin to identify the Cr-DNA adducts processed by NER, the incision of CrCl(3) [Cr(III)]-damaged plasmid DNA was studied using a thermal-resistant UvrABC NER endonuclease from Bacillus caldotenax (Bca). Treatment of plasmid DNA with Cr(III) (as CrCl(3)) increased DNA binding as a function of dose. For example, at a Cr(III) concentration of 1 microM we observed approximately 2 Cr(III)-DNA adducts per plasmid. At this same concentration of Cr(III) we found that approximately 17% of the plasmid DNA contained ICLs ( approximately 0.2 ICLs/plasmid). When plasmid DNA treated with Cr(III) (1 microM) was incubated with Bca UvrABC we observed approximately 0.8 incisions/plasmid. The formation of endonuclease IV-sensitive abasic lesions or Fpg-sensitive oxidized DNA bases was not detected suggesting that the incision of Cr(III)-damaged plasmid DNA by UvrABC was not related to the generation of oxidized DNA damage. Taken together, our data suggest that a sub-fraction of Cr(III)-DNA adducts is recognized and processed by the prokaryotic NER machinery and that ICLs are not necessarily the sole lesions generated by Cr(III) that are substrates for NER.


Assuntos
Compostos de Cromo/toxicidade , Dano ao DNA , Reparo do DNA , Endodesoxirribonucleases/metabolismo , Proteínas de Escherichia coli/metabolismo , Plasmídeos/metabolismo , Bacillus/enzimologia , Cromo/química , Cromo/toxicidade , Compostos de Cromo/química , Adutos de DNA/química , Adutos de DNA/metabolismo , Plasmídeos/efeitos dos fármacos , Plasmídeos/genética
13.
Cell Mol Life Sci ; 63(13): 1553-63, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16786225

RESUMO

The identification of 2-hydroxyphytanoyl-CoA lyase (2-HPCL), a thiamine pyrophosphate (TPP)-dependent peroxisomal enzyme involved in the alpha-oxidation of phytanic acid and of 2-hydroxy straight chain fatty acids, pointed towards a role of TPP in these processes. Until then, TPP had not been implicated in mammalian peroxisomal metabolism. The effect of thiamine deficiency on 2-HPCL and alpha-oxidation has not been studied, nor have possible adverse effects of deficient alpha-oxidation been considered in the pathogenesis of diseases associated with thiamine shortage, such as thiamine-responsive megaloblastic anemia (TRMA). Experiments with cultured cells and animal models showed that alpha-oxidation is controlled by the thiamine status of the cell/tissue/organism, and suggested that some pathological consequences of thiamine starvation could be related to impaired alpha-oxidation. Whereas accumulation of phytanic acid and/or 2-hydroxyfatty acids or their alpha-oxidation intermediates in TRMA patients given a normal supply of thiamine is unlikely, this may not be true when malnourished.


Assuntos
Anemia Megaloblástica/metabolismo , Ácidos Graxos/metabolismo , Ácido Fitânico/metabolismo , Deficiência de Tiamina/metabolismo , Tiamina Pirofosfato/metabolismo , Animais , Carbono-Carbono Liases/metabolismo , Fibroblastos/citologia , Fibroblastos/metabolismo , Hepatócitos/citologia , Hepatócitos/metabolismo , Humanos , Camundongos , Oxirredução , Ratos , Ratos Wistar , Tiamina/metabolismo
14.
Biochem Pharmacol ; 70(12): 1814-22, 2005 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-16242673

RESUMO

The metabolic reduction of hexavalent chromium [Cr(VI)] in the presence of DNA generates several lesions which impede DNA replication and gene transcription. However, the relative contribution of molecular oxygen to Cr-induced genetic damage is unclear. To elucidate the role of dioxygen in Cr genotoxicity, we studied the formation of Cr-induced lesions in DNA treated with either Cr(VI) and the physiological reductant, ascorbic acid (Asc), or Cr(III), under ambient and hypoxic (<1% oxygen) conditions. We found that hypoxia did not impede the reduction of Cr(VI) by Asc throughout a 2 h treatment. In contrast, Cr-DNA binding under these conditions was reduced up to 70% by hypoxia, and a 50-90% decrease in the frequency of Cr-induced Taq polymerase-arresting DNA adducts was also observed. In the presence of Cr(VI)/Asc, formation of Cr-DNA interstrand crosslinks (ICLs) under hypoxia was 50% or less of that under ambient conditions. Kinetic studies found that hypoxia reduced the rate at which Cr interacted with DNA, but not the ultimate steady state level of Cr-DNA binding. The inhibitory effect of hypoxia on Cr(VI)/Asc genotoxicity could not be explained solely by alterations in the reactivity of intermediate Cr(V) species because Cr(III)-DNA binding and Cr(III)-induced ICL formation were also impaired by hypoxia. Moreover, Cr(V) was generated to similar levels in ambient and hypoxic reactions. Hypoxia did not affect ICL formation by the inorganic chemotherapeutic agent cisplatin, suggesting that these effects were specific for Cr(III). Taken together, these results support a role for dioxygen in facilitating the formation of Cr-DNA coordination complexes.


Assuntos
Cromo/metabolismo , Adutos de DNA/metabolismo , DNA/metabolismo , Oxigênio/fisiologia , Sistema Livre de Células , Células Cultivadas , Cromo/toxicidade , Cisplatino/metabolismo , Humanos , Plasmídeos
15.
J Inherit Metab Dis ; 28(4): 467-72, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15902549

RESUMO

A 19-month-old girl with developmental delay was found to have moderately elevated plasma citrulline and mildly elevated plasma arginine concentrations. Dietary history revealed that she consumed large quantities of watermelon (Citrullus vulgaris), a fruit containing high free citrulline and arginine concentrations. In order to determine whether the patient's high watermelon intake could account for her elevated plasma citrulline and arginine concentrations, we studied the response of plasma citrulline and arginine to ingestion of watermelon in six healthy adult volunteers. All developed markedly elevated plasma citrulline (mean maximum 593 micromol/L, range 386-1069) and moderately elevated plasma arginine (mean maximum 199 micromol/L, range 128-251). Physicians and laboratory personnel performing metabolic investigations should be aware of watermelon-induced citrullinaemia. Its hallmarks are elevated plasma citrulline, and to a lesser extent arginine, in the absence of orotic or arginosuccinic aciduria or hyperammonaemia. This phenomenon has implications for the management of patients with urea cycle and related disorders.


Assuntos
Arginina/sangue , Citrulina/sangue , Citrulinemia/etiologia , Deficiências do Desenvolvimento/diagnóstico , Deficiências do Desenvolvimento/etiologia , Adulto , Arginina/efeitos adversos , Citrulina/efeitos adversos , Citrulinemia/sangue , Citrullus , Deficiências do Desenvolvimento/sangue , Dieta , Feminino , Humanos , Lactente , Modelos Biológicos , Necessidades Nutricionais , Ureia/metabolismo
16.
J Clin Invest ; 114(7): 872-6, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15467821

RESUMO

Despite great advances in health-related research and health care, major challenges remain regarding the causes and cures of many diseases; these may be overcome with further research. Our society is enthusiastic about fostering such investigations. However, available federal funds limit many such projects. Previously there have been sizable increases in the NIH budget, but because of the escalating cost of scientific investigation and the pressures of financing other much-needed governmental programs, recent growth in biomedical research funding has barely kept up with inflation. This article focuses on select attempts to sustain the record of scientific achievement enabled in the past by continued increasing investment and also suggests some solutions.


Assuntos
Pesquisa Biomédica/economia , Financiamento Governamental , National Institutes of Health (U.S.)/economia , Apoio à Pesquisa como Assunto , Orçamentos , Humanos , Revisão por Pares , Estados Unidos
17.
Pediatr Cardiol ; 24(5): 476-81, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-14627317

RESUMO

Thiamine-responsive megaloblastic anemia (TRMA) syndrome is a rare autosomal recessive disorder defined by the occurrence of megaloblastic anemia, diabetes mellitus, and sensorineural deafness, responding in varying degrees to thiamine treatment. Other features of this syndrome gradually develop. We describe three TRMA patients with heart rhythm abnormalities and structural cardiac anomalies. Eight other reported TRMA patients also had cardiac anomalies. Recently, the TRMA gene, SLC19A2, was identified, encoding a functional thiamine transporter. Characterization of the metabolic defect of TRMA may shed light on the role of thiamine in common cardiac abnormalities.


Assuntos
Anemia Megaloblástica/complicações , Cardiopatias/complicações , Tiamina/uso terapêutico , Adolescente , Anemia Megaloblástica/tratamento farmacológico , Criança , Cardiopatias/tratamento farmacológico , Humanos , Masculino , Síndrome
18.
J Inherit Metab Dis ; 25(5): 399-410, 2002 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-12408190

RESUMO

A biochemical variant of argininosuccinate lyase deficiency, found in five individuals, is introduced. In comparison to classical patients, the variant cases of argininosuccinate lyase deficiency were characterized by residual enzyme activity as measured by the incorporation of [14C]citrulline into proteins. The five patients of different ethnic backgrounds presented with relatively mild clinical symptoms, variable age of onset, marked argininosuccinic aciduria and severe, but not complete, deficiency of argininosuccinate lyase. [14C]Citrulline incorporation into proteins, which is completely blocked in classical argininosuccinic aciduria, was only partially reduced in fibroblasts of these patients. Further investigation showed that previous standard conditions of the assay were not optimal. Higher concentrations of citrulline in the incubation medium strongly stimulated 14C incorporation in normal cells, but not in the patients; as a result, the relative incorporation level in the patients dropped to 6-28% compared to 18-75% of normal in the original procedure. Prenatal diagnosis was successfully performed in three of the families. Affected pregnancies were indicated by (partial) deficiency of [14C]citrulline incorporation in chorionic villi and/or increased levels of argininosuccinate in amniotic fluid. Analysis of the ASL gene in the five patients revealed a considerable allelic heterogeneity. Three novel mutations--R385C (2 patients), V178M and R379C--were detected in homozygous states, whereas one patient was compound heterozygous for the known mutations R193Q and Q286R. In conclusion, there are patients of different ethnic backgrounds who are characterized by residual activity of argininosuccinate lyase and who present with less severe clinical courses. In addition, we present an improved biochemical assay for accurate prenatal and postnatal diagnosis.


Assuntos
Ácido Argininossuccínico/urina , Citrulinemia/diagnóstico , Adulto , Argininossuccinato Liase/análise , Criança , Citrulina/metabolismo , Citrulinemia/genética , Fibroblastos/enzimologia , Humanos , Lactente , Recém-Nascido , Masculino , Diagnóstico Pré-Natal
19.
Biochemistry ; 41(41): 12529-37, 2002 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-12369844

RESUMO

The genotoxicity associated with the metabolic reduction of hexavalent chromium [Cr(VI)] is complex and can impede DNA polymerase-mediated replication in vitro. The exact biochemical nature of Cr-induced polymerase arresting lesions (PALs) is not understood, but is believed to involve the formation of Cr-DNA interstrand cross-links (ICLs). The aim of this investigation was to determine the dependence of direct Cr-DNA interactions on the development of PALs in DNA treated with trivalent Cr [Cr(III)] or with Cr(VI) in the presence of ascorbic acid (Asc), a major intracellular reductant, using an in vitro, acellular system. The formation of Cr-DNA adducts, ICLs, and PALs was maximal at Asc:Cr(VI) molar ratios of 0.5-2, but gradually decreased at higher ratios. EDTA, a Cr(III) chelator, significantly decreased Cr-DNA binding and ICL and PAL formation. Co-treatment of DNA with Cr(VI)/Asc and mannitol, a Cr(V) chelator, selectively inhibited the formation of mono/bifunctional DNA adducts and PALs produced by Cr(VI) reduction, but had no effect on Cr(III)-DNA binding or Cr(III)-induced polymerase arrest. Blocking Cr-DNA phosphate interaction by preincubation of DNA with MgCl(2) abrogated DNA binding and ICL and PAL production. DNA strand breaks and abasic sites may lead to the in vitro arrest of DNA polymerases; however, we failed to detect significant increases in the frequency of these lesions following Cr(VI)/Asc treatment. These data indicate that the bifunctional adduction of Cr to DNA phosphates (ICLs) constitutes a major PAL. Furthermore, the generation of DNA strand breaks and abasic sites by Cr(VI) reduction is insufficient to explain PALs observed in vitro.


Assuntos
Cromo/química , Dano ao DNA , DNA Polimerase Dirigida por DNA/química , DNA Polimerase Dirigida por DNA/metabolismo , DNA/química , Fibroblastos/enzimologia , Pulmão/enzimologia , Ácido Ascórbico/química , Sítios de Ligação/efeitos dos fármacos , Carbono-Oxigênio Liases/química , Carcinógenos/antagonistas & inibidores , Carcinógenos/química , Carcinógenos/toxicidade , Linhagem Celular , Cromo/antagonistas & inibidores , Cromo/toxicidade , Adutos de DNA/antagonistas & inibidores , Adutos de DNA/metabolismo , DNA Bacteriano/química , DNA Super-Helicoidal/química , DNA Liase (Sítios Apurínicos ou Apirimidínicos) , Relação Dose-Resposta a Droga , Ácido Edético/farmacologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/patologia , Humanos , Pulmão/efeitos dos fármacos , Pulmão/patologia , Cloreto de Magnésio/farmacologia , Manitol/farmacologia , Oxirredução
20.
Food Chem Toxicol ; 40(9): 1231-4, 2002 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-12204386

RESUMO

This report represents a current summary of the caffeine contents of various commercial products, and provides data on the spectrum of caffeine intake levels in man. A summary of the substance's pharmacokinetics describes information on its disposition in the body. The effects of caffeine are related to its interaction with adenosine receptors.


Assuntos
Cafeína/farmacologia , Cafeína/farmacocinética , Estimulantes do Sistema Nervoso Central/farmacologia , Estimulantes do Sistema Nervoso Central/farmacocinética , Animais , Cafeína/análise , Estimulantes do Sistema Nervoso Central/análise , Análise de Alimentos , Humanos , Inibidores de Fosfodiesterase/farmacologia , Distribuição Tecidual
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