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  1. Siddique MR, Shynder S, Ashraf MA, Yusoff I, Wajid A
    Chem Cent J, 2012 Jul 18;6(1):69.
    PMID: 22809083 DOI: 10.1186/1752-153X-6-69
    BACKGROUND: Renilla Luciferase reporter gene (rLuc) GL4.82 and GL4.13 promoter are key player in transfection, but precise knowledge of its targets in colon cancer remains limited. The aim of this study was to characterize the best transfection technique to produce a stable transfected colon DLD1 (colorectal adenocarcinoma cell line), therefore imaging based approaches were employed.

    RESULTS: DLD1 cells were transfected with a Plasmid (SV40-RLuc) carrying Renilla luciferase under the control of the SV-40 promoter, by using two different transfection techniques. Cells expressing the required DNA were isolated after antibiotic (Puramycin) selection. Clones of DLD-1/SV40-RLuc were produced using two different techniques (96 well plates and Petri dish) and their florescence intensity was recorded using IVIS machine (Calliper Life Sciences, Hopkinton, USA). Both techniques were characterized with the help of serial dilution technique. Results from this study substantiated that electroporation is the best. As expected, clones varied in their specific luciferase activity along with the dilutions. With the increase in cell concentration increase in intensity of florescence was recorded.

    CONCLUSIONS: Based on the results we are confident that this transfected cell line DLD-1/SV40-RLuc (colorectal adenocarcinoma cell line) is the best for further Orthotopic Xenotransplantation Studies and in-vivo experiments as well. Investigation shows that DLD1/SV-rLuc cells have gained little bit resistance against both drugs therefore further study is suggested to know the reasons.

  2. Algin S, Sajib MWH, Ahmed SN, Siddique MR, Reza MM, Tanzilla NJ, et al.
    Cureus, 2025 Feb;17(2):e79667.
    PMID: 40017580 DOI: 10.7759/cureus.79667
    Introduction Obsessive-compulsive disorder (OCD) is a persistent psychiatric condition that causes significant clinical and functional impairments. Recent research suggests a link between deficiencies in micronutrients, particularly vitamin B12, folic acid, and elevated homocysteine, and the development of OCD. This study investigates the blood levels of these micronutrients and their relationship to OCD severity, with an emphasis on potential gender differences. Methods This cross-sectional study included 300 drug-free OCD patients. Serum levels of vitamin B12, folic acid, and homocysteine were measured using established biochemical methods. The Yale-Brown Obsessive Compulsive Scale (Y-BOCS) was used to assess clinical severity. Data were examined to determine relationships between micronutrient levels and OCD severity and differences between male and female patients. Results This study found that women had higher levels of vitamin B12 (405.3 ± 15.1 vs. 360.4 ± 14.3) and folic acid (7.18 ± 0.27 vs. 5.76 ± 0.25) but lower levels of homocysteine (9.35 ± 0.64 vs. 14.4 ± 0.60) compared to men. Higher folic acid levels were linked to study participants having higher levels of education (at a college or university, where subjects are studied at an advanced level) compared to those with primary-level education. Lower vitamin B12 levels were linked to family mental health history and noncommunicable diseases. Women exhibited lower levels of substance use but higher rates of self-harm and suicide attempts. Elevated homocysteine levels were linked to longer illness duration and more severe OCD symptoms. Conclusion These findings suggest that imbalances in micronutrients, particularly vitamin B12, folic acid, and homocysteine, may contribute to OCD severity and treatment resistance. Gender-specific variations in micronutrient levels could provide valuable insights into personalized OCD therapy choices. Further longitudinal studies are needed to understand these relationships and their potential as therapeutic targets.
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