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1.
J Hosp Infect ; 96(4): 366-370, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28571763

RESUMO

The US Centers for Disease Control and Prevention recommends the initial use of rapid antigen influenza diagnostic test (RIDT) for the detection of influenza A (H1N1-09). Nasopharyngeal samples were tested from 246 patients for H1N1-09 using target-enriched multiplex polymerase chain reaction (TEM-PCR), of which 163 were additionally tested via RIDT. RIDTs had a sensitivity of 18.7% compared with TEM-PCR as the reference standard. Patients with false-negative RIDTs were withheld from 111 days of oseltamivir and 65 days of isolation. Patients negative for H1N1 via TEM-PCR had antiviral therapy immediately stopped, thereby evading 408 days of oseltamivir and 315 days of unnecessary isolation. This cost avoidance saved US$208,982.


Assuntos
Antivirais/economia , Influenza Humana/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Nasofaringe/virologia , Oseltamivir/economia , Antivirais/uso terapêutico , Custos de Cuidados de Saúde , Humanos , Influenza Humana/tratamento farmacológico , Oseltamivir/uso terapêutico , Isolamento de Pacientes/economia , Isolamento de Pacientes/métodos , Estudos Retrospectivos , Resultado do Tratamento , Estados Unidos
2.
J Hosp Infect ; 95(3): 286-291, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28131641

RESUMO

BACKGROUND: Cardiovascular implantable electronic device (CIED) infections are associated with morbidity and mortality. Peri-operative systemic intravenous antibiotic prophylaxis reduces the rate of CIED infections. AIGISRx, a polymer envelope implanted with the CIED, releases minocycline and rifampin, and has been introduced to reduce infections. METHODS: Retrospective review of 184 patients who underwent CIED implantation was conducted. Ninety-two patients were implanted with an AIGISRx envelope (AIGISRx group) and 92 patients were not implanted with an AIGISRx envelope (control group). Data were collected on demographics and risk factors for CIED infections (i.e. congestive heart failure, renal insufficiency, chronic kidney disease, oral anticoagulant use, chronic steroid use, need for lead replacement or revision, temporary pacing, early re-intervention, and having more than two leads in place). Rates of implantation success, major infections and mortality were compared between the AIGISRx group and the control group. RESULTS: The AIGISRx group had longer hospitalizations (6.8±10.7 days vs 3.1±5.2 days; P=0.001), higher chronic corticosteroid use, higher rates of replacement or revision (51.1% vs 8.7%; P=0.001), and a greater proportion of devices with more than two intracardiac leads (42.4% vs 29.3%; P=0.03) than the control group. Successful implantation occurred in 97% of patients in both groups. Major infection was seen in 5.4% of cases in the AIGISRx group and 1.1% of cases in the control group (P=0.048). Device removal was conducted in 3.3% of cases in the AIGISRx group compared with 1.1% of cases in the control group (P=0.16). There were two deaths in the AIGISRx group. Organisms cultured were meticillin-resistant Staphylococcus aureus, meticillin-susceptible S. aureus and Enterococcus faecalis. CONCLUSION: The AIGISRx group had higher rates of major infection but also higher risk factors compared with the control group. The rate of device extraction and CIED-related mortality was higher in the AIGISRx group than in the control group.


Assuntos
Antibacterianos/administração & dosagem , Antibioticoprofilaxia/métodos , Desfibriladores Implantáveis/efeitos adversos , Minociclina/administração & dosagem , Marca-Passo Artificial/efeitos adversos , Infecções Relacionadas à Prótese/epidemiologia , Infecções Relacionadas à Prótese/mortalidade , Idoso , Remoção de Dispositivo , Feminino , Humanos , Masculino , Estudos Retrospectivos , Análise de Sobrevida
3.
Int J Obes (Lond) ; 39(4): 665-70, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25298276

RESUMO

BACKGROUND/OBJECTIVES: Cesarean section (CS) and antibiotic use during pregnancy may alter normal maternal-offspring microbiota exchange, thereby contributing to aberrant microbial colonization of the infant gut and increased susceptibility to obesity later in life. We hypothesized that (i) maternal use of antibiotics in the second or third trimester of pregnancy and (ii) CS are independently associated with higher risk of childhood obesity in the offspring. SUBJECTS/METHODS: Of the 727 mothers enrolled in the Northern Manhattan Mothers and Children Study, we analyzed the 436 mother-child dyads followed until 7 years of age with complete data. We ascertained prenatal antibiotic use by a questionnaire administered late in the third trimester, and delivery mode by medical record. We derived age- and sex-specific body mass index (BMI) z-scores using the CDC SAS Macro, and defined obesity as BMI z⩾95th percentile. We used binary regression with robust variance and linear regression models adjusted for maternal age, ethnicity, pre-gravid BMI, maternal receipt of public assistance, birth weight, sex, breastfeeding in the first year and gestational antibiotics or delivery mode. RESULTS: Compared with children not exposed to antibiotics during the second or third trimester, those exposed had 84% (33-154%) higher risk of obesity, after multivariable adjustment. Second or third trimester antibiotic exposure was also positively associated with BMI z-scores, waist circumference and % body fat (all P<0.05). Independent of prenatal antibiotic usage, CS was associated with 46% (8-98%) higher offspring risk of childhood obesity. Associations were similar for elective and non-elective CS. CONCLUSIONS: In our cohort, CS and exposure to antibiotics in the second or third trimester were associated with higher offspring risk of childhood obesity. Future studies that address the limitations of our study are warranted to determine if prenatal antibiotic use is associated with offspring obesity. Research is also needed to determine if alterations in neonatal gut microbiota underlie the observed associations.


Assuntos
Antibacterianos/efeitos adversos , Cesárea/efeitos adversos , Mães , Obesidade Infantil/etiologia , Obesidade Infantil/prevenção & controle , Efeitos Tardios da Exposição Pré-Natal/prevenção & controle , Antibacterianos/administração & dosagem , Peso ao Nascer , Cesárea/estatística & dados numéricos , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Gravidez , Fatores de Risco , Inquéritos e Questionários
4.
Int J Clin Pract ; 68(4): 503-11, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24471972

RESUMO

BACKGROUND: The prevalence of type 2 diabetes is increasing worldwide, but developing nations will bear a disproportionate share of this burden. Countries in the Middle East and Africa are in a state of transition, where marked disparities of income and access to education and healthcare exist, and where the relatively young populations are being exposed increasingly to processes of urbanisation and adverse changes in diet that are fuelling the diabetes epidemic. Optimising diabetes care in these nations is crucial, to minimise the future burden of complications of diabetes. METHODS: We have reviewed the barriers to effective diabetes care with special relevance to countries in this region. RESULTS: The effects of antidiabetic treatments themselves are unlikely to differ importantly in the region compared with elsewhere, but economic inequalities within countries restrict access to newer treatments, in particular. Values relating to family life and religion are important modifiers of the physician-patient interaction. Also, a lack of understanding of diabetes and its treatments by both physicians and patients requires more and better diabetes education, delivered by suitably qualified health educators. Finally, sub-optimal processes for delivery of care have contributed to a lack of proper provision of testing and follow-up of patients in many countries. CONCLUSION: Important barriers to the delivery of optimal diabetes care exist in the Middle East and Africa.


Assuntos
Diabetes Mellitus Tipo 2/terapia , Acessibilidade aos Serviços de Saúde , África/epidemiologia , Cultura , Diabetes Mellitus Tipo 2/prevenção & controle , Escolaridade , Feminino , Humanos , Masculino , Oriente Médio/epidemiologia , Pobreza , Fatores Sexuais , Fatores Socioeconômicos
5.
Int J Clin Pract ; 67(11): 1144-50, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24165428

RESUMO

AIMS: Developing countries face a high and growing burden of type 2 diabetes. We surveyed physicians in a diverse range of countries in the Middle East and Africa (Egypt, Kingdom of Saudi Arabia, United Arab Emirates, South Africa and Lebanon) with regard to their perceptions of barriers to type 2 diabetes care identified as potentially important in the literature and by the authors. METHODS: One thousand and eighty-two physicians completed a questionnaire developed by the authors. RESULTS: Most physicians enrolled in the study employed guideline-driven care; 80-100% of physicians prescribed metformin (with lifestyle intervention, where there are no contraindications) for newly diagnosed type 2 diabetes, with lifestyle intervention alone used where metformin was not prescribed. Sulfonylureas were prescribed widely, consistent with the poor economic status of many patients. About one quarter of physicians were not undertaking any form of continuing medical education, and relatively low proportions of practices had their own diabetes educators, dieticians or diabetic foot specialists. Physicians identified the deficiencies of their patients (unhealthy lifestyles, lack of education and poor diet) as the most important barriers to optimal diabetes care. Low-treatment compliance was not ranked highly. Access to physicians did not appear to be a problem, as most patients were seen multiple times per year. CONCLUSIONS: Physicians in the Middle East and South Africa identified limitations relating to their patients as the main barrier to delivering care for diabetes, without giving high priority to issues relating to processes of care delivery. Further study would be needed to ascertain whether these findings reflect an unduly physician-centred view of their practice. More effective provision of services relating to the prevention of complications and improved lifestyles may be needed.


Assuntos
Atitude do Pessoal de Saúde , Atenção à Saúde/estatística & dados numéricos , Diabetes Mellitus Tipo 2/tratamento farmacológico , Competência Clínica/estatística & dados numéricos , Diabetes Mellitus Tipo 2/diagnóstico , Educação Médica/estatística & dados numéricos , Humanos , Hipoglicemiantes/uso terapêutico , Oriente Médio , Percepção , Padrões de Prática Médica/estatística & dados numéricos , Qualidade da Assistência à Saúde , África do Sul , Inquéritos e Questionários
6.
Int J Clin Pract ; 67(10): 957-63, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24001317

RESUMO

AIMS: To assess, in a real-world setting, the effect of vildagliptin compared with sulphonylurea (SU) treatment on hypoglycaemia in Muslim patients with type 2 diabetes mellitus (T2DM) fasting during Ramadan. METHODS: This multinational, non-interventional study, conducted in Asia and the Middle East, included Muslim adult patients with T2DM who received treatment with vildagliptin or SU as add-on to metformin or monotherapy. During a ~16-week observation period, data were collected up to 6 weeks before and 6 weeks after Ramadan fasting. The primary study objective was to compare the proportion of patients with ≥ 1 hypoglycaemic event (HE) during fasting. RESULTS: Of > 1300 patients enrolled in the study, 684 were treated with vildagliptin and 631 with SUs. Significantly fewer patients experienced ≥ 1 HE with vildagliptin compared with those receiving SUs (5.4% vs. 19.8%, respectively; p < 0.001); no vildagliptin-treated patients reported a grade 2 HE, vs. 4 SU-treated patients (p = 0.053). Mean HbA1c changes from baseline were vildagliptin: -0.24%, SUs: +0.02% (p < 0.001). Mean body weight reductions from baseline were vildagliptin: -0.76 kg, SUs: -0.13 kg (p < 0.001). A higher proportion of SU-treated patients experienced adverse events (AEs) compared with vildagliptin (22.8% vs. 10.2%). This difference was driven by hypoglycaemia as the most common AE. CONCLUSIONS: In this real-world study of fasting Muslim patients with T2DM, vildagliptin was associated with significantly fewer hypoglycaemic episodes compared with SU therapy. This outcome is particularly meaningful when viewed in the context of good glycaemic and weight control observed in vildagliptin-treated patients. Vildagliptin was well tolerated in this patient population.


Assuntos
Adamantano/análogos & derivados , Diabetes Mellitus Tipo 2/tratamento farmacológico , Jejum/fisiologia , Hipoglicemiantes/uso terapêutico , Islamismo , Nitrilas/uso terapêutico , Pirrolidinas/uso terapêutico , Compostos de Sulfonilureia/uso terapêutico , Adamantano/uso terapêutico , Idoso , Diabetes Mellitus Tipo 2/etnologia , Feminino , Hemoglobinas Glicadas/metabolismo , Humanos , Hipoglicemia/induzido quimicamente , Masculino , Adesão à Medicação/etnologia , Pessoa de Meia-Idade , Estudos Prospectivos , Vildagliptina , Redução de Peso/efeitos dos fármacos
7.
Haemophilia ; 18(4): 503-9, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22044794

RESUMO

Nonacog alfa, a recombinant factor IX (FIX) used for the treatment of haemophilia B, was approved in Europe in 1998. In accordance with European Medicines Agency requirements, a registry study was conducted from 2002 to 2009. A reformulated iso-osmotic version was approved for European use in 2007. This study was conducted to evaluate the safety of nonacog alfa in a usual care setting, and provide clinical trial and postmarketing surveillance data support. This open-label, non-interventional, prospective observational cohort study (registry) comprised 52 sites in nine European countries. Patients with haemophilia B receiving nonacog alfa in either formulation for prevention or treatment were followed on a usual care schedule. A total of 218 patients were enrolled, of whom 66 (30.3%) were <18 years of age. Haemophilia severity was evenly distributed, with baseline FIX activity of <1%, 1-5% and >5% in 33.3%, 36.6% and 30.1% of patients, respectively. One hundred thirty-eight patients received the original formulation alone; 80 switched to or received only the new formulation. There was a low incidence of events of special interest (ESIs), with less-than-expected therapeutic effect in five patients (2.2%), inhibitor development in two (0.9%), thrombosis in one (0.5%) and allergic events in eight (3.7%). These accounted for the majority of the 15 serious AEs reported in six patients. Six patients discontinued because of AEs, primarily related to hypersensitivity. Nonacog alfa was shown to be safe for the treatment of haemophilia B, with a low incidence of serious AEs and ESIs.


Assuntos
Coagulantes/uso terapêutico , Fator IX/uso terapêutico , Hemofilia B/tratamento farmacológico , Sistema de Registros , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Coagulantes/efeitos adversos , Europa (Continente) , Fator IX/efeitos adversos , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Proteínas Recombinantes/efeitos adversos , Proteínas Recombinantes/uso terapêutico , Adulto Jovem
8.
J Int Med Res ; 39(2): 366-77, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21672340

RESUMO

The prevalence of painful diabetic peripheral neuropathy (DPN) was evaluated in type 1 or type 2 diabetes mellitus patients (n = 4097) attending outpatient clinics across the Middle East. Overall, 53.7% of 3989 patients with DN4 data met the criteria for painful DPN (Douleur Neuropathique-4 [DN4] scores ≥ 4). Significant predictors of painful DPN included long history (≥ 10 years) of diabetes (odds ratio [OR] 2.43), age ≥ 65 years (OR 2.13), age 50 - 64 years (OR 1.75), presence of type 1 versus type 2 diabetes (OR 1.59), body mass index > 30 kg/m(2) (OR 1.35) and female gender (OR 1.27). Living in one of the Gulf States was associated with the lowest odds of having painful DPN (OR 0.44). The odds of painful DPN were highest among patients with peripheral vascular disease (OR 4.98), diabetic retinopathy (OR 3.90) and diabetic nephropathy (OR 3.23). Because of the high prevalence and associated suffering, disability and economic burden of painful DPN, it is important that diabetic patients are periodically screened, using a simple instrument such as the DN4, and receive appropriate treatment if symptoms develop.


Assuntos
Diabetes Mellitus/epidemiologia , Neuropatias Diabéticas/complicações , Neuropatias Diabéticas/epidemiologia , Idoso , Demografia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Oriente Médio/epidemiologia , Análise Multivariada , Razão de Chances , Prevalência , Inquéritos e Questionários
9.
Horm Res Paediatr ; 73(2): 102-7, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20190546

RESUMO

BACKGROUND: With the epidemic of childhood obesity, it is crucial to devise a simple screening protocol to predict impaired glucose tolerance (IGT) or pre-diabetes. The oral glucose tolerance test (OGTT), which is the gold standard for the diagnosis of IGT, is impractical for screening purposes. This pilot study was designed to formulate a simple, sensitive algorithm to predict IGT using clinical and laboratory parameters. METHODS: Ethnicity, family history of diabetes, pubertal status, BMI z-score, blood pressure, lipids, hemoglobin A1c (HbA1c) and OGTT data were retrospectively collected from 209 overweight multi-ethnic subjects aged 3-21 years. Multivariate logistic regression was used to determine independent predictors of IGT. RESULTS: HbA1c was the only significant predictor of IGT (p = 0.001), whereas fasting glucose was not. A cut-off of 5.5% had the best combined sensitivity (85.7%) and specificity (56.9%) with an odds ratio of 7.9 of having IGT when HbA1c is > or =5.5%. The remaining clinical parameters were not significant predictors of IGT. CONCLUSION: While fasting blood glucose does not seem to be a predictor of IGT, we propose that HbA1c > or =5.5% can be used as a screening test to assess the risk of IGT and to determine who should undergo diagnostic OGTT. Large prospective studies validating our findings are warranted.


Assuntos
Intolerância à Glucose/diagnóstico , Teste de Tolerância a Glucose/normas , Hemoglobinas Glicadas/análise , Adolescente , Adulto , Algoritmos , Biomarcadores , Criança , Pré-Escolar , Diabetes Mellitus/diagnóstico , Diabetes Mellitus/metabolismo , Feminino , Glucose/metabolismo , Intolerância à Glucose/complicações , Intolerância à Glucose/epidemiologia , Hemoglobinas Glicadas/metabolismo , Humanos , Masculino , Obesidade/complicações , Projetos Piloto , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Adulto Jovem
10.
Int J Clin Pract ; 64(2): 149-59, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20089006

RESUMO

AIMS: Increases in the prevalence of type 2 diabetes will likely be greater in the Middle East and other developing countries than in most other regions during the coming two decades, placing a heavy burden on regional healthcare resources. METHODOLOGY: Medline search, examination of data from major epidemiological studies in the Middle Eastern countries. RESULTS: The aetiology and pathophysiology of diabetes appears comparable in Middle Eastern and other populations. Lifestyle intervention is key to the management of diabetes in all type 2 diabetes patients, who should be encouraged strongly to diet and exercise. The options for pharmacologic therapy in the management of diabetes have increased recently, particularly the number of potential antidiabetic combinations. Metformin appears to be used less frequently to initiate antidiabetic therapy in the Middle East than in other countries. Available clinical evidence, supported by current guidelines, strongly favours the initiation of antidiabetic therapy with metformin in Middle Eastern type 2 diabetes patients, where no contraindications exist. This is due to its equivalent or greater efficacy relative to other oral antidiabetic treatments, its proven tolerability and safety profiles, its weight neutrality, the lack of clinically significant hypoglycaemia, the demonstration of cardiovascular protection for metformin relative to diet in the UK Prospective Diabetes Study and in observational studies, and its low cost. Additional treatments should be added to metformin and lifestyle intervention as diabetes progresses, until patients are receiving an intensive insulin regimen with or without additional oral agents. CONCLUSIONS: The current evidence base strongly favours the initiation of antidiabetic therapy with metformin, where no contraindications exist. However, metformin may be under-prescribed in the Middle East.


Assuntos
Diabetes Mellitus Tipo 2/tratamento farmacológico , Hiperglicemia/dietoterapia , Hipoglicemiantes/administração & dosagem , Metformina/administração & dosagem , Administração Oral , Adulto , Distribuição por Idade , Idoso , Efeitos Psicossociais da Doença , Diabetes Mellitus Tipo 2/epidemiologia , Diabetes Mellitus Tipo 2/etiologia , Humanos , Hiperglicemia/etiologia , Pessoa de Meia-Idade , Oriente Médio/epidemiologia , Prevalência , Adulto Jovem
12.
Haemophilia ; 13(4): 357-60, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17610548

RESUMO

A recent multicentre collaborative study showed higher estimates of ReFacto potency when assayed with ReFacto Laboratory Standard(TM) (RLS) in comparison when standards consisting of full-length factor VIII (FVIII) were used. The RLS was hence recalibrated, leading to a 20% increase in the amount of ReFacto per vial without change in the labelled potency. The primary objective of this study was to determine the incremental and in vivo recovery of the recalibrated ReFacto in patients with severe haemophilia A. Fourteen male severe haemophilia A patients (FVIII < 1 IU dL(-1)) with a cumulative previous exposure days to any FVIII product >150 were administered an intravenous infusion 50 +/- 5 IU kg(-1) of ReFacto over a 5-min period. Blood samples were collected before infusion and after 15, 30 and 60 min. FVIII clotting activity (FVIII:C) was assessed in a central laboratory by the chromogenic substrate assay. After ReFacto infusion, peak FVIII:C was obtained within 15 min for 10 patients and within 30 min for the remaining four. Mean FVIII:C at peak was 117.7 +/- 17.3 IU dL(-1). Mean incremental recovery was 2.22 +/- 0.27 IU dL(-1) per IU kg(-1) while mean in vivo recovery was 105.9 +/- 14.6%. One patient reported three mild adverse events rated as 'unrelated' to the study drug. FVIII recovery after recalibrated ReFacto infusion falls within the expected range and is similar to the values reported for other FVIII concentrates.


Assuntos
Inibidores dos Fatores de Coagulação Sanguínea/administração & dosagem , Fator VIII/administração & dosagem , Hemofilia A/tratamento farmacológico , Adolescente , Adulto , Inibidores dos Fatores de Coagulação Sanguínea/farmacocinética , Criança , Relação Dose-Resposta a Droga , Fator VIII/farmacocinética , Humanos , Infusões Intravenosas/métodos , Masculino , Pessoa de Meia-Idade , Padrões de Referência , Equivalência Terapêutica
13.
Forensic Sci Int ; 128(1-2): 35-40, 2002 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-12208019

RESUMO

Bile is, in certain cases, collected together with blood from different sites (heart, brain, femoral), urine and other organs or matrices. This study reports comparative results obtained from the analysis of blood and bile for different drugs found: acetaminophen, amphetamine and related compounds, several antidepressants, several benzodiazepines, cocaine and its metabolites, dextropropoxyphene and its metabolite, hydroxyzine, methadone and metabolite, morphine and codeine, levomepromazine, thioridazine, propranolol, tramadol and its metabolite. Several findings are presented: (1) There were no significant differences in the levels of the compounds among the samples of blood obtained from different sites. (2) Levels in bile are generally several fold higher than those in blood. The mean bile to blood ratios vary from about 1 (for acetaminophen, amphetamine) to about 2000 (for desmethylclobazam). (3) In certain cases (16 over 44), although the drug or its metabolite was not detected in blood from different sites, it was detected in bile. As other authors had advocated, it is very useful to ask the pathologist to take the gall bladder with its contents together with the other samples, in order that the sample of bile can be used in the comprehensive toxicological analysis and therefore be complementary to the other fluids or matrices. An additional advantage for using bile is that the concentrations of drugs or their metabolites are generally several fold higher than their blood concentrations.


Assuntos
Bile/química , Medicina Legal/métodos , Preparações Farmacêuticas/sangue , Intoxicação/metabolismo , Humanos , Preparações Farmacêuticas/análise , Farmacocinética , Distribuição Tecidual
14.
Blood ; 97(10): 2972-8, 2001 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-11342419

RESUMO

The properties of 9 delta-aminolevulinate dehydratase (ALAD) mutants from patients with ALAD porphyria (ADP) were examined by bacterial expression of their complementary DNAs and by enzymologic and immunologic assays. ALADs were expressed as glutathione-S-transferase (GST) fusion proteins in Escherichia coli and purified by glutathione-affinity column chromatography. The GST-ALAD fusion proteins were recognized by anti-ALAD antibodies and were enzymatically active as ALAD. The enzymatic activities of 3 ALAD mutants, K59N, A274T, and V153M, were 69.9%, 19.3%, and 41.0% of that of the wild-type ALAD, respectively, whereas 6 mutants, G133R, K59N/G133R, F12L, R240W, V275M, and delTC, showed little activity (< 8%). These variations generally reflect the phenotype of ALAD in vivo in patients with ADP and indicate that GST-ALAD fusion protein is indeed useful for predicting of the phenotype of ALAD mutants. The location of F12L mutation in the enzyme's molecular structure indicates that its disturbance of the quaternary contact of the ALAD dimer appears to have a significant influence on the enzymatic activity. Mouse monoclonal antibodies to human ALAD were developed that specifically recognized a carboxy terminal portion of ALAD, or other regions in the enzyme. This study represents the first complete analysis of 9 mutants of ALAD identified in ADP and indicates the highly heterogeneous nature of mutations in this disorder.


Assuntos
Mutação , Sintase do Porfobilinogênio/deficiência , Sintase do Porfobilinogênio/genética , Porfirias/enzimologia , Adolescente , Animais , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos , Escherichia coli/genética , Feminino , Expressão Gênica , Glutationa Transferase/genética , Humanos , Immunoblotting , Recém-Nascido , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pessoa de Meia-Idade , Fenótipo , Sintase do Porfobilinogênio/metabolismo , Porfirias/genética , Proteínas Recombinantes de Fusão/análise , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/metabolismo
15.
Blood ; 96(10): 3618-23, 2000 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-11071662

RESUMO

Cloning, expression, and genotype studies of the defective gene for delta-aminolevulinate dehydratase (ALAD) in a patient with an unusual late onset of ALAD deficiency porphyria (ADP) were carried out. This patient was unique in that he developed the inherited disease, together with polycythemia, at the age of 63. ALAD activity in erythrocytes of the patient was less than 1% of the normal control level. ALAD complementary DNA (cDNA) isolated from the patient's Epstein-Barr virus (EBV)-transformed lymphoblastoid cells had 2 base transitions in the same allele, G(177) to C and G(397) to A, resulting in amino acid substitutions K59N and G133R, respectively. It has been verified that the patient had no other ALAD mutations in this and in the other allele. By restriction fragment length polymorphism (RFLP) analysis, all family members of the proband who had one-half ALAD activity compared with the ALAD activity of the healthy control were shown to have the same set of base transitions. Expression of ALAD cDNA in CHO cells revealed that K59N cDNA produced a protein with normal ALAD activity, while G133R and K59N/G133R cDNA produced proteins with 8% and 16% ALAD activity, respectively, compared with that expressed by the wild type cDNA. These findings indicate that while the proband was heterozygous for ALAD deficiency, the G(397) to A transition resulting in the G133R substitution is responsible for ADP, and the clinical porphyria developed presumably due to an expansion of the polycythemic clone in erythrocytes that carried the mutant alad allele.


Assuntos
Sintase do Porfobilinogênio/deficiência , Porfirias/enzimologia , Idade de Início , Alelos , DNA Complementar/química , DNA Complementar/genética , Saúde da Família , Feminino , Genótipo , Heterozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Mutação de Sentido Incorreto , Mutação Puntual , Sintase do Porfobilinogênio/genética , Porfirias/genética , Análise de Sequência de DNA
16.
Transplantation ; 70(5): 771-4, 2000 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-11003355

RESUMO

BACKGROUND: The two-layer method [University of Wisconson solution (UW)/perfluorochemical plus O2] for pancreas preservation has been demonstrated to be superior to simple UW storage alone in the canine model. For the first time, we applied the two-layer method to clinical whole-pancreas transplantation. METHODS: Pancreases were placed in the two-layer method in 10 cases and UW alone in 44 cases before transplant. The mean cold ischemic time was 16.5 hr in the two-layer group versus 18.1 hr in the UW group (P=NS). We compared the condition of graft at the time of reperfusion, and then 3 months posttransplant graft function and complications. RESULTS: At the time of reperfusion, no grafts in the two-layer group were edematous, compared with 10(23.3%) of 43 in the UW group (P=0.18). Seven (70%) of 10 grafts in the two-layer group obtained the best overall quality score, compared with 24(57.1%) of 42 in the UW group (P=0.72). Nine (90%) of 10 recipients in the two-layer group became insulin-independent during hospitalization, compared with 31(70.5%) of 44 in the UW group (P=0.26). Time to insulin independence was no different between the two groups. No pancreas grafts preserved by the two-layer method suffered acute rejection. Conclusions. The two-layer preservation method is feasible in human clinical transplantation. It was at least equivalent and may be superior to UW alone in both morphologic and functional assessment of the transplanted pancreas.


Assuntos
Soluções para Preservação de Órgãos , Preservação de Órgãos/métodos , Pâncreas , Adenosina/farmacologia , Adolescente , Adulto , Alopurinol/farmacologia , Índice de Massa Corporal , Feminino , Fluorocarbonos/farmacologia , Glutationa/farmacologia , Humanos , Insulina/farmacologia , Masculino , Pessoa de Meia-Idade , Oxigênio/farmacologia , Transplante de Pâncreas , Rafinose/farmacologia , Fatores de Tempo
17.
Acta Clin Belg ; 53 Suppl 1: 74-8, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-10216988

RESUMO

The toxicological screening, using the combination of high performance liquid chromatography with diode array ultraviolet detector and ion-pairing technique in liquid-liquid extraction, is an effective tool in the identification and quantification of the acidic and basic substances in a single run. The use of an ion-pairing technique in the conventional extraction shows the co-extraction of the uncharged and charged form of the analytes present in a serum sample. The stationary phase used is C-18-bonded phase. The mobile phase is acetonitrile--phosphate buffer (pH 3; 25 mM) containing 25 mM triethylammonium as ion-pairing agent. The analytical validation shows reproducible recoveries, good day-to-day repeatability and sensible results compatible with clinical and forensic use.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Cromatografia por Troca Iônica/métodos , Cromatografia Líquida/métodos , Intoxicação/sangue , Intoxicação/diagnóstico , Detecção do Abuso de Substâncias/métodos , Doença Aguda , Humanos , Concentração de Íons de Hidrogênio , Reprodutibilidade dos Testes
19.
Transplantation ; 68(12): 1842-6, 1999 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-10628761

RESUMO

BACKGROUND: Both antigen-dependent (immunologic) and non-antigen-dependent (nonimmunologic) factors have been implicated in long-term renal allograft loss. Differentiating between these two factors is important because prevention strategies differ. METHODS: To isolate the importance of these 2 factors, we studied long-term actuarial graft survival in a cohort of adult kidney recipients who underwent transplants at a single institution between January 1, 1984 and October 31, 1998. Excluded were recipients with graft loss as a result of death with function, technical failure, primary nonfunction, and recurrent disease, leaving 1587 recipients (757 cadaver [CAD], 830 living donor [LD]) who would be at risk for graft loss secondary to both immunologic and nonimmunologic factors. These recipients were analyzed in the following 2 groups: those treated for a previous episode of acute rejection (AR) (Group1; n = 588; 328 CAD, 260 LD) and those with no AR (Group 2: n = 999; 429 CAD, 570 LD). Actuarial graft survival and causes of graft loss were determined for each group. Presumably, graft loss in Group 1 would be caused by immunologic and nonimmunologic factors; graft loss in Group 2 would be caused primarily by nonimmunologic factors. RESULTS: The 10-year graft survival rate (censored for death with function, technical failure, primary nonfunction, and recurrent disease) in Group 2 was 91%. In contrast, the 10-year graft survival rate in Group 1 was 45% (P<0.001 vs. Group 2). Causes of graft loss in Group 2 were chronic rejection in 1.8% (3.0% CAD, 0.9% LD), de novo disease, 0.4%; sepsis, 0.2%; discontinuation of immunosuppressive therapy, 0.3%; and unknown, 0.6%. In contrast, 23.8% (29.9% CAD, 16.2% LD) of recipients in Group 1 had graft loss caused by chronic rejection (P = 0.001 vs. Group 2). CONCLUSIONS: This very low incidence of chronic rejection in recipients without previous AR suggests that immunologic factors are the main determinants of long-term kidney transplant outcome; nonimmunologic factors in isolation may have only a minimal impact on long-term graft survival.


Assuntos
Rejeição de Enxerto/etiologia , Transplante de Rim/imunologia , Adulto , Rejeição de Enxerto/epidemiologia , Humanos , Incidência , Doadores Vivos , Recidiva , Fatores de Risco , Análise de Sobrevida , Fatores de Tempo
20.
Int Orthop ; 22(4): 252-4, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9795814

RESUMO

The penetration rate of ethanol in human tendons was studied to in order to define the conditions which were necessary to achieve an inactivating concentration against the Human Immunodeficiency Virus (HIV) within the tendon. The rate of alcohol penetration was found to be slow and did not differ with different types of tendons. An average ethanol concentration of 14% (v/v) was measured in human tendons after they had been immersed for 2 h in 70% (v/v) ethanol, and a concentration of 19% (v/v) was reached after 3 h. Ethanol immersion of human tendons may represent an additional safety procedure in inactivating the HIV virus provided the duration is at least 3 h.


Assuntos
Anti-Infecciosos Locais , Etanol , HIV/efeitos dos fármacos , Tendões/transplante , Tendões/virologia , Preservação de Tecido/métodos , Anti-Infecciosos Locais/análise , Anti-Infecciosos Locais/farmacocinética , Avaliação Pré-Clínica de Medicamentos , Etanol/análise , Etanol/farmacocinética , Humanos , Controle de Infecções/métodos , Tendões/química , Tendões/efeitos dos fármacos , Fatores de Tempo , Distribuição Tecidual , Transplante Homólogo
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