Genetic skeletal disorders (GSDs) constitute a rare, heterogeneous, unique group of bone growth disorders affecting the homeostasis, development of bones, and resulting in anomalous size and shape of the skeleton. Syndromic and non-syndromic skeletal disorders epitomize a public health problem that affects 1/4000 individuals worldwide, thus leading to high health cost and poor quality of life. Detailed information about pathophysiologic mechanisms and disease-causing genetic defects is prerequisite in order to provide proper clinical intervention for different GSDs. With this inkling in mind, the present study was planned to investigate sixteen families manifested with GSDs from different populations at clinical and molecular levels. A total of sixteen families (A-P) segregating different forms of skeletal disorders were genetically and clinically characterized in the present study. The present study was performed in different steps, which included visit to remote areas in Pakistan to construct family pedigree, collection of blood samples, clinical (radiological) examination of at least two affected individuals in each family, genetic linkage analysis using STS microsatellite markers/SNPs microarray, whole exome sequencing (WES) and Sanger sequencing. The identified pathogenic variants were also analyzed for their pathogenicity using in-silico and in-vitro approaches. In the four families (A-D), after failing to establish linkage to the known genes/loci responsible for pre-axial and post axial polydactyly, WES was performed to identify the candidate causing gene. In family A, WES identified a homozygous splice acceptor site variant (c.395-1G>A) in intron 5 of IQCE gene on chromosome 7p22.3. In-vitro analysis using mini-gene splice assay in the family revealed a frameshift variant (p.Gly132Valfs*22). In family B, WES revealed a homozygous missense variant (c.223G>A; p.Asp75Asn) in a potential novel gene GLT8D1 (3p21.1). In family C, a nonsense mutation (c.84C>A: p.Tyr28*) was identified in the C9orf96 (STKLD1) gene mapped on chromosome 9q34.2. The C9orf96 is the first candidate gene identified to cause autosomal recessive non-syndomic pre-axial polydactyly. In family D with autosomal recessive uni-lateral pre-axial polydactyly, WES identified a Abstract XXVI novel biallelic deletion of ZNF468 and ZNF28 genes located on chromosome 19q13.41. In three families, segregating osteogenesis imperfecta, scanning human genome using SNP markers, mapped the causative homozygous region on chromosome 17q21.1- q21.31 in family E and F. Sequence analysis of the previously reported gene FKPB10 on 17q21.1-q21.31 led to the identification of a novel nonsense mutation (c.1490G>A; p. Trp497*) in family E and a previously reported missense variant (c.344G>A; p.Arg115Gln) in family F. In family G, using WES a homozygous splice acceptor site variant (c.359-3C>G) in the intron 2 of the WNT1 gene was identified, located on chromosome 12q13.12. Affected individuals in family H and I were diagnosed with acromesomelic dysplasia type Grebe (AMDG) phenotypes, and those in family J with acromesomelic dysplasia type maroteaux (AMDM). Linkage in families (H and I) was established to the GDF5 locus on chromosome 20q11.22, and in family J to the NPR2 locus on chromosome 9p13-q12. Subsequently, Sanger sequencing of GDF5 gene identified two novel homozygous variants, (c.157_158dupC and c.872G>A) in the family H and I. In family J, an already known homozygous splice donor site variant (c.2986+2T>G) was detected in intron 20 of the NPR2 gene. Three families (K-M), segregating split hand/foot malformation (SHFM) were investigated in the present study as well. Family K and L were subjected to whole genome SNP array analysis. In family K, SNP microarray identified two potential homozygous regions including a 35Mb on chromosome 12 and 11Mb on chromosome 4. This family will be subjected to WES upon availability of funds. Whole genome SNP array following WES in family L revealed a novel frameshift variant (c.409delA; p.Ser137Alafs*19) in the EPS15L1 gene located on chromosome 19p13.11. After establishing linkage in family M to WNT10B on chromosome 12q13.12, Sanger sequencing identified previously reported 7bp duplication (c.300_306dupAGGGCGG) in the WNT10B gene. Clinical and radiological examination of family N and family O showed typical Ellisvan Creveld syndrome (EvC) phenotypes. In family N, WES revealed two homozygous variants in the EVC2 (c.30dupC; p.Thr11Hisfs*45) and TMC1 gene Abstract XXVII (c.1696-1G>A). In family O, WES identified novel compound heterozygous variants (c.919T>C; p.Ser307Pro, c.2894+3A>G) in exon 7 and 20 of the EVC gene. Finally, clinical examination of affected members in family P displayed major features of Bardet-Biedl syndrome (BBS). DNA from an affected individual of the family was subjected to WES. A novel nonsense variant (c.119C>G; p.Ser40*) was identified in exon 1 of MKKS (BBS6) gene located on chromosome 20p12.2. In conclusion, the study, presented here, identified four novel candidate genes, novel and recurrent mutations in few previously reported genes causing different skeletal deformities. The identification of novel skeletal related genes not only improves the overall understanding of skeletal development system but also helps in creating new research dimensions such as understanding different pathways. GSDs results from mutations in different genes that encode different transcription factors (TFs), extracellular matrix proteins, signal transducers (channel proteins, receptors, ligands), RNA processing molecules, tumor suppressors, cellular transporters, enzymes (chaperones), binding proteins, cilia proteins, and numerous proteins having unknown function. The novel gene identified here, such as the IQCE share a common Hedgehog signaling pathway including the EVC/EVC2, thus helping in further understanding there important role in limb patterning, and skeletal developmental. The novel and recurrent mutations might help in the proper genotype–phenotype correlation which will help in prenatal testing and genetic counseling of the affected families. Further functional characterization of the genes, discovered here, is required to elucidate their roles in skeletal development and pathophysiology of myriad skeletal disorders.
Islamic religious militancy is a matter of great concern for the Muslim and the non-Muslim world today. The analysis of the ideology of the militants reveals that they find the legitimacy of their military activities in the ideal of the establishment of an Islamic state to establish the universal rule of Islām, and in the specific interpretations of some Qur’ānic verses, Aḥādīth of the Prophet (r), and also from the establishment of the Islamic state in Madīnah by the Prophet (r), his the military expeditions and those of his companions against their opponents and from the treatment of our historians of the individual military campaigns against the Muslim regimes of their times. The Muslim militants also fight against their Muslim governments on the grounds that they are not the true Islamic governments. The militants do not bother to kill the common Muslim masses, who vote and support such rulers. They take it as collateral damage. The world naturally reacts to this cult, especially the west, being at the helm of the world politics. Not only do the West tries to crush the Islamic militants, across the world, but also, topple the Muslim democratic governments having any ideal of an Islamic Khilāfah. This frustrates the peaceful political activists and strengthens the military activists, further. To end this ongoing and mounting cult of religious militancy, it is necessary to review the specific and traditional interpretations of the academic sources of Islām: Qur’ān, Ḥadīth and Fiqh, regarding the legitimacy of militancy in Islām. Secondly, to remove their misconceptions, it is necessary to engage the militants in dialogue through a counter narrative, which the author tried to present here.
Rare earth-based materials with pyrochlore structure have shown a potential for practical applications due to their stable crystal structure with tunable lattice parameters. Pyrochlore structure-based compounds have shown valuable interest in practical applications because of their structure tunability and stability. In this context, the stability of pyrochlore structure in a particular phase is quite essential for practical optimization of valuable dielectric, impedance and other multifunctional characteristics of rare-earth pyrochlores. This work contains a successful synthesis of pure phase pyrochlore based compounds including zirconates and niobates, with A and B-site substitution to determine their structural tunability and its effects on dielectric and other multifunctional properties. These rare-earth based pyrochlores not only show practically viable dielectric characteristics but also have significant ferroelectric and magnetic features. Most of the pyrochlores show disordered magnetic structure and have not been investigated properly yet. In this context, following detailed study of these thermally stable pyrochlores has been accomplished for their practical applications. A wet chemical sol-gel technique was utilized to synthesize pure phase pyrochlores. This method was found feasible, energy efficient and cost effective. Diffraction studies using X-rays revealed their stable pyrochlore structure with Fd ̅m space group. Structural tunability through A and B-site substitution was confirmed through Rietveld refinement. Perfect fitting and low residual factors from refinement prove the synthesis of pure phase pyrochlores. Calcination temperature of 1200 °C for 1 h was found to be the most favorable condition for the synthesis of these highly complex metal oxides. Field emission scanning electron microscopy operated in different operating modes at several voltages has confirmed the growth of nanoparticles with substitution, promoting the densification of the prepared ceramics. This densification led us to study the energy storage capability of these materials as they have already shown good ferroelectric properties. To check the variation in recoverable energy density of the material under consideration, the charge-discharge curves were plotted at different field strengths to study the recoverable energy density, energy loss density and energy storage efficiency as a function of applied field. Furthermore, magnetic properties of these pyrochlores were determined through vibrating sample magnetometer. Superparamagnetic effect was found in these pyrochlores with reasonably good magnetic susceptibility and small coercive fields. In addition, the variation in magnetic properties was also related to the particle size which showed that the maximum magnetization increases with decrease in particle size. Lastly, the magneto-dielectric study has been performed. The study of relative change in dielectric properties of the material with change in external magnetic field is called magneto-dielectric analysis. This analysis is actually the investigation of coupling between the magnetic properties of the material with its dielectric characteristics. Magneto-dielectric analysis exposed that dielectric behavior of the samples is influenced inversely with the application of external magnetic field. Negative behavior of magneto-dielectric coefficient can be understood on the basis of application of external magnetic field randomizes the electric dipoles present in the material which usually follow the external applied electric field. This study was also explained on the basis of Maxwell-Wagner‟s model.