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Introduction: Colorectal cancer (CRC) is a prevalent malignancy with significant morbidity and mortality worldwide. A deeper understanding of the interaction of cancer cells with other cells in the tumor microenvironment is crucial to devise effective therapeutic strategies. MUC2, a major component of the protective mucus layer in the gastrointestinal tract, has been implicated in CRC progression and immune response regulation. Method: In this study, we sought to elucidate the relationship between MUC2 expression and immune infiltration within CRC using in vitro models involving two well-established cell lines, HT-29 and LS-174T. By employing CRISPR-mediated MUC2 knockout, we investigated the influence of MUC2 on tumor immune infiltration and its interplay with T cells and NK cells enriched peripheral blood mononuclear cells (PBMCs) in 3D spheroid cultures. Results: While MUC2 was more abundant in LS-174T cell line compared to HT-29, its knockout resulted in increased immune infiltration solely in the HT-29 cell line, but not in the LS-174T cell line. We revealed that the removal of MUC2 protein was compensated in LS-174T by the expression of other gel-forming mucin proteins (MUC6, MUC5B) commonly expressed in the gastrointestinal epithelium, while this was not observed in HT-29 cell line. Conclusion: Our study is the first to demonstrate that MUC2 functions as a physical barrier to immune infiltration in colorectal cancer (CRC) in vitro. In HT-29 cells, MUC2 knockout increased immune infiltration, while in LS-174T cells, compensatory expression of other mucins (MUC6, MUC5B) maintained the barrier. These findings reveal the complexity of mucin biology in CRC and suggest that targeting mucin pathways could be a novel therapeutic approach. [ABSTRACT FROM AUTHOR]

Background: FFPE tissue samples are commonly used in biomedical research and are a valuable source for next-generation sequencing in oncology, however, extracting RNA from these samples can be difficult the quantity and quality achieved can impact the downstream analysis. This study compared the effectiveness of seven different commercially available RNA extraction kits specifically designed for use with FFPE samples in terms of the quantity and quality of RNA recovered. Methods: This study used 9 samples of FFPE tissue from three different types of tissue (Tonsil, Appendix and lymph node of B-cell lymphoma) to evaluate RNA extraction methods. Three sections of 20 µm of each sample were combined per sample. The slices were distributed in a systematic manner to prevent any biases. Each of the 7 commercially available RNA extraction kits were used according to manufacturer's instructions, with each sample being tested in triplicate resulting in a total of 189 extractions. The concentration, RNA quality score (RQS) and DV200 of each extraction was analysed using a nucleic acid analyser to determine the quantity and quality of the recovered RNA. Results: This study found that despite processing the FFPE samples in the same standardized way, there were disparities in the quantity and quality of RNA recovered across the different tissue types. Additionally, the study found notable differences in the quantity of RNA recovered when using different extraction kits. In terms of quality, three of the kits performed better than the other four in terms of RQS and DV200 values. Conclusion: Though many laboratories have developed their own protocols for specific tissue types, using commercially available kits is still a popular option. Although these kits use similar processes and extraction procedures, the amount and quality of RNA obtained can vary greatly between kits. In this study, among the kits tested, while the Roche kit, provided a nearly systematic better-quality recovery than other kits, the ReliaPrep FFPE Total RNA miniprep from Promega yielded the best ratio of both quantity and quality on the tested tissue samples. [ABSTRACT FROM AUTHOR]

The current study aimed to explore the experience, expression and manifestation of parentification among adolescents experiencing parental discord in Pakistan. Interpretative phenomenological analysis design (IPA) was used for the study, and purposive sampling strategy was employed. The sample included eight boys and girls of class 9th, 10th, 11th and 12th of government secondary and higher secondary schools of Lahore, Pakistan, and they were selected by non‐probability purposive sampling. The mean age of the participants was 16.37 and SD was 1.11. In depth, interviews were conducted with each participant. The interviews were audio recorded and transcribed, and IPA analysis was used for data analysis and interpretation. Three group experiential themes were identified: functional maturation, emotional ramifications and stifling self. The responses revealed that parentified adolescents were assumed accountable for a wide range of responsibilities, including financial contribution, managing household, maintaining familial relationships as well as bringing up younger siblings. This study will help understand the indigenous experience of parentification of Pakistani adolescents, and it will facilitate further elaboration of understanding the nature of the common practice of parentification, as well as its impact upon the adolescents. [ABSTRACT FROM AUTHOR]

Background: Intolerance of uncertainty is a dispositional characteristic, which arises from negative beliefs about uncertainty and shows negative reactionary inclination to uncertain situations. Objective: To develop an indigenous valid and reliable scale in order to explore intolerance of uncertainty among the university students in cultural context of Pakistan. Method: A cross sectional study design was applied and it was conducted in public as well as private sector universities situated in Lahore city. The data was collected from November, 2020 to January 2021. A standardized three step model was used to develop a reliable scale to measure Intolerance of Uncertainty in university students. The interviews were conducted with 10 university students and initially 108 items were generated. The dubious items were eliminated and a list of 56 items was generated. Moreover, a panel comprising experts had reviewed these items for validating empirically and finalized 41 items were given to a group of 14 undergraduate students as pilot project. Hence, IUSS scale with an annotated performa acquiring the age, gender, class, parental educational status etc was provided to 310 students (44.80% male and 55.20% female) through a stratified sampling technique. Result: A four (04) factor structure of Intolerance of Uncertainty scale was finalized through Exploratory Factor Analysis and the factors were named as apprehension (9 items), lack of Decisiveness (9 items), lack of Stability (11 items) and lack of Acceptance (7 items). Results showed that the indigenously developed selfreport measure has good internal consistency (Chronbach's Alpha=.81), test re-test reliability r= .88 (P <.001), concurrent validity r= .63 (P <.001). Results also showed that 55.2% of participants have mild, 28.4% have moderate and 15.8% have severe level of intolerance of uncertainty. Conclusion: An indigenous measuring tool containing 36 items with self-reporting features for ascertaining Intolerance of uncertainty in students enrolled in Universities was developed which possess sound psychometric properties. [ABSTRACT FROM AUTHOR]

Objective: To find out the experience and expression of academic stressors in college students and to develop a reliable and valid measure. Methodology: A sample of 244 college students with age range 15-20 (M = 17.38, SD = 1.22) were selected through the convenient sampling technique. Both boys and girls living in intact families, having no physical or psychological disorder studying in different government and private institutes of city Lahore were included in the study. A three-step model (item generation, expert validation, and pilot study) was used to develop the scale. Results: Descriptive statistics revealed n=97(39%) 1st year and 147 (61%) 2nd year students with highest number living in nuclear family system (n=150(61%). Exploratory factor analysis using Principal Component factor analysis revealed two factors: sustaining performance and psychosocial stressors. The value of Kaiser-Meyer-Olkin was .87 and the Bartlett test was 929.90 with p=0.000. Student Problem Checklist was used to establish the concurrent validity of the scale and was found to be r=.92. The internal consistency was 0.92 (p<0.001) and the test-retest reliability of ASS was r=.67 (p<0.001). Girls experienced more academic stressor (M=35.53, SD=19.43, p<0.001) as compared to boys (M=31.72, SD=13.45). Conclusion: The Academic Stressor Scale is an indigenous scale with established psychometrics that provides a depiction of academic stressors faced by Students in Pakistani culture. [ABSTRACT FROM AUTHOR]

Trisomy 21 is one of the commonly identified genetic causes of Intellectual Disability. Such children experience a lack of attention and neglect by parents and teachers due to their slow response to intervention, which sometimes becomes demotivating for caregivers. However, with the recent medical advances and institutional support, avenues have been opened to help overcome the challenges related to Down's Syndrome. The present case study highlights the life skills training of a 7 year old child with Down's Syndrome, studying in a special education institute of Lahore. The post-assessment showed that consistent practice of behavior modification procedures led to significant improvements in the adaptive functioning of the child. [ABSTRACT FROM AUTHOR]