Longitudinal Natural History of Pediatric Subjects Affected with Mucopolysaccharidosis IIIB.

OBJECTIVE: To characterize the longitudinal natural history of disease progression in pediatric subjects affected with mucopolysaccharidosis (MPS) IIIB. STUDY DESIGN: Sixty-five children with a confirmed diagnosis of MPS IIIB were enrolled into 1 of 2 natural history studies and followed for up to 4 years. Cognitive and adaptive behavior functions were analyzed in all subjects, and volumetric magnetic resonance imaging analysis of liver, spleen, and brain, as well as levels of heparan sulfate (HS) and heparan sulfate nonreducing ends (HS-NRE), were measured in a subset of subjects. RESULTS: The majority of subjects with MPS IIIB achieved an apex on both cognition and adaptive behavior age equivalent scales between age 3 and 6 years. Development quotients for both cognition and adaptive behavior follow a linear trajectory by which subjects reach a nadir with a score <25 for an age equivalent of 24 months by age 8 years on average and by 13.5 years at the latest. All tested subjects (n = 22) had HS and HS-NRE levels above the normal range in cerebrospinal fluid and plasma, along with signs of hepatomegaly. Subjects lost an average of 26 mL of brain volume (-2.7%) over 48 weeks, owing entirely to a loss of cortical gray matter (32 mL; -6.5%). CONCLUSIONS: MPS IIIB exists along a continuum based on cognitive decline and cortical gray matter atrophy. Although a few individuals with MPS IIIB have an attenuated phenotype, the majority follow predicted trajectories for both cognition and adaptive behavior. TRIAL REGISTRATION: ClinicalTrials.gov identifiers NCT02493998, NCT03227042, and NCT02754076.

Purinergic modulation of neuronal gap junction circuits in the CNS of the leech.

Gap junctions (GJs) are widely distributed in brains across the animal kingdom. To visualize the GJ- coupled networks of two major mechanosensory neurons in the ganglia of medicinal leeches, we injected these cells with the GJ-permeable tracer Neurobiotin. When diffusion time was limited to only 30 min, tracer coupling was highly variable for both cells, suggesting a possible modulation of GJ permeability. In invertebrates the innexins (homologs of vertebrate pannexins) form the GJs. Because extracellular adenosine triphosphate (ATP) modulates pannexin and leech innexin hemichannel permeability and is released by leech glial cells following injury, we tested the effects of bath application of ATP after the injection of Neurobiotin and observed a significant increase in the number of neurons tracer coupled to the sensory neurons. This effect required the elevation of intracellular Ca2+ and could be produced by bath application of caffeine. Conversely, scavenging endogenous extracellular ATP with the ATPase apyrase decreased the number of coupled cells. ATP also increased electrical conductance and tracer permeability between the bilateral Retzius neurons. This modulatory effect of ATP on GJ coupling was blocked by siRNA knockdown of a P1-like adenosine receptor. Finally, exposure of leech ganglia to extracellular ATP induced a characteristic low frequency (<0.3 Hz) rhythmic bursting activity that was roughly synchronous among multiple neurons, a behavior that was significantly attenuated by the GJ blocker octanol. These findings highlight the mediation by ATP of a robust physiological mechanism for modifying neuronal circuits by rapidly recruiting neurons into active networks and entraining synchronized bursting activity.

Diagnostic evaluation, monitoring, and perioperative management of spinal cord compression in patients with Morquio syndrome.

Mucopolysaccharidosis IVA is an autosomal recessive condition caused by mutations in the GALNS gene, which encodes N-acetylgalactosamine-6-sulfatase, also called galactosamine-6-sulfatase (GALNS). A reduction in or absence of effective GALNS leads to faulty catabolism of keratan sulfate and chondroitin-6-sulfate within the lysosome; their accumulation causes cell, tissue, and organ dysfunction. The connective tissue, cartilage, ligaments, and bone of patients with Morquio A syndrome are particularly affected. Patients with Morquio A syndrome are at high risk of neurological complications because of their skeletal abnormalities; many patients are in danger of cervical myelopathy due to odontoid hypoplasia and ligamentous laxity leading to atlantoaxial subluxation. The multisystemic involvement of patients with Morquio A syndrome requires treatment by multidisciplinary teams; not all members of these teams may be aware of the potential for subluxation and quadriparesis. A multinational, multidisciplinary panel of 10 skeletal dysplasia or Morquio A syndrome specialists convened in Miami, FL on December 7 and 8, 2012 to develop consensus recommendations for early identification and effective management of spinal cord compression, for anesthesia and surgical best practices, and for effectual cardiac and respiratory management in patients with Morquio A syndrome. The target audience for these recommendations includes any physician who may encounter a patient with Morquio A syndrome, however doctors who do not have access to the full spectrum of specialists and resources needed to support patients with Morquio A syndrome should attempt to refer patients to a center that does. Physicians who manage Morquio A syndrome or comorbid conditions within specialty centers should review these expert panel recommendations and fully understand the implications of spinal cord instability for their own practices.

Bioengineered skin constructs based on mesenchymal stromal cells and acellular dermal matrix exposed to inflammatory microenvironment releasing growth factors involved in skin repair.

BACKGROUND: Skin tissue engineering is a rapidly evolving field of research that effectively combines stem cells and biological scaffolds to replace damaged tissues. Human Wharton's jelly mesenchymal stromal cells (hWJ-MSCs) are essential to generate tissue constructs, due to their potent immunomodulatory effects and release of paracrine factors for tissue repair. Here, we investigated whether hWJ-MSC grown on human acellular dermal matrix (hADM) scaffolds and exposed to a proinflammatory environment maintain their ability to produce in vitro growth factors involved in skin injury repair and promote in vivo wound healing. METHODS: We developed a novel method involving physicochemical and enzymatic treatment of cadaveric human skin to obtain hADM scaffold. Subsequently, skin bioengineered constructs were generated by seeding hWJ-MSCs on the hADM scaffold (construct 1) and coating it with human platelet lysate clot (hPL) (construct 2). Either construct 1 or 2 were then incubated with proinflammatory cytokines (IL-1α, IL-1ß, IL-6, TNF-α) for 12, 24, 48, 72 and 96 h. Supernatants from treated and untreated constructs and hWJ-MSCs on tissue culture plate (TCP) were collected, and concentration of the following growth factors, bFGF, EGF, HGF, PDGF, VEGF and Angiopoietin-I, was determined by immunoassay. We also asked whether hWJ-MSCs in the construct 1 have potential toward epithelial differentiation after being cultured in an epithelial induction stimulus using an air-liquid system. Immunostaining was used to analyze the synthesis of epithelial markers such as filaggrin, involucrin, plakoglobin and the mesenchymal marker vimentin. Finally, we evaluated the in vivo potential of hADM and construct 1 in a porcine full-thickness excisional wound model. RESULTS: We obtained and characterized the hADM and confirmed the viability of hWJ-MSCs on the scaffold. In both constructs without proinflammatory treatment, we reported high bFGF production. In contrast, the levels of other growth factors were similar to the control (hWJ-MSC/TCP) with or without proinflammatory treatment. Except for PDGF in the stimulated group. These results indicated that the hADM scaffold maintained or enhanced the production of these bioactive molecules by hWJ-MSCs. On the other hand, increased expression of filaggrin, involucrin, and plakoglobin and decreased expression of vimentin were observed in constructs cultured in an air-liquid system. In vivo experiments demonstrated the potential of both hADM and hADM/hWJ-MSCs constructs to repair skin wounds with the formation of stratified epithelium, basement membrane and dermal papillae, improving the appearance of the repaired tissue. CONCLUSIONS: hADM is viable to fabricate a tissue construct with hWJ-MSCs able to promote the in vitro synthesis of growth factors and differentiation of these cells toward epithelial lineage, as well as, promote in a full-thickness skin injury the new tissue formation. These results indicate that hADM 3D architecture and its natural composition improved or maintained the cell function supporting the potential therapeutic use of this matrix or the construct for wound repair and providing an effective tissue engineering strategy for skin repair.

A phase I/II study on intracerebroventricular tralesinidase alfa in patients with Sanfilippo syndrome type B.

BackgroundSanfilippo type B is a mucopolysaccharidosis (MPS) with a major neuronopathic component characterized by heparan sulfate (HS) accumulation due to mutations in the NAGLU gene encoding alfa-N-acetyl-glucosaminidase. Enzyme replacement therapy for neuronopathic MPS requires efficient enzyme delivery throughout the brain in order to normalize HS levels, prevent brain atrophy, and potentially delay cognitive decline.MethodsIn this phase I/II open-label study, patients with MPS type IIIB (n = 22) were treated with tralesinidase alfa administered i.c.v. The patients were monitored for drug exposure; total HS and HS nonreducing end (HS-NRE) levels in both cerebrospinal fluid (CSF) and plasma; anti-drug antibody response; brain, spleen, and liver volumes as measured by MRI; and cognitive development as measured by age-equivalent (AEq) scores.ResultsIn the Part 1 dose escalation (30, 100, and 300 mg) phase, a 300 mg dose of tralesinidase alfa was necessary to achieve normalization of HS and HS-NRE levels in the CSF and plasma. In Part 2, 300 mg tralesinidase alfa sustained HS and HS-NRE normalization in the CSF and stabilized cortical gray matter volume (CGMV) over 48 weeks of treatment. Resolution of hepatomegaly and a reduction in spleen volume were observed in most patients. Significant correlations were also established between the change in cognitive AEq score and plasma drug exposure, plasma HS-NRE levels, and CGMV.ConclusionAdministration of tralesinidase alfa i.c.v. effectively normalized HS and HS-NRE levels as a prerequisite for clinical efficacy. Peripheral drug exposure data suggest a role for the glymphatic system in altering tralesinidase alfa efficacy.Trial registrationClinicaltrials.gov NCT02754076.FUNDINGBioMarin Pharmaceutical Inc. and Allievex Corporation.

Ethical and economic considerations of rare diseases in ethnic minorities: the case of mucopolysaccharidosis VI in Colombia.

Mucopolysaccharidosis VI is an autosomal recessive lysosomal storage disorder associated with severe disability and premature death. The presence of a mucopolysaccharidosis-like disease in indigenous ethnic groups in Colombia can be inferred from archaeological findings. There are several indigenous patients with mucopolysaccharidosis VI currently receiving enzyme replacement therapy. We discuss the ethical and economic considerations, regarding both direct and indirect costs, of a high-cost orphan disease in a marginalised minority population in a developing country.

Determination of genotypic and clinical characteristics of Colombian patients with mucopolysaccharidosis IVA.

BACKGROUND: As mucopolysaccharidosis IVA (MPS IVA) is the most frequent MPS in Colombia, this paper aims to describe its clinical and mutational characteristics in 32 diagnosed patients included in this study. METHODS: Genotyping was completed by amplification and Sanger sequencing of the GALNS gene. The SWISS-model platform was used for bioinformatic analysis, and mutant proteins were generated by homology from the wild-type GALNS code 4FDI template from the Protein Data Bank (PDB) database. Docking was performed using the GalNAc6S ligand (PubChem CID: 193456) by AutoDock Vina 1.0 and visualized in PyMOL and LigPlot+. RESULTS: Eleven variants were identified, and one new pathogenic variant was described in the heterozygous state, which is consistent with genotype c. 319 G> T or p.Ala107Ser. The pathogenic variant c.901G>T or p.Gly301Cys was the most frequent mutation with 51.6% of alleles. Docking revealed affinity energy of -5.9 Kcal/mol between wild-type GALNS and the G6S ligand. Some changes were evidenced at the intermolecular interaction level, and affinity energy for each mutant decreased. CONCLUSION: Clinical variables and genotypic analysis were similar to those reported for other world populations. Genotypic data showed greater allelic heterogeneity than those previously reported. Bioinformatics tools showed differences in the binding interactions of mutant proteins with the G6S ligand, in regard the wild-type GALNS.

Recommendations for Evaluation and Management of Pain in Patients With Mucopolysaccharidosis in Latin America.

The mucopolysaccharidosis (MPS) constitutes a heterogeneous group of rare genetic disorders caused by enzymatic deficiencies that lead to the accumulation of glycosaminoglycans. Several types of MPS are described, historically numbered from I to IX. Clinical observations strongly suggest the presence of chronic pain in patients with all types of MPS. There are few data in the literature on the evaluation and management of pain in these patients, a fact that can compromise the quality of life even more. Professionals with extensive experience in the care for patients with MPS held a meeting in April 2017 to discuss and propose recommendations for the evaluation and management of pain in patients with MPS in Latin America. This article summarizes the content of the discussions and presents the recommendations produced at the meeting. Patients with MPS present joint, bone, and muscle pain, as well as entrapment syndromes (spinal, optic nerve, carpal tunnel). The panel suggests the use of the following instruments for pain assessment: Face, Legs, Activity, Cry and Consolability Scale for children of up to four years of age and patients unable to communicate their pain; Child Health Assessment Questionnaire Scale; Facial Pain Scale and Numerical Pain Scale for patients of five to <18 years of age; Brief Pain Inventory and Short Form Health Survey 36 scales for patients aged 18 years or older. Based on the scores verified in these scales, the panel proposes pharmacological interventions for pain relief in this population of patients.

Development of a Clinical Algorithm for the Early Diagnosis of Mucopolysaccharidosis III

Abstract Mucopolysaccharidosis III (MPS III) is a rare inherited metabolic disease primarily affecting the central nervous system, leading to developmental and/or speech regression. Early diagnosis of the disease is important to introduce appropriate management measures and to optimize therapeutic outcomes. The diagnosis of MPS III is often significantly delayed due to the rarity of the disease, the more attenuated somatic presentation compared to other MPS types, and the symptom overlap with other developmental disorders. To shorten the time to diagnosis, a list of eight early signs and symptoms was identified through an expert system approach by a global, multidisciplinary working group of 13 specialists with expertise in various aspects of MPS and developmental disorders and three parents of MPS III patients. Coarse facial features and persistent hirsutism or prominent, thick eyebrows were identified as the most important MPS III early signs. The list of eight early MPS III signs and symptoms is the first step towards the development of a clinical algorithm aiming to identify neonates and infants with MPS III before the onset of neurocognitive damage, ultimately shortening the diagnostic journey of MPS III patients.

[Intrauterine growth retardation, low birth weight and prematurity in neonates of pregnant women with malaria in Colombia]. / Retardo no crescimento intrauterino, baixo peso ao nascer e prematuridade em recém-nascidos de grávidas com malária, na Colômbia.

INTRODUCTION: Association between malaria and pregnancy complications, such as prematurity, intrauterine growth restriction, low birthweight and infant mortality has been reported. These effects have been studied widely in areas hyperendemic for malaria, but studies in low-endemic areas are scarce. The study investigated the relation between gestational malaria and low birthweight and intrauterine growth retardation in neonates of a malarial endemic region in Colombia, between 1993 and 2007. METHODS: The pattern of development in 1,716 neonates of women with and without malaria infection during pregnancy was evaluated in a cohort study. A total of 394 infected (27% by P. falciparum and 73% by P. vivax) and 1,322 noninfected pregnant women were followed. RESULTS: Exposure to gestational malaria was associated with increased risk of low birth weight (RR = 1.37; 1.03-1.83), short height (RR = 1.52; 1.25-1.85), intrauterine growth retardation (RR = 1.29; 1.0-1.66) and prematurity (RR = 1.68; 1.3-2.17). Prematurity was 77% higher in infants of mothers with malaria by P. falciparum than infants of mothers with malaria by P. vivax (RR = 1.77; 1.2-2.6). CONCLUSIONS: Low birth weight and intrauterine growth retardation were associated with malaria during pregnancy. Infection with P. vivax was related with adverse effects on the newborn, similar to that reported for P. falciparum.

Retardo no crescimento intrauterino, baixo peso ao nascer e prematuridade em recém-nascidos de grávidas com malária, na Colômbia / Intrauterine growth retardation, low birth weight and prematurity in neonates of pregnant women with malaria in Colombia

INTRODUÇÃO: É frequente a associação da malária com complicações como prematuridade, retardo no crescimento intrauterino, baixo peso ao nascer e mortalidade infantil, efeitos pouco estudados em áreas hipoendêmicas para malaria. O objetivo deste estudo foi analisar a relação da malária gestacional com estes efeitos em recém-nascidosnuma região endêmica para malária na Colômbia, entre 1993 e 2007. MÉTODOS: Foram estudadas as características em 1.716 recém-nascidos num estudo de coorte. Fez-se seguimento em 394 gestantes com malária (27 por cento por Plasmodium falciparum e 73 por cento por P. vivax) e 1.322 sem malária. RESULTADOS: Foi encontrada uma relação entre a exposição à malária na gestação e o risco maior de baixo peso ao nascer (RR = 1,37; 1,03-1,83), assim como estatura baixa (RR = 1,52; 1,25-1,85), retardo no crescimento intrauterino (RR = 1,29; 1,0-1,66) e prematuridade (RR = 1,68; 1,3-2,17). A frequência de nascimentos prematuros foi maior nas mães com malária por P. falciparum (77 por cento) que aquelas com P. vivax (RR = 1,77; IC 95 por cento: 1,2-2,6). CONCLUSÕES: O baixo peso ao nascer e o retardo no crescimento foi associado com malária na gestação na Colômbia. A infecção por P. vivax foi relacionada com efeitos adversos sobre o recém-nascido, de modo semelhante em relação ao P. falciparum.

INTRODUCTION: Association between malaria and pregnancy complications, such as prematurity, intrauterine growth restriction, low birthweight and infant mortality has been reported. These effects have been studied widely in areas hyperendemic for malaria, but studies in low-endemic areas are scarce. The study investigated the relation between gestational malaria and low birthweight and intrauterine growth retardation in neonates of a malarial endemic region in Colombia, between 1993 and 2007. METHODS: The pattern of development in 1,716 neonates of women with and without malaria infection during pregnancy was evaluated in a cohort study. A total of 394 infected (27 percent by P. falciparum and 73 percent by P. vivax) and 1,322 noninfected pregnant women were followed. RESULTS: Exposure to gestational malaria was associated with increased risk of low birth weight (RR = 1.37; 1.03-1.83), short height (RR = 1.52; 1.25-1.85), intrauterine growth retardation (RR = 1.29; 1.0-1.66) and prematurity (RR = 1.68; 1.3-2.17). Prematurity was 77 percent higher in infants of mothers with malaria by P. falciparum than infants of mothers with malaria by P. vivax (RR = 1.77; 1.2-2.6). CONCLUSIONS: Low birth weight and intrauterine growth retardation were associated with malaria during pregnancy. Infection with P. vivax was related with adverse effects on the newborn, similar to that reported for P. falciparum.

Tamizaje para enfermedades neurovegetativas: resultados de cuatro años de investigación / Screening for neurological degenerative diseases. Results of four years of research

El objetivo de la investigación fue promover y desarrollar un protocolo para detectar enfermedades neurodegenerativas, asociadas a errores innatos del metabolismo, mediante la cuantificación de la actividad enzimática de arilsulfatasa A, de hexosaminidadas totales y de hexosaminidasa A y B, de betagalactosidasa en suero y de la detección de oligosacáridos y ácido sialico en orina. La confirmación de la alteración metabólica se realizó mediante la determinación de las enzimas en leucocitos. el estudio se realizó mediante la aplicación de un tamiz bioquímico preestablecido, entre enero de 1997 y junio de 2001, a 165 pacientes con características clínicas que hacian sospechar un desorden neurodegenerativo. Se hallaron 38 pacientes con anormalidades enzimátcas 23/100 mediante el tamiz bioquímico y cromatográfico. Sometidos a la cuantificación enzimática en leucocitos, el defecto se confirmó en solo nueve pacientes 4.2/100. El tamizaje de las enfermedades neurodegenerativas es una ayuda diagnóstica útil en pacientes con involución de su desarrollo psicomotor. Conviene resaltar que la aplicación de un tamiz bioquímico de estas caracterisitcas no tiene antecedente en Colombia ni en Latinoamérica.