Large Scale Genomic Investigation of Pediatric Cholestasis Reveals a Novel Hepatorenal Ciliopathy Caused by PSKH1 Mutations.

PURPOSE: Pediatric cholestasis is the phenotypic expression of clinically and genetically heterogeneous disorders of bile acid synthesis and flow. Although a growing number of monogenic causes of pediatric cholestasis have been identified, the majority of cases remain undiagnosed molecularly. METHODS: In a cohort of 299 pediatric participants (279 families) with intrahepatic cholestasis, we performed exome sequencing as a first-tier diagnostic test. RESULTS: A likely causal variant was identified in 135 families (48.56%). These comprise 135 families that harbor variants spanning 37 genes with established or tentative links to cholestasis. In addition, we propose a novel candidate gene (PSKH1) (HGNC:9529) in 4 families. PSKH1 was particularly compelling because of strong linkage in three consanguineous families who shared a novel hepatorenal ciliopathy phenotype. Two of the four families shared a founder homozygous variant while the third had a different homozygous variant in PSKH1. PSKH1 encodes a putative protein serine kinase of unknown function. Patient fibroblasts displayed abnormal cilia that are long and show abnormal transport. A homozygous Pskh1 mutant mouse faithfully recapitulated the human phenotype and displayed abnormally long cilia. The phenotype could be rationalized by the loss of catalytic activity observed for each recombinant PSKH1 variant using in vitro kinase assays. CONCLUSION: Our results support the use of genomics in the workup of pediatric cholestasis and reveal PSKH1-related hepatorenal ciliopathy as a novel candidate monogenic form.

Loss of function mutation in LARP7, chaperone of 7SK ncRNA, causes a syndrome of facial dysmorphism, intellectual disability, and primordial dwarfism.

Primordial dwarfism (PD) is a clinically and genetically heterogeneous condition. Various molecular mechanisms are known to underlie the disease including impaired mitotic mechanics, abnormal IGF2 expression, perturbed DNA damage response, defective spliceosomal machinery, and abnormal replication licensing. Here, we describe a syndromic form of PD associated with severe intellectual disability and distinct facial features in a large multiplex Saudi family. Analysis reveals a novel underlying mechanism for PD involving depletion of 7SK, an abundant cellular noncoding RNA (ncRNA), due to mutation of its chaperone LARP7. We show that 7SK levels are tightly linked to LARP7 expression across cell lines, and that this chaperone is ubiquitously expressed in the mouse embryo. The 7SK is known to influence the expression of a wide array of genes through its inhibitory effect on the positive transcription elongation factor b (P-TEFb) as well as its competing role in HMGA1-mediated transcriptional regulation. This study documents a critical role played by ncRNA in human development and adds to the growing list of molecular mechanisms that, when perturbed, converge on the PD phenotype.

Structural, Thermal, and Optical Studies of Gamma Irradiated Polyvinyl Alcohol-, Lignosulfonate-, and Palladium Nanocomposite Film.

Nanocomposite (NC) films of polyvinyl alcohol (PVA), lignosulfonate (Lg), and nanosized palladium (Pd) were synthesized by ex-situ casting method. Samples from the synthesized PVA-Lg/Pd NC films were irradiated with 5-100 kGy γ doses. The effect of γ doses on the structural, thermal, and optical characteristics of the NC films were studied using different characterization techniques. The results indicated that the γ irradiation improves the decomposition temperature from 227 to 239 °C, signifying an increase in the thermal stability of the NC films. This was accompanied by a reduction of the melting temperature due to the increase of the amorphous phase. This can be attributed to the dominance of crosslinking. On the other hand, the refractive index increased from 2.21 to 2.32 while increasing the γ dose up to 100 kGy. This was associated with a reduction of the optical bandgap from 3.49 to 3.30 eV, which could be attributed to the increase in the amorphous phase as a result of crosslinking. This indicates an enhancement of the spreading of the NPs inside the blend matrix due to γ irradiation. This results in a more compacted construction of the PVA-Lg/Pd NC films. Furthermore, we used the Commission Internationale de E'Claire (CIE) method to estimate the change in color among the irradiated NC films and the pristine film. The PVA-Lg/Pd NC attained a significant color difference value greater than five, meaning permanent color changes.

Role of copper and alumina for heat transfer in hybrid nanofluid by using Fourier sine transform.

The convection, thermal conductivity, and heat transfer of hybrid nanofluid through nanoparticles has become integral part of several natural and industrial processes. In this manuscript, a new fractionalized model based on hybrid nanofluid is proposed and investigated by employing singular verses and non-singular kernels. The mathematical modeling of hybrid nanofluid is handled via modern fractional definitions of differentiations. The combined Laplace and Fourier Sine transforms have been configurated on the governing equations of hybrid nanofluid. The analytical expression of the governing temperature and velocity equations of hybrid nanofluid have been solved via special functions. For the sake of thermal performance, dimensional analysis of governing equations and suitable boundary conditions based on Mittage-Leffler function have been invoked for the first time in literature. The comparative analysis of heat transfer from hybrid nanofluid has been observed through Caputo-Fabrizio and Atangana-Baleanu differential operators. Finally, our results suggest that volume fraction has the decelerated and accelerated trends of temperature distribution and inclined and declined profile of heat transfer is observed copper and alumina nanoparticles.

Analysis of transcript-deleterious variants in Mendelian disorders: implications for RNA-based diagnostics.

BACKGROUND: At least 50% of patients with suspected Mendelian disorders remain undiagnosed after whole-exome sequencing (WES), and the extent to which non-coding variants that are not captured by WES contribute to this fraction is unclear. Whole transcriptome sequencing is a promising supplement to WES, although empirical data on the contribution of RNA analysis to the diagnosis of Mendelian diseases on a large scale are scarce. RESULTS: Here, we describe our experience with transcript-deleterious variants (TDVs) based on a cohort of 5647 families with suspected Mendelian diseases. We first interrogate all families for which the respective Mendelian phenotype could be mapped to a single locus to obtain an unbiased estimate of the contribution of TDVs at 18.9%. We examine the entire cohort and find that TDVs account for 15% of all "solved" cases. We compare the results of RT-PCR to in silico prediction. Definitive results from RT-PCR are obtained from blood-derived RNA for the overwhelming majority of variants (84.1%), and only a small minority (2.6%) fail analysis on all available RNA sources (blood-, skin fibroblast-, and urine renal epithelial cells-derived), which has important implications for the clinical application of RNA-seq. We also show that RNA analysis can establish the diagnosis in 13.5% of 155 patients who had received "negative" clinical WES reports. Finally, our data suggest a role for TDVs in modulating penetrance even in otherwise highly penetrant Mendelian disorders. CONCLUSIONS: Our results provide much needed empirical data for the impending implementation of diagnostic RNA-seq in conjunction with genome sequencing.