Soluble and insoluble expression of recombinant human interleukin-2 protein using pET expression vector in Escherichia coli.

Interleukin-2 has emerged as a potent protein-based drug to treat various cancers, AIDS, and autoimmune diseases. Despite its immense requirement, the production procedures are inefficient to meet the demand. Therefore, efficient production procedures must be adopted to improve protein yield and decrease procedural loss. This study analyzed cytoplasmic and periplasmic IL-2 expression for increased protein yield and significant biological activity. The study is focused on cloning IL-2 into a pET-SUMO and pET-28a vector that expresses IL-2 in soluble form and inclusion bodies, respectively. Both constructs were expressed into different E. coli expression strains, but the periplasmic and cytoplasmic expression of IL-2 was highest in overnight culture in Rosetta 2 (DE3). Therefore, E. coli Rosetta 2 (DE3) was selected for large-scale production and purification. Purified IL-2 was characterized by SDS-PAGE and western blotting, while its biological activity was determined using MTT bioassay. The results depict that the periplasmic and cytoplasmic IL-2 achieved adequate purification, yielding 0.86 and 0.51 mg/mL, respectively, with significant cytotoxic activity of periplasmic and cytoplasmic IL-2. Periplasmic IL-2 has shown better yield and significant biological activity in vitro which describes its attainment of native protein structure and function.

Single nucleotide polymorphism within chromosome 8q24 is associated with prostate cancer development in Saudi Arabia.

Objective: Genome-wide association studies have demonstrated that single nucleotide polymorphisms (SNPs) are important risk factors for the development of prostate cancer (PCa). Preliminary studies have suggested that the incidence of PCa in Saudi males is low but is probably familial or genetically related. Methods: To identify any possible association of SNP with PCa development in Saudi patients, we investigated a group of SNPs in Saudi PCa patients (n=85) and compared the outcomes to healthy normal controls (n=115) and nodular hyperplasia patients (n=120). DNA was extracted from paraffin-embedded formalin fixed tissue or whole blood from both patients' groups and healthy control group. A total of thirteen SNPs were genotyped using TaqMan® minor groove binder polymerase chain reaction assay. Results: The rs16901979A, s629242T and rs1447295A alleles were found at significantly higher frequency in PCa patients than controls (p<0.05). The rs16901979 CA genotype was found at significantly greater frequency in PCa patients than in healthy controls (43% vs. 14%, odds ratio=4.6, p=0.0001) and benign hyperplasia group (43% vs. 25%, odds ratio=2.2, p=0.009). Conclusion: Our study has highlighted the association of rs16901979 SNP with PCa in Saudi males. Such findings have important implications in the PCa diagnosis and in screening unaffected family members of Saudi patients.

Evaluation of Hi-C sequencing for the detection of gene fusions in hematologic and solid pediatric cancer samples.

HiC sequencing is a DNA-based next-generation sequencing method that preserves the 3D conformation of the genome and has shown promise in detecting genomic rearrangements in translational research studies. To evaluate HiC as a potential clinical diagnostic platform, analytical concordance with routine laboratory testing was assessed using primary pediatric leukemia and sarcoma specimens previously positive for clinically significant genomic rearrangements. Archived specimen types tested included viable and nonviable frozen leukemic cells, as well as formalin-fixed paraffin-embedded (FFPE) tumor tissues. Initially, pediatric acute myeloid leukemia (AML) and alveolar rhabdomyosarcoma (A-RMS) specimens with known genomic rearrangements were subjected to HiC analysis to assess analytical concordance. Subsequently, a discovery cohort consisting of AML and acute lymphoblastic leukemia (ALL) cases with no known genomic rearrangements based on prior clinical diagnostic testing were evaluated to determine whether HiC could detect rearrangements. Using a standard sequencing depth of 50 million raw read-pairs per sample, or approximately 5X raw genomic coverage, 100% concordance was observed between HiC and previous clinical cytogenetic and molecular testing. In the discovery cohort, a clinically relevant gene fusion was detected in 45% of leukemia cases (5/11). This study demonstrates the value of HiC sequencing to medical diagnostic testing as it identified several clinically significant rearrangements, including those that might have been missed by current clinical testing workflows. Key points: HiC sequencing is a DNA-based next-generation sequencing method that preserves the 3D conformation of the genome, facilitating detection of genomic rearrangements.HiC was 100% concordant with clinical diagnostic testing workflows for detecting clinically significant genomic rearrangements in pediatric leukemia and rhabdomyosarcoma specimens.HiC detected clinically significant genomic rearrangements not previously detected by prior clinical cytogenetic and molecular testing.HiC performed well with archived non-viable and viable frozen leukemic cell samples, as well as archived formalin-fixed paraffin-embedded tumor tissue specimens.

Levels of apolipoproteins as risk factors for coronary artery disease / Níveis de apolipoproteínas como fatores de risco para doença arterial coronariana

INTRODUCTION: The Saudi population is renowned for their unhealthy diet and physical inactivity. OBJECTIVE:To investigate apolipoproteins B (apo B), A-I (apo A-I) and B/A-I as risk factors that might be associated with increased incidence of the coronary artery disease. METHODS: Two hundred and twenty subjects suspected of having the coronary artery disease underwent coronary angiography and blood draw following a 12-hour fast. Apolipoproteins B and A-I were both measured by turbidimetric methods. RESULTS: One hundred and forty subjects were positive and 80 subjects were negative for the coronary artery disease. Both apolipoproteins were found to be statistically significant as risk factors for the coronary artery disease: apolipoprotein B (105.33±29.22 versus 94.56±24.35 mg/dL, p<0.003), apolipoprotein A-I (123.98±25.6 versus 133.5±24.1 mg/dL, p<0.004) and apolipoproteins B/A-I (0.88±0.28 versus 0.72±0.2, p<0.0001). CONCLUSIONS:Measurements of apolipoproteins B, A-I and calculation of apolipoproteins B/A-I ratio either instead of or in addition to the customary measurements of lipoprotein cholesterol may significantly add to predicting and assessing the coronary risk factors in the Saudi population.

INTRODUÇÃO: A população da Arábia Saudita é conhecida por sua dieta não-saudável e inatividade física. OBJETIVO: Investigar as apolipoproteínas B (apo B), AI (apo AI) e B/AI como fatores de risco que podem estar associados ao aumento da incidência da doença arterial coronariana. MÉTODOS: Duzentos e vinte pacientes com suspeita de doença art RESULTADOS: Cenerial coronariana foram submetidos à angiografia coronária e extração de sangue após jejum de 12 horas. As apolipoproteínas B e AI foram medidas por métodos turbidímetros.to e quarenta pacientes foram positivos e 80 foram negativos para a doença arterial coronariana. Ambas apolipoproteínas foram estatisticamente significativas como fatores de risco para doença arterial coronariana: apolipoproteínas B (105,33±29,22 versus 94,56±24,35 mg/dL, p<0,003), AI (123,98±25,6 versus 133,5±24,1, p<0,004) e B/A-I (0,88±0,28 versus 0,72±0,2, p<0,0001). CONCLUSÕES: As medidas das apolipoproteínas B, AI e o cálculo da relação B/AI tanto por ou em adição às medidas habituais de colesterol das lipoproteínas podem aumentar significativamente a previsão e avaliação dos fatores de risco coronariano na população saudita.

Association of lipids with coronary heart disease in a saudi population / Associação de lípidos com doença cardíaca coronariana em população saudita

BACKGROUND: The Saudi population is known to have an unhealthy diet in addition to physical inactivity. OBJECTIVE: To investigate the lipid-mediated risk factors that might be associated with increased incidence of coronary heart diseasein the Saudi population as this was found in Western populations. MATERIALS AND METHODS: Two hundred and twenty subjects suspected of having coronary heart disease underwent coronary angiography and blood draw following a 12-hour fast. Total and HDL cholesterol, triglycerides, Lp(a) and lipoprotein lipase were measured by standard methods. Small, dense LDL was measured by the iodixanol method with an ultracentrifugation of only 2.5 hours. RESULTS: One hundred and forty subjects were found to be positive for coronary heart disease while 80 subjects were shown to be negative for this disease. Statistically significant risk factors for coronary heart disease in the Saudi population were hypertriglyceridemia (1.93±0.95 versus1.45±0.16 mmol/L;p<0.0001); low HDL cholesterol (1.09±0.55 vs 1.33±0.63 mmol/L, p=0.0001); high Lp(a) (46.8±45.58 versus 29.06±17.03 mg/dL;p=0.019); and the presence of small, dense LDL (1.0314±0.0028 versus 1.0300±0.0003 g/kg;p=0.0099). Total cholesterol (4.99±1.11 versus 4.75±1.11 mmol/L;p=0.099), LPL (35.56±26.6 versus 27.89±11.96 IU/L;p<0.059), and LDL cholesterol (3.06±1.12 versus 2.79±1.08 mmol/L;p=0.08) were not found to be statistically significant coronary heart disease risk factors. CONCLUSIONS: This study indicates that high TG, low HDL, high Lp(a) and the presence of small, dense LDL may contribute to the incidence of coronary heart disease and that TC was not significantly associated with incidence of coronary heart disease in the Saudi population.

CONTEXTO: A população saudita é conhecida por apresentar dietas não saudáveis, além de inatividade física. OBJETIVO: Investigar os lipídeos como fatores de risco que podem associar-se com o aumento da incidência de doença cardíaca coronariana na população saudita, uma vez que isso foi encontrado na população ocidental. MATERIAIS E MÉTODOS: Duzentos e vinte indivíduos com suspeita de doença cardíaca coronariana submeteram-se a angiografia coronária e coleta de sangue, após jejum de 12 horas. Colesterol total e HDL, triglicerídeos, Lp(a) e lipase lipoproteica foram calculados por métodos padrão. LDL pequena e densa foi medida pelo método do iodixanol com ultracentrifugação por apenas 2,5 horas. RESULTADOS: Cento e quarenta indivíduos apresentaram resultado positivo para doença cardíaca coronariana, enquanto 80 pacientes mostraram resultado negativo. Os fatores de risco estatisticamente significantes para doença cardíaca coronariana na população saudita foram hipertrigliceridemia (1,93±0,95 versus 1,45±0,16 mmol/L; p<0,0001); baixo colesterol HDL (1,09±0,55 versus 1,33±0,63 mmol/L; p=0,0001); Lp(a) elevada (46,8±45,58 versus 29,06±17,03 mg/dL; p=0,019) e a presença de LDL pequena e densa (1,0314±0,0028 versus 1,0300±0,0003 g/kg; p=0,0099). Colesterol total (4,99±1,11 versus 4,75±1,11 mmol/L; p=0,099), colesterol LPL (35,56±26,6 versus 27,89±11,96 IU/L; p<0,059) e LDL (3,06±1,12 versus 2,79±1,08 mmol/L; p=0,08) não foram fatores de risco estatisticamente significantes para doença cardíaca coronariana. CONCLUSÕES: Este estudo indica que elevados triglicerídeos, baixo HDL, elevada Lp(a) e a presença de LDL pequena e densa podem contribuir para a incidência de doença cardíaca coronariana. O colesterol total não foi significantemente associado à incidência de doença cardíaca coronariana na população saudita.