Selenoprotein M is expressed during bone development

25 selenoproteins that contain selenium, incorporated as selenocysteine (Sec), have been identified to date. Selenoprotein M (SELM) is one of seven endoplasmic reticulum (ER)-resident, Sec-containing proteins that may be involved in posttranslational processing of proteins and maintenance of ER function. Since SELM was overrepresented in a cartilage- and bone- specific expressed sequence tag (EST) library, we further investigated the expression pattern of Selm and its possible biological function in the skeleton. RNA in situ hybridization of Selm in chicken and mice of different developmental stages revealed prominent expression in bones, specifically in osteoblast, and in tendons. This result suggests that SELM functions during bone development, where it is possibly involved in the processing of secreted proteins

Genetic association study of UCMA/GRP and OPTN genes (PDB6 locus) with Paget's disease of bone

We performed a genetic association study of rare variants and single nucleotide polymorphisms (SNPs) of UCMA/GRP and OPTN genes, in French-Canadian patients with Paget's disease of bone (PDB) and in healthy controls from the same population. We reproduced the variant found in the UCMA/GRP basal promoter and tested its functionality using in vitro transient transfection assays. Interestingly, this SNP rs17152980 appears to affect the transcription level of UCMA/GRP. In addition, we have identified five rare genetic variants in UCMA/GRP gene, four of them being population-specific, although none were found to be associated with PDB. Six Tag SNPs of UCMA/GRP gene were associated with PDB, particularly the SNP rs17152980 (uncorrected P = 3.8 x 10(-3)), although not significant after Bonferroni's correction. More importantly, we replicated the strong and statistically significant genetic association of two SNPs of the OPTN gene, the rs1561570 (uncorrected P = 5.7 x 10(-7)) and the rs2095388 (uncorrected P = 4.9 x 10(-3)), With PDB. In addition, we identified a very rare variant found to be located close to the basal promoter of the OPTN gene, at -232 bp from its distal transcription start site. Furthermore, depending on the type of allele present (G or A), the binding of several important nuclear factors such as the vitamin D or the retinoic acid receptors is predicted to be altered at this position, suggesting a significant effect in the regulation of transcription of the OPTN gene. In conclusion, we identified a functional SNP located in the basal promoter of the UCMA/GRP gene which provided a weak genetic association with PDB. In addition, we replicated the strong genetic association of two already known SNPs of the OPTN gene, with PDB in a founder effect population. We also identified a very rare variant in the promoter of OPTN, and through bioinformatic analysis, identified putative transcription factor binding sites likely to affect OPTN gene transcription. (C) 2012 Elsevier Inc. All rights reserved.Fonds de la Recherche du Quebec - Sante (FRQS), Canada; Portuguese Science and Technology Foundation, Portugal [SFRH/BPD/48206/2008]; Catalyst Grant (Bone Health) from the Canadian Institutes of Health Research (Canada); CHUQ Foundation (Canada); Groupe de Recherche en Maladies Osseuses (Canada); Canadian Foundation for Innovation (Canada); FRSQ (Canada); Laval University (Canada); CHUQ (CHUL) Research Centre (Canada); Centre of Marine Sciences (CCMAR) (Portugal)info:eu-repo/semantics/publishedVersio

Correlation between FIX genotype and pharmacokinetics of Nonacog alpha according to a multicentre Italian study

Pharmacokinetic (PK) studies on recombinant FIX concentrate, Nonacog alpha, were conducted with different sampling time designs which gave rise to not complete and homogenous outcomes. In addition, patient's FIX genotype/PK relationship has never been investigated

The relationship of the knowledge of letter names and reading achievement in grade one.

Thesis (Ed.M.)--Boston Universit

Identifizierung und Charakterisierung von Genen mit Einfluss auf Entwicklung und Erhalt des Knorpel-/Knochen-Systems

Defects in formation, growth and homeostasis of the skeleton are responsible for various human monogenic and multigenic heritable disorders as skeletal dysplasias, craniosynostosis and ostheoarthritis. Thus, the present dissertation aims at the identification of novel genes and pathways involved in the complex processes of cartilage/bone formation, growth, differentiation and homeostasis, which may serve as candidate genes for the above mentioned disorders. In the first part of my work, I analysed 4748 clones from a human growth plate cartilage cDNA library generated from 20 weeks prenatal- to 2 years postnatal specimens (Tagariello et al., 2005a). In silico analysis of these sequences revealed 1688 individual transcription units, corresponding to known (1274) and to novel, yet uncharacterised genes (414). The EST profile representing with a total of approximately 10% proteins already shown to be involved in cartilage/bone development or homeostasis, reflect the tissue specifity of the library. The EST profile also reflects the developmental stage of the tissue with significant differences in the expression of matrix known compounds compared to corresponding EST profiles from 8-12 and 12-20 week human fetal cartilage. Calculation of the relative frequency of transcripts in our cDNA library, as compared to their abundance in other EST databases, revealed a set of approximately 200 genes, including 81 novel, yet uncharacterised genes, showing increased expression. These genes represent candidates for the large number of osteochondrodysplasias for which the causative gene defects have not yet been identified. In the present thesis, I described the further characterization of a putative candidate gene (ECM3: extracellular matrix protein 3). The predicted protein has a length of 583 amino acids with 12 leucine-rich repeats, a signal peptide and a high homology to ECM2 and BGN. The expression of ECM3 is restricted to a few tissues including placenta as well as normal and osteoarthritic cartilage. Investigation of the high expression level of ECM3 in several specimens of cartilage from osteoarthritis and rheumtatoid arthritis patients in different stages of disease progression by real-time PCR, indicated a function of ECM3 in the pathogenesis of these two forms of chronic arthropathies. Furthermore ECM3 turned out to be an excellent positional candidate gene for a patient with an interesting combination of frontometaphyseal dysplasia and periventricular nodular heterotopia. However, in collaboration with Dr. Zenker (Department of Human Genetics, Erlangen) ECM3 was excluded to be involved in this disease. Instead, an unusual mutation in the Filamin A (FLNA) gene was shown to be the basic cause of the clinical phenotype (Zenker et al., 2004). In the second part of my work I focused on novel genes involved in the process of desmal ossification. I analysed a patient with a craniosynostosis and a de novo balanced translocation. Craniosynostosis is a congenital developmental disorder involving premature fusion of cranial sutures resulting in an abnormal shape of the skull. Little is known about the various forms of craniosynostosis. At birth the patient presented with Crouzon-like brachycephaly, proptosis, midfacial hypoplasia and low set ears. Three-dimensional cranial computer tomography revealed fusion of the lambdoid sutures and distal part of the sagittal suture with a gaping anterior fontanelle. Mutations in the genes for FGFR1-3 were excluded. Standard chromosome analysis revealed a de novo balanced translocation t(9;11)(q33;p15). The identified breakpoint on chromosome 11p15 disrupts the SOX6 gene, a gene known to be involved in skeletal growth and differentiation processes. Screening of the SOX6 gene in 104 patients with various forms of craniosynostosis revealed a missense mutation in one particular patient with an apparently isolated complex craniosynostosis involving the coronal and sagital suture. Interestingly, an evolutionary conserved aspartate residue at position 68 was exchanged against asparagine as a result of the mutation. The affected amino acid is located in the N-terminal part of SOX6 and has so far not been assigned to any functional domain. Since the mutation was also found in the patient´s apparently non-affected mother, it remains open, whether the mutation represents a rare polymorphism or is associated with a craniosynostosis form with reduced penetrance. The breakpoint on chromosome 9 could be located in a region without any known or predicted genes, but interestingly, disrupts patches of evolutionary high conserved non-genic sequences (CNGs) and may thus led to a dysregulation of flanking genes on chromosome 9 or 11 involved in skull vault development. Two interesting target genes of this dysregulation are TLR4 (previously shown to be involved in bone development) and ARM1 a novel gene, identified in the context of my present study.Defekte bei Bildung, Wachstum und Homöostase des Skeletts sind verantwortlich für eine Reihe menschlicher Erkrankungen, zu denen neben zahlreichen Skelettdysplasien und Kraniosynostosen auch Osteoarthrose und rheumatoide Arthritis gehören. Da die diesen Erkrankungen zugrunde liegenden molekularen Prozesse nur in Ansätzen aufgeklärt sind, war es Ziel der Arbeit, neue Gene, die in die komplexen Prozesse der Knorpel-/Knochenbildung, -differenzierung und -homöostase beim Menschen involviert sind, zu identifizieren und molekular zu charakterisieren. Im Rahmen der vorliegenden Doktorarbeit wurden zunächst insgesamt 4748 ESTs aus humanem fetalen Wachstumsfugenknorpel (20.SSW – 2.Lebensjahr), die im Rahmen eines Kooperationsprojektes mit der Universität Mainz und der Fa. GENterprise generiert worden waren, bioinformatisch ausgewertet und zur Erstellung eines Expressionsprofils verwendet (Tagariello et al., 2005a). Die 4748 ESTs repräsentieren 1688 unterschiedliche Transkripte, die sich auf 1247 bekannte und 414 unbekannte, putative Gene verteilen. Die Spezifität der cDNA-Bibliothek, sowie der gewonnenen Daten, zeigte sich dadurch, dass für ca. 10% aller bekannten Gene in dem ausgewerteten Datensatz bereits eine Funktion in der Knorpel-/Knochenentwicklung und -homöostase nachgewiesen war. Im Vergleich mit entsprechenden Datensätzen anderer Entwicklungsstadien konnten entwicklungsspezifische Unterschiede im Expressionsprofil identifiziert werden. Die Berechnung der relativen Frequenz der ausgewerteten Transkripte, im Vergleich zu der Verteilung der Transkripte in anderen cDNA-Bibliotheken, ergab einen Satz von 81 Genen, die entweder völlig uncharakterisiert sind oder für die bisher keine Funktion bei der Skelettentwicklung beschrieben wurde. Diese Gene stellen mögliche Kandidatengene für Skeletterkrankungen dar. Eines der identifizierten Gene (ECM3: extracellular matrix 3) kodiert für ein putatives Protein mit einem Signalpeptid und 12 „Leucin-reichen Repeats“, das Ähnlichkeit zu den Proteinen BGN, ECM1 und ECM2 aufweist. Die Expression von ECM3 ist auf wenige Gewebe (u.a. Plazenta, sowie normalen und osteoarthrotischen Knorpel) begrenzt. Quantitative Expressionsanalysen mittels „Real-Time“-PCR ergaben eine signifikant höhere Expression des ECM3-Gens in Kniegelenkknorpel-Resektaten einzelner Patienten mit Osteoarthrose oder rheumatoider Arthritis, so dass vermutet werden kann, dass ECM3 in die Pathogenese der beiden chronischen Gelenkerkrankungen involviert ist. Darüber hinaus stellte ECM3 zu Beginn der vorliegenden Arbeit auch ein exzellentes positionelles Kandidatengen für die Frontometaphysäre Dysplasie (FMD) dar. Durch Untersuchung eines seltenen Patienten mit bilateraler Periventrikulärer Nodulärer Heterotopie und einer FMD, die in Zusammenarbeit mit Dr. Zenker (Institut für Humangenetik, Erlangen) durchgeführt wurde, konnte ECM3 im Rahmen der vorliegenden Arbeit als ursächliches Gen ausgeschlossen und stattdessen eine Mutation im Filamin A (FLNA)-Gen nachgewiesen werden (Zenker et al., 2004). Neben der breit angelegten, grundlegenden Strategie zur Genidentifizierung wurde in der vorliegenden Arbeit auch versucht, über einen Patienten-basierten Ansatz Kandidatengene für die Knorpel/Knochenentwicklung zu identifizieren. Hierzu wurden die Bruchpunkte bei einem Patienten mit Kraniosynostose und einer balancierten Chromosomentranslokation (t(9;11) (q33;p15)) kloniert und sequenziert (Tagariello et al., 2005b). Bei den Kraniosynostosen handelt es sich um die frühzeitige Verknöcherung einer oder mehrerer Schädelnähte, deren Ursachen bisher nur in Ansätzen bekannt sind. Der Bruchpunkt auf Chromosom 11p15 unterbricht das SOX6-Gen, das in Skelettentwicklung und -wachstum involviert ist. Obwohl in der vorliegenden Arbeit eine Missens-Mutation im SOX6-Gen bei einem weiteren Patienten mit Kraniosynostose identifiziert werden konnte, ließ sich ein direkter Nachweis der SOX6-Veränderung als Ursache für das Krankheitsbild nicht erbringen. Der Bruchpunkt auf Chromosom 9q33 ist in einer Region ohne bekannte oder vorhergesagte Gene lokalisiert. Hier unterbricht die Translokation interessanterweise eine Reihe von evolutionär hoch konservierten, nicht-kodierenden und nicht-transkribierten Elementen (CNGs = highly conserved non-genic sequences), was zu möglichen Positionseffekten auf flankierende Gene führen könnte. Zwei interessante Kandidatengene, die den Bruchpunkt flankieren, stellen hierbei die Gene TLR4 (für das eine Funktion bei der Knochenentwicklung bereits nachgewiesen wurde) und das in der vorliegenden Arbeit neu identifizierte und in der Schädeldecke exprimierte Gen ARM1 dar

Ucma/GRP inhibits phosphate-induced vascular smooth muscle cell calcification via SMAD-dependent BMP signalling

Vascular calcification (VC) is the process of deposition of calcium phosphate crystals in the blood vessel wall, with a central role for vascular smooth muscle cells (VSMCs). VC is highly prevalent in chronic kidney disease (CKD) patients and thought, in part, to be induced by phosphate imbalance. The molecular mechanisms that regulate VC are not fully known. Here we propose a novel role for the mineralisation regulator Ucma/GRP (Upper zone of growth plate and Cartilage Matrix Associated protein/Gla Rich Protein) in phosphate-induced VSMC calcification. We show that Ucma/GRP is present in calcified atherosclerotic plaques and highly expressed in calcifying VSMCs in vitro. VSMCs from Ucma/GRP(-/-) mice showed increased mineralisation and expression of osteo/chondrogenic markers (BMP-2, Runx2, beta-catenin, p-SMAD1/5/8, ALP, OCN), and decreased expression of mineralisation inhibitor MGP, suggesting that Ucma/GRP is an inhibitor of mineralisation. Using BMP signalling inhibitor noggin and SMAD1/5/8 signalling inhibitor dorsomorphin we showed that Ucma/GRP is involved in inhibiting the BMP-2-SMAD1/5/8 osteo/chondrogenic signalling pathway in VSMCs treated with elevated phosphate concentrations. Additionally, we showed for the first time evidence of a direct interaction between Ucma/GRP and BMP-2. These results demonstrate an important role of Ucma/GRP in regulating osteo/chondrogenic differentiation and phosphate-induced mineralisation of VSMCs.NWO ZonMw [MKMD 40-42600-98-13007]; FCT [SFRH/BPD/70277/2010]info:eu-repo/semantics/publishedVersio

A Novel Assessment and Profiling of Multidimensional Apathy in Alzheimer's Disease

BACKGROUND: Apathy is a complex multidimensional syndrome frequently reported in Alzheimer's disease (AD) and is associated with impaired awareness. Here we present a psychometrically robust method to profile apathy in AD.  OBJECTIVES: To determine the validity and reliability of a multidimensional apathy measure, the Dimensional Apathy Scale (DAS), and explore the apathy subtype profile and its associations in AD.  METHODS: 102 people with AD and 55 healthy controls were recruited. Participants completed the DAS, the Apathy Evaluation Scale (AES), Geriatric Depression Short form (GDS-15), and Lawton Instrumental Activities of Daily Living (LIADL). Psychometric properties of the DAS were determined. AD-Control comparison was performed to explore group differences on the DAS. Latent Class Analysis (LCA) was used to explore the profile of apathy in AD.  RESULTS: The DAS had a good to excellent Cronbach's standardized alpha (self-rated = 0.85, informant/carer-rated = 0.93) and good convergent and divergent validity against standard apathy (AES) and depression (GDS-15) measures. Group comparison showed people with AD were significantly higher for all apathy subtypes than controls (p < 0.001), and lacking in awareness over all apathy subtype deficits. LCA showed three distinct AD subgroups, with 42.2% in the Executive-Initiation apathy, 28.4% in the Global apathy, and 29.4% in the Minimal apathy group.  CONCLUSIONS: The DAS is a psychometrically robust method of assessing multidimensional apathy in AD. The apathy profiles in AD are heterogeneous, with additional specific impairments relating to awareness dependent on apathy subtype

Acquired factor VIII deficiency after consuming the dried gallbladder of a cobra, Naja naja

Acquired factor VIII deficiency is very rare, often fatal. It is associated with pregnancy, autoimmune diseases, malignancy, and drugs, although no underlying cause is found in 50%. A 49-year-old male was referred with right shoulder bruising. The coagulation test showed a prolonged activated partial thromboplastin time. The factor VIII level was less than 1%, and the factor VIII inhibitor antibody titer was 246 Bethesda units/mL. The findings were compatible with acquired factor VIII deficiency. He had consumed the dried gallbladder of a cobra, Naja naja, for two weeks, it contained venom. After the initial treatment with factor VIII, he did not take supplemental coagulation factor VIII. The patient was readmitted with left forearm swelling. He lost consciousness suddenly and brain computed tomography (CT) revealed a subdural hematoma. Despite administering recombinant factor VII, his bleeding was not controlled and he died

Effect of Chronic Obstructive Pulmonary Disease (COPD) on Biventricular Mechanics in Patients Without Severe Airflow Obstruction

Background: Over the last 15 years, few echocardiographic studies have examined the biventricular mechanics by speckle tracking echocardiography (STE) in patients affected by chronic obstructive pulmonary disease (COPD) without advanced lung disease. We aimed to summarize the main findings of these studies and quantify the overall effect of COPD on biventricular mechanics in patients without severe airflow obstruction. Methods: Eligible studies assessing cardiac function by conventional transthoracic echocardiography (TTE), implemented with a STE analysis of left ventricular (LV)-global longitudinal strain (GLS) and/or right ventricular (RV)-GLS in COPD patients without severe airflow obstruction vs. healthy controls, were selected from the PubMed, Embase and Scopus databases. The primary endpoint was to quantify the effect of COPD on LV-GLS and RV-GLS in individuals without advanced lung disease. Continuous data [LV-GLS, RV-GLS, left ventricular ejection fraction (LVEF) and tricuspid annular plane systolic excursion (TAPSE)] were pooled as the standardized mean difference (SMD) comparing COPD cohorts with healthy controls. Results: Ten studies were included, totaling 682 COPD patients and 316 healthy controls. Overall, COPD showed a large effect on LV-GLS (SMD −1.296; 95%CI −2.010, −0.582, p &lt; 0.001) and RV-GLS (SMD −1.474; 95% CI −2.142, −0.805, p &lt; 0.001), a medium-to-large effect on TAPSE (SMD −0.783, 95% CI −0.949, −0.618, p &lt; 0.001) and a small effect on LVEF (SMD −0.366, 95% CI −0.659, −0.074, p = 0.014). The I2 statistic value for the LV-GLS (91.1%), RV-GLS (88.2%) and LVEF (76.7%) studies suggested a high between-study heterogeneity, while that for the TAPSE (38.1%) studies was compatible with a low-to-moderate between-study heterogeneity. Egger’s test yielded a p-value of 0.16, 0.48, 0.58 and 0.50 for LV-GLS, RV-GLS, LVEF and TAPSE studies, respectively, indicating an absence of publication bias. Meta-regression analyses excluded that the effect of COPD on biventricular mechanics might be influenced by potential confounders (all p &gt; 0.05). Sensitivity analysis confirmed the robustness of the LV-GLS, RV-GLS and TAPSE studies’ results. Conclusions: COPD appears to be independently associated with a mild attenuation of biventricular mechanics in patients with moderate airflow limitations, despite a preserved LVEF and TAPSE on conventional TTE. STE analysis may allow clinicians to identify COPD patients with subclinical myocardial dysfunction and an increased risk of heart failure and cardiovascular complications early

Effects of early rehabilitation on motor function, dyspnoea intensity, respiratory muscle performance and handgrip strength in patients with COVID-19: an observational study

Background/aims Although an increasing volume of research is emerging, rehabilitative treatment of patients with COVID-19 still continues to be a matter of great importance that must be explored further. The purpose of the present study was to describe the effects of inpatient rehabilitation in acute patients treated in a sub-intensive hospital setting during the COVID-19 pandemic.Methods A retrospective analysis was conducted based on the prospectively collected data of 192 patients with COVID-19 undergoing a physiotherapeutic regimen during their hospitalisation. Patients were admitted because of COVID-19-related pneumonia from the periods of 25 March-12 June 2020 and 2 November 2020-9 June 2021. This study investigated dyspnoea intensity using the modified Borg scale, motor function through the 1-minute sit-to-stand test, and daily walked distance. In a subset of 57 patients, handgrip strength and respiratory muscle function was also evaluated. Measurements were taken at baseline and discharge.Results Patients were classified according to the severity of their ratio of arterial oxygen partial pressure to fractional inspired oxygen (mean 225 +/- 82 mmHg). At discharge to home or to another hospital facility, patients performed a mean of 12 repetitions (1-minute sit-to-stand test); dyspnoea intensity was 1.4 (modified Borg scale), and they were able to walk a mean distance of 266.7 metres. The mean handgrip strength of the dominant hand was 29.3 kg, the maximal inspiratory pressure was 43.5 cmH(2)O, and the maximal expiratory pressure was 59.1 cmH(2)O. Overall, significant differences before and after treatment were detected for all clinical variables. Dyspnoea improved by 0.7 points; walked distance by 200 metres; the number of repetitions at the 1-minute sit-to-stand test by 5.6; the handgrip strength by 1.2 kg (right hand) and 1.7 kg (left hand); maximal inspiratory pressure by 7.7 cmH(2)O; and maximal expiratory pressure by 9.5 cmH(2)O.Conclusions Patients obtained significant improvements in functional capacity, dyspnoea perception, handgrip strength and respiratory muscle function. In addition, the treatment was feasible and well tolerated by patients, and no adverse related events were observed in a sub-intensive care setting