Statistical groupings of mental and social health measurements correlate with musculoskeletal capability - A cross sectional study.

OBJECTIVE: A better understanding of the degree to which social health factors contribute uniquely to statistical clusters associated with variation in levels of capability might inform targeted whole person care strategies for more comprehensive management of musculoskeletal health. Therefore, we asked: (1) What are the statistical groupings of social and mental health measurements in patients seeking specialty care for musculoskeletal conditions? (2) Do identified psychosocial groupings correspond with different mean magnitudes of incapability accounting for demographic and clinical factors? METHODS: We included 158 patients seeking musculoskeletal specialty care and collected measures of magnitude of incapability, unhelpful thoughts and distress regarding symptoms, symptoms of depression, symptoms of anxiety, and social health. A k-means clustering algorithm was fit to the data and a linear regression model compared mean PROMIS-PF CAT scores for grouping. RESULTS: A quantitative social health measure contributed to 4 statistical clusters as follows: 1) relatively low levels of all mental health measures and high social health; 2) greater unhelpful thoughts and distress regarding symptoms, average symptoms of general anxiety and depression, and average social health; 3) higher levels of all mental health measures and severely compromised social health; and 4) severely compromised mental health and lower social health. Magnitude of incapability was significantly greater for groups with worse mental and social health. CONCLUSION: The finding of a relatively independent association of social and mental health factors with greater incapability supports the importance of introducing comprehensive health strategies in musculoskeletal specialty care. Strategies may include mindset training and case management of social unmet needs. LEVEL OF EVIDENCE: Level III; Cross-sectional study.

Respiratory diseases in Mexico: analysis from the Global Burden of Disease study 2021.

BACKGROUND: Respiratory diseases (RD) are often analyzed separately rather than collectively, possibly leading to an underestimation of their total burden. OBJECTIVE: To analyze the burden of RD in Mexico for population aged 20 or older from 1990 to 2021. MATERIAL AND METHODS: We present the burden of RD in Mexico based on estimates of the Global Burden of Disease study for mortality and disability-adjusted life years (DALYs), comprising counts, rates per 100,000, as well as age-standardized rates. RDs were categorized into three key groups: chronic respiratory diseases (CRD), respiratory infections (RI), and respiratory cancers. RESULTS: In 2021, among those aged 20+, RDs were responsible for 336,728 deaths, which accounts for 30.5% of total deaths -a nearly threefold increase since 2019, primarily due to the COVID-19 pandemic. CRDs contributed with 3.4% of total deaths; RIs, with 25.9%; and respiratory cancers, with 1.2%. CRDs showed a continuous rise in deaths, crude mortality, and DALY rates across genders, with no signs of leveling. RD burden varied widely across Mexican states. Age-standardized CRD mortality rates have generally declined since 1990, except for interstitial lung diseases, which have consistently increased. CONCLUSION: The significant burden of mortality and disability due to RDs in Mexico underscores the n|ecessity for enhanced prevention, research, and for addressing risk factors such as smoking and pollution. Ongoing healthcare training can help reduce RD burden.

ANTECEDENTES: Las enfermedades respiratorias (ER) se analizan individualmente, posiblemente con subestimación de su carga total. OBJETIVO: Analizar la carga de las ER en México para población de 20 años o más de 1990 a 2021. MATERIAL Y MÉTODOS: Se presenta la carga de ER en México a partir de estimaciones del estudio Global Burden of Disease en cuanto a mortalidad y años de vida saludable (AVISA) perdidos que comprenden recuentos, tasas por 100 000 y tasas estandarizadas por edad. Las ER se categorizaron en enfermedades respiratorias crónicas (ERC), infecciones respiratorias y cánceres respiratorios. RESULTADOS: En 2021, las ER causaron la muerte de 336 728 adultos mayores de 20 años, lo que representó 30.5 % del total de defunciones, incremento cercano al triple respecto a 2019, principalmente debido a COVID-19. Las ERC contribuyeron con 3.4 % del total de muertes, las infecciones respiratorias con 25.9 % y los cánceres respiratorios con 1.2 %. La mortalidad y AVISA perdidos por ERC se incrementaron persistentemente, con variaciones entre los estados. Las tasas de mortalidad ajustadas por edad de las ERC disminuyeron desde 1990, excepto las enfermedades pulmonares intersticiales, que se incrementaron constantemente. CONCLUSIÓN: Los significativos niveles de mortalidad y discapacidad debidos a enfermedades respiratorias en México exigen mejorar la prevención, investigación y abordar factores de riesgo como tabaquismo y contaminación, además de fomentar la capacitación médica continua.

A Fokker-Planck Framework for Parameter Estimation and Sensitivity Analysis in Colon Cancer.

A new stochastic framework for parameter estimation and uncertainty quantification in colon cancer-induced immune response is presented. The dynamics of colon cancer is given by a stochastic process that captures the inherent randomness in the system. The stochastic framework is based on the Fokker-Planck equation that represents the evolution of the probability density function corresponding to the stochastic process. An optimization problem is formulated that takes input individual patient data with randomness present, and is solved to obtain the unknown parameters corresponding to the individual tumor characteristics. Furthermore, sensitivity analysis of the optimal parameter set is performed to determine the parameters that need to be controlled, thus, providing information of the type of drugs that can be used for treatment.

Liver transplant with controlled donors after circulatory death with normothermic regional perfusion and brain dead donors: A multicenter cohort study of transfusion, one-year graft survival and mortality.

BACKGROUND: Controlled donation after circulatory death (cDCD) has expanded the donor pool for liver transplantation (LT). However, transfusion requirements and perioperative outcomes should be elucidated. The aim of this multicenter study was to assess red blood cell (RBC) transfusions, one-year graft and patient survival after LT after cDCD with normothermic regional perfusion (NRP) compared with donors after brain death (DBD). METHODS: 591 LT carried out in ten centers during 2019 were reviewed. Thromboelastometry was used to manage coagulation and blood product transfusion in all centers. Normothermic regional perfusion was the standard technique for organ recovery. RESULTS: 447 patients received DBD and 144 cDCD with NRP. Baseline MCF Extem was lower in the cDCD group There were no differences in the percentage of patients (63% vs. 61% p = 0.69), nor in the number of RBC units transfused (4.7 (0.2) vs 5.5 (0.4) in DBD vs cDCD, p = 0.11. Twenty-six patients (6%) died during admission for LT in the DBD group compared with 3 patients (2%) in the cDCD group (p = 0.15). To overcome the bias due to a worse coagulation profile in cDCD recipients, matched samples were compared. No differences in baseline laboratory data, or in intraoperative use of RBC or one-year outcome data were observed between DBD and cDCD recipients. CONCLUSIONS: cDCD with NRP is not associated with increased RBC transfusion. No differences in graft and patient survival between cDCD and DBD were found. Donors after controlled circulatory death with NRP can increasingly be utilized with safety, improving the imbalance between organ donors and the ever-growing demand.

When genetic and surname analyses meet historical sources: The C56R mutation associated with factor XI deficiency as a marker of human migration during the Spanish Reconquista.

The C56R mutation associated with factor XI deficiency has been first evidenced in individuals from the French Basque Country. Genetic investigations revealed that this mutation occurred about 5400 years ago as a founder effect in this zone. Other cases were subsequently described in Southwestern Europe. Noticeably a cluster of cases was evidenced in Yecla, a small city from the province of Murcia, in Southeastern Spain. In correlation with historical sources our genetic data and surname analysis argue for associating this mutation with the migration of people from Western Pyrenees (and more probably from the Navarra province) toward Southeastern Spain during the Reconquista period.

A dedicated revision total knee service: a surgeon's perspective.

AIMS: Revision total knee arthroplasty (rTKA) accounts for approximately 5% to 10% of all TKAs. Although the complexity of these procedures is well recognized, few investigators have evaluated the cost and value-added with the implementation of a dedicated revision arthroplasty service. The aim of the present study is to compare and contrast surgeon productivity in several differing models of activity. MATERIALS AND METHODS: All patients that underwent primary or revision TKA from January 2016 to June 2018 were included as the primary source of data. All rTKA patients were categorized by the number of components revised (e.g. liner exchange, two or more components). Three models were used to assess the potential surgical productivity of a dedicated rTKA service : 1) work relative value unit (RVU) versus mean surgical time; 2) primary TKA with a single operating theatre (OT) versus rTKA with a single OT; and 3) primary TKA with two OTs versus rTKA with a single OT. RESULTS: In total, 4570 procedures were performed: 4128 primary TKAs, 51 TKA liner exchanges, and 391 full rTKAs. Surgical time was significantly different between the primary TKA, liner exchange, and rTKA cohorts (100.6, 97.1, and 141.7 minutes, respectively; p < 0.001). Primary TKA yielded a mean of 7.1% more RVU/min per procedure than rTKA. Our one-OT model demonstrated that primary TKA (n = 4) had a 1.9% RVU/day advantage over rTKA (n = 3). If two OTs are used for primary TKA (n = 6), the outcome strongly favours primary TKA by an added 34.6% RVUs/day. CONCLUSION: Our results suggest that a dedicated rTKA service would lead to lower surgeon remuneration based on the current RVU paradigm. Revision arthroplasty specialists may need additional or alternative incentives to promote the development of a dedicated revision service. Through such an approach, healthcare organizations could enhance the quality of care provided, but surgeon productivity measures would need to be adjusted to reflect the burden of these cases. Cite this article: Bone Joint J 2019;101-B:675-681.

Image Registration Measures and Chronic Osteoarthritis Knee Pain Prediction: Data from the Osteoarthritis Initiative / Métricas de Registro de Imágenes y Predicción de Dolor de Rodilla por Osteoartritis Crónica: Datos de la Osteoarthritis Initiative

Abstract Osteoarthritis (OA) is the most common type of arthritis, is a growing disease in the industrialized world. OA is an incapacitate disease that affects more than 1 in 10 adults over 60 years old. X-ray medical imaging is a primary diagnose technique used on staging OA that the expert reads and quantify the stage of the disease. Some Computer-Aided Diagnosis (CADx) efforts to automate the OA detection have been made to aid the radiologist in the detection and control, nevertheless, the pain inherits to the disease progression is left behind. In this research, it's proposed a CADx system that quantify the bilateral similarity of the patient's knees to correlate the degree of asymmetry with the pain development. Firstly, the knee images were aligned using a B-spline image registration algorithm, then, a set of similarity measures were quantified, lastly, using this measures it's proposed a multivariate model to predict the pain development up to 48 months. The methodology was validated on a cohort of 131 patients from the Osteoarthritis Initiative (OAI) database. Results suggest that mutual information can be associated with K&L OAI scores, and Multivariate models predicted knee chronic pain with: AUC 0.756, 0.704, 0.713 at baseline, one year, and two years' follow-up.

Resumen La osteoartritis (OA) es el tipo de artritis más común. OA es una enfermedad limitante que afecta a 1 de 10 adultos con 60 años o más. Las imágenes de rayos-x son una técnica de diagnóstico primario que permite conocer el estado de OA, las cuales el experto lee y cuantifica así la etapa de la enfermedad. El Diagnóstico Asistido por Computadora (CADx, por sus siglas en inglés) ha buscado automatizar el diagnóstico de OA para ayudar al radiólogo en la detección y control; sin embargo, el dolor provocado por la progresión de la enfermedad es dejado atrás. En este trabajo se propone un sistema de CADx que cuantifica la similitud bilateral de las rodillas de los pacientes, con el fin de correlacionar el grado de asimetría con el dolor. Inicialmente, las imágenes de las rodillas fueron alineadas usando el algoritmo B-spline para su registro, después, un conjunto de métricas estándar fue cuantificado; finalmente, con estas métricas se propone un modelo multivariado para predecir el dolor de rodilla desarrollado en 48 meses. La metodología fue validada con 131 pacientes obtenidos de la base de datos de la Osteoarthritis Initiative (OAI). Los resultados sugieren que las métricas pueden ser asociadas con los puntajes de KellgrenLawrence; además, los modelos predicen significativamente el dolor crónico de rodilla con: AUC 0.756, 0.704 y 0.7113, al inicio, un año y dos años después, respectivamente.

Consensus on the diagnosis, treatment and follow-up of patients with Duchenne muscular dystrophy. / Consenso para el diagnóstico, tratamiento y seguimiento del paciente con distrofia muscular de Duchenne.

INTRODUCTION: Duchenne muscular dystrophy (DMD) is the most common myopathy in children, with a worldwide prevalence of approximately 0.5 cases per 10,000 male births. It is characterised by a progressive muscular weakness manifesting in early childhood, with the subsequent appearance of musculoskeletal, respiratory, and cardiac complications, causing disability, dependence, and premature death. Currently, DMD is mainly managed with multidisciplinary symptomatic treatment, with favourable results in terms of the progression of the disease. It is therefore crucial to establish clear, up-to-date guidelines enabling early detection, appropriate treatment, and monitoring of possible complications. DEVELOPMENT: We performed a literature search of the main biomedical databases for articles published in the last 10years in order to obtain an overview of the issues addressed by current guidelines and to identify relevant issues for which no consensus has yet been established. The degree of evidence and level of recommendation of the information obtained were classified and ordered according to the criteria of the American Academy of Neurology. CONCLUSIONS: DMD management should be multidisciplinary and adapted to the patient's profile and the stage of clinical progression. In addition to corticotherapy, treatment targeting gastrointestinal, respiratory, cardiac, and orthopaedic problems, as well as physiotherapy, should be provided with a view to improving patients' quality of life. Genetic studies play a key role in the management of the disease, both in detecting cases and potential carriers and in characterising the mutation involved and developing new therapies.

Optimization of Regioselective α-Glucosylation of Hesperetin Catalyzed by Cyclodextrin Glucanotransferase.

The regioselective α-glucosylation of hesperetin was achieved by a transglycosylation reaction catalyzed by cyclodextrin glucanotransferase (CGTase) from Thermoanaerobacter sp. using soluble starch as glucosyl donor. By combining mass spectrometry (ESI-TOF) and 2D-NMR analysis, the main monoglucosylated derivative was fully characterized (hesperetin 7-O-α-d-glucopyranoside). In order to increase the yield of monoglucoside, several reaction parameters were optimized: Nature and percentage of cosolvent, composition of the aqueous phase, glucosyl donor, temperature, and the concentrations of hesperetin and soluble starch. Under the optimal conditions, which included the presence of 30% of bis(2-methoxyethyl) ether as cosolvent, the maximum concentration of monoglucoside was approximately 2 mM, obtained after 24 h of reaction. To our knowledge, this is the first report of direct glucosylation of hesperetin employing free enzymes instead of whole cells.

General Linear Models for Pain Prediction in Knee Osteoarthritis: Data from the Osteoarthritis Initiative / Modelos Lineales Generales para Predecir Dolor en Osteoartritis de Rodilla: Datos de la "Osteoarthritis Initiative"

Abstract: Knee pain is the most common and disabling symptom in Osteoarthritis (OA). Joint pain is a late manifestation of the OA. In earlier stages of the disease changes in joint structures are shown. Also, formation of bony osteophytes, cartilage degradation, and joint space reduction which are some of the most common, among others. The main goal of this study is to associate radiological features with the joint pain symptom. Univariate and multivariate studies were performed using Bioinformatics tools to determine the relationship of future pain with early radiological evidence of the disease. All data was retrieved from the Osteoarthritis Initiative repository (OAI). A case-control study was done using available data from participants in OAI database. Radiological data was assessed with different OAI radiology groups. We have used quantitative and semi-quantitative scores to measure two different relations between radiological data in three different time points. The goal was to track the appearance and prevalence of pain as a symptom. All predictive models were statistically significant (P ≤ 0,05), obtaining the receiving operating characteristic (ROC) curves with their respective area under the curves (AUC) of 0.6516, 0.6174, and 0.6737 for T-0, T-1 and T-2 in quantitative analysis. For semi-quantitative an AU C of 0.6865, 0.6486, and 0.6406 for T-0, T-1 and T-2. The models obtained in the Bioinformatics study suggest that early joint structure changes can be associated with future joint pain. An image-based biomarker that could predict future pain, measured in early OA stages, could become a useful tool to improve the quality of life of people dealing OA.

Resumen: El dolor de rodilla es el síntoma más común y limitante de la Osteoartritis (OA), además de presentarse como una manifestación tardía de la enfermedad. Los cambios que ocurren en las estructuras de las articulaciones se presentan en las primeras etapas de la OA. Algunos de los cambios más comunes son la formación de osteofitos óseos, degradación del cartílago, y la reducción del espacio en la articulación, entre otros. El principal objetivo de este estudio es la asociación de características radiológicas con el síntoma de dolor de las articulaciones, para lo que fueron realizados dos estudios: univariado y multivariado, usando herramientas bioinformáticas para determinar la relación de futuro dolor con la evidencia radiológica temprana de la enfermedad. Todos los datos fueron recuperados de la Osteoarthritis Initiative repository (OAI). Este estudio de caso-control se llevó a cabo utilizando los datos disponibles de los participantes de la base de datos de la OAI. Los datos radiológicos fueron evaluados con diferentes grupos de radiología de la OAI. Fueron usadas puntuaciones cuantitativas y semi- cuantitativas para medir las dos diferentes relaciones entre los datos radiológicos en tres diferentes puntos de tiempo. El objetivo fue seguir la trayectoria de la aparición y prevalencia del dolor como síntoma. Todos los modelos predictivos fueron estadísticamente significativos (P ≤ 0,05). Para el análisis cuantitativo se calcularon las áreas bajo la curva (AUC): 0.6516, 0.6174, y 0.6737 para T-0, T-1 y T-2, y para el análisis semi-cuantitativo se calcularon las AU C: 0.6865, 0.6486, y 0.6406 para T-0, T-1 y T-2. Los modelos obtenidos en el estudio bioinformático sugieren que los cambios tempranos en la estructura de las articulaciones pueden estar asociados con el futuro dolor de rodilla. Un biomarcador basado en imágenes que pueda predecir el futuro dolor, medido en las primeras etapas de OA, podría convertirse en una herramienta útil para mejorar la calidad de vida de la gente que padece OA.

Current model systems for the study of preeclampsia.

Preeclampsia (PE) is a pregnancy complex disease, distinguished by high blood pressure and proteinuria, diagnosed after the 20th gestation week. Depending on the values of blood pressure, urine protein concentrations, symptomatology, and onset of disease there is a wide range of phenotypes, from mild forms developing predominantly at the end of pregnancy to severe forms developing in the early stage of pregnancy. In the worst cases severe forms of PE could lead to systemic endothelial dysfunction, eclampsia, and maternal and/or fetal death. Worldwide the fetal morbidity and mortality related to PE is calculated to be around 8% of the total pregnancies. PE still being an enigma regarding its etiology and pathophysiology, in general a deficient trophoblast invasion during placentation at first stage of pregnancy, in combination with maternal conditions are accepted as a cause of endothelial dysfunction, inflammatory alterations and appearance of symptoms. Depending on the PE multifactorial origin, several in vitro, in vivo, and in silico models have been used to evaluate the PE pathophysiology as well as to identify or test biomarkers predicting, diagnosing or prognosing the syndrome. This review focuses on the most common models used for the study of PE, including those related to placental development, abnormal trophoblast invasion, uteroplacental ischemia, angiogenesis, oxygen deregulation, and immune response to maternal-fetal interactions. The advances in mathematical and computational modeling of metabolic network behavior, gene prioritization, the protein-protein interaction network, the genetics of PE, and the PE prediction/classification are discussed. Finally, the potential of these models to enable understanding of PE pathogenesis and to evaluate new preventative and therapeutic approaches in the management of PE are also highlighted. Impact statement This review is important to the field of preeclampsia (PE), because it provides a description of the principal in vitro, in vivo, and in silico models developed for the study of its principal aspects, and to test emerging therapies or biomarkers predicting the syndrome before their evaluation in clinical trials. Despite the current advance, the field still lacking of new methods and original modeling approaches that leads to new knowledge about pathophysiology. The part of in silico models described in this review has not been considered in the previous reports.

High incidence of FXI deficiency in a Spanish town caused by 11 different mutations and the first duplication of F11: Results from the Yecla study.

INTRODUCTION: Factor XI (FXI) deficiency is a rare disorder with molecular heterogeneity in Caucasians but relatively frequent and molecularly homogeneous in certain populations. AIM: To characterize FXI deficiency in a Spanish town of 60 000 inhabitants. METHODS: A total of 324 764 APTT tests were screened during 20 years. FXI was evaluated by FXI:C and by Western blot. Genetic analysis of F11 was performed by sequencing, multiplex ligation-dependent probe amplification and genotyping. RESULTS: Our study identified 46 unrelated cases and 170 relatives with FXI deficiency carrying 12 different genetic defects. p.Cys56Arg, described as founder mutation in the French-Basque population, was identified in 109 subjects from 24 unrelated families. This mutation was also identified in 2% of the general population. p.Cys416Tyr, c.1693G>A and p.Pro538Leu were identified in 7, 6 and 2 unrelated families, respectively. NGS analysis of the whole F11 gene revealed a common haplotype for each of the four recurrent mutations, suggesting a founder effect. The analysis of plasma FXI of four p.Pro538Leu homozygous carriers revealed that this variant was not activated by FXIIa. We identified four mutations previously described in other Caucasian subjects with FXI deficiency (p.Lys536Asn; p.Thr322Ile, p.Arg268Cys and c.325G>A) and four new gene defects: p.(Cys599Tyr) potentially causing a functional deficiency, p.(Ile426Thr), p.(Ile592Thr) and the first worldwide duplication of 1653 bp involving exons 8 and 9. Bleeding was rare and mild. CONCLUSIONS: Our population-cohort study supplies new evidences that FXI deficiency in Caucasians is more common than previously thought and confirmed the wide underlying genetic heterogeneity, caused by both recurrent and sporadic mutations.

Evolvability meets biogeography: evolutionary potential decreases at high and low environmental favourability.

Understanding and forecasting the effects of environmental change on wild populations requires knowledge on a critical question: do populations have the ability to evolve in response to that change? However, our knowledge on how evolution works in wild conditions under different environmental circumstances is extremely limited. We investigated how environmental variation influences the evolutionary potential of phenotypic traits. We used published data to collect or calculate 135 estimates of evolvability of morphological traits of European wild bird populations. We characterized the environmental favourability of each population throughout the species' breeding distribution. Our results suggest that the evolutionary potential of morphological traits decreases as environmental favourability becomes high or low. Strong environmental selection pressures and high intra-specific competition may reduce species' evolutionary potential in low- and high- favourability areas, respectively. This suggests that species may be least able to adapt to new climate conditions at their range margins and at the centre. Our results underscore the need to consider the evolutionary potential of populations when studying the drivers of species distributions, particularly when predicting the effects of environmental change. We discuss the utility of integrating evolutionary dynamics into a biogeographical perspective to understand how environmental variation shapes evolutionary patterns. This approach would also produce more reliable predictions about the effect of environmental change on population persistence and therefore on biodiversity.

Epitaxial Growth of SrTiO3 Films on Cube-Textured Cu-Clad Substrates by PLD at Low Temperature Under Reducing Atmosphere.

The growth of epitaxial {001}<100> SrTiO3 (STO) on low-cost cube-textured Cu-based clad substrate at low temperature was carried out by means of pulsed laser deposition (PLD). STO film was deposited in one step under a reducing atmosphere (5% H2 and 95% Ar mixture) to prevent the oxidation of the metal surface. The optimization of PLD parameters leads to a sharpest biaxial texture at a temperature as low as 500 °C and a thickness of 500 nm with a (100) STO layer. The upper limit of highly textured STO thickness was also investigated. The maximum thickness which retains the best quality {001}<100> texture is 800 nm, since the texture is preserved not only through the layer but also on the surface. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) measurements showed that STO films are continuous, dense, and smooth with very low roughness (between 5 and 7 nm). This paper describes the development of STO layer by means of PLD in absence of oxygen throughout the process, suggesting an alternative and effective method for growing highly {001}<100> textured STO layer on low-cost metal substrates.

Uniparental disomy causes deficiencies of vitamin K-dependent proteins.

Essentials Vitamin K-dependent coagulant factor deficiency (VKCFD) is a rare autosomal recessive disorder. We describe a case of inherited VKCFD due to uniparental disomy. The homozygous mutation caused the absence of GGCX isoform 1 and overexpression of Δ2GGCX. Hepatic and non-hepatic vitamin K-dependent proteins must be assayed to monitor VKCFD treatment. SUMMARY: Background Inherited deficiency of all vitamin K-dependent coagulant factors (VKCFD) is a rare autosomal recessive disorder caused by mutations in the γ-glutamyl carboxylase gene (GGCX) or the vitamin K epoxide reductase gene (VKORC1), with great heterogeneity in terms of both clinical presentation and response to treatment. Objective To characterize the molecular basis of VKCFD in a Spanish family. Methods and Results Sequencing of candidate genes, comparative genomic hybridization and massive sequencing identified a new mechanism causing VKCFD in the proband. Uniparental disomy (UPD) of chromosome 2 caused homozygosity of a mutation (c.44-1G>A) resulting in aberrant GGCX splicing. This change contributed to absent expression of the mRNA coding for the full-length protein, and to four-fold overexpression of the smaller mRNA isoform lacking exon 2 (Δ2GGCX). Δ2GGCX might be responsible for two unexpected clinical observations in the patient: (i) increased plasma osteocalcin levels following vitamin K1 supplementation; and (ii) a mild non-bleeding phenotype. Conclusions Our study identifies a new autosomal disease, VKCFD1, caused by UPD. These data suggest that the Δ2GGCX isoform may retain enzymatic activity, and strongly encourage the evaluation of both hepatic and non-hepatic vitamin K-dependent proteins to assess differing responses to vitamin K supplementation in VKCFD patients.

Molecular characterization and developmental expression patterns of apolipoprotein A-I in Senegalese sole (Solea senegalensis Kaup).

The apolipoprotein A-I (ApoA-I) is an essential component of the high density lipoproteins (HDL). In this study, the cDNA and genomic sequences of this apolipoprotein were characterized for first time in Solea senegalensis. The predicted polypeptide revealed conserved structural features including ten repeats in the lipid-binding domain and some residues involved in cholesterol interaction and binding. The gene structure analysis identified four exons and three introns. Moreover, the synteny analysis revealed that apoA-I did not localize with other apolipoproteins indicating a divergent evolution with respect to the apoA-IV and apoE cluster. The phylogenetic analyses identified two distinct apoA-I paralogs in Ostariophysi (referred to as Ia and Ib) and only one (Ib) in Acanthopterygii. Whole-mount in situ hybridization located the apoA-I signal mainly in the yolk syncytial layer in lecitotrophic larval stages. Later at mouth opening, the mRNA signals were detected mainly in liver and intestine compatible with its role in the HDL formation. Moreover, a clear signal was detected in some regions of the brain, retina and neural cord suggesting a role in local regulation of cholesterol homeostasis. After metamorphosis, apoA-I was also detected in other tissues such as gills, head kidney and spleen suggesting a putative role in immunity. Expression analyses in larvae fed two diets with different triacylglycerol levels indicated that apoA-I mRNA levels were more associated to larval size and development than dietary lipid levels. Finally, qPCR analyses of immature and mature transcripts revealed distinct expression profiles suggesting a posttranscriptional regulatory mechanism.

Genomic characterization and expression analysis of four apolipoprotein A-IV paralogs in Senegalese sole (Solea senegalensis Kaup).

The apolipoprotein A-IV (ApoA-IV) plays a key role in lipid transport and feed intake regulation. In this work, four cDNA sequences encoding ApoA-IV paralogs were identified. Sequence analysis revealed conserved structural features including the common 33-codon block and nine repeated motifs. Gene structure analysis identified four exons and three introns except for apoA-IVAa1 (with only 3 exons). Synteny analysis showed that the four paralogs were structured into two clusters (cluster A containing apoA-IVAa1 and apoA-IVAa2 and cluster B with apoA-IVBa3 and apoA-IVBa4) linked to an apolipoprotein E. Phylogenetic analysis clearly separated the paralogs according to their cluster organization as well as revealed four subclades highly conserved in Acanthopterygii. Whole-mount analyses (WISH) in early larvae (0 and 1day post-hatch (dph)) showed that the four paralogs were mainly expressed in yolk syncytial layer surrounding the oil globules. Later, at 3 and 5dph, the four paralogs were mainly expressed in liver and intestine although with differences in their relative abundance and temporal expression patterns. Diet supply triggered the intensity of WISH signals in the intestine of the four paralogs. Quantification of mRNA abundance by qPCR using whole larvae only detected the induction by diet at 5dph. Moreover, transcript levels increased progressively with age except for apoA-IVAa2, which appeared as a low-expressed isoform. Expression analysis in juvenile tissues confirmed that the four paralogs were mainly expressed in liver and intestine and secondary in other tissues. The role of these ApoA-IV genes in lipid transport and the possible role of apoA-IVAa2 as a regulatory form are discussed.