There have been incredible breakthroughs in the field of human genetics in the last few decades. Phenotypic diversity and genome evolution takes place due to genetic variation. Localization and identification of these genetic variants associated with disease is possible with the help of techniques used for manipulation of normal and mutant genes. Linkage analysis, haplotype analysis and DNA sequencing are used for the identification of genomic regions associated with genes and mutational analysis of genes. Pakistani population is a good candidate for genetic linkage analysis due to the large family size and higher rate of consanguineous marriages leading to higher risk of inherited diseases in this population. These large inbred families are very good candidates for mapping the genes behind the disease. The aim of this thesis is to identify the genes, mutations and regions underlying some inherited skin disorders i.e Ectodermal Dysplasia (ED), Alopecia (AP), Nail dysplasia (ND), Mal de Meleda (MDM), and Icthyosis. This Study will contribute to our understanding of human genetics and will shed light to dysfunction of genes in development and disease as the first step is always been the identification of tentative genes affecting the normal development, in this case Skin and Nails. ED comprises a heterogeneous group of developmental abnormalities constituting about 200 clinical entities. All EDs are characterized by impaired development of at least two of the epidermal appendages hair, nails, teeth and sweat glands. Approximately 30 EDs have been characterised at the molecular level. Odonto-onycho-dermal dysplasia (OODD) is a rare syndrome characterized by severe hypodontia, nail dystrophy, smooth tongue, dry skin, keratoderma and hyperhydrosis of palms and soles. A large consanguineous Pakistani pedigree comprising six individuals affected by a complete OODD syndrome was investigated. Autozygosity mapping using SNP array analysis showed that the affected individuals are homozygous for the Wnt10A gene region. Subsequent mutation screening showed a homozygous c.392C4T transition in exon 3 of Wnt10A, which predicts a p.A131V substitution in a conserved a-helix domain. First inherited missense mutation in Wnt10A with associated ectodermal features was observed. Mutations in EDA1 are underlying cause of inherited X-linked hypohidrotic ectodermal dysplasia (XLHED). A large consanguineous Pakistani kindred was investigated with hypohidrotic ED. Careful pedigree analysis suggested X-linked recessive inheritance. There were five affected male and four carrier females in the family. They presented with classical phenotype of disorder. Sequencing revealed a 4-bp insertion at nucleotide position 913 (913_914insTATA) in all affected family members investigated. The mutation is located in exon 8 and results in frameshift and a premature stop codon 2-bp downstream in the same exon. Another type of ED is “Pure hair-nail ectodermal dysplasia”: a rare subgroup with variable expression. A multigenerational consanguineous Pakistani family with four members affected by “pure hair-nail ectodermal dysplasia” was identified. Linkage was confirmed with microsatellite markers on chromosome 12 with maximum LOD score 2.92 (θ=0.0) at marker locus D12S368 suggesting linkage to this region. Further investigations associate this hair-nail type ectodermal dysplasia with novel gene present on chromosome 12p11.1-q14.3. Atrichia with papular lesions (APL) is a rare autosomal recessive form of congenital Alopecias characterized by onset of complete or near to complete irreversible alopecia or loss of hair soon after birth or in first few months of life. Diagnosis of APL involves autosomal recessive form of transmittance of disease with possible consanguinity of parents, hair either present at birth or not that never re-grow and presence of papules. APL has been mapped to 8p12 and identification of Human Hairless (HR) gene mutations are associated with its pathogenesis. Atrichea with papules has equal prevalence in both men and women but exact prevalence is still unknown. A novel homozygous missense c.2427 C>T transition in exon10 of HR gene was identified that results in p.A765V substitution and small variation of phenotype in a consanguineous Pakistani family in all affected individuals. Developmental abnormalities of the nails constitute a large and extremely heterogeneous group of disorders. These abnormalities have a wide range of appearance in form of slight and hardly noticeable change to the complete absence of nails. These can be inherited as isolated abnormality or part of complex syndromes with associated abnormalities of other ectodermal appendages. A large family with isolated congenital anonychia as sole phenotype and consanguineous pedigree with autosomal recessive inheritance was ascertained. Linkage was confirmed to 20p13 to RSPO4 gene using multi allelic microsatellite markers. A maximum two point LOD score was obtained at marker locus D20S199 (θ =0.00) 3.92 suggestive of linkage. RSPO4 gene was sequenced in two affected individual of the family, which revealed G to A transversion at nucleotide position 353 in exon 3, resulting in a (p.Cys118Tyr) amino acid substitution. Another consanguineous Pakistani family with four members affected by isolated congenital autosomal recessive nail dysplasia was investigated. Claw-like pachonychia, nail clubbing and onycholysis of both finger and toenails was present in affected individuals but no other ectodermal symptoms. Genetic analysis on the family using a 250K SNP array (Affymetrix) revealed a genomic region spanning more than 90 homozygous SNPs on chromosome 8 found to be shared by affected individuals. Linkage analysis with microsatellite markers revealed a maximum LOD score of 2.96 (θ=0) at polymorphic microsatellite marker D8S1122 chromosome 8. Recombination events restricted the candidate region to 18 Mb which spans approximately 100 genes. Sequencing revealed a nonsense c.1750G>T (p.Gly584) substitution in homozygous form in Frizzled 6 gene in exon 6. MDM or keratosis palmoplantaris transgrediens of Siemens is a rare genodermatosis which belongs to Palmo planter keratoderma PPK; a heterogeneous group of disorders by clinical and genetic definitions. Characteristic features are abnormal thickening of skin of palms and soles soon after birth and progresses to the dorsal epidermis of hands and feet with age. A consanguineous Pakistani family appeared with MDM. ARS component B gene associated with MDM phenotype found mutated in affected members. Linkage was confirmed to 8qter regions using multi allelic microsatellite marker and maximum two point LOD score was obtained at marker locus D8S161 (θ =0.00) 2.41 suggestive for linkage. Sequencing revealed a homozygous c.256 G>A transition in all 4 affected family members investigated. The mutation is located in exon 3 and results in a p.G86R substitution. Nonbullous congenital ichthyosiform erythroderma (NBCIE) is autosomal recessive congenital ichthyosis with prominent features of generalized severe white scaling all over the body and erythrodermic skin without blister formation. ABC12, causal gene is localized on chromosome 2q33-35 belongs to a subfamily of ATP-binding cassette (ABC) transporters which implicate in some autosomal recessive disorders pertaining to lipid metabolism. An extended consanguineous Pakistani family exhibiting autosomal recessive inheritance and classic phenotype of NBCIE was investigated. Sequencing revealed a novel homozygous c.4896 G>T transition in all 4 affected family members investigated. The mutation is located in exon 31 and results in a p.G1241V substitution.
The learning of Arabic language like any other foreign language contains four main aspects; reading, writing, speaking and understanding while listening.[i] This learning process can be enhanced if the most appropriate Learning Strategy is used. In this paper the most appropriate Learning Strategy of Arabic Language is suggested. The course outlines for Arabic language are thoroughly studied and several professors and experts of Arabic Language from Pakistan, Egypt, Saudi Arabia and Sudan are interviewed. The author, who himself has vast experience in teaching Arabic language, also had the opportunity to sit in the Arabic language classes to observe various strategies and methodologies adopted by different professors while teaching Arabic. In this paper the time spent on teaching Arabic to the students is also discussed. The appropriate size of the class room i.e, the number of students in Arabic language class also matters in improving the quality of Arabic among the students. The matter of teaching Arabic in Arabic only or in the native language of the students will also be touched in here. As the time has changed and the world is moving ahead on a fast pace, it seems necessary to apply the “Direct Method” while teaching Arabic or any foreign language.[ii] This paper will shed light on what is meant by “Direct Method”. The idea of making the student sit and memorize the dry rules of grammar has become obsolete. The idea of telling the student what part of the phrase is subject or predicate, or what is object and what is a noun or verb, may come later. The idea of memorizing the bulk of new vocabulary in the beginning can also be postponed. Hence a paradigm shift is needed here while talking about the Methodology of Teaching Arabic Language, under the heading of “Direct Method”. [i] Muhammad Abdul Khaliq, Professor of Arabic and co-author of 'al-Arabia baina Yadaik'. The author of this research paper had a personal interview with him on 21.03.2014, in the Institute of Arabic Language, King Saud University, Riyadh, Saudi Arabia. [ii] This method is adopted roughly by some great scholars of Arabic language like Dr. V. Abdur Rahim who taught Arabic language for decades in the Islamic University of Madina, Kingdom of Saudi Arabia. The author was fortunate to meet with him many times and get benefitted from his experience. See for details: Abdurrahim, V. (1999), Arabic Course for English-Speaking Students, Leicester: UK Islamic Academy. See also: Abdullah, F. Ibrahim. (1999), Iqra Arabic Reader. Chicago: Iqra International Educational Foundation. Moreover see: Fawzan, Abdurrahman and others. (2004), Al-Arabia Baina Yadaik, Riyadh: Ministry of Education.
In nature, biotic and abiotic stresses impose negative effects on all life forms. Being a sessile life habitat, plant always exposed to these stresses. Plant when subjected to stress condition, always resulted in a reduced growth. Among the abiotic stresses, salinity is a major cause often hindering plant growth which inturn curtails the crops yield. To lessen the impacts of salinity i.e. alleviation for the betterment of plant growth and crop yield is the matter of interest in the current research scenario. Plant-derived smoke obtaining from burning vegetation have been using in agricultural and horticultural since long. Being a cheap and environment friendly source, smoke play an important role in the enhancement of crops yield. Plant derived smoke possesses alleviation potential for number of stresses. The present research work was conducted to investigate the effect of Cymbopogon jawaracusa smoke extracts (1:100, 1:200,1:300,1:400 and 1:500 dilutions) on physiological and biochemical aspects of maize (Zea mays L.) under different concentrations of NaCl (50,100, 150, 200 and 250 mM). To investigate the alleviation potential of smoke, seeds were primed for 24 h with smoke solution and then subjected to salt stress. Obtained results showed that seed germination percentage was improved up to 93% with smoke as compared to control (70%). Similarly, seedling vigor in term of root and shoot length, fresh and dry weights were also significantly increased in plants raised from seeds primed with smoke extracts. Priming of seeds with smoke solution alleviated the adverse effects of salt stress. Ions analysis indicated that priming with smoke solution increased the level of potassium and calcium while reduced the level of sodium in the exposed plants. In addition, the levels of photosynthetic pigments, total nitrogen and protein contents were also improved with the application of seed priming techniques by using plant smoke solution as compared to salt stressed plants. Salt stress increased the level of Reactive Oxygen Species (ROS) as well as antioxidants in the exposed maize seedlings. Seed priming with smoke solution showed alleviation of stress and the level of ROS as well as antioxidants remained lowered although the seedlings were exposed to salt stress. Findings of this study showed that smoke solution has the potential to alleviate the toxic effects of saline condition as well as can increase the productivity in plants