Reservoir host immune responses to emerging zoonotic viruses.

Zoonotic viruses, such as HIV, Ebola virus, coronaviruses, influenza A viruses, hantaviruses, or henipaviruses, can result in profound pathology in humans. In contrast, populations of the reservoir hosts of zoonotic pathogens often appear to tolerate these infections with little evidence of disease. Why are viruses more dangerous in one species than another? Immunological studies investigating quantitative and qualitative differences in the host-virus equilibrium in animal reservoirs will be key to answering this question, informing new approaches for treating and preventing zoonotic diseases. Integrating an understanding of host immune responses with epidemiological, ecological, and evolutionary insights into viral emergence will shed light on mechanisms that minimize fitness costs associated with viral infection, facilitate transmission to other hosts, and underlie the association of specific reservoir hosts with multiple emerging viruses. Reservoir host studies provide a rich opportunity for elucidating fundamental immunological processes and their underlying genetic basis, in the context of distinct physiological and metabolic constraints that contribute to host resistance and disease tolerance.

MicroRNA-181 in cardiovascular disease: Emerging biomarkers and therapeutic targets.

Cardiovascular disease (CVD) is the leading cause of death worldwide. MicroRNAs (MiRNAs) have attracted considerable attention for their roles in several cardiovascular disease states, including both the physiological and pathological processes. In this review, we will briefly describe microRNA-181 (miR-181) transcription and regulation and summarize recent findings on the roles of miR-181 family members as biomarkers or therapeutic targets in different cardiovascular-related conditions, including atherosclerosis, myocardial infarction, hypertension, and heart failure. Lessons learned from these studies may provide new theoretical foundations for CVD.

FAM222A, Part of the BET-Regulated Basal Endothelial Transcriptome, Is a Novel Determinant of Endothelial Biology and Angiogenesis-Brief Report.

BACKGROUND: BETs (bromodomain and extraterminal domain-containing epigenetic reader proteins), including BRD4 (bromodomain-containing protein 4), orchestrate transcriptional programs induced by pathogenic stimuli, as intensively studied in cardiovascular disease and elsewhere. In endothelial cells (ECs), BRD4 directs induced proinflammatory, proatherosclerotic transcriptional responses; BET inhibitors, like JQ1, repress these effects and decrease atherosclerosis. While BET effects in pathogenic conditions have prompted therapeutic BET inhibitor development, BET action under basal conditions, including ECs, has remained understudied. To understand BET action in basal endothelial transcriptional programs, we first analyzed EC RNA-Seq data in the absence versus presence of JQ1 before using BET regulation to identify novel determinants of EC biology and function. METHODS: RNA-Seq datasets of human umbilical vein ECs without and with JQ1 treatment were analyzed. After identifying C12orf34, also known as FAM222A (family with sequence similarity 222 member A), as a previously unreported, basally expressed, potently JQ1-induced EC gene, FAM222A was studied in endothelial and angiogenic responses in vitro using small-interference RNA silencing and lentiviral overexpression, in vitro, ex vivo and in vivo, including aortic sprouting, matrigel plug assays, and murine neonatal oxygen-induced retinopathy. RESULTS: Resting EC RNA-Seq data indicate BETs direct transcriptional programs underlying core endothelial properties including migration, proliferation, and angiogenesis. BET inhibition in resting ECs also significantly induced a subset of mRNAs, including FAM222A-a unique BRD4-regulated gene with no reported EC role. Silencing endothelial FAM222A significantly decreased cellular proliferation, migration, network formation, aorta sprouting, and Matrigel plug vascularization through coordinated modulation of VEGF (vascular endothelial growth factor) and NOTCH mediator expression in vitro, ex vivo, in vivo; lentiviral FAM222A overexpression had opposite effects. In vivo, siFAM222A significantly repressed retinal revascularization in neonatal murine oxygen-induced retinopathy through similar angiogenic signaling modulation. CONCLUSIONS: BET control over the basal endothelial transcriptome includes FAM222A, a novel, BRD4-regulated, key determinant of endothelial biology and angiogenesis.

Injectable Self-Oxygenating Cardio-Protective and Tissue Adhesive Silk-Based Hydrogel for Alleviating Ischemia After Mi Injury.

Myocardial infarction (MI) is a significant cardiovascular disease that restricts blood flow, resulting in massive cell death and leading to stiff and noncontractile fibrotic scar tissue formation. Recently, sustained oxygen release in the MI area has shown regeneration ability; however, improving its therapeutic efficiency for regenerative medicine remains challenging. Here, a combinatorial strategy for cardiac repair by developing cardioprotective and oxygenating hybrid hydrogels that locally sustain the release of stromal cell-derived factor-1 alpha (SDF) and oxygen for simultaneous activation of neovascularization at the infarct area is presented. A sustained release of oxygen and SDF from injectable, mechanically robust, and tissue-adhesive silk-based hybrid hydrogels is achieved. Enhanced endothelialization under normoxia and anoxia is observed. Furthermore, there is a marked improvement in vascularization that leads to an increment in cardiomyocyte survival by ≈30% and a reduction of the fibrotic scar formation in an MI animal rodent model. Improved left ventricular systolic and diastolic functions by ≈10% and 20%, respectively, with a ≈25% higher ejection fraction on day 7 are also observed. Therefore, local delivery of therapeutic oxygenating and cardioprotective hydrogels demonstrates beneficial effects on cardiac functional recovery for reparative therapy.

Association between tissue loss type and amputation risk among Medicare patients with concomitant diabetes and peripheral arterial disease.

OBJECTIVE: Prior studies have described risk factors associated with amputation in patients with concomitant diabetes and peripheral arterial disease (DM/PAD). However, the association between the severity and extent of tissue loss type and amputation risk remains less well-described. We aimed to quantify the role of different tissue loss types in amputation risk among patients with DM/PAD, in the context of demographic, preventive, and socioeconomic factors. METHODS: Applying International Classification of Diseases (ICD)-9 and ICD-10 codes to Medicare claims data (2007-2019), we identified all patients with continuous fee-for-service Medicare coverage diagnosed with DM/PAD. Eight tissue loss categories were established using ICD-9 and ICD-10 diagnosis codes, ranging from lymphadenitis (least severe) to gangrene (most severe). We created a Cox proportional hazards model to quantify associations between tissue loss type and 1- and 5-year amputation risk, adjusting for age, race/ethnicity, sex, rurality, income, comorbidities, and preventive factors. Regional variation in DM/PAD rates and risk-adjusted amputation rates was examined at the hospital referral region level. RESULTS: We identified 12,257,174 patients with DM/PAD (48% male, 76% White, 10% prior myocardial infarction, 30% chronic kidney disease). Although 2.2 million patients (18%) had some form of tissue loss, 10.0 million patients (82%) did not. The 1-year crude amputation rate (major and minor) was 6.4% in patients with tissue loss, and 0.4% in patients without tissue loss. Among patients with tissue loss, the 1-year any amputation rate varied from 0.89% for patients with lymphadenitis to 26% for patients with gangrene. The 1-year amputation risk varied from two-fold for patients with lymphadenitis (adjusted hazard ratio, 1.96; 95% confidence interval, 1.43-2.69) to 29-fold for patients with gangrene (adjusted hazard ratio, 28.7; 95% confidence interval, 28.1-29.3), compared with patients without tissue loss. No other demographic variable including age, sex, race, or region incurred a hazard ratio for 1- or 5-year amputation risk higher than the least severe tissue loss category. Results were similar across minor and major amputation, and 1- and 5-year amputation outcomes. At a regional level, higher DM/PAD rates were inversely correlated with risk-adjusted 5-year amputation rates (R2 = 0.43). CONCLUSIONS: Among 12 million patients with DM/PAD, the most significant predictor of amputation was the presence and extent of tissue loss, with an association greater in effect size than any other factor studied. Tissue loss could be used in awareness campaigns as a simple marker of high-risk patients. Patients with any type of tissue loss require expedited wound care, revascularization as appropriate, and infection management to avoid amputation. Establishing systems of care to provide these interventions in regions with high amputation rates may prove beneficial for these populations.

Children's empathy moderates the association between perceived interparental conflict and child health.

Interparental conflict is known to negatively impact child well-being, including behavioral and physiological well-being. Children's empathy - that is, vicariously experiencing others' emotions - may increase children's sensitivity to and the biological repercussions of interparental conflict. Although empathy represents a valued trait and is an important part of socioemotional development, its influence on children's physical health is unknown. This study examined whether empathy moderates the association between perceived interparental conflict and both child systemic inflammation and parent-rated overall child health in a sample of children between the ages of seven to nine. Children and their parents participating in the long-term evaluation of the Family Foundations program, a randomized trial of a perinatal preventative intervention, provided data approximately eight years following enrollment into the program. We collected peripheral blood samples via dried blood spots, anthropometric measurements, and child and parent psychosocial questionnaires. Results indicated significant positive main effects of child empathy on both C-reactive protein (CRP; B = 0.26, SE = 0.11, p =.026) and Interleukin-6 (IL-6; B = 0.20, SE = 0.10, p =.045) levels. Further, child affective empathy moderated the associations between perceived interparental conflict and both CRP (B = 0.39, SE = 0.19, p =.050) and parent-reported child health (B = 0.30, SE = 0.13, p =.021), such that greater empathy strengthened the negative associations between interparental conflict and child health. Overall, findings suggests that there may be a biological cost of being more empathic in high-conflict environments and highlight the need for tools to help more empathic children appropriately manage vicarious emotions.

Perivascular Fibrosis Is Mediated by a KLF10-IL-9 Signaling Axis in CD4+ T Cells.

BACKGROUND: Perivascular fibrosis, characterized by increased amount of connective tissue around vessels, is a hallmark for vascular disease. Ang II (angiotensin II) contributes to vascular disease and end-organ damage via promoting T-cell activation. Despite recent data suggesting the role of T cells in the progression of perivascular fibrosis, the underlying mechanisms are poorly understood. METHODS: TF (transcription factor) profiling was performed in peripheral blood mononuclear cells of hypertensive patients. CD4-targeted KLF10 (Kruppel like factor 10)-deficient (Klf10fl/flCD4Cre+; [TKO]) and CD4-Cre (Klf10+/+CD4Cre+; [Cre]) control mice were subjected to Ang II infusion. End point characterization included cardiac echocardiography, aortic imaging, multiorgan histology, flow cytometry, cytokine analysis, aorta and fibroblast transcriptomic analysis, and aortic single-cell RNA-sequencing. RESULTS: TF profiling identified increased KLF10 expression in hypertensive human subjects and in CD4+ T cells in Ang II-treated mice. TKO mice showed enhanced perivascular fibrosis, but not interstitial fibrosis, in aorta, heart, and kidney in response to Ang II, accompanied by alterations in global longitudinal strain, arterial stiffness, and kidney function compared with Cre control mice. However, blood pressure was unchanged between the 2 groups. Mechanistically, KLF10 bound to the IL (interleukin)-9 promoter and interacted with HDAC1 (histone deacetylase 1) inhibit IL-9 transcription. Increased IL-9 in TKO mice induced fibroblast intracellular calcium mobilization, fibroblast activation, and differentiation and increased production of collagen and extracellular matrix, thereby promoting the progression of perivascular fibrosis and impairing target organ function. Remarkably, injection of anti-IL9 antibodies reversed perivascular fibrosis in Ang II-infused TKO mice and C57BL/6 mice. Single-cell RNA-sequencing revealed fibroblast heterogeneity with activated signatures associated with robust ECM (extracellular matrix) and perivascular fibrosis in Ang II-treated TKO mice. CONCLUSIONS: CD4+ T cell deficiency of Klf10 exacerbated perivascular fibrosis and multi-organ dysfunction in response to Ang II via upregulation of IL-9. Klf10 or IL-9 in T cells might represent novel therapeutic targets for treatment of vascular or fibrotic diseases.

Risk of new-onset diabetes and efficacy of pharmacological weight loss therapy.

AIMS: To develop a clinical risk model to identify individuals at higher risk of developing new-onset diabetes and who might benefit more from weight loss pharmacotherapy. MATERIALS AND METHODS: A total of 21 143 patients without type 2 diabetes at baseline from two TIMI clinical trials of stable cardiovascular patients were divided into a derivation (~2/3) and validation (~1/3) cohort. The primary outcome was new-onset diabetes. Twenty-seven candidate risk variables were considered, and variable selection was performed using multivariable Cox regression. The final model was evaluated for discrimination and calibration, and for its ability to identify patients who experienced a larger benefit from the weight loss medication lorcaserin in terms of risk of new-onset diabetes. RESULTS: During a median (interquartile range) follow-up of 2.3 (1.8-2.7) years, new-onset diabetes occurred in 1013 patients (7.7%). The final model included five independent predictors (glycated haemoglobin, fasting glucose, age, body mass index, and triglycerides/high-density lipoprotein). The clinical risk model showed good discrimination (Harrell's C-indices 0.802, 95% confidence interval [CI] 0.788-0.817 and 0.807, 95% CI 0.788-0.826) in the derivation and validation cohorts. The calibration plot demonstrated adequate calibration (2.5-year area under the curve was 81.2 [79.1-83.5]). While hazard ratios for new-onset diabetes with a weight-loss therapy were comparable across risk groups (annual risks of <1%, 1%-5%, and >5%), there was a sixfold gradient in absolute risk reduction from lowest to highest risk group (p = 0.027). CONCLUSIONS: The developed clinical risk model effectively predicts new-onset diabetes, with potential implications for personalized patient care and therapeutic decision making.

Promoting toddlers' self-regulation and healthy eating habits among families living in poverty: A randomized controlled trial of Recipe 4 Success.

The Recipe 4 Success preventive intervention targeted multiple factors critical to the health and well-being of toddlers living in poverty. This randomized controlled trial, which was embedded within Early Head Start home visits for 12 weeks, included 242 racially and ethnically diverse families (51% girls; toddler mean age = 2.58 years; data collected 2016-2019). Compared to parents in usual practice home visits, parents in Recipe 4 Success displayed greater sensitive scaffolding of toddlers' learning and more responsive food parenting practices (Cohen's d = .21-.30). Toddlers in Recipe 4 Success exhibited greater self-regulation and had healthier eating habits (Cohen's d = |.16-.35|). Results highlight the value of Recipe 4 Success in promoting parent and toddler behavior change that could have life-long benefits.

Examining profiles of convergence and divergence in reports of parental warmth: Links to adolescent developmental problems.

Parental warmth during the transition from childhood to adolescence is a key protective factor against a host of adolescent problems, including substance use, maladjustment, and diminished well-being. Moreover, adolescents and parents often disagree in their perceptions of parenting quality, and these discrepancies may confer risk for problem outcomes. The current study applies latent profile analysis to a sample of 687 mother-father-6th grade adolescent triads to identify patterns of adolescent-parent convergence and divergence in perceptions of parental warmth. Five profiles were identified, and associations with adolescent positive well-being, substance use, and maladjustment outcomes in 9th grade were assessed. Patterns of divergence in which adolescents had a pronounced negative perception of parental warmth compared to parents, as well as those wherein pronounced divergence was present in only one adolescent-parent dyad, were associated with diminished positive well-being compared to adolescents who had more positive perceptions of warmth than parents. Having more negative perceptions of warmth compared to parents was also associated with elevated risk for alcohol and marijuana initiation, but only when the divergence was pronounced rather than more moderate. These findings add nuance to findings from previous between-family investigations of informant discrepancies, calling for further family-centered methods for investigating multiple perspectives.

A case study in developmental discontinuity: PROSPER Interventions and adolescent substance use trajectories shape young adult substance use and mental health problems.

The loss of John Schulenberg reverberates across the developmental and prevention sciences. In honor of his many contributions, this paper applies his ideas of developmental continuity and discontinuity to understand the process by which PROSPER delivered universal prevention programs (delivered in Grades 6 and 7) affect young adult outcomes. Guided by these developmental models, we deconstructed adolescent substance use initiation trajectories into two discrete phases-early and late adolescence, demarcated by substance use initiation levels at the end of 9th grade. We evaluated the effects of PROSPER interventions on these phases, and in turn, the effects of adolescent substance use initiation on young adult antisocial behavior, alcohol and drug use consequences, and depression symptoms. This sample included 1,984 young adults who participated in the PROSPER intervention trial in Grade 6 (two cohorts, 2002 and 2003), followed over 8 adolescent measurement occasions (Fall and Spring of Grade 6; Spring of Grades 7-12). Young adult outcomes were averaged across three waves (collected at ages 20, 23, and 25). PROSPER interventions were associated with reduced substance use initiation in early adolescence, but not escalation during late adolescence. In turn, substance use in both early and late adolescence was uniquely associated with young adult antisocial behavior, depression symptoms, and substance use consequences. PROSPER interventions were associated with young adult antisocial behavior and problematic substance use via reduced risk for early initiation status. Findings are discussed in terms of developmental continuity and discontinuity.

Couple relationship functioning and social adjustment during the transition to parenthood among fathers with a history of maltreatment.

Study of fathers has gained significant traction over recent decades. However, the experience for men over the transition to parenthood remains focused on high-socioeconomic and socially advantaged fathers. Researchers have yet to thoroughly investigate how fathers may uniquely experience this transition period with a history of childhood maltreatment, given that childhood abuse is known to impact several components of development and relationship functioning into adulthood. The current study endeavored to fill this gap by evaluating the associations between fathers' childhood experiences of physical and emotional abuse and their relationship functioning over the transition to parenthood in terms of both the couple relationship and social adjustment in relationships with others. Using data from 399 fathers who participated in a randomized control trial during pregnancy, the results from stepwise regressions indicate fathers with a history of emotional abuse experience particular declines in their external relationships (reductions in social support and increases in social stress) from prenatal (Wave 1) to postpartum (Wave 2) reports. However, no significant association emerged between fathers' history of maltreatment and their relationship functioning with their partners. These results underscore the importance of investigating the impact of different types of abuse on men in fatherhood. Moreover, we emphasize the need to study further fathers' social adjustment over the transition to parenthood beyond the couple relationship and broad social support to address the needs of men with a history of maltreatment in their new role as fathers.

Daily implementation of health-protective behaviors and family life during the early months of the COVID-19 pandemic.

The coronavirus disease (COVID-19) pandemic has necessitated the use of health-protective behaviors (HPB), such as social distancing, staying at home, frequent handwashing, and wearing facemasks to mitigate the transmission of disease. An investigation of interpersonal costs associated with the use of HPB can help inform strategies to promote their sustained implementation. This study examined the daily associations between the implementation of HPB and family functioning and assessed moderation by coparenting quality, economic strain, and the number of days that state-level stay-at-home policies had been in effect, during the early days of the pandemic. Mothers and fathers from 155 families with children who were 9 years old, on average, completed daily reports of HPB, parental stress, and family relationship quality over eight consecutive days in April or May of 2020. Hierarchal linear models showed that HPB was associated with increased levels of parental stress and interparental conflict. Negative coparenting relations exacerbated the next-day association between HPB and interparental conflict. HPB was also associated with increased levels of parent-child and interparental closeness, but these linkages dissipated for families who had spent more days under state-level stay-at-home policies. Although crucial for public health, the implementation of HPB may have detrimental short-term effects on daily family life. Family support and interventions are necessary to minimize the psychosocial burden of these important public health measures and increase their sustained adherence.

MicroRNA-375 repression of Kruppel-like factor 5 improves angiogenesis in diabetic critical limb ischemia.

Peripheral artery disease (PAD) is an occlusive disease of limb arteries. Critical limb ischemia (CLI) is an advanced form of PAD that is prognostically worse in subjects with diabetes and can result in limb loss, gangrene, and death, although the underlying signaling mechanisms that contribute to its development remain poorly understood. By comparing plasma samples from diabetic humans with PAD and mouse models of PAD, we identified miR-375 to be significantly downregulated in humans and mice during progression to CLI. Overexpression of miR-375 was pro-angiogenic in endothelial cells in vitro and induced endothelial migration, proliferation, sprouting, and vascular network formation, whereas miR-375 inhibition conferred anti-angiogenic effects. Intramuscular delivery of miR-375 improved blood flow recovery to diabetic mouse hindlimbs following femoral artery ligation (FAL) and improved neovessel growth and arteriogenesis in muscle tissues. Using RNA-sequencing and prediction algorithms, Kruppel-like factor 5 (KLF5) was identified as a direct target of miR-375 and siRNA knockdown of KLF5 phenocopied the effects of miR-375 overexpression in vitro and in vivo through regulatory changes in NF-kB signaling. Together, a miR-375-KLF5-NF-kB signaling axis figures prominently as a potential therapeutic pathway in the development CLI in diabetes.

miR-181b regulates vascular endothelial aging by modulating an MAP3K3 signaling pathway.

Endothelial cell (EC) aging plays a vital role in the pathogenesis of cardiovascular disease (CVD). MicroRNAs have emerged as crucial regulators of target gene expression by inhibiting mRNA translation and/or promoting mRNA degradation. We identify an aging-related and oxidative stress-responsive microRNA, miR-181b, that inhibits endothelial cell apoptosis and senescence. In gain- or loss-of-function studies, miR-181b regulated the expression of key apoptosis markers (Bcl2, Bax, cleaved-Caspase3) and senescence markers (p16, p21, γH2AX) and the ratio of apoptotic cells (TUNEL-positive) and senescent cells (SA-ßgal-positive) in H2 O2 -induced ECs. Mechanistically, miR-181b targets MAP3K3 and modulates a MAP3K3/MKK/MAPK signaling pathway. MAP3K3 knockdown recapitulated the phenotype of miR-181b overexpression and miR-181b was dependent on MAP3K3 for regulating EC apoptosis and senescence. In vivo, miR-181b expression showed a negative correlation with increasing age in the mouse aorta. Endothelial-specific deficiency of miR-181a2b2 increased the target MAP3K3, markers of vascular senescence (p16, p21), and DNA double-strand breaks (γH2AX) in the aorta of aged mice. Collectively, this study unveils an important role of miR-181b in regulating vascular endothelial aging via an MAP3K3-MAPK signaling pathway, providing new potential therapeutic targets for antiaging therapy in CVD.

A miRNA cassette reprograms smooth muscle cells into endothelial cells.

Cellular reprogramming through targeting microRNAs (miRNAs) holds promise for regenerative therapy due to their profound regulatory effects in proliferation, differentiation, and function. We hypothesized that transdifferentiation of vascular smooth muscle cells (SMCs) into endothelial cells (ECs) using a miRNA cassette may provide a novel approach for use in vascular disease states associated with endothelial injury or dysfunction. miRNA profiling of SMCs and ECs and iterative combinatorial miRNA transfections of human coronary SMCs revealed a 4-miRNA cassette consisting of miR-143-3p and miR-145-5p inhibitors and miR-146a-5p and miR-181b-5p mimics that efficiently produced induced endothelial cells (iECs). Transcriptome profiling, protein expression, and functional studies demonstrated that iECs exhibit high similarity to ECs. Injected iECs restored blood flow recovery even faster than conventional ECs in a murine hindlimb ischemia model. This study demonstrates that a 4-miRNA cassette is sufficient to reprogram SMCs into ECs and shows promise as a novel regenerative strategy for endothelial repair.

Parental warmth and young adult depression: A comparison of enduring effects and revisionist models.

Guided by a novel analytic framework, this study investigates the developmental mechanism through which parental warmth is related to young adult depression. Data were from a large sample of participants followed from early adolescence to young adulthood (N = 1,988; 54% female). Using structural equation modeling, we estimated and compared competing developmental models - enduring effects vs. revisionist models - to assess whether parental warmth during adolescence had enduring or transient effects on depression in young adulthood. We also examined whether contemporaneous experiences of parental warmth in young adulthood were more salient than parental warmth in adolescence. Results supported the revisionist model: early intergenerational experiences in adolescence predicted psychopathology early in young adulthood, but their unique effects gradually diminished; whereas parental warmth in young adulthood continued to be protective of young adult depression. Effects of mother and father warmth on young adult depression were similar in pattern and magnitude. Results were held when accounting for covariates such as adolescent sex, family income status, and family structure. Young adult mental health interventions may consider targeting maintenance or improvement in parental warmth to help offset the long-term impact of adversity early in life.

Trajectories of parent and child well-being across the pandemic year: Role of financial strain, social distancing, and COVID-19 prevalence.

Existing research demonstrated large deteriorations in parent, child, and family well-being within 2 months after the onset of the COVID-19 pandemic. Yet, little is known about the trajectories of families' adjustment in the following months, including what risk factors are associated with changes in families' adjustment. The current study examined (1) change in the parent, child, and family well-being over time; (2) associations of pandemic-related stressors, financial and social distancing-associated stress, with well-being between and within families; and (3) the role of local COVID-19 prevalence, prior participation in family-focused prevention, and parent gender. From April 2020 to January 2021, 393 parents from 235 families reported five times on parent mental health, child behavior problems, family relationships, and pandemic-related stressors. Findings indicate that, across all domains of well-being, there was either little change across the 8 months or a small degree of recovery followed by a shift to further deterioration. On average, parents experiencing greater pandemic-related stressors also reported poorer functioning in all domains; monthly fluctuations in pandemic-related stressors were also associated with fluctuations in parent mental health and child behavior problems. In some domains, the links between pandemic-related stressors and parent and child well-being were stronger among families living in areas with overall higher COVID-19 prevalence rates. Parents' experiences during the pandemic did not differ systematically across prior intervention participation or parent gender. Taken together, findings suggest a need for supportive interventions to help families navigate extended periods of crisis.

LncRNA-MAP3K4 regulates vascular inflammation through the p38 MAPK signaling pathway and cis-modulation of MAP3K4.

Chronic vascular inflammation plays a key role in the pathogenesis of atherosclerosis. Long non-coding RNAs (lncRNAs) have emerged as essential inflammation regulators. We identify a novel lncRNA termed lncRNA-MAP3K4 that is enriched in the vessel wall and regulates vascular inflammation. In the aortic intima, lncRNA-MAP3K4 expression was reduced by 50% during the progression of atherosclerosis (chronic inflammation) and 70% during endotoxemia (acute inflammation). lncRNA-MAP3K4 knockdown reduced the expression of key inflammatory factors (eg, ICAM-1, E-selectin, MCP-1) in endothelial cells or vascular smooth muscle cells and decreased monocytes adhesion to endothelium, as well as reducing TNF-α, IL-1ß, COX2 expression in macrophages. Mechanistically, lncRNA-MAP3K4 regulates inflammation through the p38 MAPK signaling pathway. lncRNA-MAP3K4 shares a bidirectional promoter with MAP3K4, an upstream regulator of the MAPK signaling pathway, and regulates its transcription in cis. lncRNA-MAP3K4 and MAP3K4 show coordinated expression in response to inflammation in vivo and in vitro. Similar to lncRNA-MAP3K4, MAP3K4 knockdown reduced the expression of inflammatory factors in several different vascular cells. Furthermore, lncRNA-MAP3K4 and MAP3K4 knockdown showed cooperativity in reducing inflammation in endothelial cells. Collectively, these findings unveil the role of a novel lncRNA in vascular inflammation by cis-regulating MAP3K4 via a p38 MAPK pathway.

Rapid cGMP manufacturing of COVID-19 monoclonal antibody using stable CHO cell pools.

Therapeutic proteins, including monoclonal antibodies, are typically manufactured using clonally derived, stable host cell lines, since consistent and predictable cell culture performance is highly desirable. However, selecting and preparing banks of stable clones takes considerable time, which inevitably extends overall development timelines for new therapeutics by delaying the start of subsequent activities, such as the scale-up of manufacturing processes. In the context of the coronavirus disease 2019 (COVID-19) pandemic, with its intense pressure for accelerated development strategies, we used a novel transposon-based Leap-In Transposase® system to rapidly generate high-titer stable pools and then used them directly for large scale-manufacturing of an anti-severe acute respiratory syndrome coronavirus 2 monoclonal antibody under cGMP. We performed the safety testing of our non-clonal cell bank, then used it to produce material at a 200L-scale for preclinical safety studies and formulation development work, and thereafter at 2000L scale for supply of material for a Phase 1 clinical trial. Testing demonstrated the comparability of critical product qualities between the two scales and, more importantly, that our final clinical trial product met all pre-set product quality specifications. The above expediated approach provided clinical trial material within 4.5 months, in comparison to 12-14 months for production of clinical trial material via the conventional approach.