Epiphyseal Fusion of Iliac Crests in Male and Female Adolescents: An Age Estimation Criterion

Background: Determination of age depends upon physical examination, dental assessment, and skeletal evaluation. The radiological examination of bone for appearance and fusion of ossification centers helps in the assessment of skeletal maturity as the process occurs in a particular sequence which is almost constant for that particular bone. Objectives: The objective of this study was to determine the age of fusion of iliac crest by radiological examination of subjects of age bracket 17-25 years coming to Shalamar Hospital Lahore Methods: In this cross-sectional study, radiological examinations (Digital X-Rays) were performed to evaluate the fusion of Iliac Crest in 200 subjects of both genders of 17 – 25 years. Data analysis was done using SPSS Version 23. Conclusions were drawn and compared with available results of previous work done in this field. Results: Out of 200 subjects, there were 132 males (66 %) and 68 females (34%). The mean ± SD age of both genders was 20.41± 2.55. There were 93 cases (70.45%) of complete fusion among males, showing 100 % union in the age groups of 21-25 years, while 40 cases (58.83%) of complete union among females were observed during 20-25 year of age groups. The mean ± SD age of complete union for males was 20.67± 2.61 years and for females 19.90 ± 2.38 years, with a significant p value of <0.05. Similarly, a statistically significant difference was observed among people of different socio-economic statuses. No difference was observed among different ethnic groups. Conclusions: The fusion of the iliac crest is not affected by ethnicity. Factors like diet and nutrition directly affect bone growth and hence bone age. More studies should be conducted across the country to formulate a standard in setting up a uniform criterion for assessing the age of adolescents

Isolation, Identification and Characterization of Osrglp2 Promoter Binding Protein S

Germin and germin-like proteins (GLPs) belong to cell wall glycoproteins that have shown asignificant resistance to detergent action, denaturation by heating, and degradation by proteases. GLPs expression is reportedly modulated during exposure to pathogens and abiotic stresses. Yet, little is identified about the regulatory mechanism of the GLP genes. The promoter of OsRGLP2 gene was isolated and cloned by Mahmood et al. (2007). This promoter showed strong expression ofthe GUS gene in transgenic tobacco during salinity, dehydration and wounding stresses. In this study, the regulatory cis-elements and their binding proteins for OsRGLP2 promoter are characterized. Various putative stress responsive cis-regulatory motifs and their specific binding proteins were identified by in silico analysis. DNA binding domains of OsWRKY71, OsDOF18 and OsMYB1 were cloned, overexpressed in E. coli and then purified by affinity chromatography using Glutathione Sepharose resin followed by cationic exchange chromatography. Electrophoretic mobility shift assays (EMSAs) have shown that recombinant OsWRKY71, OsDOF18 and OsMYB1 proteins were capable to interact with DIG labeled fragments of OsRGLP2 promoter containing W-box, AAAG and WAACCA motifs respectively. Binding was further confirmed by competitor EMSA and EMSA with mutant oligonucleotides. These regulatory elements were also active in binding with nuclear factors from rice nuclear proteins extract in vitro as confirmed by competitive EMSA. Expression analysis was performed to check the level of OsWRKY71, OsDOF18 and OsMYB1 against salt, cold, heat, wounding and drought stresses. Expression of OsWRKY71 was found to be induced in case of salt and cold stresses while OsDOF1 was expressed at relatively high level in response to salinity and drought. OsMYB1 expression was 23 fold higher in response to wounding which demonstrates the valueofOsMYB1 up-regulation in response to wounding stress inrice. In order to investigatethe functionof OsWRKY71, OsMYB1 and OsDOF18againstdiverse abiotic stresses, recombinant plasmids were subjected to transformationin E. coli and their effect on E. coli growth was analyzed. The E. colicells containing pGEX-OsWRKY71, pGEX-OsDOF18 and pGEX-OsMYB1 has shown different levels of tolerance against salt, drought, cold and heat stresses as compared to empty pGEX-4T1 vector. In silico characterization suggested the involvement of these proteins in protein-protein interaction. In conclusion, the positive response of OsWRKY71, OsDOF18 and OsMYB1 in abiotic stresses suggests their association with OsRGLP2 promoter and the importance of these proteins in providing protection to plants under abiotic stresses. Overexpression of OsWRKY71, OsDOF18 and OsMYB1 genes in crop plants may help in obtaining stress tolerant lines.