Homology-directed gene-editing approaches for hematopoietic stem and progenitor cell gene therapy

AbstractThe advent of next-generation genome engineering tools like CRISPR-Cas9 has transformed the field of gene therapy, rendering targeted treatment for several incurable diseases. Hematopoietic stem and progenitor cells (HSPCs) continue to be the ideal target cells for gene manipulation due to their long-term repopulation potential. Among the gene manipulation strategies such as lentiviral gene augmentation, non-homologous end joining (NHEJ)-mediated gene editing, base editing and prime editing, only the homology-directed repair (HDR)-mediated gene editing provides the option of inserting a large transgene under its endogenous promoter or any desired locus. In addition, HDR-mediated gene editing can be applied for the gene knock-out, correction of point mutations and introduction of beneficial mutations. HSPC gene therapy studies involving lentiviral vectors and NHEJ-based gene-editing studies have exhibited substantial clinical progress. However, studies involving HDR-mediated HSPC gene editing have not yet progressed to the clinical testing. This suggests the existence of unique challenges in exploiting HDR pathway for HSPC gene therapy. Our review summarizes the mechanism, recent progresses, challenges, and the scope of HDR-based gene editing for the HSPC gene therapy.</jats:p

SMALL Molecules Mediated Hematopoietic STEM and Progenitor CELLS Expansion for GENE Editing Application

Abstract Genome editing of Hematopoietic stem Cells has revolutionized the treatment strategies for genetic disorders. Despite this, it still remains a great challenge as hematopoietic stem cells tend to lose its stem-ness during the ex vivo culture and gene editing process. The need for large dose of CD34+ HSPCs for manipulation makes it a seemingly difficult strategy. Recent works suggest that the potential effects of small molecules in expanding cord blood HSPCs ex vivo promoting self-renewal and delaying differentiation. We screened several reported small molecules to identify a condition that promotes the expansion of adult HSPCs for gene manipulation process. The mobilized Peripheral blood HSPCs are purified and cultured with a cytokine cocktail. Along with the cytokine cocktail, we tested several small molecules and in different combinations. Expression of cell surface receptors were analysed by FACS after 12 days of ex vivo culture. Our screening identified a unique culture condition that expanded the primitive stem cell population (CD34+/CD133+/CD90+cells) along with the early progenitors (CD34+/CD133+) and the progenitors (CD34+). Our culture conditions expanded the primitive cells by 20 folds compared to the mock treated cells. Our treatment release experiments suggested that the expansion is due to our culture conditions and are reversible.The colony forming cell (CFC) assay showed about 30 fold increase in the numbers of multilineage colony forming cell (CFU-GEMM) thereby ensuring the proliferation and differentiation capacity of expanded HSPCs. Their differentiation ability was also confirmed by ex vivo differentiation into Megakaryocytes. Our treatment conditions reduced the apoptosis rate during the ex vivo culture and improved their cell migration response towards SDF. The reduced reactive oxygen species levels and increased CXCR4 expression were observed in our expanded HSPCs and these might be the possible reasons for the low apoptosis and better cell migration respectively. Disclosures No relevant conflicts of interest to declare. </jats:sec

School-Based Exercise and Life Style Motivation Intervention (SEAL.MI) on Adolescent’s Cardiovascular Risk Factors and Academic Performance: Catch Them Young

There are shreds of evidence of shared biological mechanisms between obesity and hypertension during childhood intoadulthood, and loads of research literature has proven that it will profoundly cost nations’ economies and health if neglected. The prevention and early diagnosis of cardiovascular risk factors such as overweight and hypertension is an essential strategy for control, effective treatment and prevention of its’ complications. The study aims to assess the effect of school-based Exercise and Lifestyle Motivation Intervention (SEAL-MI) on adolescents’ cardiovascular risk factors and academic performance. An experimental study was conducted among 1005 adolescents—520 and 485 were randomly selected for the control and study groups, respectively.A structured interview questionnaire was used to collect demographic details and data related to dietary habits, physical activity, sleep qualityand academic performance. The study group adolescents were given the SEAL-MI for six months, including a school-based rope exercise for 45 min per day for 5 days a week and a motivation intervention related to dietary habits, physical activity, and sleep. Post tests-1 and 2 were done after 3 and 6 months of intervention.The prevalence of overweight among adolescents was 28.73%, and prehypertension was 9.26%. Among overweight adolescents, the prevalence of prehypertension was found to be very high (32.25%). There was a significant weight reduction in post-intervention B.P. (p = 0.000) and improvement in dietary habits, physical activity, sleep (p = 0.000), and academic performance. A significant positive correlation was found between BMI and SBP (p = 0.000) and BMI and academic performance (p = 0.003). The linear regression analyses revealed that the gender (ß: 0.47, 95% CI: 0.39, 0.81), age (ß: 0.39, 95% CI: 0.17, 0.46), family income (ß: 0.2, 95% CI: 0.41, 0.5), residence (ß: 0.19, 95% CI: 0.01, 0.27), and type of family (ß: 0.25, 95% CI: 0.39, 0.02) had the strongest correlation with the BMI of the adolescents. Additionally, Mother’s education (ß: 0.35, 95% CI: 0.18, 0.59) had the strongest correlation with the SBP of the adolescents. In contrast, the DBP was negatively persuaded by age (ß: −0.36, 95% CI: 1.54, 0.29) and gender (ß: −0.26, 95% CI: 1.34, 0.12) of the adolescents. Regular practice of rope exercise and lifestyle modification such as diet, physical activity, and quality sleep among adolescents prevent and control childhood CVD risk factors such asoverweight and hypertension. The SEAL-MI may lead to age-appropriate development of adolescents as well as improve their academic performance and quality of life. Giving importance to adolescents from urban habitats, affluent, nuclear families, and catching them young will change the disease burden significantly

School-Based Exercise and Life Style Motivation Intervention (SEAL.MI) on Adolescent’s Cardiovascular Risk Factors and Academic Performance: Catch Them Young

There are shreds of evidence of shared biological mechanisms between obesity and hypertension during childhood intoadulthood, and loads of research literature has proven that it will profoundly cost nations’ economies and health if neglected. The prevention and early diagnosis of cardiovascular risk factors such as overweight and hypertension is an essential strategy for control, effective treatment and prevention of its’ complications. The study aims to assess the effect of school-based Exercise and Lifestyle Motivation Intervention (SEAL-MI) on adolescents’ cardiovascular risk factors and academic performance. An experimental study was conducted among 1005 adolescents—520 and 485 were randomly selected for the control and study groups, respectively.A structured interview questionnaire was used to collect demographic details and data related to dietary habits, physical activity, sleep qualityand academic performance. The study group adolescents were given the SEAL-MI for six months, including a school-based rope exercise for 45 min per day for 5 days a week and a motivation intervention related to dietary habits, physical activity, and sleep. Post tests-1 and 2 were done after 3 and 6 months of intervention.The prevalence of overweight among adolescents was 28.73%, and prehypertension was 9.26%. Among overweight adolescents, the prevalence of prehypertension was found to be very high (32.25%). There was a significant weight reduction in post-intervention B.P. (p = 0.000) and improvement in dietary habits, physical activity, sleep (p = 0.000), and academic performance. A significant positive correlation was found between BMI and SBP (p = 0.000) and BMI and academic performance (p = 0.003). The linear regression analyses revealed that the gender (ß: 0.47, 95% CI: 0.39, 0.81), age (ß: 0.39, 95% CI: 0.17, 0.46), family income (ß: 0.2, 95% CI: 0.41, 0.5), residence (ß: 0.19, 95% CI: 0.01, 0.27), and type of family (ß: 0.25, 95% CI: 0.39, 0.02) had the strongest correlation with the BMI of the adolescents. Additionally, Mother’s education (ß: 0.35, 95% CI: 0.18, 0.59) had the strongest correlation with the SBP of the adolescents. In contrast, the DBP was negatively persuaded by age (ß: −0.36, 95% CI: 1.54, 0.29) and gender (ß: −0.26, 95% CI: 1.34, 0.12) of the adolescents. Regular practice of rope exercise and lifestyle modification such as diet, physical activity, and quality sleep among adolescents prevent and control childhood CVD risk factors such asoverweight and hypertension. The SEAL-MI may lead to age-appropriate development of adolescents as well as improve their academic performance and quality of life. Giving importance to adolescents from urban habitats, affluent, nuclear families, and catching them young will change the disease burden significantly.</jats:p

School Based Exercise and Life Style Motivation Intervention (SEAL.MI) on Adolescent's Cardio Vascular Risk Factors and Academic Performance: Catch Them Young

There are evidences of shared biological mechanisms between obesity and hypertension during childhood in the adulthood, and loads of research literatures have proven that, it will cost the economies and health of nations profoundly if neglected. The Prevention and early diagnosis of cardiovascular risk factors such as overweight and hypertension is an essential strategy for control, effective treatment and prevention of its&amp;rsquo; complications. The aim of the study is to assess the effect of school based Exercise and Life style Motivation Intervention (SEAL-MI) on adolescent's cardiovascular risk factors and academic performance. An experimental study was conducted among 1005 adolescents - 520 and 485 adolescents were randomly selected for control and study group, respectively. Demographic details and the data related to dietary habits, physical activity and sleep quality were collected by a structured interview questionnaire. The study group adolescents were given the SEAL-MI for six months which includes a school based rope exercise for 45 minutes per day for 5 days a week and a motivation intervention related to dietary habits, physical activity and sleep. Post-test 1 and 2 were done after 3 and 6 months of intervention. The prevalence of overweight among adolescents was 28.73% and prehypertension was 9.26%. Among overweight adolescents, the prevalence of prehypertension was found to be very high (32.25%). In post intervention, there was a significant reduction in weight, BP (p=0.000) and improvement in dietary habits, physical activity, sleep (p=0.000) and academic performance. A significant positive correlation was found between BMI and SBP (p=0.000) and BMI and academic performance (p=0.003). The linear regression analyses revealed that the gender (&amp;szlig;: 0.47, 95% CI: 0.39, 0.81), age (&amp;szlig;: 0.39, 95% CI: 0.17, 0.46), family income (&amp;szlig;: 0.2, 95% CI: 0.41, 0.5), residence (&amp;szlig;: 0.19, 95% CI: 0.01, 0.27) and type of family (&amp;szlig;: 0.25, 95% CI: 0.39, 0.02) had the strongest correlate with the BMI of the adolescents. Also, Mother&amp;rsquo;s education Mother&amp;rsquo;s education (&amp;szlig;: 0.35, 95% CI: 0.18, 0.59) had the strongest correlate with the SBP of the adolescents. In contrast, the DBP was negatively persuaded by age (&amp;szlig;: -0.36, 95% CI: 1.54, 0.29) and gender (&amp;szlig;: -0.26, 95% CI: 1.34, 0.12) of the adolescents. Regular practice of rope exercise and lifestyle modification such as diet, physical activity and sleep quality among adolescents prevent and control childhood CVD risk factors like overweight, hypertension. The SEAL-MI may lead to age appropriate development of adolescents and improves their academic performance and quality of life. Giving importance to the adolescents from urban habitat, from affluent nuclear family and catching them young will bring significant change in the disease burden.</jats:p

Knowledge, attitude and preparedness of healthcare students toward basic life support at King Khalid University, Abha, Kingdom of Saudi Arabia

Background Worldwide, millions of people die of sudden cardiac arrest every year. A well-timed cardiopulmonary resuscitation (CPR) increases the possibility of survival by two- to fourfolds. This study aimed to assess the knowledge, attitude, and preparedness of health care students toward basic life support (BLS) at King Khalid University. Methods A cross-sectional study was conducted among the health care students of King Khalid University from August to October 2020. Data were collected using a pretested, semi-structured questionnaire and the data were analyzed using Statistical Package for the Social Sciences. Resulrs The total number of participants was 346. Overall, the participant’s knowledge regarding the BLS was inadequate. Majority of the participants were not aware of the acronyms used in BLS. The level of education has a significant impact on the knowledge, whereas gender has no significant impact on the knowledge. The answers to the attitude and the preparedness items were also not satisfying. Lack of knowledge is one of the common reasons for not performing BLS. Periodical training program and refresher courses were the most recommended methods to increase the knowledge toward the BLS. Conclusion It is evident from the current study that there is a lack of knowledge and preparedness toward BLS among most health care students. It is recommended to incorporate more BLS training and refresher courses in the health care college curricula

Association of Maternal Observation and Motivation (MOM) Program with M-Health Support on Maternal and Newborn Health

Maternal and child nutrition has been a critical component of health, sustainable development, and progress in low- and middle-income countries (LMIC). While a decrement in maternal mortality is an important indicator, simply surviving pregnancy and childbirth does not imply better maternal health. One of the fundamental obligations of nations under international human rights law is to enable mothers and teenage girls to endure pregnancy and delivery as an aspect of their enjoyment of reproductive and sexual health and rights and live a dignified life. The aim of this study was to discover the correlation between the Maternal Observation and Motivation (MOM) program and m-Health support for maternal and newborn health. A Comparative study was done among 109 pregnant mothers (study group-94; control group-102 mothers) with not less than 20 weeks of gestation. Maternal outcomes such as Hb, weight gain and newborn results like birth weight and crown- heel length was obtained on the baseline, 28 and 36 weeks of gestation. Other secondary data collected were abortion, stillbirth, low birth weight, major congenital malformations, twin or triplet pregnancies, physical activity and maternal wellbeing. The MOM intervention included initial face to face education, three in-person visits and eight virtual health coaching by WhatsApp. The baseline data on Hb of the mothers show that 31(32.98%) vs 27(28.72%) of the study and control group had anaemia, which improved to 27.66% and 14.98% among study group mothers at 28 and 36 weeks of gestation (p&amp;lt;0.001). The weight gain (p&amp;lt; 0.001), level of physical activity (p&amp;lt; 0.001), and maternal wellbeing (p&amp;lt; 0.01) also had significant differences after the Intervention. Even after controlling for potentially confounding variables, the maternal food practices regression model revealed that birth weight was directly correlated with consumption of milk (p 0.001), fruits (p 0.01), and green vegetables (p 0.05).As per the physical activity and maternal wellbeing regression model, the birth weight and crown heel length were strongly related with the physical activity and maternal wellbeing of mothers at 36 weeks of gestation (p &amp;lt;0.05). Combining the MOM intervention with standard antenatal care is a safe and effective way to improve maternal welfare while upholding pregnant mothers' human rights.</jats:p