A new case of platelet-type von Willebrand disease supports the recent findings of gain-of-function GP1BA variants outside the C-terminal disulphide loop enhances affinity for von Willebrand factor.

Platelet-type von Willebrand disease (PT-VWD) is a rare autosomal dominant bleeding disorder characterized by an increased ristocetin-induced platelet aggregation (RIPA) and enhanced affinity of platelet glycoprotein Ibα (GPIbα) to von Willebrand factor (VWF). To date, only seven variants have been described with this gain-of-function effect, most of them located in the C-terminal disulphide loop of the VWF-binding domain of GPIbα. We herein describe a patient with moderate bleeding symptoms, mild thrombocytopenia and increased RIPA. By direct sequencing of GP1BA, a novel leucine-rich repeat heterozygous variant was identified (c.580C>T; predictably p.Leu194Phe), strongly suggestive as being the underlying cause for the PT-VWD phenotype of our patient.

Novel manifestations of immune dysregulation and granule defects in gray platelet syndrome.

Gray platelet syndrome (GPS) is a rare recessive disorder caused by biallelic variants in NBEAL2 and characterized by bleeding symptoms, the absence of platelet α-granules, splenomegaly, and bone marrow (BM) fibrosis. Due to the rarity of GPS, it has been difficult to fully understand the pathogenic processes that lead to these clinical sequelae. To discern the spectrum of pathologic features, we performed a detailed clinical genotypic and phenotypic study of 47 patients with GPS and identified 32 new etiologic variants in NBEAL2. The GPS patient cohort exhibited known phenotypes, including macrothrombocytopenia, BM fibrosis, megakaryocyte emperipolesis of neutrophils, splenomegaly, and elevated serum vitamin B12 levels. Novel clinical phenotypes were also observed, including reduced leukocyte counts and increased presence of autoimmune disease and positive autoantibodies. There were widespread differences in the transcriptome and proteome of GPS platelets, neutrophils, monocytes, and CD4 lymphocytes. Proteins less abundant in these cells were enriched for constituents of granules, supporting a role for Nbeal2 in the function of these organelles across a wide range of blood cells. Proteomic analysis of GPS plasma showed increased levels of proteins associated with inflammation and immune response. One-quarter of plasma proteins increased in GPS are known to be synthesized outside of hematopoietic cells, predominantly in the liver. In summary, our data show that, in addition to the well-described platelet defects in GPS, there are immune defects. The abnormal immune cells may be the drivers of systemic abnormalities such as autoimmune disease.

Thrombocytopenia-Absent Radius Syndrome: Descriptions of Three New Cases and a Novel Splicing Variant in RBM8A That Expands the Spectrum of Null Alleles.

Thrombocytopenia-absent radius (TAR) syndrome is a rare congenital disorder characterized by the bilateral absence of the radius and thrombocytopenia, and sometimes by other skeletal, gastrointestinal, cardiac, and renal abnormalities. The underlying genetic defect is usually the compound inheritance of a microdeletion in 1q21.1 (null allele) and a low-frequency, non-coding single nucleotide variant (SNV) in the RBM8A gene (hypomorphic allele). We report three new cases from two unrelated families. The two siblings presented the common genotype, namely the compound heterozygosity for a 1q21.1 microdeletion and the hypomorphic SNV c.-21G>A in RBM8A, whereas the third, unrelated patient presented a rare genotype comprised by two RBM8A variants: c.-21G>A (hypomorphic allele) and a novel pathogenic variant, c.343-2A>G (null allele). Of the eight documented RBM8A variants identified in TAR syndrome patients, four have hypomorphic expression and four behave as null alleles. The present report expands the RBM8A null allele spectrum and corroborates the particularities of RBM8A involvement in TAR syndrome pathogenesis.

Can increased load carriage affect lower limbs kinematics during military gait?

The aim of this study was to investigate if increased load carriage, in male military personnel, can affect the lower limbs kinematics. Twelve male military volunteers from the Portuguese Army were recruited and evaluated in an unloaded and loaded gait condition. Linear kinematics and lower limbs joint angle at heel strike, midstance and toe off were calculated. The stance, swing and double support times were found to be different between load conditions (p < 0.05). There was an interaction between load and limb (p < 0.05) for joint angles, during midstance, with limbs performing different movements in the frontal plane during loaded gait. Load increase had a different effect on the right knee, with a reduction in the abduction (valgus). This study may be beneficial in offering suggestion to improve the performance of gait with load and in an attempt to help prevent possible injuries. Practitioner summary: Increased load can affect lower limbs of male soldiers at the pelvic, hip and knee angles on the frontal plane, which can alter the joint force distribution. While these alterations may indicate protective mechanics, load management procedures should be implemented along with gait monitoring to avoid negative effects in performance.

Increasing involvement of CAPN1 variants in spastic ataxias and phenotype-genotype correlations.

Spastic ataxias are rare neurogenetic disorders involving spinocerebellar and pyramidal tracts. Many genes are involved. Among them, CAPN1, when mutated, is responsible for a complex inherited form of spastic paraplegia (SPG76). We report the largest published series of 21 novel patients with nine new CAPN1 disease-causing variants and their clinical characteristics from two European university hospitals (Paris and Stockholm). After a formal clinical examination, causative variants were identified by next-generation sequencing and confirmed by Sanger sequencing. CAPN1 variants are a rare cause (~ 1.4%) of young-adult-onset spastic ataxia; however, together with all published cases, they allowed us to better describe the clinical and genetic spectra of this form. Truncating variants are the most frequent, and missense variants lead to earlier age at onset in favor of an additional deleterious effect. Cerebellar ataxia with cerebellar atrophy, dysarthria and lower limb weakness are often associated with spasticity. We also suggest that cognitive impairment and depression should be assessed specifically in the follow-up of SPG76 cases.

Congenital ataxia due to novel variant in ATP8A2.

Congenital ataxias are a heterogeneous group of disorders characterized by congenital or early-onset ataxia. Here, we describe two siblings with congenital ataxia, who acquired independent gait by age 4 years. After 16 years of follow-up they presented near normal cognition, cerebellar ataxia, mild pyramidal signs, and dystonia. On exome sequencing, a novel homozygous variant (c.1580-18C > G - intron 17) in ATP8A2 was identified. A new acceptor splice site was predicted by bioinformatics tools, and functionally characterized through a minigene assay. Minigene constructs were generated by PCR-amplification of genomic sequences surrounding the variant of interest and cloning into the pCMVdi vector. Altered splicing was evaluated by expressing these constructs in HEK293T cells. The construct with the c.1580-18C > G homozygous variant produced an aberrant transcript, leading to retention of 17 bp of intron 17, by the use of an alternative acceptor splice site, resulting in a premature stop codon by insertion of four amino acids. These results allowed us to establish this as a disease-causing variant and expand ATP8A2-related disorders to include less severe forms of congenital ataxia.

DFT study of H2 adsorption at a Cu-SSZ-13 zeolite: a cluster approach.

In this work the H2 adsorption at a Cu(i)-SSZ-13 exchanged zeolite was theoretically investigated. A systematic cluster approach was used and different density functionals (B3LYP, B3LYP-D3(BJ), M06L, PBE, PBE-D3(BJ) and ωB97XD) and a def2-SVP basis set were benchmarked. In order to select the best approach to the H2 adsorption over a Cu(i)-SSZ-13 cluster with 78 atoms (16 T-sites), two main tasks were performed: (1) a comparison between theoretical and experimental structures and (2) a comparison between theoretical and experimental adsorption enthalpies. By employing the most suitable functional - the ωB97X-D - the H2 interaction with the zeolite structure was studied by means of NBO, NCI, AIM and DLPNO-CCSD(T)/LED analyses.

CalDAG-GEFI Deficiency in a Family with Symptomatic Heterozygous and Homozygous Carriers of a Likely Pathogenic Variant in RASGRP2.

RASGRP2 encodes the calcium and diacylglycerol (DAG)-regulated guanine nucleotide exchange factor I (CalDAG-GEFI) identified as a Rap1-activating molecule. Pathogenic variants previously identified in RASGRP2 allowed the characterization of CalDAG-GEFI deficiency as a non-syndromic, autosomal recessive platelet function disease. We report on the clinical manifestations and laboratory features of a Portuguese family with a likely pathogenic variant in RASGRP2 (c.999G>C leading to a p.Lys333Asn change in the CDC25 catalytic domain of CalDAG-GEFI) and discuss the contribution of this variant to the disease manifestations. Based on the study of this family with one homozygous patient and five heterozygous carriers and on a critical analysis of the literature, we challenge previous knowledge that CalDAG-GEFI deficiency only manifests in homozygous patients. Our data suggest that at least for the RASGRP2 variant reported herein, there is a phenotypic expression, albeit milder, in heterozygous carriers.

Bulk pH and Carbon Source Are Key Factors for Calcium Phosphate Granulation.

Recovery of calcium phosphate granules (CaP granules) from high-strength wastewater is an opportunity to reduce the natural phosphorus (P) scarcity, geographic imbalances of P reserves, and eutrophication. Formation of CaP granules was previously observed in an upflow anaerobic sludge bed (UASB) reactor treating source separated black water and is enhanced by Ca2+ addition. However, the required operating conditions and influent composition for CaP granulation are still unknown. In this study, we have experimentally demonstrated that the carbon source and bulk pH are crucial parameters for the formation and growth of CaP granules in a UASB reactor, operating at relatively low upflow velocity (<1 cm h-1). Degradation of glucose yielded sufficient biomass (microbial cells and extracellular biopolymers) to cover crystal and amorphous calcium phosphate [Ca x(PO4) y], forming CaP granules. Influent only containing volatile fatty acids as the carbon source did not generate CaP granules. Moreover, bulk pH between 7.0 and 7.5 was crucial for the enrichment of Ca x(PO4) y in the granules over bulk precipitation. Bulk pH 8 reduced the Ca x(PO4) y enrichment in granules of >1.4 mm diameter from 9 to 5 wt % P. Moreover, for bulk pH 7.5, co-precipitation of CaCO3 with Ca x(PO4) y was reduced.

Update on the Genetics of Spastic Paraplegias.

PURPOSE OF REVIEW: Hereditary spastic paraplegias are a genetically heterogeneous group of neurological disorders. Patients present lower limb weakness and spasticity, complicated in complex forms by additional neurological signs. We review here the major steps toward understanding the molecular basis of these diseases made over the last 10 years. RECENT FINDINGS: Our perception of the intricate connections between clinical, genetic, and molecular aspects of neurodegenerative disorders has radically changed in recent years, thanks to improvements in genetic approaches. This is particularly true for hereditary spastic paraplegias, for which > 60 genes have been identified, highlighting (i) the considerable genetic heterogeneity of this group of clinically diverse disorders, (ii) the fuzzy border between recessive and dominant inheritance for several mutations, and (iii) the overlap of these mutations with other neurological conditions in terms of their clinical effects. Several hypotheses have been put forward concerning the pathophysiological mechanisms involved, based on the genes implicated and their known function and based on studies on patient samples and animal models. These mechanisms include mainly abnormal intracellular trafficking, changes to endoplasmic reticulum shaping and defects affecting lipid metabolism, lysosome physiology, autophagy, myelination, and development. Several causative genes affect multiple of these functions, which are, most of the time, interconnected. Recent major advances in our understanding of these diseases have revealed unifying pathogenic models that could be targeted in the much-needed development of new treatments.

Rare Neurodegenerative Diseases: Clinical and Genetic Update.

More than 600 human disorders afflict the nervous system. Of these, neurodegenerative diseases are usually characterised by onset in late adulthood, progressive clinical course, and neuronal loss with regional specificity in the central nervous system. They include Alzheimer's disease and other less frequent dementias, brain cancer, degenerative nerve diseases, encephalitis, epilepsy, genetic brain disorders, head and brain malformations, hydrocephalus, stroke, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS or Lou Gehrig's Disease), Huntington's disease, and Prion diseases, among others. Neurodegeneration usually affects, but is not limited to, the cerebral cortex, intracranial white matter, basal ganglia, thalamus, hypothalamus, brain stem, and cerebellum. Although the majority of neurodegenerative diseases are sporadic, Mendelian inheritance is well documented. Intriguingly, the clinical presentations and neuropathological findings in inherited neurodegenerative forms are often indistinguishable from those of sporadic cases, suggesting that converging genomic signatures and pathophysiologic mechanisms underlie both hereditary and sporadic neurodegenerative diseases. Unfortunately, effective therapies for these diseases are scarce to non-existent. In this chapter, we highlight the clinical and genetic features associated with the rare inherited forms of neurodegenerative diseases, including ataxias, multiple system atrophy, spastic paraplegias, Parkinson's disease, dementias, motor neuron diseases, and rare metabolic disorders.

Biomechanics, energetics and coordination during extreme swimming intensity: effect of performance level.

The present study aimed to examine how high- and low-speed swimmers organise biomechanical, energetic and coordinative factors throughout extreme intensity swim. Sixteen swimmers (eight high- and eight low-speed) performed, in free condition, 100-m front crawl at maximal intensity and 25, 50 and 75-m bouts (at same pace as the previous 100-m), and 100-m maximal front crawl on the measuring active drag system (MAD-system). A 3D dual-media optoelectronic system was used to assess speed, stroke frequency, stroke length, propelling efficiency and index of coordination (IdC), with power assessed by MAD-system and energy cost by quantifying oxygen consumption plus blood lactate. Both groups presented a similar profile in speed, power output, stroke frequency, stroke length, propelling efficiency and energy cost along the effort, while a distinct coordination profile was observed (F(3, 42) = 3.59, P = 0.04). Speed, power, stroke frequency and propelling efficiency (not significant, only a tendency) were higher in high-speed swimmers, while stroke length and energy cost were similar between groups. Performing at extreme intensity led better level swimmers to achieve superior speed due to higher power and propelling efficiency, with consequent ability to swim at higher stroke frequencies. This imposes specific constraints, resulting in a distinct IdC magnitude and profile between groups.

Alteration of ornithine metabolism leads to dominant and recessive hereditary spastic paraplegia.

Hereditary spastic paraplegias are heterogeneous neurological disorders characterized by a pyramidal syndrome with symptoms predominantly affecting the lower limbs. Some limited pyramidal involvement also occurs in patients with an autosomal recessive neurocutaneous syndrome due to ALDH18A1 mutations. ALDH18A1 encodes delta-1-pyrroline-5-carboxylate synthase (P5CS), an enzyme that catalyses the first and common step of proline and ornithine biosynthesis from glutamate. Through exome sequencing and candidate gene screening, we report two families with autosomal recessive transmission of ALDH18A1 mutations, and predominant complex hereditary spastic paraplegia with marked cognitive impairment, without any cutaneous abnormality. More interestingly, we also identified monoallelic ALDH18A1 mutations segregating in three independent families with autosomal dominant pure or complex hereditary spastic paraplegia, as well as in two sporadic patients. Low levels of plasma ornithine, citrulline, arginine and proline in four individuals from two families suggested P5CS deficiency. Glutamine loading tests in two fibroblast cultures from two related affected subjects confirmed a metabolic block at the level of P5CS in vivo. Besides expanding the clinical spectrum of ALDH18A1-related pathology, we describe mutations segregating in an autosomal dominant pattern. The latter are associated with a potential trait biomarker; we therefore suggest including amino acid chromatography in the clinico-genetic work-up of hereditary spastic paraplegia, particularly in dominant cases, as the associated phenotype is not distinct from other causative genes.

An alternative interpretation of the ultracold methylhydroxycarbene rearrangement mechanism: cooperative effects.

Recent studies have reported surprising results related to the rearrangement of carbenes under ultracold conditions, making use of sophisticated models of quantum tunnelling to explain the observed phenomena. Here, we demonstrate that a methylhydroxycarbene (H3C-C-OH) rearrangement is possible by making changes in molecularity (i.e., through cooperative effects), owing to intermolecular hydrogen bond/H-transfer. The model used for accomplishing these changes in molecularity suggests the occurrence of two chemical species during the rearrangement and preferential formation of acetaldehyde. We propose an alternative interpretation for the methylhydroxycarbene rearrangement, as well as for a bimolecular isomerization mechanism for acetaldehyde formation with an activation barrier, Ea, of +0.25 kcal mol(-1), relative to 1a' (−8.06 kcal mol(-1) relative to 1a); this barrier is lower than that required by H-tunnelling as proposed by Schreiner et al. We also note that the mechanism for obtaining vinyl alcohol leads to the simultaneous formation of acetaldehyde through an Ea of +13.53 kcal mol(-1), relative to 1a (+0.93 kcal mol(-1) relative to 1b), again confirming the predominant presence of acetaldehyde.

Blood transfusion after primary total knee arthroplasty can be significantly minimised through a multimodal blood-loss prevention approach.

PURPOSE: Our aim was to clarify the effective decrease in blood transfusion after primary total knee arthroplasty (TKA) from a multimodal blood-loss prevention approach (MBLPA) and the related risk factors of blood transfusion. METHODS: We retrospectively compared the rate of postoperative blood transfusion in 418 cases of primary TKA during 2010 from a single institution with two different groups of patients, allocating cases to the group with MBLPA (group 1, study group, N = 71) and controls to the group without MBLPA (group 2, standard group, N = 347). MBLPA procedure included pre-operative haemoglobin (Hb) optimisation; femoral canal obturation; limited incision and release; peri- and intra-articular use of saline with adrenalin, morpheic chloride, tobramycin, betamethasone and ropivacaine; tourniquet release after skin closure; 24 hour drain under atmospheric pressure; and two doses of tranexamic acid (TXA) i.v.. In the control group, surgeons followed the standard procedure without blood-saving techniques. Case-control comparison and blood transfusion risk factors were analysed. RESULTS: Group 1 had a zero transfusion rate (0/71), whereas 27.4% of patients (95/347) in group 2 received allogenic blood transfusion. Significant transfusion risk factors were pre-operative Hb <12 g/dl), American Society of Anesthesiologists (ASA) status III and nonobese body mass index (BMI); Age and gender were not significant risk factors. CONCLUSIONS: MBLPA in primary TKA was highly effective, with a zero transfusion rate. Risk factors for transfusion were determined, and eliminating them contributed to the avoidance of allogeneic blood transfusion in our study series.

Drain use can be avoided in reverse shoulder arthroplasty.

BACKGROUND: Drains have demonstrated no clear benefits and some potentially harmful effects in hip and knee replacements. There is little evidence about the effects of its use in shoulder arthroplasty. We hypothesized that drain use would increase postoperative blood loss without reducing wound complications. METHODS: We included 103 reverse shoulder arthroplasties (RSA), 71 were operated for degenerative pathology, 32 due to a fracture. All complications were recorded. Hemoglobin (Hb) and hematocrit (Htc.) level were collected and compared to postoperative data. Length of hospitalization and volume output were also noted. RESULTS: 45 patients received a closed-suction drain. Patients with coagulopathy had significant higher bleeding and were excluded (p = 0.03). Patients operated for a fracture were older (80.1y.o vs 72.1 p < 0.01) and had higher blood drop (∆Hb p = 0.01; ∆Htc p = 0.03). There were neither differences between drain and control group in ∆Hb or ∆Htc in the degenerative RSA group (1.84+/-0.89 vs 1.68+/-0.84, p = 0.36; 5.78+/-2.89 vs 5.53+/-2.87 p = 0.50) nor in the fracture RSA group (2.65+/-0.94 vs 2.65+/-1.01, p = 0.90; 7.91+/-2.99 vs. 7.09+/-4.21, p = 0.56). There were neither differences in complications (degenerative p = 0.33; fracture p = 0.21). Drain use was related to a longer hospital stay in elective surgery (2.6 vs 1.8 days; p < 0.01). DISCUSSION: The rate of complication is similar between patients with and without drain use. Drain use after shoulder arthroplasty does not affect postoperative bleeding but increases the length of hospital stay. Drains seems to be an unnecessary intervention after RSA that may increase associated costs and can be safely abandoned. LEVEL OF EVIDENCE: Level III retrospective comparative study.

A coumarin based Schiff Base: An effective colorimetric sensor for selective detection of F- ion in real samples and DFT studies.

Chemosensors are molecular devices which react with target and give a visible signal, which is a degree of its sensitivity. Herein, a novel coumarin based Schiff Base has been synthesized for F- ions detection. The chemosensor showed an intense color change upon the addition of F- ions (light yellow to purple). The chemosensor has fewer effects of competing anions. The limit of detection is calculated as low as 1.1 × 10-6 and the binding constant was determined as 1.61 × 104. The job's plot confirmed 1:1 stoichiometry between chemosensor and F- ion. The reverse reaction of chemosensor with MeOH is useful to construct a combinatorial logic circuit gates. The interaction mechanism of chemosensor was deliberated by 1H NMR, FTIR, and DFT studies. Finally, the chemosensor was useful to detect F- ions in tooth-paste sample and test strip is prepared for F- ions detection.

Therapeutic Ultrasound Alone and Associated with Lymphatic Drainage in Women with Breast Engorgement: A Clinical Trial.

Introduction: Breast engorgement (BE) is a problem that affects many women, especially in the first days of breastfeeding, producing inflammatory symptoms. Nonpharmacological therapies are inexpensive, safe, and can produce symptom relief. Objective: This study aims to analyze the safety of therapeutic ultrasound regarding possible risks of overheating and the effects of its use alone and associated with lymphatic drainage (LD) in women. Material and Methods: Effectiveness is measured through thermography, visual analog scale, and six-point scale of BE. This is a nonrandomized clinical trial with a sample of 34 in the ultrasound group (G1), 28 in the ultrasound and LD group (G2), and 37 in the control group (G3). Results: The mean reduction for engorgement was 1.3 ± 0.8 to G1, 1.4 ± 1.0 to G2, and 1.2 ± 0.9 to G3 according to the six-point scale. The mean reduction for pain level was 3.6 ± 2.1 to G1, 4.0 ± 3.1 to G2, and 4.0 ± 2.2 to G3 according to the visual analogue scale. Conclusion: It was observed that all therapies were effective in reducing the level of engorgement, according to the six-point scale. However, combined ultrasound and LD therapy has been shown to be more effective in reducing the level of pain. Brazilian Registry of Clinical Trials (RBR-6btb6zz).

Intestinal Intussusception: A Shocking Diagnosis.

Intussusception is a rare condition diagnosed in adults, with few cases reported as idiopathic. It is defined as the invagination of an intestinal segment into another adjacent one, due to the presence of a lead point, or in some cases, without identifiable causative lesions. The presentation is non-specific, even with careful evaluation, and most of the time, the diagnosis is made during surgery. We hereby present the case of a 73-year-old woman with idiopathic intussusception who presented in the emergency room. She was taken to the operating theatre, where intestinal resection was performed. Few cases of true idiopathic intestinal intussusception in adults are seen, and literature on this topic remains scarce. We discuss diagnostic and therapeutic options and do a brief review of the literature.