Novel pathogenic variants in an Indian cohort with epidermolysis bullosa: Expanding the genotypic spectrum

BackgroundEpidermolysis bullosa (EB) is a genodermatosis characterized by skin fragility and blisters with variable severity. Patients with Dystrophic EB (DEB) or Junctional EB (JEB) mainly present to clinic due to greater functional impairment. Pathogenic sequence variations in COL7A1 are implicated in DEB.ObjectiveWe have tried to decipher the molecular spectrum and genotype phenotype correlation of 21 Indian patients with EB.MethodsNext generation sequencing (NGS) was performed to determine the pathogenic variants. Sanger sequencing was also done for validation of the variants in eleven individuals.ResultsPathogenic variants were detected in 20 individuals (diagnostic yield of 95%). Majority of them (90%) had sequence variation in COL7A1 while two had pathogenic variants in ITGB4 and KRT14 respectively. Out of the 18 patients confirmed to have DEB, 3 had Dominant DEB (DDEB) whereas 15 patients had Recessive DEB (RDEB). Amongst 23 sequence variations identified, 12 were found to be novel (3 were missense, 5 were premature termination codon variants while 4 were splice-site changes).ConclusionGenotype phenotype correlation was noted with milder manifestations in those with dominant inheritance types. Exact molecular diagnosis can be ascertained by NGS in majority of cases.     

Alteration of lumbar muscle morphology and composition in relation to low back pain: a systematic review and meta-analysis

BACKGROUND CONTEXTPrevious studies have proposed that there is a relationship between low back pain (LBP) and morphology and composition of paraspinal muscles. However, results have been conflicting, especially regarding fatty infiltration of muscles.PURPOSEThe primary goal of this study was to review and analyze results from imaging studies which investigated morphological and composition changes in the multifidus, erector spinae and psoas major muscles in people with LBP.STUDY DESIGN/SETTINGSystematic review with meta-analysis.PATIENT SAMPLEA patient sample was not requiredOUTCOME MEASURESThis review did not have outcome measures.METHODSPubMed, Scopus, Web of Sciences, EMBASE and ProQuest were searched for eligible studies up to 31st July 2020 (all languages). A systematic search of electronic databases was conducted to identify studies investigating the association between the morphology and fat content of lumbar muscles in people with LBP compared with a (no LBP) control group. 13,795 articles were identified. Based on the screening for inclusion/ exclusion, 25 were included. The quality of the studies was evaluated using the Newcastle-Ottawa Scale. From the 25 articles, 20 were included in the meta-analysis.RESULTSResults showed that the total cross-sectional area of the multifidus was smaller in people with LBP (Standardized mean difference, SMD = -0.24, 95% CI = -0.5 to 0.03). Combined SMDs showed a medium effect of LBP on increasing multifidus muscle fat infiltration (SMD = 0.61, 95% CI = 0.30 to 0.91). There were no LBP related differences identified in the morphology or composition of the lumbar erector spine and psoas major muscles.CONCLUSIONSPeople with LBP were found to have somewhat smaller multifidus muscles with a significant amount of intramuscular fat infiltration. Varying sample size, age and BMI of participants, quality of studies and the procedures used to measure fat infiltration are possible reasons for inconsistencies in results of previous studies.     

Bibliometric analysis of orthopaedic related publications by Indian authors from the last decade

PurposeBibliometric studies have been established methods of analysing publications on a particular topic. These studies have been done on various orthopaedic topics and are increasing. The advantages of these studies have been highlighted in previous publications. Although some studies have been done on Indian publications from other specialties, those analysing Indian Orthopaedic Publications are lacking.MethodsWe performed a search in Scopus to look for all publications related to orthopaedics from India. Our search strategy in Scopus included ((TITLE-ABS-KEY(Orthopaedics OR Orthopaedics) AND AFFIL(India)) AND PUBYEAR > 2009 AND PUBYEAR < 2020) which resulted in 3270 articles on 02/11/2021. We analyzed the most publishing universities, city, state, specialty, authors, and anatomic location of these publications. We also mined the data to draw word clouds based on data obtained from the titles of articles, keywords and the affiliations of each of the articles published.ResultsTamil Nadu and New Delhi and their institutes appear to be the epicenter of publication activities in Orthopaedics in India. There has been a healthy trend of growth of articles in the orthopaedic specialty. Since there is a significant overlap of technology and engineering, it is not surprising to see engineering and technology institutes among the top 10 published institutes and even journals for the publications on orthopaedics.ConclusionThere has been a steady increase in the number of publications in the last decade. New Delhi and its Universities and Institutes appear to contribute the majority of citations and publications related to orthopaedics. Journal of Clinical Orthopaedics and Trauma was the most publishing journal for Indian authors on Orthopaedic related articles.     

Connectivity measures suggest a sub-cortical generator of myoclonus in Angelman syndrome

ObjectiveThe clinical and neurophysiological characteristics of myoclonus in Angelman syndrome (AS) have been evaluated in single case or small cohorts, with contrasting results. We evaluated the features of myoclonus in a wide cohort of AS patients.MethodsWe performed polygraphic EEG-EMG recording in 24 patients with genetically confirmed AS and myoclonus. Neurophysiological investigations included jerk-locked back-averaging (JLBA), cortico-muscular coherence (CMC) and generalised partial directed coherence (GPDC). CMC and GPDC analyses were compared to those obtained from 10 healthy controls (HC).ResultsTwenty-four patients (aged 3–35 years, median 20) were evaluated. Sequences of quasi-continuous rhythmic jerks mostly occurred at alpha frequency or just below (mean 8.4 ± 1.4 Hz), without EEG correlate. JLBA did not show any clear transient preceding the jerks. CMC showed bilateral over-threshold CMC in alpha band that was prominent on the contralateral hemisphere in the patient group as compared to HC group. GPDC showed a significantly higher alpha outflow from both hemispheres toward activated muscles in the patient group, and a significantly higher beta outflow from contralateral hemisphere in the HC group.ConclusionsThese neurophysiological findings suggest a subcortical generator of myoclonus in AS.SignificanceMyoclonus in AS has not a cortical origin as previously hypothesised.     

Engraftable neural crest stem cells derived from cynomolgus monkey embryonic stem cells

Neural crest stem cells (NCSCs), a population of multipotent cells that migrate extensively and give rise to diverse derivatives, including peripheral and enteric neurons and glia, craniofacial cartilage and bone, melanocytes and smooth muscle, have great potential for regenerative medicine. Non-human primates provide optimal models for the development of stem cell therapies. Here, we describe the first derivation of NCSCs from cynomolgus monkey embryonic stem cells (CmESCs) at the neural rosette stage. CmESC-derived neurospheres replated on polyornithine/laminin-coated dishes migrated onto the substrate and showed characteristic expression of NCSC markers, including Sox10, AP2α, Slug, Nestin, p75, and HNK1. CmNCSCs were capable of propagating in an undifferentiated state in vitro as adherent or suspension cultures, and could be subsequently induced to differentiate towards peripheral nervous system lineages (peripheral sympathetic neurons, sensory neurons, and Schwann cells) and mesenchymal lineages (osteoblasts, adipocytes, chondrocytes, and smooth muscle cells). CmNCSCs transplanted into developing chick embryos or fetal brains of cynomolgus macaques survived, migrated, and differentiated into progeny consistent with a neural crest identity. Our studies demonstrate that CmNCSCs offer a new tool for investigating neural crest development and neural crest-associated human disease and suggest that this non-human primate model may facilitate tissue engineering and regenerative medicine efforts.     

Genomic and phenotypic delineation of congenital microcephaly

PurposeCongenital microcephaly (CM) is an important birth defect with long term neurological sequelae. We aimed to perform detailed phenotypic and genomic analysis of patients with Mendelian forms of CM.MethodsClinical phenotyping, targeted or exome sequencing, and autozygome analysis.ResultsWe describe 150 patients (104 families) with 56 Mendelian forms of CM. Our data show little overlap with the genetic causes of postnatal microcephaly. We also show that a broad definition of primary microcephaly —as an autosomal recessive form of nonsyndromic CM with severe postnatal deceleration of occipitofrontal circumference—is highly sensitive but has a limited specificity. In addition, we expand the overlap between primary microcephaly and microcephalic primordial dwarfism both clinically (short stature in >52% of patients with primary microcephaly) and molecularly (e.g., we report the first instance of CEP135-related microcephalic primordial dwarfism). We expand the allelic and locus heterogeneity of CM by reporting 37 novel likely disease-causing variants in 27 disease genes, confirming the candidacy of ANKLE2, YARS, FRMD4A, and THG1L, and proposing the candidacy of BPTF, MAP1B, CCNH, and PPFIBP1.ConclusionOur study refines the phenotype of CM, expands its genetics heterogeneity, and informs the workup of children born with this developmental brain defect.     

Sequential delivery of immunomodulatory cytokines to facilitate the M1-to-M2 transition of macrophages and enhance vascularization of bone scaffolds

In normal tissue repair, macrophages exhibit a pro-inflammatory phenotype (M1) at early stages and a pro-healing phenotype (M2) at later stages. We have previously shown that M1 macrophages initiate angiogenesis while M2 macrophages promote vessel maturation. Therefore, we reasoned that scaffolds that promote sequential M1 and M2 polarization of infiltrating macrophages should result in enhanced angiogenesis and healing. To this end, we first analyzed the in vitro kinetics of macrophage phenotype switch using flow cytometry, gene expression, and cytokine secretion analysis. Then, we designed scaffolds for bone regeneration based on modifications of decellularized bone for a short release of interferon-gamma (IFNg) to promote the M1 phenotype, followed by a more sustained release of interleukin-4 (IL4) to promote the M2 phenotype. To achieve this sequential release profile, IFNg was physically adsorbed onto the scaffolds, while IL4 was attached via biotin-streptavidin binding. Interestingly, despite the strong interactions between biotin and streptavidin, release studies showed that biotinylated IL4 was released over 6 days. These scaffolds promoted sequential M1 and M2 polarization of primary human macrophages as measured by gene expression of ten M1 and M2 markers and secretion of four cytokines, although the overlapping phases of IFNg and IL4 release tempered polarization to some extent. Murine subcutaneous implantation model showed increased vascularization in scaffolds releasing IFNg compared to controls. This study demonstrates that scaffolds for tissue engineering can be designed to harness the angiogenic behavior of host macrophages towards scaffold vascularization.