Multiplex Imaging for Cell Phenotyping of Early Human Atherosclerosis.

BACKGROUND: Previous studies using animal models and cultured cells suggest that vascular smooth muscle cells (SMCs) and inflammatory cytokines are important players in atherogenesis. Validating these findings in human disease is critical to designing therapeutics that target these components. Multiplex imaging is a powerful tool for characterizing cell phenotypes and microenvironments using biobanked human tissue sections. However, this technology has not been applied to human atherosclerotic lesions and needs to first be customized and validated. METHODS AND RESULTS: For validation, we created an 8-plex imaging panel to distinguish foam cells from SMC and leukocyte origins on tissue sections of early human atherosclerotic lesions (n=9). The spatial distribution and characteristics of these foam cells were further analyzed to test the association between SMC phenotypes and inflammation. Consistent with previous reports using human lesions, multiplex imaging showed that foam cells of SMC origin outnumbered those of leukocyte origin and were enriched in the deep intima, where the lipids accumulate in early atherogenesis. This new technology also found that apoptosis or the expression of pro-inflammatory cytokines were not more associated with foam cells than with nonfoam cells in early human lesions. More CD68+ SMCs were present among SMCs that highly expressed interleukin-1ß. Highly inflamed SMCs showed a trend of increased apoptosis, whereas leukocytes expressing similar levels of cytokines were enriched in regions of extracellular matrix remodeling. CONCLUSIONS: The multiplex imaging method can be applied to biobanked human tissue sections to enable proof-of-concept studies and validate theories based on animal models and cultured cells.

A Deep Learning Approach for Improving Two-Photon Vascular Imaging Speeds.

A potential method for tracking neurovascular disease progression over time in preclinical models is multiphoton fluorescence microscopy (MPM), which can image cerebral vasculature with capillary-level resolution. However, obtaining high-quality, three-dimensional images with traditional point scanning MPM is time-consuming and limits sample sizes for chronic studies. Here, we present a convolutional neural network-based (PSSR Res-U-Net architecture) algorithm for fast upscaling of low-resolution or sparsely sampled images and combine it with a segmentation-less vectorization process for 3D reconstruction and statistical analysis of vascular network structure. In doing so, we also demonstrate that the use of semi-synthetic training data can replace the expensive and arduous process of acquiring low- and high-resolution training pairs without compromising vectorization outcomes, and thus open the possibility of utilizing such approaches for other MPM tasks where collecting training data is challenging. We applied our approach to images with large fields of view from a mouse model and show that our method generalizes across imaging depths, disease states and other differences in neurovasculature. Our pretrained models and lightweight architecture can be used to reduce MPM imaging time by up to fourfold without any changes in underlying hardware, thereby enabling deployability across a range of settings.

Chemoproteomics identifies proteoform-selective caspase-2 inhibitors.

Caspases are a highly conserved family of cysteine-aspartyl proteases known for their essential roles in regulating apoptosis, inflammation, cell differentiation, and proliferation. Complementary to genetic approaches, small-molecule probes have emerged as useful tools for modulating caspase activity. However, due to the high sequence and structure homology of all twelve human caspases, achieving selectivity remains a central challenge for caspase-directed small-molecule inhibitor development efforts. Here, using mass spectrometry-based chemoproteomics, we first identify a highly reactive non-catalytic cysteine that is unique to caspase-2. By combining both gel-based activity-based protein profiling (ABPP) and a tobacco etch virus (TEV) protease activation assay, we then identify covalent lead compounds that react preferentially with this cysteine and afford a complete blockade of caspase-2 activity. Inhibitory activity is restricted to the zymogen or precursor form of monomeric caspase-2. Focused analogue synthesis combined with chemoproteomic target engagement analysis in cellular lysates and in cells yielded both pan-caspase reactive molecules and caspase-2 selective lead compounds together with a structurally matched inactive control. Application of this focused set of tool compounds to stratify caspase contributions to initiation of intrinsic apoptosis, supports compensatory caspase-9 activity in the context of caspase-2 inactivation. More broadly, our study highlights future opportunities for the development of proteoform-selective caspase inhibitors that target non-conserved and non-catalytic cysteine residues.

Diamond Raman laser and Yb fiber amplifier for in vivo multiphoton fluorescence microscopy.

Here we introduce a fiber amplifier and a diamond Raman laser that output high powers (6.5 W, 1.3 W) at valuable wavelengths (1060 nm, 1250 nm) for two-photon excitation of red-shifted fluorophores. These custom excitation sources are both simple to construct and cost-efficient in comparison to similar custom and commercial alternatives. Furthermore, they operate at a repetition rate (80 MHz) that allows fast image acquisition using resonant scanners. With our system we demonstrate compatibility with fast resonant scanning, the ability to acquire neuronal images, and the capability to image vasculature at deep locations (>1 mm) within the mouse cerebral cortex.

Evaluation of resonant scanning as a high-speed imaging technique for two-photon imaging of cortical vasculature.

We demonstrate a simple, low-cost two-photon microscope design with both galvo-galvo and resonant-galvo scanning capabilities. We quantify and compare the signal-to-noise ratios and imaging speeds of the galvo-galvo and resonant-galvo scanning modes when used for murine neurovascular imaging. The two scanning modes perform as expected under shot-noise limited detection and are found to achieve comparable signal-to-noise ratios. Resonant-galvo scanning is capable of reaching desired signal-to-noise ratios using less acquisition time when higher excitation power can be used. Given equal excitation power and total pixel dwell time between the two methods, galvo-galvo scanning outperforms resonant-galvo scanning in image quality when detection deviates from being shot-noise limited.

Caregiver status and illness self-efficacy during the COVID-19 pandemic among older adults with chronic conditions.

OBJECTIVES: Older adults providing unpaid care to a relative or friend during the COVID-19 pandemic may have diminished self-efficacy in managing their own chronic illness, especially in the context of more complex self-management. We evaluated whether adults aged 50 and older with caregiving roles are more likely to report reduced illness self-efficacy since the pandemic, and whether this link is exacerbated by a higher number of conditions. METHODS: Participants (105 caregivers and 590 noncaregivers) residing in Michigan (82.6%) and 33 other U.S. states completed one online survey between May 14 and July 9, 2020. RESULTS: Controlling for sociodemographic and health characteristics, stressors related to COVID-19, and behavioral and psychosocial changes since the pandemic, caregivers were more likely than noncaregivers to report reduced illness self-efficacy when they had a higher number of chronic conditions. CONCLUSION: These findings highlight the importance of maintaining caregivers' self-care during the COVID-19 pandemic and in future public health crises.

Social Contact, Emotional Support, and Anxiety during the COVID-19 Pandemic among Older Adults with Chronic Conditions.

OBJECTIVES: The COVID-19 pandemic may contribute to heightened anxiety among older adults with chronic conditions, which might be attenuated by social resources. This study examined how social contact and emotional support were linked to anxiety symptoms among adults aged 50 and older with chronic conditions, and whether these links varied by age. METHODS: Participants included 705 adults (M = 64.61 years, SD = 8.85, range = 50- 94) from Michigan (82.4%) and 33 other U.S. states who reported at least one chronic condition and completed an anonymous online survey between May 14 and July 9, 2020. RESULTS: Multiple regression models revealed among younger people, those reporting more frequent social contact had significantly lower anxiety symptoms. Emotional support was not significantly associated with anxiety symptoms. CONCLUSIONS: More frequent social contact was linked to lower anxiety symptoms for younger but not older individuals. Emotional support was not significantly associated with anxiety symptoms. CLINICAL IMPLICATIONS: Interventions to manage anxiety during the pandemic among older adults with chronic conditions may benefit from strategies to safely increase social contact, especially for middle-aged adults.

Dyadic associations between relationship quality and risk of opioid use among couples receiving methadone for opioid use disorder.

BACKGROUND: Social relationships may serve as both protective factors and risk factors for opioid use (nonmedical prescription opioid or illicit opioid use) among patients receiving methadone for opioid use disorder (OUD). Yet little is known about how relationship quality is linked to outcomes among couples receiving methadone. We evaluated the links between relationship quality and risk of opioid use among couples in which both partners received methadone. METHODS: Participants included 53 heterosexual married or cohabiting couples aged 18 and older who were drawn from two opioid treatment programs in Rhode Island and Massachusetts. Both members of the couple completed a self-administered survey assessing their sociodemographic information, relationship and treatment characteristics, and risk of opioid use. RESULTS: Roughly half of women (47.2%) and men (52.8%) had a moderate to high risk of nonmedical prescription opioid use and almost two-thirds (64.2%) had a moderate to high risk of street opioid use. Risk of street opioid use was highly correlated within couples. Actor-partner interdependence models revealed that when women reported higher positive relationship quality, they had a lower risk of nonmedical prescription opioid use and their partners had a lower risk of street opioid use. Negative relationship quality was not significantly linked to risk of opioid use. CONCLUSIONS: Couples in which both partners receive methadone for OUD may be at risk of return to use, and positive partner relationships may play a role in lowering this risk. Women's perceptions of relationship quality might be a particularly important target for clinical care and interventions.

Severe acute malnutrition treatment delivered by low-literate community health workers in South Sudan: A prospective cohort study.

BACKGROUND: Community health worker (CHW)-delivered acute malnutrition treatment programs have been tested previously, but not with low-literate/-numerate cadres who operate in areas with the highest malnutrition burden and under-five mortality rates. The International Rescue Committee developed low-literacy-adapted tools and treatment protocol to enable low-literate/-numerate community-based distributors (CBD, the CHW cadre in South Sudan) to treat children for severe acute malnutrition (SAM) in their communities. METHODS: We conducted a prospective cohort study in March-September 2017, with 44 CBDs enrolling a total of 308 SAM children into treatment in their communities. Child treatment outcomes and length of treatment were documented. Uncomplicated SAM cases, defined for our study as children with mid-upper arm circumference (MUAC) of 90 to <115 mm or bilateral pitting oedema, without any medical complications, were treated for up to 16 weeks, and were considered fully recovered when they reached MUAC≥125 mm for two consecutive weeks. RESULTS: The recovery rate from the severe to the moderate acute malnutrition (MAM) cut-off of MUAC 115 mm was 91% (95% confidence interval (CI) = 88%-95%). The median length of treatment was five weeks. The recovery rate of children from SAM to full recovery was 75% (95% CI = 69%-81%). The median time to full recovery was eight weeks. The recovery rates reported here exclude children referred for care from the denominator, per standard reporting of acute malnutrition treatment recovery rates. When the data were compared against routine monitoring and evaluation data from nearby static clinics, children treated by CBDs appeared to have improved continuity of care and shorter time to recovery. CONCLUSIONS: The recovery rate for SAM children enrolled in acute malnutrition treatment by low-literate CBDs shows promise that deploying CHWs to treat SAM in areas with high prevalence and low treatment access may lead to higher recovery, better continuity of care in the transition between SAM and MAM, and shorter treatment time. Proper adaptations of tools and protocols can empower CHW cadres with low literacy and numeracy to successfully complete treatment steps. Key questions of scalability and cost-effectiveness remain.

Performance of low-literate community health workers treating severe acute malnutrition in South Sudan.

Previous studies have described barriers to access of childhood severe acute malnutrition (SAM) treatment, including long travel distances and high opportunity costs. To increase access in remote communities, the International Rescue Committee developed a simplified SAM treatment protocol and low-literacy-adapted tools for community-based distributors (CBD, the community health worker cadre in South Sudan) to deliver treatment in the community. A mixed-methods pilot study was conducted to assess whether low-literate CBDs can adhere to a simplified SAM treatment protocol and to examine the community acceptability of CBDs providing treatment. Fifty-seven CBDs were randomly selected to receive training. CBD performance was assessed immediately after training, and 44 CBDs whose performance score met a predetermined standard were deployed to test the delivery of SAM treatment in their communities. CBDs were observed and scored on their performance on a biweekly basis through the study. Immediately after training, 91% of the CBDs passed the predetermined 80% performance score cut-off, and 49% of the CBDs had perfect scores. During the study, 141 case management observations by supervisory staff were conducted, resulting in a mean score of 89.9% (95% CI: 86.4%-96.0%). For each performance supervision completed, the final performance score of the CBD rose by 2.0% (95% CI: 0.3%-3.7%), but no other CBD characteristic was associated with the final performance score. This study shows that low-literate CBDs in South Sudan were able to follow a simplified treatment protocol for uncomplicated SAM with high accuracy using low-literacy-adapted tools, showing promise for increasing access to acute malnutrition treatment in remote communities.

The effects of noise on key workplace skills.

This study explored the effect on memory and psychomotor performance of wideband noise (simulated in-cabin aircraft noise) at 75 dBA, which is similar to that experienced during the cruise phase of a commercial flight. The results from the tests were compared to the effects of a widely known and common metric on the same skills, namely, blood alcohol concentration (BAC). All 32 participants, half non-native English speakers, completed three different tests (recognition memory, working memory, and reaction time) presented in counterbalanced order, either in the presence of noise, with or without noise attenuation headphones, and without noise but with a BAC of 0.05 or 0.10. Simulated aircraft noise was found to affect recognition memory but not working memory or reaction time. These effects were more pronounced for non-native speakers and reflected performance similar to that for BAC of 0.05 or 0.10.

Temporal associations of popularity and alcohol use among middle school students.

PURPOSE: The goal of this study is to better understand the longitudinal cross-lagged associations between popularity, assessed through self-rating and peer nominations, and alcohol use among middle school students. METHODS: The analytical sample comprises 1,835 sixth- to eighth-grade students who were initially recruited from three California middle schools and surveyed in the fall and spring semesters of 2 academic years. Students reported on their background characteristics, past-month alcohol use, and perceived popularity. Additionally, students provided school-based friendship nominations, which were used to calculate peer-nominated popularity. A cross-lagged regression approach within a structural equation modeling framework was used to examine the longitudinal relationship between popularity (self-rated and peer-nominated) and alcohol use. RESULTS: There was a statistically significant (p = .024) association between peer-nominated popularity and the probability of alcohol consumption at the subsequent survey, but not vice versa. Our results suggest that in a scenario where 8% of students are past-month drinkers, each increase of five friendship nominations is associated with a 30% greater risk of being a current drinker at the next wave. We found no evidence of longitudinal associations between past-month alcohol consumption and self-rated popularity. CONCLUSIONS: Popularity is a risk factor for drinking during the middle school years, with peer-nominated popularity being more predictive of use than self-perceptions of popularity. To inform alcohol prevention efforts for middle school students, additional research is needed to better understand why adolescents with a larger number of school-based friendship ties are more inclined to drink.

Do client attributes moderate the effectiveness of a group cognitive behavioral therapy for depression in addiction treatment?

The study goal was to determine whether client attributes were associated with outcomes from group cognitive behavioral therapy for depression (GCBT-D) as delivered in community-based addiction treatment settings. Data from 299 depressed residential clients assigned to receive either usual care (N = 159) or usual care plus GCBT-D (N = 140) were examined. Potential moderators included gender, race/ethnicity, education, referral status, and problem substance use. Study outcomes at 6 months post-baseline included changes in depressive symptoms, mental health functioning, negative consequences from substance use, and percentage of days abstinent. Initial examination indicated that non-Hispanic Whites had significantly better outcomes than other racial/ethnic groups on two of the four outcomes. After correcting for multiple testing, none of the examined client attributes moderated the treatment effect. GCBT-D appears effective; however, the magnitude and consistency of treatment effects indicate that it may be less helpful among members of racial/ethnic minority groups and is worthy of future study.