Congenital disorder of glycosylation type Id (CDG Id) : phenotypic, biochemical and molecular characterization of a new patient

Rapport de synthèseLes troubles de la glycosylation (Congenital Disorders of Glycosylation, CDG) regroupent une famille de maladies multi-systémiques héréditaires causées par des défauts dans la synthèse de glycoconjugés. La glycosylation est une réaction enzymatique consistant à lier de façon covalente un glucide à une chaîne peptidique ou une protéine. Il existe deux types de glycosylation. La N-gjycosylation est l'addition de glucides aux chaînes peptidiques en croissance dès leur entrée dans la lumière du réticulum endoplasmique. Elle s'effectue sur les futures glycoprotéines membranaires et conduit à des chaînes de sucres courtes et ramifiées. La O-glycosylation est l'addition de glucides au niveau des résidus hydroxylés des acides aminés sérine et thréonine des chaînes peptidiques déjà présentes dans la lumière de l'appareil de Golgi. Elle est, dans la plupart des cas, effectuée sur îes protéoglycanes et conduit à des chaînes de sucres longues et non ramifiées. La classification des CDG repose sur le niveau de l'étape limitante de la glycosylation. Les CDG de type 1, plus fréquents, regroupent les déficits enzymatiques se situant en amont du transfert de Poligosaccharide sur la chaîne peptidique. Les CDG de type 2 regroupent ceux ayant lieu en aval de ce transfert. Parmi les nombreux différents sous-types de CDG, le CDG de type ld est causé par une anomalie de la mannosyltransferase, enzyme codée par le gène ALG3 (chromosome 3q27). Jusqu'à ce jour, six patients atteints de CDG ld ont été reportés dans la littérature. Notre travail a permis de décrire un septième patient et d'affiner les caractéristiques cliniques, biologiques, neuroradiologiques et moléculaires du CDG ld. Notre patient est notamment porteur d'une nouvelle mutation de type missense sur le gène ALG3. Tous les patients atteints de CDG ld présentent une encéphalopathie progressive avec microcéphalie, retard psychomoteur sévère et épilepsie. Une ostéopénie marquée est présente chez certains patients. Elle est parfois sous diagnostiquée et révélée uniquement lors de fracture pathologique. Les patients atteints de CDG ld présentent également des traits dysmorphiques typiques, mais aucune atteinte multi-systémique ou anomalie biologique spécifique n'est retrouvée telle que dans les autres types de CDG. Le dépistage biochimique des troubles de la glycosylation se fait par une analyse simple et peu coûteuse qui est l'analyse de la transferrine sérique par isoelectrofocusing ou par électrophorèse capillaire. Un tel dépistage devrait être effectué chez tout patient présentant une encéphalopathie d'origine indéterminée, et cela même en l'absence d'atteinte multi- systémique. Notre travail a été publié sous forme d'article de type « short report », peer-reviewed, dans le Journal of Inherited Metabolic Diseases. Le Journal est une révue spécialisée du domaine des erreirs innées du métabolisme. S'agissant d'un seul patient rapporté, l'article ne montre que très synthétiquement le travail effectué, Pour cette raison un complément à l'article avec matériel, méthodes et résultats figure ci-après et concerne la partie de recherche moléculaire de notre travail. La doctorante a non seulement encadré personnellement le patient au niveau clinique et biochimique, mais a plus particulièrement mis au point l'analyse moléculaire du gène ALG3 dans le laboratoire de Pédiatrie Moléculaire pour la première fois ; cela a impliqué l'étude du gène, le choix des oligonucleotides et l'optimisation des réactions d'amplification et séquençage.

Bone metabolism and risk of secondary hyperparathyroidism 12 months after gastric banding in obese pre-menopausal women.

OBJECTIVE: To evaluate, during the first postoperative year in obese pre-menopausal women, the effects of laparoscopic gastric banding on calcium and vitamin D metabolism, the potential modifications of bone mineral content and bone mineral density, and the risk of development of secondary hyperparathyroidism. SUBJECTS: Thirty-one obese pre-menopausal women aged between 25 and 52 y with a mean body mass index (BMI) of 43.6 kg/m(2), scheduled for gastric banding were included. Patients with renal, hepatic, metabolic and bone disease were excluded. METHODS: Body composition and bone mineral density (BMD) were measured at baseline, 6 and 12 months after gastric banding using dual-energy X-ray absorptiometry. Serum calcium, phosphate, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, bilirubin, urea, creatinine, uric acid, proteins, parathormone, vitamin D(3), IGF-1, IGF-BP3 and telopeptide, as well as urinary telopeptide, were measured at baseline and 1, 3, 6, 9 and 12 months after surgery. RESULTS: After 1 y vitamin D3 remained stable and PTH decreased by 12%, but the difference was not significant. Serum telopeptide C increased significantly by 100% (P<0.001). There was an initial drop of the IGF-BP3 during the first 6 months (P<0.05), but the reduction was no longer significant after 1 y. The BMD of cortical bone (femoral neck) decreased significantly and showed a trend of a positive correlation with the increase of telopeptides (P<0.06). The BMD of trabecular bone, at the lumbar spine, increased proportionally to the reduction of hip circumference and of body fat. CONCLUSION: There is no evidence of secondary hyperparathyroidism 1 y after gastric banding. Nevertheless biochemical bone markers show a negative remodelling balance, characterized by an increase of bone resorption. The serum telopeptide seems to be a reliable parameter, not affected by weight loss, to follow up bone turnover after gastroplasty.

A genetic and biochemical investigation of malondialdehyde production and turnover in Arabidopsis

Malondialdehyde (MDA) is a small reactive molecule which occurs ubiqui¬tous among eukaryotes. Interest in this molecule stems from the fact that it can be highly reactive. In green tissues of plants it is apparently formed pre¬dominantly by reactive oxygen species (ROS)-mediated non-enzymatic oxi¬dation (nLPO) of triunsaturated fatty acids (TFAs). MDA which is formed by nLPO is widely used as a disease marker and is regarded to be a cel-lular toxin. Surprisingly, sites of ROS production like mitochondria and chloroplasts possess membranes which are enriched in nLPO-prone polyun¬saturated fatty acids (PUFAs). In this work we showed that chloroplasts are the major site of MDA production in leaves of adult Arabidopsis thaliana plants, whereas analyses in seedlings revealed accumulation in meristematic tissues like the root tip, lateral roots and the apical meristem region. Char-acterizing the MDA pools in more detail, we could show that MDA in plants was predominantly present in a free, non-reactive enolate form. This might explain why it is tolerated in sites where its protonated form could poten¬tially damage the genome and proteome. Analyzing the biological fate of MDA in leaves using labeled MDA-isotopes. we were able to show that MDA is metabolized and used to assemble lipids. The major end-point metabolite was identified as 18:3-16:3-monqgalactosyldiacylglycerol (MGDG), which is the most abundant lipid in chloroplasts. We hypothesize that PUFAs in sites of ROS production, like at PS II in chloroplasts, might act as buffers pre¬venting damage of proteins, thereby generating molecules such as MDA. The MDA produced in this way appears predominantly in a non-reactive enolate form in the cell until it fulfills a biological function or until it is metabo¬lized in order to assemble polyunsaturated MGDGs. Additionally, nLPO has been reported to increase in pathogenesis and we challenged seedlings and adult plants with necrotrophic fungi. Monitoring MDA during the in¬fections, we found MDA pools in seedlings were highly inducible although they were tightly controlled in the leaves of adult plants. - Malondialdehyde (MDA) est une petite molecule réactive présente de manière ubiquitaire dans les eucaryotes. L'intérêt de cette molécule vient du fait que celle-ci pourrait être très réactive. Dans les tissus verts des plantes, la majorité du MDA est apparement formée par l'oxydation non-enzymatique (nLPO) des acides gras polyinsaturés (PUFAs) transmis par des espèces ac¬tives d'oxygène (ROS). Le MDA formé par nLPO est souvent utilisé comme marqueur de maladies et il est considéré comme une toxine cellulaire. Etonnament, les sites de production comme les mitochondries et les chloro- plastes sont riches en PUFAs qui sont sensibles à la nLPO. Dans cette thèse nous montrons que les chloroplastes répresentent le site de production de MDA dans les feuilles adultes d'Arabidopsis thaliana. Les analyses de MDA dans les plantules ont révélé que le MDA s'accumule dans les tissus meris- tematiques comme celles de la pointe de la racine, des racines latéralles et du meristème apical. Par la caractérisation du MDA présent nous avons pu montrer que la majorité du MDA était présent sous la forme d'un énolate non-réactif. Ceci pourrait expliquer pourquoi le MDA est toléré dans les sites où il pourrait casser le genome ou le protéome s'il est présent sous sa forme protonée. Les analyses du devenir du MDA dans les feuilles par des isotopes du MDA ont montré que celui-ci est metabolisé et utilisé pour assembler des lipides. Le lipide majoritairement métabolisé a été identifié comme étant le 18:3-16:3-monogalactosyldiacylglycerole (MGDG); le lipide le plus abondant dans les chloroplastes. Nous supposons que la présence des PUFAs dans les sites de production du ROS, tout comme le PS II dans les chloroplastes, pourrait jouer un rôle de tampon pour prevenir les protéines de différentes dégradations et ainsi générer des molécules telle que le MDA. La majorité du MDA produit par cette réaction est présente dans la cellule sous la forme d'énolate non-réactif, jusqu'au moment de son utilisation ou lorsqu'il serra metabolisé pour produire des MGDGs polyinsaturés. De plus, il a été décrit que nLPO pourait augmenter dans la pathogenèse, et nous avons testé des plantes adultes et des plantules en présence de champignons nécrotrophiques. L'observation du MDA pendant les infections a montré que les concentrations en MDA sont fortement induites dans les plantules mais contrôlées dans les plantes adultes.

Hyperthermia and postmortem biochemical investigations.

The postmortem diagnosis of heat-related deaths presents certain difficulties. Firstly, preterminal or terminal body temperatures are often not available. Additionally, macroscopic and microscopic findings are nonspecific or inconclusive and depend on survival duration after exposure. The diagnosis of hyperthermia is therefore essentially based on scene investigation, the circumstances of death, and the reasonable exclusion of other causes of death. Immunohistochemistry and postmortem biochemical investigations have been performed by several authors in order to better circumstantiate the physiopathology of hyperthermia and provide further information to confirm or exclude a heat-related cause of death. Biochemical markers, such as electrolytes, hormones, blood proteins, enzymes, and neurotransmitters, have been analyzed in blood and other biological fluids to improve the diagnostic potential of autopsy, histology, and immunohistochemistry. The aim of this article is to present a review of the medicolegal literature pertaining to the postmortem biochemical investigations that are associated with heat-related deaths.

Body composition using bio-impedance analysis in pediatric patients with inflammatory bowel disease. Corcordance with dual energy X-ray absorptiometry and comparison with healthy controls

Introduction: Growth is a central process in paediatrics. Weight and height evaluation are therefore routine exams for every child but in some situation, particularly inflammatory bowel disease (IBD), a wider evaluation of nutritional status needs to be performed. Objectives: To assess the accuracy of bio-impedance analysis (BIA) compared to the gold standard dual energy X-ray absorptiometry (DEXA) in estimating percentage body fat (fat mass; FM) and lean body mass (fat free mass; FFM) in children with inflammatory bowel disease (IBD). To compare FM and FFM levels between patients with IBD and healthy controls. Methods: Twenty-nine healthy controls (12 females; mean age: 12.7 ± 1.9 years) and 21 patients (11 females; 14.3 ± 1.3 years) were recruited from August 2011 to October 2012 at our institution. BIA was performed in all children and DEXA in patients only. Concordance between BIA and DEXA was assessed using Lin's concordance correlation and the Bland-Altman method. Between-group comparisons were made using analysis of variance adjusting for age. Results: BIA-derived FM% showed a good concordance with DEXA-derived values, while BIA-derived FFM% tended to be slightly higher than DEXA-derived values (table). No differences were found between patients and controls regarding body mass index (mean ± SD: 19.3 ± 3.3 vs. 20.1 ± 2.8 kg/m2, respectively; age-adjusted P = 0.08) and FM% (boys: 25.3 ± 10.2 vs. 22.6 ± 7.1%, for patients and controls, respectively; P = 0.20; girls: 28.2 ± 5.7 vs. 26.4 ± 7.7%; P = 0.91). Also, no differences were found regarding FFM% in boys (74.9 ± 10.2 vs. 77.4 ± 7.1%; P = 0.22) and girls (71.8 ± 5.6 vs. 73.5 ± 7.7%; P = 0.85). Conclusion: BIA adequately assesses body composition (FM%) in children with IBD and could advantageously replace DEXA, which is more expensive and less available. No differences in body composition were found between children with IBD and healthy controls.

Postmortem biochemical investigations in hypothermia fatalities.

Despite the progress made during the past several decades in forensic pathology, the possibilities for the postmortem diagnosis of hypothermia remains relatively limited. Aside from histology and immunohistochemistry, numerous authors have investigated the postmortem biochemistry of hypothermia fatalities. Several biochemical markers (e.g., glucose, electrolytes, hormones, ketone bodies, and neurotransmitters) and various biological samples (e.g., blood, urine, heart, liver, skeletal muscle as well as pericardial and cerebrospinal fluids) have been proposed as potentially useful markers to improve the insufficient diagnostic efficacy of macroscopic and microscopic findings. The aim of this article is to review the medicolegal literature covering the postmortem biochemical investigations that are associated with hypothermia cases as well as report our own research results on this topic where possible.

Potential influence of the chemical composition of water on the stable oxygen isotope composition of continental ostracods

Many studies in continental areas have successfully used the oxygen isotope composition of fossil ostracod valves to reconstruct past hydrological conditions associated with large changes in climate. Yet, ostracods are known to crystallise their valves out of isotopic equilibrium for oxygen and they generally have higher 18O contents compared to inorganic calcite grown at equilibrium under the same condi- tions. A review of vital offsets determined for continental ostracods indicates that vital offsets might change from site to site, questioning a potential influence of environmental conditions on oxygen isotope fractionation in ostracods. Results from the literature suggest that pH has no influence on ostracod vital offset. A re-evaluation of results from Li and Liu (J Paleolimnol 43:111-120, 2010) suggests that salin- ity may influence oxygen isotope fractionation in ostracods, with lower vital offsets for higher salinities. Such a relationship was also observed for the vital offsets determined by Chivas et al. (The ostracoda- applications in quaternary research. American Geo- physical Union, Washington, DC, 2002). Yet, when results of all studies are compiled, the correlation between vital offsets and salinity is low while the correlation between vital offsets and host water Mg/Ca is higher, suggesting that ionic composition of water and/or relative abundance of major ions may also control oxygen isotope fractionation in ostracods. Lack of data on host water ionic composition for the different studies precludes more detailed examination at this stage. Further studies such as natural or laboratory cultures done under strictly controlled conditions are needed to better understand the potential influence of varying environmental condi- tions on oxygen isotope compositions of ostracod valves.

Serum-free aggregate cultures of rat CNS glial cells: biochemical, immunocytochemical and morphological characterization.

Aggregate cultures of mixed glial cells, as well as of enriched astrocytes and oligodendrocytes were prepared, and maintained in serum-free medium for up to 25 days. Biochemical measurements of both neuron-specific and glia-specific enzyme activities showed that these three types of aggregate cultures were virtually devoid of neurons. Astrocyte-enriched cultures were greater than 95% pure, with oligodendrocytes as the only apparent contaminant, whereas oligodendrocyte-enriched cultures still contained a considerable proportion of astrocytes. In all these neuron-free aggregate cultures both astrocytes and oligodendrocytes attained a high degree of maturation. These findings were confirmed by morphological examinations, and by immunofluorescence studies. Furthermore, ultrastructural as well as immunocytochemical investigations using antibodies to myelin basic protein revealed that all three types of glial cell aggregate cultures contained myelin membranes, indicating that the presence of axons is not a prerequisite for the formation of myelin.

Clay mineral and geochemical composition of Cenozoic paleosol in the Eastern Alps (Austria)

Red clays from Cenozoic palaeosols of the Eastern Alps record periods of stagnating uplift and decrease of relief. Tropical to sub-tropical weathering of a crystalline substratum formed dominant or abundant kaolinite, reflecting Paleogene and Early Miocene conditions, respectively. Abundant illite and chlorite, but a lack of kaolinite in red clays on the plateaus of the Northern Calcareous Alps reflects feldspar-poor compositons of the Cenozoic siliciclastic cover. The presence of high Ba/Sr and Rb/Sr ratios and vermiculite in these red clays indicates high precipitation and temperate weathering conditions, respectively, during the Late Miocene and Early Pilocene on the uplifting plateaus of the Northern Calcareous Alps.

Biochemical properties of RuvBD113N: a mutation in helicase motif II of the RuvB hexamer affects DNA binding and ATPase activities.

Many DNA helicases utilise the energy derived from nucleoside triphosphate hydrolysis to fuel their actions as molecular motors in a variety of biological processes. In association with RuvA, the E. coli RuvB protein (a hexameric ring helicase), promotes the branch migration of Holliday junctions during genetic recombination and DNA repair. To analyse the relationship between ATP-dependent DNA helicase activity and branch migration, a site-directed mutation was introduced into the helicase II motif of RuvB. Over-expression of RuvBD113N in wild-type E. coli resulted in a dominant negative UVs phenotype. The biochemical properties of RuvBD113N were examined and compared with wild-type RuvB in vitro. The single amino acid substitution resulted in major alterations to the biochemical activities of RuvB, such that RuvBD113N was defective in DNA binding and ATP hydrolysis, while retaining the ability to form hexameric rings and interact with RuvA. RuvBD113N formed heterohexamers with wild-type RuvB, and could inhibit RuvB function by affecting its ability to bind DNA. However, heterohexamers exhibited an ability to promote branch migration in vitro indicating that not all subunits of the ring need to be catalytically competent.

Frailty and risk for heart failure in older adults: The health, aging, and body composition study.

OBJECTIVE: The aim of this study was to assess the association between frailty and risk for heart failure (HF) in older adults. BACKGROUND: Frailty is common in the elderly and is associated with adverse health outcomes. Impact of frailty on HF risk is not known. METHODS: We assessed the association between frailty, using the Health ABC Short Physical Performance Battery (HABC Battery) and the Gill index, and incident HF in 2825 participants aged 70 to 79 years. RESULTS: Mean age of participants was 74 ± 3 years; 48% were men and 59% were white. During a median follow up of 11.4 (7.1-11.7) years, 466 participants developed HF. Compared to non-frail participants, moderate (HR 1.36, 95% CI 1.08-1.71) and severe frailty (HR 1.88, 95% CI 1.02-3.47) by Gill index was associated with a higher risk for HF. HABC Battery score was linearly associated with HF risk after adjusting for the Health ABC HF Model (HR 1.24, 95% CI 1.13-1.36 per SD decrease in score) and remained significant when controlled for death as a competing risk (HR 1.30; 95% CI 1.00-1.55). Results were comparable across age, sex, and race, and in sub-groups based on diabetes mellitus or cardiovascular disease at baseline. Addition of HABC Battery scores to the Health ABC HF Risk Model improved discrimination (change in C-index, 0.014; 95% CI 0.018-0.010) and appropriately reclassified 13.4% (net-reclassification-improvement 0.073, 95% CI 0.021-0.125; P = .006) of participants (8.3% who developed HF and 5.1% who did not). CONCLUSIONS: Frailty is independently associated with risk of HF in older adults.

The carbon isotope composition of natural SiC (moissanite) from the Earth's mantle: New discoveries from ophiolites

Moissanite (natural SiC) has been recovered from podiform chromitites of several ophiolite complexes, including the Luobusa and Donqiao ophiolites in Tibet, the Semail ophiolite in Oman and the United Arab Emirates, and the Ray-Iz ophiolite of the Polar Urals, Russia. Taking these new occurrences with the numerous earlier reports of moissanite in diamondiferous kimberlites leads to the conclusion that natural SiC is a widespread mineral in the Earth's mantle, which implies at least locally extremely low redox conditions. The ophiolite moissanite grains are mostly fragments (20 to 150 mu m) with one or more crystal faces, but some euhedral hexagonal grains have also been recovered. Twinned crystals are common in chromitites from the Luobusa ophiolite. The moissanite is rarely colorless, more commonly light bluish-gray to blue or green. Many grains contain inclusions of native Si and Fe-Si alloys (FeSi(2), Fe(3)Si(7)). Secondary ion mass spectrometric (SIMS) analysis shows that the ophiolite-hosted moissanite has a distinctive (13)C-depleted isotopic composition (delta(13)C from -18 to -35 parts per thousand, n=36), much lighter than the main carbon reservoir in the upper mantle (delta(13)C near -5 parts per thousand). The compiled data from moissanite from kimberlites and other mantle settings share the characteristic of strongly (13)C-depleted isotopic composition. This suggests that moissanite originates from a separate carbon reservoir in the mantle or that its formation involved strong isotopic fractionation. The degree of fractionation needed to produce the observed moissanite compositions from the main C-reservoir would be unrealistically large at the high temperatures required for moissanite formation. Subduction of biogenic carbonaceous material could potentially satisfy both the unusual isotopic and redox constraints on moissanite formation, but this material would need to stay chemically isolated from the upper mantle until it reached the high-T stability field of moissanite. The origin of moissanite in the mantle is still unsolved, but all evidence from the upper mantle indicates that it cannot have formed there, barring special and local redox conditions. We suggest, alternatively, that moissanite may have formed in the lower mantle, where the existence of (13)C-depleted carbon is strongly supported by studies of extraterrestrial carbon (Mars, Moon, meteorites). (C) 2009 Elsevier B. V. All rights reserved.

Congenital disorder of glycosylation type Id (CDG Id): phenotypic, biochemical and molecular characterization of a new patient.

Congenital disorders of glycosylation (CDG) are a family of multisystem inherited disorders caused by defects in the biosynthesis of N- or O-glycans. Among the many different subtypes of CDG, the defect of a mannosyltransferase encoded by the human ALG3 gene (chromosome 3q27) is known to cause CDG Id. Six patients with CDG Id have been described in the literature so far. We further delineate the clinical, biochemical, neuroradiological and molecular features of CDG Id by reporting an additional patient bearing a novel missense mutation in the ALG3 gene. All patients with CDG Id display a slowly progressive encephalopathy with microcephaly, severe psychomotor retardation and epileptic seizures. They also share some typical dysmorphic features but they do not present the multisystem involvement observed in other CDG syndromes or any biological marker abnormalities. Unusually marked osteopenia is a feature in some patients and may remain undiagnosed until revealed by pathological fractures. Serum transferrin screening for CDG should be extended to all patients with encephalopathy of unknown origin, even in the absence of multisystem involvement.