Unhealthy habits and comorbidities associated with uncontrolled asthma in young people.

Introduction: Asthma is a chronic inflammatory disease of the lower airways that affects more than 260 million people worldwide and has been related to more than 460,000 deaths a year. It is estimated that in 60% of asthma cases, the symptoms are not adequately controlled. The objective of this study was to determine the association between some comorbidities, habits, and health risk behaviors with uncontrolled asthma in a sample of young people with asthma.Methods: Through a cross-sectional study, data from 1,078 young people aged 17 to 19 years were analysed. Information was collected through physical examination, direct questioning, and the application of a self-administered questionnaire.Results: In the group of young people with asthma, the prevalence of uncontrolled asthma was 20.6%, of which 53.8% were women, 76.9% suffered from rhinitis, 46.2% were overweight and 23.1% were obese. In the group of young with uncontrolled asthma, gingivitis was detected in 53.8% and alcohol consumption in 84.6%. Logistic regression analysis showed a significant association between allergic rhinitis, gingivitis, carbohydrate intake, alcohol consumption, overweight, and obesity with uncontrolled asthma.Conclusions: Parents and members of the health team need to identify on time the risk factors associated with uncontrolled asthma in young people with asthma to limit its development and the negative effects it generates. The results of this study should be used to strengthen programs that promote the comprehensive health of adolescents.

HEXACO personality factors as predictors of physical activity, resting heart rate, body mass index, and healthy lifestyle behaviors.

BACKGROUND: Personality traits are known factors that may influence levels of physical activity and other healthy lifestyle measures and behaviors that ultimately lead to health problems later in life. Participants And Procedure: The aim of this study was to examine the association between personality traits (HEXACO) and levels of physical activity and resting heart rate (RHR) - measured using Fitbits, BMI, and a self-reported whole-person healthy lifestyle score for N = 2580 college students. Data were collected and analyzed for students enrolled in a University Success type course from August 2017 to May 2021. The relationships between HEXACO personality traits and various physical activity and healthy lifestyle behaviors were analyzed by building several multiple regression models using R version 4.0.2. Results: In general, students who are extraverted were more physically active and students who are more open to experience had a higher RHR, even when controlling for gender. Females and males however had different profiles as to how personality influenced physical activity and other health-related measures. Male extraverts with high negative emotionality scores tend to be more physically active, whereas females tend to be more physically active when they were high in extroversion and conscientiousness, and low in openness to experience. BMI values were higher for female participants with high honesty-humility and low agreeableness and conscientiousness scores. Females also had a lower RHR for high honesty-humility and emotionality and low conscientiousness scores. CONCLUSIONS: Personality can influence levels of physical activity, RHR, and BMI. This is especially true of women. Being aware of one's personality and the relationship of personality traits to levels of physical activity and other measures of leading a healthy lifestyle can be beneficial in determining strategies to improve long-term health outcomes.

Excitoprotective effects of conditional tau reduction in excitatory neurons and in adulthood.

Tau reduction is a promising therapeutic strategy for Alzheimer's disease. In numerous models, tau reduction via genetic knockout is beneficial, at least in part due to protection against hyperexcitability and seizures, but the underlying mechanisms are unclear. Here we describe the generation and initial study of a new conditional Tau flox model to address these mechanisms. Given the protective effects of tau reduction against hyperexcitability, we compared the effects of selective tau reduction in excitatory or inhibitory neurons. Tau reduction in excitatory neurons mimicked the protective effects of global tau reduction, while tau reduction in inhibitory neurons had the opposite effect and increased seizure susceptibility. Since most prior studies used knockout mice lacking tau throughout development, we crossed Tau flox mice with inducible Cre mice and found beneficial effects of tau reduction in adulthood. Our findings support the effectiveness of tau reduction in adulthood and indicate that excitatory neurons may be a key site for its excitoprotective effects. SUMMARY: A new conditional tau knockout model was generated to study the protective effects of tau reduction against hyperexcitability. Conditional tau reduction in excitatory, but not inhibitory, neurons was excitoprotective, and induced tau reduction in adulthood was excitoprotective without adverse effects.

Healthcare Resource Utilization and Cost Associated with Allogeneic Hematopoietic Stem Cell Transplantation: A Scoping Review.

This scoping review summarizes the evidence regarding healthcare resource utilization (HRU) and costs associated with allogeneic hematopoietic stem cell transplantation (allo-HSCT). This study was conducted in accordance with the Joanne Briggs Institute methodology for scoping reviews. The PubMed, Embase, and Health Business Elite Electronic databases were searched, in addition to grey literature. The databases were searched from inception up to November 2022. Studies that reported HRU and/or costs associated with adult (≥18 years) allo-HSCT were eligible for inclusion. Two reviewers independently screened 20% of the sample at each of the 2 stages of screening (abstract and full text). Details of the HRU and costs extracted from the study data were summarized, based on the elements and timeframes reported. HRU measures and costs were combined across studies reporting results defined in a comparable manner. Monetary values were standardized to 2022 US Dollars (USD). We identified 43 studies that reported HRU, costs, or both for allo-HSCT. Of these studies, 93.0% reported on costs, 81.4% reported on HRU, and 74.4% reported on both. HRU measures and cost calculations, including the timeframe for which they were reported, were heterogeneous across the studies. Length of hospital stay was the most frequently reported HRU measure (76.7% of studies) and ranged from a median initial hospitalization of 10 days (reduced-intensity conditioning [RIC]) to 73 days (myeloablative conditioning). The total cost of an allo-HSCT ranged from $63,096 (RIC) to $782,190 (double umbilical cord blood transplantation) at 100 days and from $69,218 (RIC) to $637,193 at 1 year (not stratified). There is heterogeneity in the reporting of HRU and costs associated with allo-HSCT in the literature, making it difficult for clinicians, policymakers, and governments to draw definitive conclusions regarding the resources required for the delivery of these services. Nevertheless, to ensure that access to healthcare meets the necessary high cost and resource demands of allo-HSCT, it is imperative for clinicians, policymakers, and government officials to be aware of both the short- and long-term health resource requirements for this patient population. Further research is needed to understand the key determinants of HRU and costs associated with allo-HSCT to better inform the design and delivery of health care for HSCT recipients and ensure the quality, safety, and efficiency of care.

The epigenome under pressure: On regulatory adaptation to chronic stress in the brain.

Chronic stress (CS) can have long-lasting consequences on behavior and cognition, that are associated with stable changes in gene expression in the brain. Recent work has examined the role of the epigenome in the effects of CS on the brain. This review summarizes experimental evidence in rodents showing that CS can alter the epigenome and the expression of epigenetic modifiers in brain cells, and critically assesses their functional effect on genome function. It discusses the influence of the developmental time of stress exposure on the type of epigenetic changes, and proposes new lines of research that can help clarify these changes and their causal involvement in the impact of CS.

A phase II clinical trial of frameless, fractionated stereotactic radiation therapy for brain metastases.

Stereotactic radiation therapy yields high rates of local control for brain metastases, but patients in rural or suburban areas face geographic and socioeconomic barriers to its access. We conducted a phase II clinical trial of frameless, fractionated stereotactic radiation therapy for brain metastases in an integrated academic satellite network for patients 18 years of age or older with 4 or fewer brain metastases. Dose was based on gross tumor volume: less than 3.0 cm, 27 Gy in 3 fractions and 3.0 to 3.9 cm, 30 Gy in 5 fractions. Median follow-up was 10 months for 73 evaluable patients, with a median age of 68 years. Median intracranial progression-free survival was 7.1 months (95% confidence interval = 5.3 to not reached), and median survival was 7.2 months (95% confidence interval = 5.4 to not reached); there were no serious adverse events. Outcomes of this trial compare favorably with contemporary trials, and this treatment strategy provides opportunities to expand stereotactic radiation therapy access to underserved populations.

Association of Primary Care Risk Mitigation Visits and Nonelective Emergency Department Visits in Patients Using Long-term Opioid Therapy.

Objective: To determine risk factors for nonelective emergency department visits (NEDVs) and whether primary care visits incorporating risk mitigation tools prevented NEDVs among patients using long-term opioid therapy (LOT). Patients and Methods: We retrospectively searched the electronic health records at Mayo Clinic primary care outpatient practices in Arizona and Florida in all of 2018 and 2019 for the records of individual adult patients using LOT. Patient and clinician demographic characteristics and patient risk factors were compared between patients with and without risk mitigation visits. Univariate and multivariable logistic regression was used to determine risk factors for NEDVs. Results: Among 457 patients using LOT identified during the study period, most were women (n=266, 58.2%), and the median age was 69 years. Long-term opioid therapy risk mitigation visits were performed equally by family medicine and internal medicine clinicians and by a significantly higher proportion of Florida clinicians than Arizona clinicians (87.0% vs 70.5%; P<.001). Older age, falls, and mental health care utilization all increased the risk of NEDVs. Risk mitigation visits were protective against NEDVs (odds ratio, 0.56; 95% CI, 0.35-0.89; P=.01) after adjustment for older age, falls, and mental health care utilization. Conclusion: Risk mitigation visits are effective in preventing NEDVs, and all patients using LOT should have such visits when possible.

Association of systemic inflammation with posttraumatic stress disorder after a myocardial infarction.

Background: Adverse mental health conditions including depression, posttraumatic stress disorder (PTSD), and anxiety are prevalent among patients who survive myocardial infarctions (MI) and are associated with adverse outcomes. The mechanisms underlying these associations, however, are not well understood. Inflammatory pathways may mediate the cardiovascular outcomes of patients with mental health disorders. We examined the bidirectional association between PTSD symptoms and inflammatory biomarkers in a young/middle-aged post MI population. We further examined how this association may differ between women and men as well as between Black and non-Black individuals. Methods: Participants included individuals with early onset MI between the ages 25 and 60. Mental health scores for depression, PTSD, perceived stress, and anxiety as well as inflammatory biomarkers, interleukin-6 (IL-6) and high sensitivity C-reactive protein (hsCRP), were collected at baseline and at six-month follow up. We examined the bidirectional changes in mental health symptoms and inflammatory biomarkers between baseline and follow-up. Results: Among 244 patients in the study (mean age: 50.8, 48.4% female, 64.3% Black), the geometric means for IL-6 level and hsCRP at rest were 1.7 pg/mL and 2.76 mg/L, respectively. Mental health scores at baseline did not consistently predict changes in inflammatory biomarkers at follow-up. However, baseline levels of both IL-6 and hsCRP were robustly associated with an increase in re-experiencing PTSD symptoms at 6 months: in adjusted linear mixed models, there was a 1.58-point increase in re-experiencing PTSD symptoms per unit of baseline hsCRP (p = 0.01) and 2.59-point increase per unit of baseline IL-6 (p = 0.02). Once the analysis was stratified by race, the association was only noted in Black individuals. Baseline inflammation was not associated with change in any of the other mental health symptom scores. Conclusion: Markers of inflammation are associated with an increase in post-event PTSD symptoms in younger or middle-aged patients who experienced an MI, especially Black patients. These results suggest a mechanistic link between inflammation and the development of PTSD among individuals with cardiovascular disease.

Patients with sporadic FTLD exhibit similar increases in lysosomal proteins and storage material as patients with FTD due to GRN mutations.

Loss of function progranulin (GRN) mutations are a major autosomal dominant cause of frontotemporal dementia (FTD). Patients with FTD due to GRN mutations (FTD-GRN) develop frontotemporal lobar degeneration with TDP-43 pathology type A (FTLD-TDP type A) and exhibit elevated levels of lysosomal proteins and storage material in frontal cortex, perhaps indicating lysosomal dysfunction as a mechanism of disease. To investigate whether patients with sporadic FTLD exhibit similar signs of lysosomal dysfunction, we compared lysosomal protein levels, transcript levels, and storage material in patients with FTD-GRN or sporadic FTLD-TDP type A. We analyzed samples from frontal cortex, a degenerated brain region, and occipital cortex, a relatively spared brain region. In frontal cortex, patients with sporadic FTLD-TDP type A exhibited similar increases in lysosomal protein levels, transcript levels, and storage material as patients with FTD-GRN. In occipital cortex of both patient groups, most lysosomal measures did not differ from controls. Frontal cortex from a transgenic mouse model of TDP-opathy had similar increases in cathepsin D and lysosomal storage material, showing that TDP-opathy and neurodegeneration can drive these changes independently of progranulin. To investigate these changes in additional FTLD subtypes, we analyzed frontal cortical samples from patients with sporadic FTLD-TDP type C or Pick's disease, an FTLD-tau subtype. All sporadic FTLD groups had similar increases in cathepsin D activity, lysosomal membrane proteins, and storage material as FTD-GRN patients. However, patients with FTLD-TDP type C or Pick's disease did not have similar increases in lysosomal transcripts as patients with FTD-GRN or sporadic FTLD-TDP type A. Based on these data, accumulation of lysosomal proteins and storage material may be a common aspect of end-stage FTLD. However, the unique changes in gene expression in patients with FTD-GRN or sporadic FTLD-TDP type A may indicate distinct underlying lysosomal changes among FTLD subtypes.

Factors Associated with Early Sexual Activity in Adolescents Residing in an Urban Area of Northeastern Mexico Who Use WhatsApp or Facebook.

Early sexual activity has been associated with unplanned pregnancy and sexually transmitted infections. In recent decades, the prevalence of early sexual activity and social media use has increased. However, the relationship between these variables has been little studied. Consequently, we examined the relationships of some factors present in the family and personal environment with early sexual activity in adolescents who use and do not use the WhatsApp and Facebook platforms. Through a cross-sectional study, data from 1328 adolescents aged 13 to 19 years were analyzed. Data collection was carried out using a self-administered questionnaire. Of the total participants, 35.7% reported early sexual activity, and 37.7% reported using social networks such as WhatsApp and Facebook. The mean age of onset of sexual activity was 15.59 ± 1.56 years. Multivariate logistic regression analysis showed significant associations of not studying (adjusted OR 4.70; CI 95% 1.31-16.78), consuming alcohol (adjusted OR 3.71; CI 95% 2.44-5.65) and having parents who consumed alcoholic beverages in the home (adjusted OR 1.48; 95% CI 1.03-2.12) with ESA. In the family and personal environment, some factors favored early sexual activity in young people who used the WhatsApp and Facebook applications. This information should be used by authorities in the health and education sectors to strengthen preventive programs targeting health risk habits and behaviors in adolescents.

An observational study of the factors associated with frequency of outpatient benzodiazepine prescribing to patients receiving chronic opioid analgesic therapy in primary care at a major academic center.

BACKGROUND: Prescribing benzodiazepines to patients taking chronic opioid analgesic therapy increases risks of adverse events. In 2016, the Centers for Disease Control and Prevention recommended avoidance of benzodiazepine prescribing concurrently with opioids, and various organizations have instituted similar guidelines. We aimed to determine the frequency and patterns of benzodiazepine prescribing at Mayo Clinic primary care (Community Internal Medicine, Family Medicine) clinics for patients taking chronic opioid analgesic therapy and the characteristics of patients receiving the prescriptions and providers administering them. METHODS: This retrospective observational study included adult patients taking chronic opioid analgesic therapy for 2 full years in 2018 and 2019 at Mayo Clinic primary care practices in Arizona and Florida. We assessed electronic health records for these individual patients to determine whether they received a benzodiazepine prescription during the study period and how frequently they received a prescription. Variations in prescriptions by provider specialty, location, and sex were studied. Documented data included receipt of a benzodiazepine prescription by patients with at-risk alcohol use or alcohol use disorder, depression, anxiety, chronic obstructive pulmonary disease, falls, and psychiatric referral. Data were compared between patients who received benzodiazepines and those who did not with the Kruskal-Wallis test or χ2 test, and the Wilcoxon signed rank test was used to assess whether the change in number of benzodiazepine prescriptions (2018 vs. 2019) was different from zero. RESULTS: Study participants (N = 457) were predominantly women (n = 266, 58.2%); median age was 69 years. In total, 148 patients (32.4%) received benzodiazepine prescription. These patients were more likely to be women (P = .046) and younger (P = .02). Mean percentage change was 176.9% (P < .001) in number of benzodiazepine prescriptions provided from 2018 to 2019. Frequency of referral to mental health providers was low, as was presence of an established mental health provider despite a greater prevalence of anxiety (P < .001) and depression (P = .001) among patients receiving benzodiazepines. CONCLUSION: Benzodiazepine prescription to individual patients taking chronic opioid analgesic therapy significantly increased from 2018 to 2019 despite the documented risks and harms associated with such practice. No statistically significant difference was observed in frequency of benzodiazepine prescriptions between practice location, sex of provider, or specialty.

Prescribing patterns and attitudes of primary care providers regarding long-term opioid therapy.

OBJECTIVE: To assess prescribing and referral patterns and attitudes of primary care providers treating patients receiving long-term opioid therapy (LOT) according to recent guidelines. DESIGN, SETTING, AND PARTICIPANTS: An anonymous 25-question survey was distributed to all primary care providers at Mayo Clinic in Arizona (from April 30 through May 22, 2020). MAIN OUTCOME MEASURES: Knowledge and comfort with LOT guidelines, prescribing patterns, referral patterns to behavioral and pain specialties, patients' concomitant substance use, and response variability by provider sex and specialty. RESULTS: Most of the 31 survey respondents were familiar with LOT guidelines and were comfortable in prescribing opioids; 36 percent reported no increase in prescribing safety. Patient education on naloxone was infrequent. Access affected referral to behavioral and addiction specialties; 87.1 percent referred patients to pain medicine specialists despite reporting little long-term improvement in symptoms. For a significantly larger proportion of internal medicine and women's health (IM/WH) providers, compared with family medicine (FM) providers, Patient Health Questionnaire 9 and Generalized Anxiety Disorder 7-item scale scores were the reasons for behavioral health referral. Many providers prescribed additional substances. More female providers reported that patients used gabapentin concomitantly (p = .03). More FM providers than IM/WH providers typically referred patients receiving LOT to addiction specialists (p = .02). Most expressed a need for a multispecialty LOT clinic, and 83.9 percent supported buprenorphine prescribing. CONCLUSIONS: Despite familiarity with LOT guidelines, many providers felt that patient safety and prescribing diligence have not improved. Patient education on naloxone treatment is needed, and access to behavioral specialists is a barrier to referrals.

Predictores de potenciales reacciones adversas e interacciones medicamentosas en pacientes post accidente cerebrovascular en Perú

Objetivo Identificar los predictores clínicos y farmacoterapéuticos asociados a los niveles de severidad de las reacciones adversas (RAM) e interacciones medicamentosas (IM) en pacientes hospitalizados post accidente cerebrovascular. Métodos Estudio analítico, predictivo y transversal mediante el modelo de regresión lineal múltiple. Los niveles de severidad de las potenciales reacciones adversas e interacciones medicamentosas se evaluaron mediante Drugs.com. Resultados De la evaluación de 992 prescripciones médicas de 55 (56,7%) pacientes mujeres y 42 (43,3%) varones post accidente cerebrovascular isquémico 62 (63,9%) y hemorrágico 35 (36,1%), se identificó un total de 11 790±46,8 potenciales reacciones adversas y 1 034±9,8 interacciones medicamentosas. La hipertensión arterial se asoció a las reacciones adversas graves y moderadas, en tanto que la neumonía intrahospitalaria y alcalosis metabólica a reacciones adversas leves y moderadas. La alcalosis metabólica se asoció a las interacciones medicamentosas moderadas y leves. Los predictores farmacoterapéuticos como la prescripción en polifarmacia y el uso de antibióticos se relacionaron con reacciones adversas graves, moderadas y leves; los antidiabéticos se relacionaron con interacciones medicamentosas graves, moderadas y los fármacos para terapia cardiaca con interacciones medicamentosas leves. Conclusiones Las variables clínicas como factores de riesgo cardiovascular, presencia de comorbilidades que exacerban las enfermedades crónicas no trasmisibles, los signos y síntomas de alarma, el mayor tiempo de estancia hospitalaria y la prescripción en polifarmacia fueron predictores de mayor frecuencia de reacciones adversas e interacciones medicamentosas graves y moderadas que requieren especial vigilancia y estudio individualizado.

Objective To identify clinical and pharmacotherapeutic predictors associated with severity levels of adverse reactions and drug-drug interactions in post-stroke hospita-lized patients. Methods Analytic, predictive, cross-sectional study using multiple linear regression modeling. Severity levels of potential adverse reactions and drug-drug interactions were assessed using Drugs.com. Results From the evaluation of 992 medical prescriptions of 55 (56.7%) female and 42 (43.3%) male patients post ischemic stroke 62(63.9%) and hemorrhagic stroke 35 (36.1%); a total of 11 790±46.8 potential adverse reactions and 1 034±9.8 drug-drug interactions were identified; arterial hypertension was associated with severe and moderate adverse reactions; while in-hospital pneumonia and metabolic alkalosis with mild and moderate adverse reactions. While metabolic alkalosis was associated with moderate and mild drug-drug interactions. Pharmacotherapeutic predictors such as polypharmacy prescription and antibiotic use were related to moderate and mild severe adverse reactions; antidiabetic drugs were related to moderate and severe drug-drug interactions and cardiac therapy drugs were related to mild drug-drug interactions. Conclusions Clinical variables such as cardiovascular risk factors, presence of comorbidities that exacerbate chronic noncommunicable diseases, alarm signs and symptoms, longer hospital stay, as well as polypharmacy prescriptions, were predictors of a higher frequency of severe and moderate adverse reactions and drug-drug interactions, which require special vigilance and individualized study.

Asociación de la violencia obstétrica, control prenatal y embarazo no planeado con la prematurez / Association of obstetric violence, prenatal control, and unplanned pregnancy with prematurity

Resumen La prematurez (PM) neonatal es un problema mayor de salud pública en el mundo, debido a su frecuencia de presentación y los años de vida potencialmente perdidos; además, es la principal causa de muerte en menores de cinco años. Entre las consecuencias negativas de la PM están las discapacidades del aprendizaje, visual y auditiva. Múltiples son los factores asociados a la PM; algunos pueden ser detectados y modificados de forma oportuna en las consultas prenatales y también, en las preconcepcionales. Por otro lado, la PM no es solamente responsabilidad del prestador de los servicios de salud, sino también del sistema de salud, el cual, debe otorgar los recursos mínimos necesarios para monitorear y mantener a la embarazada en un estado óptimo de salud para evitarla, y en caso de presentarse, debe contar con la infraestructura hospitalaria básica que permita la viabilidad del producto y evitar su muerte. Aunado a lo anterior, se debe de considerar la responsabilidad de la paciente y su pareja, debido a que en ellos recae la obligación de llevar un estilo de vida saludable y acudir a las consultas preconcepcionales y control prenatal.

Abstract Neonatal prematurity (MP) is a major public health problem in the world, due to its presentation frequency and the years of life potentially lost; In addition, it is the main cause of death in children under five years of age. Among the negative consequences of MP are learning, visual and hearing disabilities. There are multiple factors associated with MP; some can be detected and modified in a timely manner in prenatal consultations and in preconception. On the other hand, MP is not only responsibility of the health service provider, but also of the health system, which must grant the minimum resources necessary to monitor and maintain the pregnant woman in an optimal state of health to avoid it, and if it occurs, the hospital must have the basic infrastructure that allows the viability of the product and prevents its death. In addition, the responsibility of the patient and her partner must be considered since they are obliged to lead a healthy lifestyle and attend preconception consultations and prenatal care.

A Cre-Dependent CRISPR/dCas9 System for Gene Expression Regulation in Neurons.

Site-specific genetic and epigenetic targeting of distinct cell populations is a central goal in molecular neuroscience and is crucial to understand the gene regulatory mechanisms that underlie complex phenotypes and behaviors. While recent technological advances have enabled unprecedented control over gene expression, many of these approaches are focused on selected model organisms and/or require labor-intensive customization for different applications. The simplicity and modularity of clustered regularly interspaced short palindromic repeats (CRISPR)-based systems have transformed genome editing and expanded the gene regulatory toolbox. However, there are few available tools for cell-selective CRISPR regulation in neurons. We designed, validated, and optimized CRISPR activation (CRISPRa) and CRISPR interference (CRISPRi) systems for Cre recombinase-dependent gene regulation. Unexpectedly, CRISPRa systems based on a traditional double-floxed inverted open reading frame (DIO) strategy exhibited leaky target gene induction even without Cre. Therefore, we developed an intron-containing Cre-dependent CRISPRa system (SVI-DIO-dCas9-VPR) that alleviated leaky gene induction and outperformed the traditional DIO system at endogenous genes in HEK293T cells and rat primary neuron cultures. Using gene-specific CRISPR sgRNAs, we demonstrate that SVI-DIO-dCas9-VPR can activate numerous rat or human genes (GRM2, Tent5b, Fos, Sstr2, and Gadd45b) in a Cre-specific manner. To illustrate the versatility of this tool, we created a parallel CRISPRi construct that successfully inhibited expression from a luciferase reporter in HEK293T cells only in the presence of Cre. These results provide a robust framework for Cre-dependent CRISPR-dCas9 approaches across different model systems, and enable cell-specific targeting when combined with common Cre driver lines or Cre delivery via viral vectors.

A set of simple methods for detection and extraction of laminarinase.

A carefully designed ammonium sulfate precipitation will simplify extraction of proteins and is considered to be a gold standard among various precipitation methods. Therefore, optimization of ammonium sulfate precipitation can be an important functional step in protein purification. The presence of high amounts of ammonium sulphate precludes direct detection of many enzymatically active proteins including reducing sugar assays (e.g. Nelson-Somogyi, Reissig and 3,5-dinitrosalicylic acid methods) for assessing carbohydrases (e.g. laminarinase (ß (1-3)-glucanohydrolase), cellulases and chitinases). In this study, a simple method was developed using laminarin infused agarose plate for the direct analysis of the ammonium sulphate precipitates from Streptomyces rimosus AFM-1. The developed method is simple and convenient that can give accurate results even in presence of ammonium sulfate in the crude precipitates. Laminarin is a translucent substrate requiring the use of a stain to visualize the zones of hydrolysis in a plate assay. A very low-cost and locally available fluorescent optical fabric brightener Tinopal CBS-X has been used as a stain to detect the zones of hydrolysis. We also report simple methods to prepare colloidal chitin and cell free supernatant in this manuscript.

Regulation of dopamine-dependent transcription and cocaine action by Gadd45b.

Exposure to drugs of abuse produces robust transcriptional and epigenetic reorganization within brain reward circuits that outlives the direct effects of the drug and may contribute to addiction. DNA methylation is a covalent epigenetic modification that is altered following stimulant exposure and is critical for behavioral and physiological adaptations to drugs of abuse. Although activity-related loss of DNA methylation requires the Gadd45 (Growth arrest and DNA-damage-inducible) gene family, very little is known about how this family regulates activity within the nucleus accumbens or behavioral responses to drugs of abuse. Here, we combined genome-wide transcriptional profiling, pharmacological manipulations, electrophysiological measurements, and CRISPR tools with traditional knockout and behavioral approaches in rodent model systems to dissect the role of Gadd45b in dopamine-dependent epigenetic regulation and cocaine reward. We show that acute cocaine administration induces rapid upregulation of Gadd45b mRNA in the rat nucleus accumbens, and that knockout or site-specific CRISPR/Cas9 gene knockdown of Gadd45b blocks cocaine conditioned place preference. In vitro, dopamine treatment in primary striatal neurons increases Gadd45b mRNA expression through a dopamine receptor type 1 (DRD1)-dependent mechanism. Moreover, shRNA-induced Gadd45b knockdown decreases expression of genes involved in psychostimulant addiction, blocks induction of immediate early genes by DRD1 stimulation, and prevents DRD1-mediated changes in DNA methylation. Finally, we demonstrate that Gadd45b knockdown decreases striatal neuron action potential burst duration in vitro, without altering other electrophysiological characteristics. These results suggest that striatal Gadd45b functions as a dopamine-induced gene that is necessary for cocaine reward memory and DRD1-mediated transcriptional activity.

Risk factors of non-adherence to guidelines for the prevention of COVID-19 among young adults with asthma in a region with a high risk of a COVID-19 outbreak.

OBJECTIVE: SARS-CoV-2 disease 2019 (COVID-19) is highly contagious and spreads rapidly. The application of preventive measures has proven to be the best strategy to minimize the number of patients and the dissemination of and deaths from COVID-19. The objective of this study was to determine the risk factors that limit the adherence of asthmatic patients to measures that prevent COVID-19 among residents of a region with a high risk of a COVID-19 outbreak. METHODS: Through a cross-sectional study, data from 2,372 participants aged 16-24 years were analyzed. To collect their information, a questionnaire was constructed using the Google Forms tool. Due to the prevention measures in place for COVID-19, the home quarantine of many people, and the lack of socialization, the questionnaire was distributed through email (Internet) and WhatsApp. A logistic regression analysis was performed to determine the relationship between the variables. RESULTS: The prevalence of asthma was 12.2%, and non-adherence to the guidelines for the prevention of COVID-19 was 53.1%. Approximately 30.8% of asthma patients did not comply with the basic prevention measures for COVID-19. The results of the logistic regression analysis showed that being male, active smoking, and believing that COVID-19 is not a more severe disease for people suffering from asthma than others were associated with non-adherence to the basic protection measures established in the guidelines for the prevention of COVID-19. CONCLUSIONS: It is important for health professionals to advise asthma patients to comply with the basic measures of protection against COVID-19 and timely use medications for asthma control.