Diagnosis of sickle cell disease by innovative PCR without DNA extraction
; BOEMER, François ;
Poster (2016, February)Detailed reference viewed: 34 (1 ULg)
Samaras and seedlings of Acer pseudoplatanus are potential sources of hypoglycin A intoxication in atypical myopathy without necessarily inducing clinical signs
Baise, Etienne ; Habyarimana, Jean ; Amory, Hélène et al
in Equine Veterinary Journal (2015)Detailed reference viewed: 61 (27 ULg)
Detection of hypoglycin A in the seeds of sycamore (Acer pseudoplatanus) and box elder (A. negundo) in New Zealand; the toxin associated with cases of equine atypical myopathy.
; ; Habyarimana, Jean et al
in New Zealand veterinary journal (2015)
CASE HISTORY AND CLINICAL FINDINGS: During April and May 2014 four horses aged between 5 months and 9 years, located in the Canterbury, Marlborough and Southland regions, presented with a variety of ... [more ▼]
CASE HISTORY AND CLINICAL FINDINGS: During April and May 2014 four horses aged between 5 months and 9 years, located in the Canterbury, Marlborough and Southland regions, presented with a variety of clinical signs including recumbency, stiffness, lethargy, dehydration, depression, and myoglobinuria suggestive of acute muscle damage. Two horses were subjected to euthanasia and two recovered. In all cases seeds of sycamore maple (Acer pseudoplatanus) or box elder (A. negundo) were present in the area where the horse had been grazing. LABORATORY INVESTIGATION: The samaras (seeds) of some Acer spp. may contain hypoglycin A, that has been associated with cases of atypical myopathy in Europe and North America. To determine if hypoglycin A is present in the samaras of Acer spp. in New Zealand, samples were collected from trees throughout the country that were associated with historical and/or current cases of atypical myopathy, and analysed for hypoglycin A. Serum samples from the four cases and four unaffected horses were analysed for the presence of hypoglycin A, profiles of acylcarnitines (the definitive diagnosis for atypical myopathy) and activities of creatine kinase and aspartate aminotransferase. Markedly elevated serum activities of creatine kinase and aspartate aminotransferase, and increased concentrations of selected acylcarnitines were found in the case horses. Hypoglycin A was detected in the serum of those horses but not in the healthy controls. Hypoglycin A was detected in 10/15 samples of samaras from sycamore maple and box elder from throughout New Zealand. DIAGNOSIS: Cases of atypical myopathy were diagnosed on properties where samaras containing hypoglycin A were also found. CLINICAL RELEVANCE: Sycamore and box elder trees in New Zealand are a source of hypoglycin A associated with the development of atypical myopathy. If pastured horses present with clinical and biochemical signs of severe muscle damage then the environment should be checked for the presence of these trees. Horses should be prevented from grazing samaras from Acer spp. in the autumn. [less ▲]Detailed reference viewed: 15 (5 ULg)
Epidemiological data on sickle cell disease in Belgium
; ; BOEMER, François et al
in Belgian Journal of Hematology (2015), 6(4), 135-141Detailed reference viewed: 16 (1 ULg)
Quantification of physiological aminoacids using aTRAQ kit: evaluation and implementation of new markers.
BOEMER, François ; SCHOOS, Roland ; DEBERG, Michelle
in Annales de biologie clinique (2015), 73(4), 427-442
Nowadays, physiological amino acids profiling is based primarily on ion exchange chromatography (IEC) coupled to a post-column derivatization with ninhydrin and UV detection at two wavelengths ... [more ▼]
Nowadays, physiological amino acids profiling is based primarily on ion exchange chromatography (IEC) coupled to a post-column derivatization with ninhydrin and UV detection at two wavelengths. Unfortunately, this technique suffers various drawbacks such as long analysis time, high sample volume and specific costs related to the maintenance of a dedicated equipment. These reasons have led us to consider a technology switch to a mass spectrometry method. We tested the kit aTRAQ amino acids analysis for physiological samples (AB Sciex), offering a selective quantification of more than 40 amino acids, and have implemented the acquisition of various original markers to the initial method. The accuracy profiles established for each amino acid show that the results are very reliable. The linearity is assured between 1 and 1.000 mumol/L for most analytes. Result comparison with IEC method showed good agreement. Reference ranges are similar to those defined for the IEC method and patients with inborn errors of metabolism were readily identified. The aTRAQ method offers a valid alternative to IEC method with several advantages: reduced sample volume, decreased run time and increased specificity. However, the procedure requires a thorough review of all chromatographic peaks, process that considerably lengthens the overall time of the procedure. Finally, financial and practical considerations of both techniques have to be counterbalanced before initiating any methodological transition. [less ▲]Detailed reference viewed: 29 (0 ULg)
Quantification of hypoglycin A in serum using aTRAQ((R)) assay.
BOEMER, François ; DEBERG, Michelle ; SCHOOS, Roland et al
in Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2015), 997
BACKGROUND: Hypoglycin A has been recently identified has the causal agent of atypical myopathy (AM) in horses. Its identification and quantification in equine's biological fluids is thus a major concern ... [more ▼]
BACKGROUND: Hypoglycin A has been recently identified has the causal agent of atypical myopathy (AM) in horses. Its identification and quantification in equine's biological fluids is thus a major concern to confirm maple poisoning and to provide insight into the poorly understood mechanism of hypoglycin A intoxication. METHODS: Quantification of hypoglycin A has been achieved with the aTRAQ kit for amino acid analysis of physiological fluids (AB Sciex). Acquisition method on mass spectrometer has been updated to record the hypoglycin A specific MRM transition. RESULTS: Outlined accuracy profiles demonstrated very reliable data. A good linearity was observed from 0.09 to 50mumol/L and precision was very good with coefficient of variation below 8%. Fifty-five samples collected from 25 confirmed AM horses revealed significant hypoglycin A concentrations, while toxin was not found in serum of 8 control animals. CONCLUSIONS: The described aTRAQ variant method has been analytically and clinically validated. The reliability of our approach is thus demonstrated into the workup of atypical myopathy. [less ▲]Detailed reference viewed: 26 (9 ULg)
Mutation of the iron-sulfur cluster assembly gene IBA57 causes fatal infantile leukodystrophy.
DEBRAY, François-Guillaume ; ; et al
in Journal of inherited metabolic disease (2015)
Leukodystrophies are a heterogeneous group of severe genetic neurodegenerative disorders. A multiple mitochondrial dysfunctions syndrome was found in an infant presenting with a progressive ... [more ▼]
Leukodystrophies are a heterogeneous group of severe genetic neurodegenerative disorders. A multiple mitochondrial dysfunctions syndrome was found in an infant presenting with a progressive leukoencephalopathy. Homozygosity mapping, whole exome sequencing, and functional studies were used to define the underlying molecular defect. Respiratory chain studies in skeletal muscle isolated from the proband revealed a combined deficiency of complexes I and II. In addition, western blotting indicated lack of protein lipoylation. The combination of these findings was suggestive for a defect in the iron-sulfur (Fe/S) protein assembly pathway. SNP array identified loss of heterozygosity in large chromosomal regions, covering the NFU1 and BOLA3, and the IBA57 and ABCB10 candidate genes, in 2p15-p11.2 and 1q31.1-q42.13, respectively. A homozygous c.436C > T (p.Arg146Trp) variant was detected in IBA57 using whole exome sequencing. Complementation studies in a HeLa cell line depleted for IBA57 showed that the mutant protein with the semi-conservative amino acid exchange was unable to restore the biochemical phenotype indicating a loss-of-function mutation of IBA57. In conclusion, defects in the Fe/S protein assembly gene IBA57 can cause autosomal recessive neurodegeneration associated with progressive leukodystrophy and fatal outcome at young age. In the affected patient, the biochemical phenotype was characterized by a defect in the respiratory chain complexes I and II and a decrease in mitochondrial protein lipoylation, both resulting from impaired assembly of Fe/S clusters. [less ▲]Detailed reference viewed: 20 (6 ULg)
Diagnostic pitfall in antenatal manifestations of CPT II deficiency.
BOEMER, François ; DEBERG, Michelle ; SCHOOS, Roland et al
in Clinical genetics (2015)
Carnitine palmitoyltransferase II (CPT2) deficiency is a rare inborn error of mitochondrial fatty acid metabolism associated with various phenotypes. Whereas most patients present with postnatal signs of ... [more ▼]
Carnitine palmitoyltransferase II (CPT2) deficiency is a rare inborn error of mitochondrial fatty acid metabolism associated with various phenotypes. Whereas most patients present with postnatal signs of energetic failure affecting muscle and liver, a small subset of patients presents antenatal malformations including brain dysgenesis and neuronal migration defects. Here, we report recurrence of severe cerebral dysgenesis with Dandy-Walker malformation in three successive pregnancies and review previously reported antenatal cases. Interestingly, we also report that acylcarnitines profile, tested retrospectively on the amniotic fluid of last pregnancy, was not sensitive enough to allow reliable prenatal diagnosis of CPT2 deficiency. Finally, because fetuses affected by severe cerebral malformations are frequently aborted, CPT2 deficiency may be underestimated and fatty acid oxidation disorders should be considered when faced with a fetus with Dandy-Walker anomaly or another brain dysgenesis. [less ▲]Detailed reference viewed: 25 (8 ULg)
Les nouveaux outils de diagnostic et de pronostic de la myopathie atypique
Habyarimana, Jean ; BOEMER, François ; Amory, Hélène et al
in Proceeding de la 41ème Journée de la Recherche équine (2015)
In equines, ingestion of hypoglycin A, a toxin produced in the seeds (samaras) of sycamore maple (Acer pseudoplatanus) tree alters the energetic metabolism of muscle cells and results in atypical myopathy ... [more ▼]
In equines, ingestion of hypoglycin A, a toxin produced in the seeds (samaras) of sycamore maple (Acer pseudoplatanus) tree alters the energetic metabolism of muscle cells and results in atypical myopathy (AM). This alterations leads to a characteristic biochemical profile of acylcarnitines (AC) that enables to confirm the diagnosis of AM. This study aims at validating a methodology for the dosage of hypoglycin A in vegetal extracts but also in blood. In addition, the biochemical profile in AC has been determined in AM cases (5 survivors and 13 deceased) and in 5 horses suffering from exercise-induced myopathy. The AC profiles of these horses have been compared to the one of healthy horses (n = 35). This study showed that hypoglycin A was present in seeds and spring seedlings of sycamore and also in blood of AM cases horses. In addition, the establishment of AC profile contributes to the diagnostic and helps to assess the prognosis of AM cases. [less ▲]Detailed reference viewed: 115 (32 ULg)
Cystic Fibrosis screening by simultaneous ELISA for IRT1 and PAP proteins. A pilot study.
Scientific conference (2014, December 11)Detailed reference viewed: 12 (1 ULg)
6-Years Experience of NBS for Hemoglobin Disorders using TMS
Conference (2014, October)Detailed reference viewed: 15 (0 ULg)
Falsos positivos de C5-carnitina elevada en cribado neonatal: A que son debidos?
; ; et al
in Medicina Clinica (2014)Detailed reference viewed: 37 (2 ULg)
Folinic acid treatment for schizophrenia associated with folate receptor autoantibodies.
RAMAEKERS, Vincent ; ; et al
in Molecular genetics and metabolism (2014), 113(4), 307-14
BACKGROUND: Auto-antibodies against folate receptor alpha (FRalpha) at the choroid plexus that block N(5)-methyltetrahydrofolate (MTHF) transfer to the brain were identified in catatonic schizophrenia ... [more ▼]
BACKGROUND: Auto-antibodies against folate receptor alpha (FRalpha) at the choroid plexus that block N(5)-methyltetrahydrofolate (MTHF) transfer to the brain were identified in catatonic schizophrenia. Acoustic hallucinations disappeared following folinic acid treatment. Folate transport to the CNS prevents homocysteine accumulation and delivers one-carbon units for methyl-transfer reactions and synthesis of purines. The guanosine derivative tetrahydrobiopterin acts as common co-factor for the enzymes producing dopamine, serotonin and nitric oxide. METHODS: Our study selected patients with schizophrenia unresponsive to conventional treatment. Serum from these patients with normal plasma homocysteine, folate and vitamin B12 was tested for FR autoantibodies of the blocking type on serial samples each week. Spinal fluid was analyzed for MTHF and the metabolites of pterins, dopamine and serotonin. The clinical response to folinic acid treatment was evaluated. RESULTS: Fifteen of 18 patients (83.3%) had positive serum FR auto-antibodies compared to only 1 in 30 controls (3.3%) (chi(2)=21.6; p<0.0001). FRalpha antibody titers in patients fluctuated over time varying between negative and high titers, modulating folate flux to the CNS, which explained low CSF folate values in 6 and normal values in 7 patients. The mean+/-SD for CSF MTHF was diminished compared to previously established controls (t-test: 3.90; p=0.0002). A positive linear correlation existed between CSF MTHF and biopterin levels. CSF dopamine and serotonin metabolites were low or in the lower normal range. Administration of folinic acid (0.3-1mg/kg/day) to 7 participating patients during at least six months resulted in clinical improvement. CONCLUSION: Assessment of FR auto-antibodies in serum is recommended for schizophrenic patients. Clinical negative or positive symptoms are speculated to be influenced by the level and evolution of FRalpha antibody titers which determine folate flux to the brain with up- or down-regulation of brain folate intermediates linked to metabolic processes affecting homocysteine levels, synthesis of tetrahydrobiopterin and neurotransmitters. Folinic acid intervention appears to stabilize the disease process. [less ▲]Detailed reference viewed: 56 (3 ULg)
Mitochondrial encephalomyopathy with cytochrome c oxidase deficiency caused by a novel mutation in the MTCO1 gene.
Debray, François-Guillaume ; ; et al
in Mitochondrion (2014)
Cytochrome c oxidase (COX) deficiency is one of the most common respiratory chain deficiencies. A woman was presented at the age of 18y with acute loss of consciousness, non-convulsive status epilepticus ... [more ▼]
Cytochrome c oxidase (COX) deficiency is one of the most common respiratory chain deficiencies. A woman was presented at the age of 18y with acute loss of consciousness, non-convulsive status epilepticus, slow neurological deterioration, transient cortical blindness, exercise intolerance, muscle weakness, hearing loss, cataract and cognitive decline. Muscle biopsy revealed ragged-red fibers, COX negative fibers and a significant decreased activity of complex IV in a homogenate. Using next generation massive parallel sequencing of the mtDNA, a novel heteroplasmic mutation was identified in MTCO1, m.7402delC, causing frameshift and a premature termination codon. Single fiber PCR showed co-segregation of high mutant load in COX negative fibers. Mutation in mitochondrially encoded complex IV subunits should be considered in mitochondrial encephalomyopathies and COX negative fibers after the common mtDNA mutations have been excluded. [less ▲]Detailed reference viewed: 33 (1 ULg)
Surprising causes of C5-carnitine false positive results in newborn screening.
BOEMER, François ; SCHOOS, Roland ; de HALLEUX, Virginie et al
in Molecular genetics and metabolism (2014), 111(1), 52-4
During an 18-month period, we noticed an alarming increase of newborn screening false positivity rate in identifying isovaleric acidemia. In 50 of 50 newborns presenting elevated C5-carnitine, we ... [more ▼]
During an 18-month period, we noticed an alarming increase of newborn screening false positivity rate in identifying isovaleric acidemia. In 50 of 50 newborns presenting elevated C5-carnitine, we confirmed the presence of pivaloylcarnitine. Exogenous pivalate administration had been previously identified as the causal agent of this concern. No pivalic-ester prodrug is commercially available in Belgium, but pivalic derivates are also used in the cosmetic industry as emollient under the term "neopentanoate". We have identified neopentanoate-esters in a nipple-fissure unguent that was provided to young mothers. Ceasing distribution of this product hugely reduced the C5-carnitine false positivity rate. [less ▲]Detailed reference viewed: 67 (21 ULg)
BLOOD GROUPS, HEMOGLOBIN PHENOTYPES AND CLINICAL DISORDERS OF CONSANGUINEOUS YANSI POPULATION
Munlemvo Mavanga, Nana ; BOEMER, François ; SEIDEL, Laurence et al
in World Journal of Hematology (2013), 2(4), 109-114
AIM To study frequency of blood groups, prevalence of sickle-cell anemia trait and glucose-6-phosphate dehydrogenase deficiency, among consanguineous Yansi tribe. METHODS A total of 525 blood samples were ... [more ▼]
AIM To study frequency of blood groups, prevalence of sickle-cell anemia trait and glucose-6-phosphate dehydrogenase deficiency, among consanguineous Yansi tribe. METHODS A total of 525 blood samples were collected, of which 256 among the Yansi population, and 269 for the unrelated control group in the Bandundu province of Democratic Republic of Congo. Blood group antigens were determined in the following systems: ABO, Rh, Kell, Duffy, Kidd and MNS. Blood grouping and extended phenotype tests were performed according to standard immunohematological procedures. Spot tests and tandem mass spectrometry were used respectively for the assessment of glucose-6-phosphate dehydrogenase deficiency and sickle-cell anemia trait. RESULTS The frequency of ABO phenotypes conformed to the following order O>A>B>AB with notably 62.5, 23.8, 12.1 and 1.6% for the Yansi, and 54.6, 27.5, 14.1 and 3.7% for the unrelated control group, respectively (P=0.19). As for the Rh phenotypes, the most frequent were ccD.ee, ccD.Ee, CcD.ee, corresponding to 71.5, 12.1 and 12.1% for the Yansi, and 70.6, 15.6 and 8.2%, for the unrelated control group (P=0.27). The frequency of MN and Ss phenotypes were statistically different between groups (P=0.0021 and P=0.0006). G6PD deficiency was observed in 11.3% of subjects in the Yansi group, and in 12.4% of controls (P = 0.74). The sickle-cell anemia trait was present in 22.4% of Yansi subjects and 17.8% in the control group (P=0.24). Miscarriages and deaths in young age were more common among Yansi people. CONCLUSION This study shows a significant difference in MNS blood group distribution between the Yansi tribe and a control population. The distribution of other blood groups and the prevalence of hemoglobinopathies did not differ in the Yansi tribe. [less ▲]Detailed reference viewed: 106 (17 ULg)
Evaluation of Physiological Amino Acids Profiling by Tandem Mass Spectrometry
; ; BOEMER, François
in Journal of Inherited Metabolic Disease Reports (2013)Detailed reference viewed: 26 (4 ULg)
HETEROGENOUS CLINICAL AND LABORATORY PRESENTATIONS IN MAD DEFICIENCY
BOEMER, François ; SCHOOS, Roland ; et al
Poster (2013, September)Detailed reference viewed: 16 (6 ULg)
Identification of methylenecyclopropyl acetic acid in serum of European horses with atypical myopathy
Votion, Dominique ; ; et al
in Equine Veterinary Journal (2013)Detailed reference viewed: 179 (35 ULg)
Enhanced interpretation of newborn screening results without analyte cutoff values
; ; et al
in Genetics in Medicine (2012), 14(7), 648-655
Purpose: To improve quality of newborn screening by tandem mass spectrometry with a novel approach made possible by the collaboration of 154 laboratories in 49 countries. Methods: A database of 767,464 ... [more ▼]
Purpose: To improve quality of newborn screening by tandem mass spectrometry with a novel approach made possible by the collaboration of 154 laboratories in 49 countries. Methods: A database of 767,464 results from 12,721 cases affected with 60 conditions was used to build multivariate pattern recognition software that generates tools integrating multiple clinically significant results into a single score. This score is determined by the overlap between normal and disease ranges, penetration within the disease range, differences between conditions, and weighted correction factors. Results: Ninety tools target either a single condition or the differential diagnosis between multiple conditions. Scores are expressed as the percentile rank among all cases with the same condition and are compared to interpretation guidelines. Retrospective evaluation of past cases suggests that these tools could have avoided at least half of 279 false-positive outcomes caused by carrier status for fatty-acid oxidation disorders and could have prevented 88% of known false-negative events. Conclusion: Application of this computational approach to raw data is independent from single analyte cutoff values. In Minnesota, the tools have been a major contributing factor to the sustained achievement of a false-positive rate below 0.1% and a positive predictive value above 60%. © 2012 American College of Medical Genetics and Genomics. [less ▲]Detailed reference viewed: 56 (10 ULg)