Impact of GSK-3ß and CK-1δ on Wnt signaling pathway in alzheimer disease: A dual target approach.

Alzheimer's disease (AD) is an enigmatic neurological illness that offers few treatment options. Recent exploration has highlighted the crucial connection of the Wnt signaling pathway in AD pathogenesis, shedding light on potential therapeutic targets. The present study focuses on the dual targeting of glycogen synthase kinase-3ß (GSK-3ß) and casein kinase-1δ (CK-1δ) within the framework of the Wnt signaling pathway as a possible technique for AD intervention. GSK-3ß and CK-1δ are multifunctional kinases known for their roles in tau hyperphosphorylation, amyloid processing, and synaptic dysfunction, all of which are major hallmarks of Alzheimer's disease. They are intricately linked to Wnt signaling, which plays a pivotal part in sustaining neuronal function and synaptic plasticity. Dysregulation of the Wnt pathway in AD contributes to cognitive decline and neurodegeneration. This review delves into the molecular mechanisms by which GSK-3ß and CK-1δ impact the Wnt signaling pathway, elucidating their roles in AD pathogenesis. We discuss the potential of small-molecule inhibitors along with their SAR studies along with the multi-targetd approach targeting GSK-3ß and CK-1δ to modulate Wnt signaling and mitigate AD-related pathology. In summary, the dual targeting of GSK-3ß and CK-1δ within the framework of the Wnt signaling pathway presents an innovative and promising avenue for future AD therapies, offering new hope for patients and caregivers in the quest to combat this challenging condition.

Missing Opportunities in the Screening of Alcohol Use and Problematic Use, and the Provision of Brief Advice and Treatment Information Among Individuals With Alcohol Use Disorder.

OBJECTIVES: The aim of this study was to identify sociodemographic and substance-related factors associated with being screened, receiving advice or treatment information from healthcare providers, among individuals who met the criteria for the past 12-month alcohol use disorder (AUD). METHODS: The 2015-2019 National Survey on Drug Use and Health data were analyzed to identify factors associated with being (1) asked about alcohol used among adults with AUD, who visited a healthcare provider within the past 12 months, and were not receiving AUD treatment (sample 1, n = 13,321); (2) asked about problematic use; (3) advised to reduce consumption; and (4) offered alcohol treatment information, among those in sample 1 who were asked about their use (n = 6,905). RESULTS: About half (52.9%) in sample 1 were asked about their alcohol use. Among them, 21.6% were asked about problematic use, 17.7% were advised to reduce alcohol consumption, and 7.6% were offered information. The odds of being asked about alcohol use among male participants were 0.72 times the odds of female participants; however once asked, male participants showed greater odds of being asked about problematic use (adjusted odds ratio [aOR] = 1.53, 95% confidence interval [CI] = 1.29-1.82), advised to reduce consumption (aOR = 1.64, 95% CI = 1.24-2.16), and offered treatment information (aOR = 1.77, 95% CI = 1.34-2.35). As compared with non-Hispanic White participants, other racial/ethnic groups were less likely to be asked about alcohol use; however, once asked, no differences were observed for other outcomes. CONCLUSIONS: Significant gaps in the screening and provision of advice or treatment information were identified, particularly for racial/ethnic and sex subgroups. Reducing barriers for effective screening could help address AUD-related disparities.

A sucrose non-fermenting-1-related protein kinase 1 gene from wheat, TaSnRK1α regulates starch biosynthesis by modulating AGPase activity.

Major portion of wheat grain consist of carbohydrate, mainly starch. The proportion of amylose and amylopectin in starch greatly influence the end product quality. Advancement in understanding starch biosynthesis pathway and modulating key genes has enabled the genetic modification of crops resulting in enhanced starch quality. However, the regulation of starch biosynthesis genes still remains unexplored. So, to expand the limited knowledge, here, we characterized a Ser/Thr kinase, SnRK1α in wheat and determined its role in regulating starch biosynthesis. SnRK1 is an evolutionary conserved protein kinase and share homology to yeast SNF1. Yeast complementation assay suggests TaSnRK1α restores growth defect and promotes glycogen accumulation. Domain analysis and complementation assay with truncated domain proteins suggest the importance of ATP-binding and UBA domain in TaSnRK1α activity. Sub-cellular localization identified nuclear and cytoplasmic localization of TaSnRK1α in tobacco leaves. Further, heterologous over-expression (O/E) of TaSnRK1α in Arabidopsis not only led to increase in starch content but also enlarges the starch granules. TaSnRK1α was found to restore starch accumulation in Arabidopsis kin10. Remarkably, TaSnRK1α O/E increases the AGPase activity suggesting the direct regulation of rate limiting enzyme AGPase involved in starch biosynthesis. Furthermore, in vitro and in vivo interaction assay reveal that TaSnRK1α interacts with AGPase large sub-unit. Overall, our findings indicate that TaSnRK1α plays a role in starch biosynthesis by regulating AGPase activity.

Exploring the associations between serious psychological distress and the quantity or frequency of tobacco, alcohol, and cannabis use among pregnant women in the United States.

Serious Psychological Distress (SPD) and prenatal exposure to substances are associated with adverse outcomes for pregnant individuals and their developing offspring. This study aims to examine the relationship between SPD and quantity, or frequency of substance use among pregnant women in the United States (US). Descriptive and negative binomial regression analyses of the 2015-2019 National Survey on Drug Use and Health (NSDUH) were conducted among 3373 pregnant women (18 to 44 years old) to examine the association between SPD and (1) average number of cigarettes smoked in the past 30 days, (2) number of days of binge drinking in the past 30 days, and (3) number of days of cannabis use in the past 30 days. About 6% of the study population experienced SPD in the past 30 days. Compared to pregnant women who did not report SPD, pregnant women experiencing SPD showed greater rates in the number of cigarettes smoked during the past 30 days (IRR = 2.1, 95%CI = 1.1, 4.5), the number of days of binge drinking in the past 30 days (IRR = 5.1, 95%CI = 1.7, 15.4), and the number of days of cannabis use in the past 30 days (IRR = 2.9, 95%CI = 1.3, 6.5). Our results extend findings from prior research by documenting an association between SPD and the quantity and frequency of substance use among pregnant women in the US. Individual and structural interventions addressing SPD and/or substance might help reduce the impact of these comorbid conditions on expectant parents and their offspring.

Rethinking underutilized cereal crops: pan-omics integration and green system biology.

MAIN CONCLUSION: Due to harsh lifestyle changes, in the present era, nutritional security is needed along with food security so it is necessary to include underutilized cereal crops (UCCs) in our daily diet to counteract the rising danger of human metabolic illness. We can attain both the goal of zero hunger and nutritional security by developing improved UCCs using advanced pan-omics (genomics, transcriptomics, proteomics, metabolomics, nutrigenomics, phenomics and ionomics) practices. Plant sciences research progressed profoundly since the last few decades with the introduction of advanced technologies and approaches, addressing issues of food demand of the growing population, nutritional security challenges and climate change. However, throughout the expansion and popularization of commonly consumed major cereal crops such as wheat and rice, other cereal crops such as millet, rye, sorghum, and others were impeded, despite their potential medicinal and nutraceutical qualities. Undoubtedly neglected underutilized cereal crops (UCCs) also have the capability to withstand diverse climate change. To relieve the burden of major crops, it is necessary to introduce the new crops in our diet in the way of UCCs. Introgression of agronomically and nutritionally important traits by pan-omics approaches in UCCs could be a defining moment for the population's well-being on the globe. This review discusses the importance of underutilized cereal crops, as well as the application of contemporary omics techniques and advanced bioinformatics tools that could open up new avenues for future study and be valuable assets in the development and usage of UCCs in the perspective of green system biology. The increased and improved use of UCCs is dependent on number of factors that necessitate a concerted research effort in agricultural sciences. The emergence of functional genomics with molecular genetics might gear toward the reawakening of interest in underutilized cereals crops. The need of this era is to focus on potential UCCs in advanced agriculture and breeding programmes. Hence, targeting the UCCs, might provide a bright future for better health and scientific rationale for its use.

Interaction between long noncoding RNA (lnc663) and microRNA (miR1128) regulates PDAT-like gene activity in bread wheat (Triticum aestivum L.).

Amylose, a starch subcomponent, can bind lipids within its helical groove and form an amylose-lipid complex, known as resistant starch type 5 (RS-5). RS contributes to lower glycaemic index of grain with health benefits. Unfortunately, genes involved in lipid biosynthesis in wheat grain remain elusive. Our study aims to characterize the lipid biosynthesis gene and its post-transcriptional regulation using the parent bread wheat variety 'C 306' and its EMS-induced mutant line 'TAC 75' varying in amylose content. Quantitative analyses of starch-bound lipids showed that 'TAC 75' has significantly higher lipid content in grains than 'C 306' variety. Furthermore, expression analyses revealed the higher expression of wheat phospholipid: diacylglycerol acyltransferase-like (PDAT-like) in the 'TAC 75' compared to the 'C 306'. Overexpression and ectopic expression of TaPDAT in yeast and tobacco leaf confirmed its ability to accumulate lipids in vivo. Enzyme activity assay showed that TaPDAT catalyzes the triacylglycerol synthesis by acylating 1,2-diacylglycerol. Interestingly, the long non-coding RNA, lnc663, was upregulated with the TaPDAT gene, while the miRNA, miR1128, downregulated in the 'TAC 75', indicating a regulatory relationship. The GFP reporter assay confirmed that the lnc663 acts as a positive regulator, and the miR1128 as a negative regulator of the TaPDAT gene, which controls lipid accumulation in wheat grain. Our findings outline TaPDAT-mediated biosynthesis of lipid accumulation and reveal the molecular mechanism of the lnc663 and miR1128 mediated regulation of the TaPDAT gene in wheat grain.

Socio-demographic Correlates of Electronic Cigarette and Cannabis Co-use Among Naïve and Tobacco Adolescent Users.

The increasing co-use of e-cigarette and cannabis among youth has become a public health challenge. The present analyses aimed to identify prevalence and correlates of past-month co-use of e-cigarettes and cannabis among adolescents with and without prior tobacco use. For this panel study, 5 years of cross-sectional data (2014-2018) were used from 8th, 10th-, and 12th-grade adolescents in the Monitoring the Future study, a nationally representative survey of U.S. students. We examined prevalence and correlates of e-cigarettes and cannabis co-use among adolescents who had ever used tobacco (n = 15,136) and among those who had never used tobacco (n = 56,525). Adolescents who had ever used tobacco showed significantly higher rates of e-cigarettes and cannabis co-use compared to adolescents who had never used tobacco (17.1% vs. 2.2%, p < 0.01). Results from adjusted multinomial regression models showed that overall, Black and Hispanic adolescents tobacco users were less likely than Whites to co-use e-cigarettes and cannabis. Black adolescents who had used tobacco previously were more likely than Whites to have used cannabis exclusively. Black and Hispanic tobacco-naïve adolescents were more likely than Whites to have used cannabis exclusively, while Black tobacco-naïve adolescents were less likely to use e-cigarettes exclusively or co-use e-cigarettes and cannabis. Overall, males and twelve graders were more likely than males and eight graders to use or co-use cannabis or e-cigarettes, respectively. Among lifetime tobacco users, higher levels of parental education were associated with co-use of cannabis and e-cigarettes. Racial/ethnic-specific patterns of e-cigarette and cannabis co-use depends on adolescents' prior experience with tobacco. The higher rates of use and co-use of e-cigarettes and cannabis among prior tobacco users suggest that targeted interventions are needed for this group. Identified socio-demographic groups at higher risk of co-use of e-cigarettes and cannabis need to be further studied.

Discovery of VEGFR inhibitors through virtual screening and energy assessment.

Vascular endothelial growth factor receptor-2 (VEGFR-2) is crucial in promoting tumor angiogenesis and cancer metastasis. Thus, inhibition of VEGFR-2 has appeared as a good tactic for cancer treatment. To find out novel VEGFR-2 inhibitors, first, the PDB structure of VEGFR-2, 6GQO, was selected based on atomic nonlocal environment assessment (ANOLEA) and PROCHECK assessment. 6GQO was then further used for structure-based virtual screening (SBVS) of different molecular databases, including US-FDA approved drugs, US-FDA withdrawn drugs, may bridge, MDPI, and Specs databases using Glide. Based on SBVS, receptor fit, drug-like filters, and absorption, distribution, metabolism, excretion, and toxicity (ADMET) analysis of 427877 compounds, the best 22 hits were selected. From the 22 hits, hit 5 complex with 6GQO was put through molecular mechanics/generalized born surface area (MM/GBSA) study and hERG binding. The MM/GBSA study revealed that hit 5 possesses lesser binding free energy with more inferior stability in the receptor pocket than the reference compound. The VEGFR-2 inhibition assay of hit 5 disclosed an IC50 of 165.23 nM against VEGFR-2, which can be possibly enhanced through structural modifications.

Oxidative stress, inflammation and hormesis: The role of dietary and lifestyle modifications on aging.

Oxidative stress (OS) is primarily caused by the formation of free radicals and reactive oxygen species; it is considered as one of the prominent factors in slowing down and degrading cellular machinery of an individual, and it eventually leads to aging and age-related diseases by its continuous higher state. The relation between molecular damage and OS should be particularized to understand the beginning of destruction at the cellular levels, extending outwards to affect tissues, organs, and ultimately to the organism. Several OS biomarkers, which are established at the biomolecular level, are useful in investigating the disease susceptibility during aging. Slowing down the aging process is a matter of reducing the rate of oxidative damage to the cellular machinery over time. The breakdown of homeostasis, the mild overcompensation, the reestablishment of homeostasis, and the adaptive nature of the process are the essential features of hormesis, which incorporates several factors, including calorie restriction, nutrition and lifestyle modifications that play an important role in reducing the OS. In the current review, along with the concept and theories of aging (with emphasis on free radical theory), various manifestations of OS with special attention on mitochondrial dysfunction and age-related diseases have been discussed. To alleviate the OS, hormetic approaches including caloric restriction, exercise, and nutrition have also been discussed.

Screening the "Invisible Population" of Older Adult Patients for Prescription Pain Reliever Non-Medical Use and Use Disorders.

Background: In the United States, the number of older adults reporting non-medical use of prescription pain relievers (NMUPPR) between 2015 and 2019 has remained constant, while those meeting criteria for opioid use disorders (OUDs) between 2013 and 2018 increased three-fold. These rates are expected to increase due to increased life expectancy among this population coupled with higher rates of substance use. However, they have consistently lower screening rates for problematic prescription pain reliever use, compared to younger cohorts. Objectives: This commentary reviewed trends in older adult NMUPPR and OUDs and reviewed several available screening tools. We then considered reasons why providers may not be screening their patients, with a focus on older adults, for NMUPPR and OUDs. Finally, we provided recommendations to increase screenings in healthcare settings. Results: Low screening rates in older adult patients may be due to several contributing factors, such as providers' implicit biases and lack of training, time constraints, and comorbid conditions that mask NMUPPR and OUD-related symptoms. Recommendations include incorporating more addiction-related curricula in medical schools, encouraging participation in CME training focused on substance use, attending implicit bias training, and breaking down the silos between pharmacy and geriatric, addiction, and family medicine. Conclusions: There is a growing need for older adult drug screenings, and we have provided several recommendations for improvement. By increasing screenings among older populations, providers will assist in the identification and referral of patients to appropriate and timely substance use treatment and resources to ultimately ameliorate the health of older adult patients.

Identification of multi-targeting natural antiviral peptides to impede SARS-CoV-2 infection.

SARS-CoV-2 and its variants cause serious health concerns throughout the world. The alarming increase in the daily number of cases has become a nightmare in many low-income countries; although some vaccines are available, their high cost and low vaccine production make them unreachable to ordinary people in developing countries. Other treatment strategies are required for novel therapeutic options. The peptide-based drug is one of the alternatives with low toxicity, more specificity, and ease of synthesis. Herein, we have applied structure-based virtual screening to identify potential peptides targeting the critical proteins of SARS-CoV-2. Non-toxic natural antiviral peptides were selected from the enormous number of peptides. Comparative modeling was applied to prepare a 3D structure of selected peptides. 3D models of the peptides were docked using the ClusPro docking server to determine their binding affinity and peptide-protein interaction. The high-scoring peptides were docked with other crucial proteins to analyze multiple targeting peptides. The two best peptides were subjected to MD simulations to validate the structure stability and evaluated RMSD, RMSF, Rg, SASA, and H-bonding from the trajectory analysis of 100 ns. The proposed lead peptides can be used as a broad-spectrum drug and potentially develop as a therapeutic to combat SARS-CoV-2, positively impacting the current pandemic. Supplementary Information: The online version contains supplementary material available at 10.1007/s11224-022-02113-9.

Mechanistic insights into granule-bound starch synthase I (GBSSI.L539P) allele in high amylose starch biosynthesis in wheat (Triticum aestivum L.).

Amylose fraction of grain starch is correlated with a type of resistant starch with better nutritional quality. Granule-bound starch synthase I (GBSSI) is the known starch synthase, responsible for elongation of linear amylose chains. GBSSI expression, activity, and binding to starch and other proteins are the key factors that can affect amylose content. Previously, a QTL, qhams7A.1 carrying GBSSI mutant allele, was identified through QTL mapping using F2 population of the high amylose mutant line, 'TAC 75'. This high amylose mutant line has >2-fold higher amylose content than wild variety 'C 306'. In this study, we characterized this novel mutant allele, GBSSI.L539P. In vitro starch synthase activity of GBSSI.L539P showed improved activity than the wild type (GBSSI-wt). When expressed in yeast glycogen synthase mutants (Δgsy1gsy2), GBSSI-wt and GBSSI.L539P partially complemented the glycogen synthase (gsy1gsy2) activity in yeast. Structural analysis by circular dichroism (CD) and homology modelling showed no significant structural distortion in the mutant enzyme. Molecular docking studies suggested that the residue Leu539 is distant from the catalytic active site (ADP binding pocket) and had no detectable conformational changes in active site. Both wild and mutant enzymes were assayed for starch binding in vitro, and demonstrating higher affinity of the GBSSI.L539P mutant for starch than the wild type. The present study indicated that distant residue (L539P) influenced GBSSI activity by affecting its starch-binding ability. Therefore, it may be a potential molecular target for enhanced amylose content in grain.

Avian Haemosporidian (Plasmodium and Haemoproteus) Status in Two Bird Groups (Old-World Flycatchers and Thrushes) of India and Their Phylogenetic Relationships with Other Lineages of the World.

PURPOSE: Avian haemosporidian may affect the host from body damage to the extinction of a population. Knowledge of their status may help in future avifauna conservation plans. Hence, their status in two bird groups of India and their phylogenetic relationships with other known lineages of the world were examined. METHODS: Cytochrome b gene sequences (479 bp) generated from India and available at MalAvi database were used to study the avian haemosporidian prevalence and phylogenetic analysis of lineages at local and world levels. RESULTS: One common (COLL2) and only once in the study (CYOPOL01, CHD01, CYORUB01, EUMTHA01, GEOCIT01) haemosporidian lineages were discovered. 5.88% prevalence of haemosporidian infection was found in 102 samples belonging to 6 host species. Haemoproteus prevalence was 4.90% across five host species (Phylloscopus trochiloides, Cyornis poliogenys, C. hainanus dialilaemus, C. rubeculoides, Eumiyas thalassinus) and Plasmodium prevalence was 0.98% in Geokichla citrina. Spatial phylogeny at the global level showed that COLL2 lineage, found in C. poliogenys in India, was genetically identical to H. pallidus lineages (COLL2) in parts of Africa, Europe, North America, Malaysia, and the Philippines. The Plasmodium lineage (GEOCIT01) was related to PADOM16 in Egypt, but the sequences were only 93.89% alike. CONCLUSIONS: Four new lineages of Haemoproteus and one of Plasmodium were reported. COLL2 similarity with other H. pallidus lineages may suggest their hosts as possible infection sources.

Comparative transcriptome analyses revealed key genes involved in high amylopectin biosynthesis in wheat.

High amylopectin starch is an important modified starch for food processing industries. Despite a thorough understanding of starch biosynthesis pathway, the regulatory mechanism responsible for amylopectin biosynthesis is not well explored. The present study utilized transcriptome sequencing approach to understand the molecular basis of high amylopectin content in three high amylopectin mutant wheat lines ('TAC 6', 'TAC 358', and 'TAC 846') along with parent variety 'C 306'. Differential scanning calorimetry (DSC) of high amylopectin starch identified a high thermal transition temperature and scanning electron microscopy (SEM) revealed more spherical starch granules in mutant lines compared to parent variety. A set of 4455 differentially expressed genes (DEGs) were identified at two-fold compared to the parent variety in high amylopectin wheat mutants. At ten-fold, 279 genes, including two starch branching genes (SBEIIa and SBEIIb), were up-regulated and only 30 genes, including the starch debranching enzyme (DBE), were down-regulated. Among the genes, different isoforms of sucrose non-fermenting-1-related protein kinase-1 (TaSnRK1α2-3B and TaSnRK1α2-3D) and its regulatory subunit, sucrose non-fermenting-4 (SNF-4-2A, SNF-4-2B, and SNF-4-5D), were found to be highly up-regulated. Further, expression of the DEGs related to starch biosynthesis pathway and TaSnRK1α2 and SNF-4 was performed using qRT-PCR. High expression of TaSnRK1α2, SNF-4, and SBEII isoforms suggests their probable role in high amylopectin starch biosynthesis in grain endosperm. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03364-3.

Quinoline-derivatives as privileged scaffolds for medicinal and pharmaceutical chemists: A comprehensive review.

The quinoline scaffolds are privileged for their numerous biological activities in the pharmaceutical field. This moiety constitutes a well-known space in several marketed preparations. The quinoline scaffolds gained attention in modern days being an important chemical moiety in the identification, designing, and synthesis of novel potent derivatives. The current review is developed to shine the light on critical and significant insights on the quinoline derivatives possessing diverse biological activities such as analgesic, anti-inflammatory, antialzheimer, anti-convulsant, anti-oxidant, antimicrobial, anti-cancer activities and so on. A detailed summary of quinoline ring from its origin to the recent advancements regarding its synthesis, green chemistry approaches, patented methods, and its marketed drugs is presented in the review. We attempted to review the literature compiling the critical information that has potential to encourage fellow researchers and scientists for the design and development of quinoline scaffold based active molecules that have improved therapeutic performance along with profound pharmacological properties.

Protein targeting to starch 1, a functional protein of starch biosynthesis in wheat (Triticum aestivum L.).

KEY MESSAGE: TaPTST1, a wheat homolog of AtPTST1 containing CBM can interact with GBSSI and regulate starch metabolism in wheat endosperm. In cereal endosperm, native starch comprising amylose and amylopectin is synthesized by the coordinated activities of several pathway enzymes. Amylose in starch influences its physio-chemical properties resulting in several human health benefits. The Granule-Bound Starch Synthase I (GBSSI) is the most abundant starch-associated protein. GBSSI lacks dedicated Carbohydrate-binding module (CBM). Previously, Protein Targeting To Starch 1 (PTST1) was identified as a crucial protein for the localization of GBSSI to the starch granules in Arabidopsis. The function of its homologous protein in the wheat endosperm is not known. In this study, TaPTST1, an AtPTST1 homolog, containing a CBM and a coiled-coil domain was identified in wheat. Protein-coding nucleotide sequence of TaPTST1 from Indian wheat variety 'C 306' was cloned and characterized. Homology modelling and molecular docking suggested the potential interaction of TaPTST1 with glucans and GBSSI. The TaPTST1 expression was higher in wheat grain than the other tissues, suggesting a grain-specific function. In vitro binding assays demonstrated different binding affinities of TaPTST1 for native starch, amylose, and amylopectin. Furthermore, the immunoaffinity pull-down assay revealed that TaPTST1 directly interacts with GBSSI, and the interaction is mediated by a coiled-coil domain. The direct protein-protein interaction was further confirmed by bimolecular fluorescence complementation assay (BiFC) in planta. Based on our findings we postulate a functional role for TaPTST1 in starch metabolism by targeting GBSSI to starch granules in wheat endosperm.

Developing a Clinic-Based, Vaccine-Promoting Intervention for African American Youth in Rural Alabama: Protocol for a Pilot Cluster-Randomized Controlled Implementation Science Trial.

BACKGROUND: African American youth in rural Alabama are clinically underserved and have limited knowledge about the human papillomavirus and the novel coronavirus 2019 (COVID-19) vaccines, including knowledge about the risk for developing cervical or oropharyngeal cancers or COVID-19. OBJECTIVE: In this 30-month study, we propose to develop an in-clinic, youth-tailored, vaccine-promoting intervention for vaccine hesitancy reduction that can be seamlessly integrated into the existing environments of pediatric and family practice settings in rural Alabama. METHODS: This exploratory, sequential mixed methods study will be conducted in 3 phases. In the first phase, we will assess stakeholders' knowledge, sentiments, and beliefs related to vaccination in general, COVID-19 vaccination, and human papillomavirus vaccination. We will also assess stakeholders' perceptions of barriers to vaccination that exist in rural Alabama. This will be followed by a second phase wherein we will use the data collected in the first phase to inform the development and finalization of a noninvasive, modular, synchronous counseling intervention that targets the behaviors of 15- to 26-year-old adolescents. In the third phase, we will conduct a pilot hybrid type 1 effectiveness-implementation cluster-randomized controlled trial to assess intervention acceptability and feasibility (clinics: N=4; African American youth: N=120) while assessing a "clinical signal" of effectiveness. We will document implementation contexts to provide real-world insight and support dissemination and scale-up. RESULTS: The study was funded at the end of December 2020. Approval from the University of Alabama at Birmingham Institutional Review Board was obtained in May 2021, and the qualitative data collection process outlined in the first phase of this project concluded in November 2021. The entire study is expected to be complete at the end of December 2023. CONCLUSIONS: The results of the trial will provide much needed information on vaccine hesitancy in rural Alabama, and if found efficacious, the intervention could notably increase rates of vaccinations in one of the most underserved parts of the United States. The results from the trial will provide information that is valuable to public health practitioners and providers in rural settings to inform their efforts in increasing vaccination rates among 15- to 26-year-old African American youth in rural southern United States. TRIAL REGISTRATION: ClinicalTrials.gov NCT04604743; https://clinicaltrials.gov/ct2/show/NCT04604743. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/33982.

Unraveling novel and rare mutations for alpha-amylase and key transcription factors in EMS-induced wheat mutants for amylose by TILLING.

BACKGROUND: TILLING (Targeting Induced Local Lesions in Genomes) is a reverse-genetic strategy that is used to locate an allelic series of induced point mutations in genes of interest. High-throughput TILLING allows the rapid and cost-effective detection of induced point mutations in populations of chemically mutagenized individuals. Grain amylose content is the major constraints for its nutritional quality and have drawn research interest. Identification of allelic variations in genes involved in starch biosynthesis in wheat endosperm is pre-requisite to amenable for nutritional quality improvement. METHODS AND RESULTS: In this study, 44 EMS-induced (M4 generation) mutant lines having variation for amylose content were used for TILLING sequencing. Overall 2098.08 kb of the sequence was analyzed, and the average mutation density was 1/65.56 kb. In analysis, at the high depth score a total of 32 variations were identified including three natural variations, 76% transitions, 10% transversions, and 14% InDels respectively. The substitutions led to intronic variants, UTRs and up-downstream gene variants in Alpha-amylase, TabZIP77.1, TabZIP1 and Myb respectively. In the Myb transcription factor two missense mutations recorded namely Myb_7B c.680G > A and c.1358 T > C led to p.Gly227Asp and p.Met453Thr and c.1390G > A one substitution in Myb_7D led to p.Val464Ile. CONCLUSION: The identified missense substitutions were predicted to affect the protein function; hence they may have a probable role in context to the amylose content in mutants. The mutations ascertained in the current study will help in gene discovery in wheat and identified mutants can be used as genetic resources to improve nutritional quality of wheat.

iNOS Deletion in Alveolar Epithelium Cannot Reverse the Elastase-Induced Emphysema in Mice.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is the third leading cause of death worldwide. In addition to chronic bronchitis and emphysema, patients often develop at least mild pulmonary hypertension (PH). We previously demonstrated that inhibition of inducible nitric oxide synthase (iNOS) prevents and reverses emphysema and PH in mice. Interestingly, strong iNOS upregulation was found in alveolar epithelial type II cells (AECII) in emphysematous murine lungs, and peroxynitrite, which can be formed from iNOS-derived NO, was shown to induce AECII apoptosis in vitro. However, the specific cell type(s) that drive(s) iNOS-dependent lung regeneration in emphysema/PH has (have) not been identified yet. AIM: we tested whether iNOS knockout in AECII affects established elastase-induced emphysema in mice. METHODS: four weeks after a single intratracheal instillation of porcine pancreatic elastase for the induction of emphysema and PH, we induced iNOS knockout in AECII in mice, and gave an additional twelve weeks for the potential recovery. RESULTS: iNOS knockout in AECII did not reduce elastase-induced functional and structural lung changes such as increased lung compliance, decreased mean linear intercept and increased airspace, decreased right ventricular function, increased right ventricular systolic pressure and increased pulmonary vascular muscularization. In vitro, iNOS inhibition did not reduce apoptosis of AECII following exposure to a noxious stimulus. CONCLUSION: taken together, our data demonstrate that iNOS deletion in AECII is not sufficient for the regeneration of emphysematous murine lungs, and suggest that iNOS expression in pulmonary vascular or stromal cells might be critically important in this regard.

Racial-ethnic Differences in Reasons for Misuse of Prescription Medications Among US Adults.

OBJECTIVES: This study aims to investigate racial-ethnic differences in reasons for misuse of prescription medications among a nationally representative sample of US adults. METHODS: We analyzed data from the 2015-2019 National Surveys on Drug Use and Health. The study population includes US adults (18-49 years old) who reported misuse of 3 types of prescription drugs (stimulants [n = 6139], sedatives and tranquilizers [n = 5643], and pain relievers [n = 8780]) for 3 reasons: medical-only (eg, to help with pain), recreational-only (eg, to get high), or combined medical and recreational reasons. Multinomial logistic regressions assessed the association between reasons of misuse of prescription medications and self-identified race-ethnicity. RESULTS: Misuse of the 4 types of prescription medications was primarily motivated by medical reasons (63%-80%). Compared to non-Hispanic Whites, non-Hispanic Blacks (nHB), and Hispanics (H) were less likely to report misuse of pain relievers for combined (nHB: adjusted relative risk ratio [aRRR] = 0.6, 95% confidence interval [CI]: 0.4, 0.7; H; aRRR = 0.7, 95% CI: 0.5, 0.9) or recreational reasons (nHB: aRRR = 0.8, 95% CI: 0.6, 1.0; H; aRRR = 0.7, 95% CI: 0.6, 0.9) rather than medical-only reasons. The odds of misuse of sedatives and tranquilizers for recreational-only reasons as opposed to medical-only reasons were higher among nHB (aRRR = 1.9, 95% CI: 1.3, 2.7) and H (aRRR = 1.9, 95% CI: 1.4, 2.4) than among non-Hispanic Whites. CONCLUSIONS: The increased misuse of prescription pain relievers for medical reasons among racial-ethnic minority groups demonstrates a continued need to investigate underlying structural factors driving these behaviors. The higher odds of sedative and tranquilizer misuse for recreational purposes among racial-ethnic minority groups warrant further investigation.