Work-related factors of mental health among Chicago residents two years into the COVID-19 pandemic.

The COVID-19 pandemic led to widespread consequences for economic, social, and general wellbeing with rates of anxiety and depression increasing across the population and disproportionately for some workers. This study explored which factors were the most salient contributors to mental health through a cross-sectional 68-item questionnaire that addressed topics related to the pandemic. Data were collected through an address-based sampling frame over the two months from April 2022 to June 2022. A total of 2,049 completed surveys were collected throughout Chicago's 77 Community Areas. Descriptive statistics including frequency and percentages were generated to describe workplace characteristics, work-related stress, and sample demographics and their relationship to psychological distress. Independent participant and workplace factors associated with the outcomes were identified using multivariable logistic regression. The weighted prevalence of persons experiencing some form of psychological distress from mild to serious was 32%. After adjusting for potential confounding factors, certain marginalized communities experienced psychological distress more than others including females, adults over the age of 25 years of age, and people with higher income levels. Those who had been laid off, lost pay, or had reduced hours had increased odds of psychological distress (aOR = 1.71, CI95% 1.14-2.56; p = 0.009) as did people that reported that their work-related stress was somewhat or much worse as compared to before the COVID-19 pandemic (aOR = 2.22, CI95% 1.02-4.82; p = 0.04, aOR = 11.0, CI95% 4.65-26.1; p < 0.001, respectively). These results warrant further investigation and consideration in developing workplace and mental health interventions.

Discovery, Optimization, and Biological Evaluation of Arylpyridones as Cbl-b Inhibitors.

Casitas B-lymphoma proto-oncogene-b (Cbl-b), a member of the Cbl family of RING finger E3 ubiquitin ligases, has been demonstrated to play a central role in regulating effector T-cell function. Multiple studies using gene-targeting approaches have provided direct evidence that Cbl-b negatively regulates T, B, and NK cell activation via a ubiquitin-mediated protein modulation. Thus, inhibition of Cbl-b ligase activity can lead to immune activation and has therapeutic potential in immuno-oncology. Herein, we describe the discovery and optimization of an arylpyridone series as Cbl-b inhibitors by structure-based drug discovery to afford compound 31. This compound binds to Cbl-b with an IC50 value of 30 nM and induces IL-2 production in T-cells with an EC50 value of 230 nM. Compound 31 also shows robust intracellular target engagement demonstrated through inhibition of Cbl-b autoubiquitination, inhibition of ubiquitin transfer to ZAP70, and the cellular modulation of phosphorylation of a downstream signal within the TCR axis.

The Effects of Interpersonal Violence on Sleep Following the COVID-19 Stay-at-Home Order.

PURPOSE: Lack of sleep is a harm that can lead to chronic diseases ranging from diabetes to heart disease. We examined the exposure to interpersonal violence and its association with sleep, following the COVID-19 stay-at-home order. DESIGN: Cross-sectional. SETTING: Surveys were completed online and via paper-and-pencil in English and Spanish (N = 2049; RR = 68.4%). SUBJECTS: Respondents were 18+ and residing in Chicago. MEASURES: The Chicago Department of Public Health's "2022 Healthy Chicago Survey COVID-19 Social Impact Survey". ANALYSIS: We developed two weighted models. Model 1 examined the effects of neighborhood violence on meeting the national sleep recommendation. Model 2 examined the effects of violence in the home among friends or family on meeting the sleep recommendation, incorporating additional predictors: victimization, stress, gender, race/ethnicity, household income, and general health. Odds ratios were estimated using multivariate logistic regression. RESULTS: Exposure to neighborhood violence and sleep was not significant, but knowing a friend or family member who experienced violence or mistreatment in their home affected the odds of meeting the sleep recommendation (OR = .61, 95% CI = .44-.84). Non-Hispanic Blacks had 52% lower odds of meeting sleep recommendations (OR = .48, 95% CI = .37-.63). CONCLUSION: Addressing the harms to sleep that followed COVID-19 should engage diverse stakeholders in implementing culturally responsive interventions to promote adequate sleep and prevent chronic disease.

Financial, Social, and Health Impacts from the COVID-19 Pandemic: Findings from the Healthy Chicago Survey.

BACKGROUND: This study examines self-reported impacts of the COVID-19 pandemic among Chicago residents and disparities of these impacts across demographic characteristics. Six logistic regression models were developed to identify demographic and socioeconomic factors associated with each COVID-19 impact. METHOD: The study used de-identified and weighted data from the 2020 Healthy Chicago Survey (HCS). HCS is an annual survey using an address-based random sampling method administered by the Chicago Department of Public Health (CDPH) across the city of Chicago on adults aged 18 or older living between July 17 and November 11, 2020, and received 4517 responses. RESULTS: Chicago residents with lower socioeconomic status had a higher likelihood of food and housing insecurity. Non-Hispanic (NH) Black residents were more likely to report being unable to obtain food (OR: 2.996; 95% CI: 2.268-2.324); being unable to pay for rent, mortgage, or bills (OR: 2.352; 95% CI: 2.325-2.380); and grief from a loss of someone to COVID-19 (OR: 2.037; 95% CI: 2.013-2.061) compared to NH White residents. NH White residents were more likely than other racial/ethnic groups to report loss of social connections, worsened mental health, and canceling or postponing medical care. CONCLUSIONS: The analysis showed higher odds of social contact loss and worsened mental health from COVID-19 in NH White and higher education populations. By implementing strategies to address specific challenges faced by different racial groups, Chicago may effectively mitigate pandemic's adverse effects. These strategies can promote a more inclusive approach to distributing COVID investments for programs and policies.

Assessment of changes in Streptococcus pyogenes levels using N-acetylgalactosamine-6-sulfatase marker and pharyngeal airway space with appliance therapy in mouth breathers - An ELISA-based study.

Background: The frequency of adenotonsillar hypertrophy in mouth-breathing children when compared to the average found in the general population is considered to be higher. Mouth breathing is considered as one of the causative factors for tonsillitis in children. Through continuous irritation on tonsillar wall, tonsils swell up and inflammation develops. Purpose: The purpose of the study is to evaluate Streptococcus pyogenes count using colony-forming units (CFUs) and N-acetylgalactosamine-6-sulfatase side chain marker on ELISA (enzyme linked immunosorbent assay) in mouth breathers and to establish its correlation with pharyngeal airway space pre- and post-oral screen appliance therapy. Materials and Methods: A total number of 24 (n) mouth breathers aged between 5 and 12 years were included in the study and given oral screen appliance therapy. The subjects were evaluated for the various parameters before the delivery of a habit-breaking appliance and then reevaluated for the same parameters (presence of S. pyogenes and its counts, size of tonsils, and pharyngeal airway space dimensions) after 6 months of appliance usage. Results: A statistically significant difference was seen in levels of S. pyogenes using ELISA and CFUs. Furthermore, statistically significant difference was observed in Friedman tonsil scoring and pharyngeal airway space and pre- and post-oral screen appliance therapy. Conclusion: Oral screen appliance therapy reduced the frequency of occurrence of tonsillitis in mouth breathers by decreasing the counts of S. pyogenes bacteria. Upper and lower pharyngeal airway space dimensions were increased after 6 months of appliance therapy in mouth breathers.

An SMC-like protein binds and regulates Caenorhabditis elegans condensins.

Structural Maintenance of Chromosomes (SMC) family proteins participate in multisubunit complexes that govern chromosome structure and dynamics. SMC-containing condensin complexes create chromosome topologies essential for mitosis/meiosis, gene expression, recombination, and repair. Many eukaryotes have two condensin complexes (I and II); C. elegans has three (I, II, and the X-chromosome specialized condensin IDC) and their regulation is poorly understood. Here we identify a novel SMC-like protein, SMCL-1, that binds to C. elegans condensin SMC subunits, and modulates condensin functions. Consistent with a possible role as a negative regulator, loss of SMCL-1 partially rescued the lethal and sterile phenotypes of a hypomorphic condensin mutant, while over-expression of SMCL-1 caused lethality, chromosome mis-segregation, and disruption of condensin IDC localization on X chromosomes. Unlike canonical SMC proteins, SMCL-1 lacks hinge and coil domains, and its ATPase domain lacks conserved amino acids required for ATP hydrolysis, leading to the speculation that it may inhibit condensin ATPase activity. SMCL-1 homologs are apparent only in the subset of Caenorhabditis species in which the condensin I and II subunit SMC-4 duplicated to create the condensin IDC- specific subunit DPY-27, suggesting that SMCL-1 helps this lineage cope with the regulatory challenges imposed by evolution of a third condensin complex. Our findings uncover a new regulator of condensins and highlight how the duplication and divergence of SMC complex components in various lineages has created new proteins with diverse functions in chromosome dynamics.

Heat-shock response transcriptional program enables high-yield and high-quality recombinant protein production in Escherichia coli.

The biosynthesis of soluble, properly folded recombinant proteins in large quantities from Escherichia coli is desirable for academic research and industrial protein production. The basal E. coli protein homeostasis (proteostasis) network capacity is often insufficient to efficiently fold overexpressed proteins. Herein we demonstrate that a transcriptionally reprogrammed E. coli proteostasis network is generally superior for producing soluble, folded, and functional recombinant proteins. Reprogramming is accomplished by overexpressing a negative feedback deficient heat-shock response transcription factor before and during overexpression of the protein-of-interest. The advantage of transcriptional reprogramming versus simply overexpressing select proteostasis network components (e.g., chaperones and co-chaperones, which has been explored previously) is that a large number of proteostasis network components are upregulated at their evolved stoichiometry, thus maintaining the system capabilities of the proteostasis network that are currently incompletely understood. Transcriptional proteostasis network reprogramming mediated by stress-responsive signaling in the absence of stress should also be useful for protein production in other cells.

Census 2: isobaric labeling data analysis.

MOTIVATION: We introduce Census 2, an update of a mass spectrometry data analysis tool for peptide/protein quantification. New features for analysis of isobaric labeling, such as Tandem Mass Tag (TMT) or Isobaric Tags for Relative and Absolute Quantification (iTRAQ), have been added in this version, including a reporter ion impurity correction, a reporter ion intensity threshold filter and an option for weighted normalization to correct mixing errors. TMT/iTRAQ analysis can be performed on experiments using HCD (High Energy Collision Dissociation) only, CID (Collision Induced Dissociation)/HCD (High Energy Collision Dissociation) dual scans or HCD triple-stage mass spectrometry data. To improve measurement accuracy, we implemented weighted normalization, multiple tandem spectral approach, impurity correction and dynamic intensity threshold features. AVAILABILITY AND IMPLEMENTATION: Census 2 supports multiple input file formats including MS1/MS2, DTASelect, mzXML and pepXML. It requires JAVA version 6 or later to run. Free download of Census 2 for academic users is available at http://fields.scripps.edu/census/index.php. CONTACT: jyates@scripps.edu SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Single-step inline hydroxyapatite enrichment facilitates identification and quantitation of phosphopeptides from mass-limited proteomes with MudPIT.

Herein we report the characterization and optimization of single-step inline enrichment of phosphopeptides directly from small amounts of whole cell and tissue lysates (100-500 µg) using a hydroxyapatite (HAP) microcolumn and Multidimensional Protein Identification Technology (MudPIT). In comparison to a triplicate HILIC-IMAC phosphopeptide enrichment study, ∼80% of the phosphopeptides identified using HAP-MudPIT were unique. Similarly, analysis of the consensus phosphorylation motifs between the two enrichment methods illustrates the complementarity of calcium- and iron-based enrichment methods and the higher sensitivity and selectivity of HAP-MudPIT for acidic motifs. We demonstrate how the identification of more multiply phosphorylated peptides from HAP-MudPIT can be used to quantify phosphorylation cooperativity. Through optimization of HAP-MudPIT on a whole cell lysate we routinely achieved identification and quantification of ca. 1000 phosphopeptides from a ∼1 h enrichment and 12 h MudPIT analysis on small quantities of material. Finally, we applied this optimized method to identify phosphorylation sites from a mass-limited mouse brain region, the amygdala (200-500 µg), identifying up to 4000 phosphopeptides per run.

Proteome analysis of Plasmodium falciparum extracellular secretory antigens at asexual blood stages reveals a cohort of proteins with possible roles in immune modulation and signaling.

The highly co-evolved relationship of parasites and their hosts appears to include modulation of host immune signals, although the molecular mechanisms involved in the host-parasite interplay remain poorly understood. Characterization of these key genes and their cognate proteins related to the host-parasite interplay should lead to a better understanding of this intriguing biological phenomenon. The malaria agent Plasmodium falciparum is predicted to export a cohort of several hundred proteins to remodel the host erythrocyte. However, proteins actively exported by the asexual intracellular parasite beyond the host red blood cell membrane (before merozoite egress) have been poorly investigated so far. Here we used two complementary methodologies, two-dimensional gel electrophoresis/MS and LC-MS/MS, to examine the extracellular secreted antigens at asexual blood stages of P. falciparum. We identified 27 novel antigens exported by P. falciparum in the culture medium of which some showed clustering with highly polymorphic genes on chromosomes, suggesting that they may encode putative antigenic determinants of the parasite. Immunolocalization of four novel secreted proteins confirmed their export beyond the infected red blood cell membrane. Of these, preliminary functional characterization of two novel (Sel1 repeat-containing) parasite proteins, PfSEL1 and PfSEL2 revealed that they down-regulate expression of cell surface Notch signaling molecules in host cells. Also a novel protein kinase (PfEK) and a novel protein phosphatase (PfEP) were found to, respectively, phosphorylate/dephosphorylate parasite-specific proteins in the extracellular culture supernatant. Our study thus sheds new light on malaria parasite extracellular secreted antigens of which some may be essential for parasite development and could constitute promising new drug targets.

Increase of metabolic activity and disruption of normal contractile protein distribution by bilirubin oxidation products in vascular smooth-muscle cells.

OBJECT: Cerebral vasospasm is a common cause of morbidity and death following aneurysmal subarachnoid hemorrhage (SAH). Previous research has shown that bilirubin oxidation products (BOXes) are present in the cerebral spinal fluid in patients with SAH-induced cerebral vasospasm and can contribute to vasoconstriction and vasospasm in vitro and in vivo. The events leading to cerebral vasospasm are not understood; however, one component of the occlusion may be due to vascular remodeling. In this study the authors have investigated the actions of BOXes, okadaic acid ([OA], a phosphatase inhibitor), and phorbol-12 myristate-13 acetate ([PMA], a protein kinase activator) on vascular smooth-muscle cell (VSMC) morphology and metabolism. METHODS: Immunohistochemical analysis was performed to assess VSMC morphology and alpha-smooth-muscle actin (alphaSMA) distribution following the application of BOXes, OA, or PMA. Changes in the level of lactate dehydrogenase (LDH) release and oxidative metabolism were also measured. The BOXes, OA, or PMA caused VSMCs to change their shape and exhibit altered alphaSMA distribution. These treatments increased LDH release (p < 0.05), which is an index of increased cell stress. Oxidative metabolism significantly increased at low and high doses of BOXes, that is, 143 +/- 8.5% and 180 +/- 11.8%, respectively (p < 0.0001). Both PMA and OA also caused a significant increase in metabolism. CONCLUSIONS: The authors concluded that BOXes, OA, and PMA alter VSMC morphology and metabolic activity, events that have been observed during vascular remodeling. Although the mechanism remains unclear, the results indicate that BOXes may play a role in the vascular remodeling that occurs following aneurysmal SAH.