72 research outputs found
Comparison of the TaqMan and LightCycler systems in pharmacogenetic testing: evaluation of the CYP2C9*2/*3 polymorphisms.
Background: Pharmacogenetic testing for drugmetabolizing enzymes is not yet widely used in clinical practice. Methods: In an attempt to facilitate the application of this procedure, we have compared two real-time PCRbased methods, the TaqMan_ and the LightCycler_ for the pharmacogenetic evaluation of CYP2C9*2/*3 polymorphisms. Results and Conclusion: Both procedures are suitable for pharmacogenetic studies. The TaqMan procedure was less expensive in terms of cost per sample, but the TaqMan apparatus is more expensive than the LightCycler apparatus
Molecular analysis has allowed the definitive diagnosis of multiple acyl-CoA dehydrogenase deficiency (MADD)
Multiple acyl-CoA dehydrogenation deficiency (MADD) is a rare autosomal recessive disorder due to defects in the electron transfer
flavoprotein (ETF) or in the electron transfer flavoprotein dehydrogenase (ETFDH) enzymes, involved in the mitochondrial electron
transport chain. Patients with MADD fall into different clinical phenotypes, ranging from a severe neonatal presentation, with metabolic
acidosis, cardiomyopathy and liver disease to a mild childhood/adult disease, with episodic metabolic decompensation, muscle weakness
and respiratory failure.Nowadays, the MADD diagnosis is established by the presence of dicarboxylic organic acids and acylglycine
derivatives in the urine and increased levels of medium-and long-chain acylcarnitines in the blood. Mutations in ETFA, ETFB, ETFDH
genes, encoding for alpha and beta subunits of ETF and for ETF-dehydrogenase are associated with MADD. We report the case of a three
years old child, affected by lethargy and asthenia associated with anorexia. Biochemical analyses showed hypoketotic hypoglycemia with
remarkable increments in transaminases, lactic dehydrogenase, aldolase and creatine kinase. The chromatographic layout of urinary
organic acids showed a typical dicarboxylic aciduria. Thus, based on these features, MADD was suspected. Fifteen years later, at the
age of 19, MADD diagnosis was confirmed by molecular analysis, showing a compound heterozygosity for the mutations c.1074G>C
(p.R358S; HGMD: CM031670 in HGMD database) and c.1073G>A (p.R358K) in the ETFDH gene. The c.1073G>A (p.R358K;
rs796051959) mutation is reported in ClinVar database as pathogenic allele, although lacking link to a specific clinical condition. However,
familial segregation study and in silico analysis, performed by bioinformatics tools, confirmed that this substitution is likely pathogenetic.
Her parents were healthy carriers of one of the two mutations. It is known that the severity of the clinical phenotype of MADD may
be related to the type of mutation in the ETFA/ETFB/ETFDH genes. Particularly, missense mutations in the ETFDH gene, leaving a
detectable residual enzyme activity, may account for the milder form of the disease, as is the case here. In conclusion we suggest that
molecular analysis is essential to the definitive diagnosis of MADD and to direct the adequate therapeutic management. Thus, through
a close nutritional follow up, a few months ago the patient gave birth to a healthy boy.
References
Olsen et al. Clear relationship between ETF/ETFDH genotype and phenotype in patients with multiple acyl-CoA dehydrogenation
deficiency. Hum Mutat. 2003; 22:12–23
The hidden fragility in the heart of the athletes: A review of genetic biomarkers
Sudden cardiac death (SCD) is a devastating event which can also affect people in apparent good health, such as young athletes. It is known that intense and continuous exercise along with a genetic background that predisposes a person to the risk of fatal arrhythmias is a trigger for SCD. Therefore, knowledge of the athlete’s genetic conditions underlying the onset of SCD must be extended, in order to develop new effective prevention and/or therapeutic strategies. Arrhythmic features occur across a broad spectrum of cardiac diseases, sometimes presenting with overlapping phenotypes. The genetic basis of arrhythmogenic disorders has been greatly highlighted in the last 30 years, and has shown marked heterogeneity. The advent of next-generation sequencing has constantly updated our understanding of the genetic basis of arrhythmogenic diseases and is laying the foundation for precision medicine. With the exception of a few clinical cases involving a single athlete showing a highly suspected phenotype for the presence of a heart disease, there are few studies to date that analysed the applicability of genetic testing on cohorts of athletes. This evidence shows that genetic testing can contribute to the diagnosis of up to 13% of athletes; however, the presence of clinical markers is essential. This review aims to provide a reference collection on current knowledge of the genetic basis of sudden cardiac death in athletes and to review updated evidence on the effectiveness of genetic testing in early identification of athletes at risk for SCD
A new case of Congenital Hyperinsulinemic Hypoglycemia due to M/SCHAD deficiency: the contribution of metabolic and molecular diagnosis for the management
Congenital Hyperinsulinemic Hypoglycemia (CHH) is a rare metabolic disease (prevalence <1/1.000.000) characterized by a persistent
hypoglycemia and high secretion of insulin in the neonatal and infancy period. An early management of patients with CHH is mandatory to
avoid brain damage. Recent advances in molecular analysis have linked CHH to mutations in nine genes: ABCC8, KCNJ11, GCK causing
either diazoxide-responsive or diazoxide-unresponsive Hyperinsulinemic Hypoglycemia, and GLUD1, HADH, SLC16A1, UCP2, HNF4A
and HNF1A, causing generally diazoxide-responsive CHH. However, HADH defect is the most common form in presence of consanguinity
and diazoxide-responsiveness. The HADH gene codifies the M/SCHAD mitochondrial enzyme, which catalyses the penultimate reaction in
the β-oxidation of medium and short-chain fatty acids, causing in some affected individuals an elevated plasmatic hydroxybutyrylcarnitine
and urinary medium-chain dicarboxylic, and 3-hydroxydicarboxylic metabolites. To date about 40 cases of M/SCHAD defect have been
reported in literature.We report here a new case of CHH due to M/SCHAD deficiency. The index case was a Pakistan infant, born
from consanguineous parents, showing a diazoxide-responsive hyperinsulinism and organic aciduria. The M/SCHAD deficiency was
confirmed by the molecular diagnosis performed by sequencing of HADH gene, which revealed the presence of the nonsense mutation
c.706C>T (p.R236*) in HADH gene, at homozygous state, while both parents were heterozygous for the mutated allele. The patient
started diazoxide treatment at the maximum dose of 10 mg/kg/day, which resulted in adverse drug reactions (hypertrichosis, peripheral
edemas and persistent hypertension) gradually solved with antihypertensive regimen. Diazoxide was progressively titrated to 2 mg/kg/
day with good results in glycemic control and no hypertensive crisis. Low organic aciduria was followed.In conclusion, when the metabolic
profile suggests a CHH disorder, the molecular analysis is necessary for the precise diagnosis and the appropriate counseling to the
parents, also for the possibility of a prenatal diagnosis. In this setting, the definitive diagnosis of CHH, due to M/SCHAD deficiency, may
suggest also the most appropriate therapeutic intervention to avoid both risk of worsening or adverse drug effect
PEGylated helper-dependent adenoviral vector expressing human Apo A-I for gene therapy in LDLR-deficient mice
Helper-dependent adenoviral (HD-Ad) vectors have great potential for gene therapy applications; however, their administration
induces acute toxicity that impairs safe clinical applications. We previously observed that PEGylation of HD-Ad vectors strongly
reduces the acute response in murine and primate models. To evaluate whether PEGylated HD-Ad vectors combine reduced
toxicity with the correction of pathological phenotypes, we administered an HD-Ad vector expressing the human apolipoprotein
A-I (hApoA-I) to low-density lipoprotein (LDL)-receptor-deficient mice (a model for familial hypercholesterolemia) fed a
high-cholesterol diet. Mice were treated with high doses of HD-Ad-expressing apo A-I or its PEGylated version. Twelve weeks later,
LDL levels were lower and high-density lipoprotein (HDL) levels higher in mice treated with either of the vectors than in untreated
mice. After terminal killing, the areas of atherosclerotic plaques were much smaller in the vector-treated mice than in the control
animals. Moreover, the increase in pro-inflammatory cytokines was lower and consequently the toxicity profile better in mice
treated with PEGylated vector than in mice treated with the unmodified vector. This finding indicates that the reduction in toxicity
resulting from PEGylation of HD-Ad vectors does not impair the correction of pathological phenotypes. It also supports the clinical
potential of these vectors for the correction of genetic diseases
Physical activity and thrombophilic risk in a short series
The role of influence on protein C anticoagulant system and PC deficiency-related thrombophilic risk due to strenuous physical exercise is still under discussion. To investigate the modification of the protein C anticoagulant pathway after vigorous exercise, we measured ProC® Global assay, a protein C activity dependent clotting time, in 20 healthy subjects before and immediately after maximal treadmill exercise, and at 5, 15, 30 and 60 min in the recovery phase. The most evident change was a shortening of ProC® Global clotting time from the average basal value of 123 sec to 84 sec at 30 min in post-exercise. Our study shows that the coagulation unbalance observed after strenuous exercise and with no consequence in healthy individuals with normal PC level, could increase the thrombophilic risk in silent carriers of significant defects of the protein C system and occasionally trigger an episode of deep vein thrombosis
Dietary thiols: A potential supporting strategy against oxidative stress in heart failure and muscular damage during sports activity
Moderate exercise combined with proper nutrition are considered protective factors against cardiovascular disease and musculoskeletal disorders. However, physical activity is known not only to have positive effects. In fact, the achievement of a good performance requires a very high oxygen consumption, which leads to the formation of oxygen free radicals, responsible for premature cell aging and diseases such as heart failure and muscle injury. In this scenario, a primary role is played by antioxidants, in particular by natural antioxidants that can be taken through the diet. Natural antioxidants are molecules capable of counteracting oxygen free radicals without causing cellular cytotoxicity. In recent years, therefore, research has conducted numerous studies on the identification of natural micronutrients, in order to prevent or mitigate oxidative stress induced by physical activity by helping to support conventional drug therapies against heart failure and muscle damage. The aim of this review is to have an overview of how controlled physical activity and a diet rich in antioxidants can represent a “natural cure” to prevent imbalances caused by free oxygen radicals in diseases such as heart failure and muscle damage. In particular, we will focus on sulfur-containing compounds that have the ability to protect the body from oxidative stress. We will mainly focus on six natural antioxidants: Glutathione, taurine, lipoic acid, sulforaphane, garlic and methylsulfonylmethane
The Biological Role of Vitamins in Athletes’ Muscle, Heart and Microbiota
Physical activity, combined with adequate nutrition, is considered a protective factor against cardiovascular disease, musculoskeletal disorders, and intestinal dysbiosis. Achieving optimal performance requires a significantly high energy expenditure, which must be correctly supplied to avoid the occurrence of diseases such as muscle injuries, oxidative stress, and heart pathologies, and a decrease in physical performance during competition. Moreover, in sports activities, the replenishment of water, vitamins, and minerals consumed during training is essential for safeguarding athletes’ health. In this scenario, vitamins play a pivotal role in numerous metabolic reactions and some muscle biochemical adaptation processes induced by sports activity. Vitamins are introduced to the diet because the human body is unable to produce these micronutrients. The aim of this review is to highlight the fundamental role of vitamin supplementation in physical activity. Above all, we focus on the roles of vitamins A, B6, D, E, and K in the prevention and treatment of cardiovascular disorders, muscle injuries, and regulation of the microbiome
Exercise, immune system, nutrition, respiratory and cardiovascular diseases during COVID-19: A complex combination
Coronaviruses (CoVs) represent a large family of RNA viruses that can infect different living species, posing a global threat to human health. CoVs can evade the immune response, replicate within the host, and cause a rapid immune compromise culminating in severe acute respiratory syndrome. In humans, the immune system functions are influenced by physical activity, nutrition, and the absence of respiratory or cardiovascular diseases. This review provides an in-depth study between the interactions of the immune system and coronaviruses in the host to defend against CoVs disease
Childhood obesity: An overview of laboratory medicine, exercise and microbiome
In the last few years, a significant increase of childhood obesity incidence unequally distributed within countries and population groups has been observed, thus representing an important public health problem associated with several health and social consequences. Obese children have more than a 50% probability of becoming obese adults, and to develop pathologies typical of obese adults, that include type 2-diabetes, dyslipidemia and hypertension. Also environmental factors, such as reduced physical activity and increased sedentary activities, may also result in increased caloric intake and/or decreased caloric expenditure. In the present review, we aimed to identify and describe a specific panel of parameters in order to evaluate and characterize the childhood obesity status useful in setting up a preventive diagnostic approach directed at improving health-related behaviors and identifying predisposing risk factors. An early identification of risk factors for childhood obesity could definitely help in setting up adequate and specific clinical treatments
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