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The COVID-19 pandemic forced many people to work from home. Even after pandemic restrictions have been lifted, many parents continue to work from home. This poses several challenges and has led to a number of issues related to sharing rooms and equipment, dealing with distractions, and dealing with feelings. This paper examines the issues that arose for parents working from home in Flanders within these three categories, and recommends potential solutions. Data were collected through a survey. The target population of the survey was a group of Flemish parents with children under the age of 18 and working from home during the lockdown. The main findings of the research are summarized as follows. Most respondents did not have a separate workspace at home. This means that parents experienced different distractions at home than while working from the office. Children were the number one distraction for parents working from home. The care of children and the lack of work-life balance were also the main sources of conflict. However, the feelings when working from home are not only negative. Parents felt frustration and powerlessness, as well as motivation and happiness. Based on the analysis of the responses, recommendations are formulated from parents to parents on how to organise working from home with children.

Minoritized racial and ethnic groups experience mental health issues yet persistently encounter systemic barriers to accessing mental health care. Disparities in mental health services are linked to structural racism, discrimination, and stigma. Social determinants of health also impact the risk of experiencing mental health issues and contribute to mental healthcare access. Community-based organizations (CBOs) can serve as vital linkages for delivering much-needed support across many domains (e.g., physical and mental health, housing, food, recreation, etc.) and provide opportunities for fostering connections with members of underserved communities. There is untapped potential in leveraging the skills and expertise offered by CBOs and their staff. Designing programs and interventions that incorporate the goals and ideals of the community served may facilitate sustainability and contribute to a reconceptualization of care delivery based on equity. Ensuring that community-informed initiatives are set up for success will require advocacy for multisector collaboration and innovative approaches and policies that facilitate community collaborations to improve mental health and well-being for communities of color to flourish.

Is noun dominance in early lexical acquisition a widespread or a language-specific phenomenon? Thirty Singaporean bilingual English–Mandarin learning toddlers and their mothers were observed in a mother-child play interaction. For both English and Mandarin, toddlers’ speech and reported vocabulary contained more nouns than verbs across book reading and toy playing. In contrast, their mothers’ speech contained more verbs than nouns in both English and Mandarin but differed depending on the context of the interaction. Although toddlers demonstrated a noun bias for both languages, the noun bias was more pronounced in English than in Mandarin. Together, these findings support early noun dominance as a widespread phenomenon in the lexical acquisition debate but also provide evidence that language specificity also plays a minor role in children’s early lexical development. Ministry of Education (MOE) Published version This research was supported by Singapore Ministry of Education’s Academic Research Fund Tier 1 and Social Science Research Thematic Grant (MOE2016-SSRTG-017) to Peipei Setoh.

Background Skin fibrosis is the clinical hallmark of systemic sclerosis (SSc), where collagen deposition and remodeling of the dermis occur over time. The most widely used outcome measure in SSc clinical trials is the modified Rodnan skin score (mRSS), which is a semi-quantitative assessment of skin stiffness at seventeen body sites. However, the mRSS is confounded by obesity, edema, and high inter-rater variability. In order to develop a new histopathological outcome measure for SSc, we applied a computer vision technology called a deep neural network (DNN) to stained sections of SSc skin. We tested the hypotheses that DNN analysis could reliably assess mRSS and discriminate SSc from normal skin. Methods We analyzed biopsies from two independent (primary and secondary) cohorts. One investigator performed mRSS assessments and forearm biopsies, and trichrome-stained biopsy sections were photomicrographed. We used the AlexNet DNN to generate a numerical signature of 4096 quantitative image features (QIFs) for 100 randomly selected dermal image patches/biopsy. In the primary cohort, we used principal components analysis (PCA) to summarize the QIFs into a Biopsy Score for comparison with mRSS. In the secondary cohort, using QIF signatures as the input, we fit a logistic regression model to discriminate between SSc vs. control biopsy, and a linear regression model to estimate mRSS, yielding Diagnostic Scores and Fibrosis Scores, respectively. We determined the correlation between Fibrosis Scores and the published Scleroderma Skin Severity Score (4S) and between Fibrosis Scores and longitudinal changes in mRSS on a per patient basis. Results In the primary cohort (n = 6, 26 SSc biopsies), Biopsy Scores significantly correlated with mRSS (R = 0.55, p = 0.01). In the secondary cohort (n = 60 SSc and 16 controls, 164 biopsies; divided into 70% training and 30% test sets), the Diagnostic Score was significantly associated with SSc-status (misclassification rate = 1.9% [training], 6.6% [test]), and the Fibrosis Score significantly correlated with mRSS (R = 0.70 [training], 0.55 [test]). The DNN-derived Fibrosis Score significantly correlated with 4S (R = 0.69, p = 3 × 10− 17). Conclusions DNN analysis of SSc biopsies is an unbiased, quantitative, and reproducible outcome that is associated with validated SSc outcomes.

The antimicrobial peptide cathelicidin inhibits development of colitis-associated colon cancer. However, the role of cathelicidin in colon cancer metastasis remains unknown. We hypothesized that cathelicidin is effective in inhibiting colon cancer metastasis. Human colon cancer HT-29 cells were injected intravenously into nude mice. Control HA-tagged adeno-associated virus (HA-AAV) or cathelicidin-overexpressing AAV (CAMP-HA-AAV) were injected intravenously into nude mice on the same day. Four weeks later, the nude mice were assessed for lung and liver metastases. Human colon cancer SW620 cells were used to study the effect of cathelicidin on cell migration and cytoskeleton. Incubation of SW620 cells with cathelicidin dose-dependently reduced cell migration, disrupted cytoskeletal structure, and reduced βIII-tubulin (TUBB3) mRNA expression. The addition of the P2RX7 antagonist KN62, but not the FPRL1 antagonist WRW4, prevented the LL-37-mediated inhibition of cell migration and TUBB3 mRNA expression. The CAMP-HA-AAV-overexpressing group showed significantly reduced human CK20 protein (by 60%) and TUBB3 mRNA expression (by 40%) in the lungs and liver of the HT-29-loaded nude mice, compared to the HA-AAV control group. Intraperitoneal injection of KN62 reversed the CAMP-HA-AAV-mediated inhibition of human CK20 and TUBB3 expression in the lungs and liver of HT-29-loaded nude mice. In conclusion, cathelicidin inhibits colon cancer metastasis via a P2RX7-dependent pathway. Keywords: colon cancer, metastasis, antimicrobial peptide

ObjectivesWe sought to assess the presence and reporting quality of peer-reviewed literature concerning the accuracy, precision, and reliability of home monitoring technologies for vital signs and glucose determinations in older adult populations.MethodsA narrative literature review was undertaken searching the databases Medline, Embase, and Compendex. Peer-reviewed publications with keywords related to vital signs, monitoring devices and technologies, independent living, and older adults were searched. Publications between the years 2012 and 2018 were included. Two reviewers independently conducted title and abstract screening, and four reviewers independently undertook full-text screening and data extraction with all disagreements resolved through discussion and consensus.ResultsTwo hundred nine articles were included. Our review showed limited assessment and low-quality reporting of evidence concerning the accuracy, precision, and reliability of home monitoring technologies. Of 209 articles describing a relevant device, only 45 percent (n = 95) provided a citation or some evidence to support their validation claim. Of forty-eight articles that described the use of a comparator device, 65 percent (n = 31) used low-quality statistical methods, 23 percent (n = 11) used moderate-quality statistical methods, and only 12 percent (n = 6) used high-quality statistical methods.ConclusionsOur review found that current validity claims were based on low-quality assessments that do not provide the necessary confidence needed by clinicians for medical decision-making purposes. This narrative review highlights the need for standardized health technology reporting to increase health practitioner confidence in these devices, support the appropriate adoption of such devices within the healthcare system, and improve health outcomes.

As our understanding of the genetic underpinnings of SSc increases, questions regarding the environmental trigger(s) that induce and propagate SSc in the genetically predisposed individual emerge. The interplay between the environment, the immune system, and the microbial species that inhabit the patient’s skin and gastrointestinal tract is a pathobiological frontier that is largely unexplored in SSc. The purpose of this review is to provide an overview of the methodologies, experimental study results and future roadmap for elucidating the relationship between the SSc host and his/her microbiome.

A major problem in human cognition is to understand how newly acquired information and long-standing beliefs about the environment combine to make decisions and plan behaviors. Over-dependence on long-standing beliefs may be a significant source of suboptimal decision-making in unusual circumstances. While the contribution of long-standing beliefs about the environment to search in real-world scenes is well-studied, less is known about how new evidence informs search decisions, and it is unclear whether the two sources of information are used together optimally to guide search. The present study expanded on the literature on semantic guidance in visual search by modeling a Bayesian ideal observer’s use of long-standing semantic beliefs and recent experience in an active search task. The ability to adjust expectations to the task environment was simulated using the Bayesian ideal observer, and subjects’ performance was compared to ideal observers that depended on prior knowledge and recent experience to varying degrees. Target locations were either congruent with scene semantics, incongruent with what would be expected from scene semantics, or random. Half of the subjects were able to learn to search for the target in incongruent locations over repeated experimental sessions when it was optimal to do so. These results suggest that searchers can learn to prioritize recent experience over knowledge of scenes in a near-optimal fashion when it is beneficial to do so, as long as the evidence from recent experience was learnable.

Chinese-speaking parents are believed to use less cognitive mental-state-talk than their English-speaking counterparts on account of their cultural goals in socializing their children to follow an interdependence script. Here, we investigated bilingual English&ndash, Mandarin Singaporean mothers who associate different functions for each language as prescribed by their government: English for school and Mandarin for in-group contexts. English and Mandarin maternal mental-state-talk from bilingual English&ndash, Mandarin mothers with their toddlers was examined. Mothers produced more &lsquo, &rsquo, cognitive&rsquo, terms in English than in Mandarin and more &lsquo, desire&rsquo, terms in Mandarin than in English. We show that mental-state-talk differs between bilingual parents&rsquo, languages, suggesting that mothers adjust their mental-state-talk to reflect the functions of each language.

Purpose To characterize and provide population-based prevalence estimates of myopic degeneration (MD) among Chinese Americans, the fastest-growing minority population in the United States in the last decade. Design Population-based, cross-sectional study. Methods A total of 1523 Chinese-American adults with myopia, aged 50 years and older, residing in the city of Monterey Park, California, underwent an interview and comprehensive eye examination, including subjective and objective refraction and stereoscopic fundus photography. For each participant, the eye with the worse myopic refractive error was included in this analysis. MD was assessed in a masked manner by an expert grader. Results The prevalence of any MD was 44.9% among myopic subjects, based on the presence of any degenerative lesion secondary to myopia. The prevalence was 32.2% when MD was defined by a modified version of the Meta-Analysis for Pathologic Myopia. The prevalence of specific lesions included tessellation (31.7%), tilted disc (28.1%), peripapillary atrophy (7.0%), staphyloma (5.7%), diffuse atrophy (6.4%), lacquer cracks (2.6%), intrachoroidal cavitation (2.2%), patchy atrophy (0.9%), and end-stage MD (0.2%). The prevalence of MD was higher among older myopic subjects and among participants with more severe myopia and longer axial length ( P Conclusions These data provide the first population-based estimates of MD in Chinese Americans and indicate that a high proportion of Chinese Americans might be at risk for MD and associated visual complications. Prevalence of MD in this cohort of Chinese Americans was higher than that observed in other East Asian populations and in people of other ethnicities.

Eosinophilic fasciitis is a rare connective tissue disorder characterized by inflammation of the fascia that leads to painful, indurated skin. Because of its variable clinical presentation and overlap with conditions, such as morphea, the diagnosis of eosinophilic fasciitis can be challenging and relies on clinical presentation, histopathologic and laboratory analysis, and response to therapy. Herein, we present an unusual, solitary, isolated plaque with pathologic features and response to therapy most consistent with eosinophilic fasciitis.

Background Brain metastases (BM) are frequently observed in advanced renal-cell carcinoma (RCC). Historically these individuals have been excluded from clinical trials, but recently, with better local control, many can receive aggressive therapy after treatment. We evaluate our single-institution experience over various treatment eras. Patients and Methods Patients undergoing evaluation for RCC BM from 2001 to 2018 were identified from our institutional database. Clinical notes, demographics, comorbidities, histology, central nervous system (CNS) treatments, systemic therapy, and outcomes were reviewed. Overall survival (OS) and CNS recurrence-free survival (RFS) were evaluated by the Kaplan-Meier method. Cumulative incidence was evaluated using a competing risk model. Results We identified 158 patients with RCC BM, of whom 94.4% had clear-cell RCC, and 90.6% had extracranial metastases at diagnosis. Of these patients, 94 (60%) developed RCC BM over time, while 46 (29.1%) had RCC BM at initial presentation. Clinical symptoms were noted in 81.9% of patients. The median OS after diagnosis of RCC BM was 8.4 months, with a 3-year OS of 28.2%. The median CNS RFS was 8.5 months overall; however, those with one and more than one lesion had median CNS RFS of 12.4 and 6 months, respectively (P Conclusion The majority of RCC patients with BM are symptomatic and had prior metastatic disease that progressed to the brain. Those with a solitary RCC BM are less likely to develop CNS recurrence after local therapy and are ideal candidates for enrollment onto clinical trials.

Aspergillus spp. are a leading cause of mortality in chronic granulomatous disease (CGD), but other fungi have emerged in the era of mould prophylaxis. Of these, Phellinus spp. are an under-recognised cause of invasive fungal infections (IFIs) in CGD, and data on their presentation and management are scarce. We present a patient with CGD who developed disseminated IFI involving the lungs and brain. Surgical specimens grew a basidiomycete which was disregarded as a contaminant. After three months of progressive disease despite antifungals, he was diagnosed with Phellinus tropicalis by internal transcribed spacer (ITS) sequencing. He improved with amphotericin B and isavuconazole but required haematopoietic stem cell transplantation (HSCT). We review the literature on Phellinus infections in CGD and conclude that: (i) these infections emerge on mould-active prophylaxis and are indolent; (ii) they typically cause locally destructive disease but can disseminate; (iii) diagnosis is delayed and requires molecular methods; (iv) amphotericin B is most active in vitro; and (v) treatment is protracted and requires surgery and possibly HSCT. In conclusion, Phellinus spp. are emerging pathogens in CGD. Every effort should be made to establish the diagnosis of non-Aspergillus IFIs in patients with CGD by sending tissue specimens for molecular diagnostics.

Social and linguistic explanations have been proposed for pronoun reversals in young typically developing (TD) children and those with autism spectrum disorders (ASD). The current study breaks new ground in investigating both explanations, comparing 18 TD toddlers and 15 children with ASD at similar language levels. Spontaneous speech was sampled every four months for six visits. Vocabulary and joint attention were also measured. Both groups produced pronoun reversals at low rates. The ASD group produced somewhat more reversals than the TD group, overall and at multiple visits. In the ASD group, early language and joint attention scores contributed significantly and independently to the incidence of reversal. Both linguistic and social factors seem implicated; moreover, reversals seem to occur when children’s language and social abilities develop asynchronously. These findings can help clinicians devise both linguistic and social interventions for the relevant children.

Background and objectivesObesity is a global epidemic which increases the risk of the metabolic syndrome. Cathelicidin (LL-37 and mCRAMP) is an antimicrobial peptide with an unknown role in obesity. We hypothesize that cathelicidin expression correlates with obesity and modulates fat mass and hepatic steatosis.Materials and methodsMale C57BL/6 J mice were fed a high-fat diet. Streptozotocin was injected into mice to induce diabetes. Experimental groups were injected with cathelicidin and CD36 overexpressing lentiviruses. Human mesenteric fat adipocytes, mouse 3T3-L1 differentiated adipocytes and human HepG2 hepatocytes were used in the in vitro experiments. Cathelicidin levels in non-diabetic, prediabetic and type II diabetic patients were measured by enzyme-linked immunosorbent assay.ResultsLentiviral cathelicidin overexpression reduced hepatic steatosis and decreased the fat mass of high-fat diet-treated diabetic mice. Cathelicidin overexpression reduced mesenteric fat and hepatic fatty acid translocase (CD36) expression that was reversed by lentiviral CD36 overexpression. Exposure of adipocytes and hepatocytes to cathelicidin significantly inhibited CD36 expression and reduced lipid accumulation. Serum cathelicidin protein levels were significantly increased in non-diabetic and prediabetic patients with obesity, compared with non-diabetic patients with normal body mass index (BMI) values. Prediabetic patients had lower serum cathelicidin protein levels than non-diabetic subjects.ConclusionsCathelicidin inhibits the CD36 fat receptor and lipid accumulation in adipocytes and hepatocytes, leading to a reduction of fat mass and hepatic steatosis in vivo. Circulating cathelicidin levels are associated with increased BMI. Our results demonstrate that cathelicidin modulates the development of obesity.

Value-based healthcare (VBHC) is a term synonymous with the pursuit of greater value in healthcare, however, the term “value” lacks conceptual clarity. Using Porter’s 2010 seminal VBHC shows promise for placing the older adult and their family caregiver at the center of care models, which is a central tenet in healthcare value. The purpose of our study is to conceptualize how Porter’s VBHC is defined, operationalized, and implemented to address the need for person-and-family centered care models that improve outcomes while being cost effective. A literature search in six academic databases was conducted to identify articles examining VBHC; specifically studies with a Porter-based patient-centric focus. 1,001 articles were retrieved for initial review. Using a consensus-based logic model for inclusion/exclusion, 802 met the inclusion criteria for full text review. Articles were examined using the following objectives: 1) conceptually map the VBHC literature, 2) identify application of Porter’s equation, and 3) identify the methodologies used to measure outcomes, costs, and value. Findings were cross-compared and emergent themes organized to Porter facets of value (outcomes, costs, and value). Our review found the three facets of Porter’s VBHC are established across the literature, however, most studies examine only one or two facets and fail to specify or define all three. Although applied in research, there is a lack of consistency in the actual use of Porter’s definition. Five recommendations for future research using Porter’s VBHC include: (1) pre-selection of outcomes/costs, (2) operationalizing outcomes/costs, (3) value framework creation, (4) data collection, and (5) value calculation.

Numerous antimicrobial peptides (AMPs) are expressed in the mucosa of the gastrointestinal tract where they are able to modulate innate immune responses and gut microbiota. Interestingly, the antimicrobial functions of AMPs are not necessarily the most important mechanism in the modulation of gastrointestinal diseases. Instead, AMPs may promote certain protective microbial species and modulate innate immune responses. The interactions of AMPs with innate immunity and gut microbiota reveal interesting drug targets. AMPs may serve as novel therapeutic approaches against gastrointestinal infection and inflammation, but the application of AMPs and their derivatives in treating gastrointestinal diseases remains at an early stage. AMPs may even directly modulate obesity and metabolic diseases. Additionally, AMPs may serve as biomarkers of gastrointestinal diseases, as expression of AMPs is often altered during the development of gastrointestinal diseases. This report summarizes the latest development of AMP-related gastrointestinal research with emphasis on innate immunity and gut microbiota.

Clostridium difficile infection (CDI) is a common, debilitating infection with high morbidity and mortality. C. difficile causes diarrhea and intestinal inflammation by releasing two toxins, toxin A and toxin B. The macrolide antibiotic fidaxomicin was recently shown to be effective in treating CDI, and its beneficial effect was associated with fewer recurrent infections in CDI patients. Since other macrolides possess anti-inflammatory properties, we examined the possibility that fidaxomicin alters C. difficile toxin A-induced ileal inflammation in mice. The ileal loops of anesthetized mice were injected with fidaxomicin (5, 10, or 20 μM), and after 30 min, the loops were injected with purified C. difficile toxin A or phosphate-buffered saline alone. Four hours after toxin A administration, ileal tissues were processed for histological evaluation (epithelial cell damage, neutrophil infiltration, congestion, and edema) and cytokine measurements. C. difficile toxin A caused histologic damage, evidenced by increased mean histologic score and ileal interleukin-1β (IL-1β) protein and mRNA expression. Treatment with fidaxomicin (20 μM) or its primary metabolite, OP-1118 (120 μM), significantly inhibited toxin A-mediated histologic damage and reduced the mean histology score and ileal IL-1β protein and mRNA expression. Both fidaxomicin and OP-1118 reduced toxin A-induced cell rounding in human colonic CCD-18Co fibroblasts. Treatment of ileal loops with vancomycin (20 μM) and metronidazole (20 μM) did not alter toxin A-induced histologic damage and IL-1β protein expression. In addition to its well known antibacterial effects against C. difficile , fidaxomicin may possess anti-inflammatory activity directed against the intestinal effects of C. difficile toxins.

Although the field of spiritual coping has greatly expanded in recent years, there is no short scale of Buddhist coping as there is for the Abrahamic religions. The goal of the present study is to create the Brief BCOPE, a 15-item scale of Buddhist coping based on a study of 860 Buddhist participants across the United States. Exploratory and confirmatory factor analyses provide preliminary evidence of the construct validity of the Brief BCOPE, and data are presented suggesting potential convergent, discriminant, criterion, and incremental validity for the scale. Although there are limitations to the study, the present research suggests that the Brief BCOPE might provide an efficient means of assessing Buddhist coping in applied and research settings.

Background The impact of having an established dermatologist on melanoma depth at diagnosis is incompletely understood. Objective We sought to determine whether having had a previous dermatologic examination (an established dermatologist), the recency of the last examination, and the wait time for the dermatology appointment are associated with melanoma invasiveness and depth. Methods This was a retrospective cross-sectional study of 388 patients with primary melanoma at an academic dermatology department. Results Patients with an established dermatologist were more likely than patients without an established dermatologist to be given a diagnosis of melanoma in situ (103/162 [63.6%] vs 69/155 [44.5%], P = .001) and to have thinner invasive melanoma (0.48 [0.30-0.71] mm vs 0.61 [0.40-1.10] mm, respectively, P = .003). These trends were observed for patients with self-detected, but not dermatologist-detected, melanoma. Patient-detected melanomas made up 184/361 (51.0%) of all melanomas, 83/199 (41.7%) of in situ melanomas, and 101/162 (62.4%) invasive melanomas. Self-detected melanomas were in situ in 36 of 61 (59.0%) patients with an established dermatologist versus 40 of 108 (37.0%) patients without an established dermatologist, P = .006. Neither time from last dermatologic examination nor wait time for an appointment was associated with melanoma invasiveness or depth. Limitations Data are retrospective and from 1 large academic health care system. Conclusion Education obtained at the dermatology appointment may improve early self-detection of melanoma, and having an established dermatologist may facilitate earlier evaluation of concerning lesions.

Misattributions people make about their own reaction to ambiguous stimuli can be used to measure personality self–concepts implicitly. On the basis of a semantic misattribution priming paradigm [semantic misattribution procedure (SMP)], we assessed the implicit personality self–concept related to three dimensions included in the Big–Five model: conscientiousness, neuroticism, and extraversion. Across three studies (N1 = 98, N2 = 140, and N3 = 135), the SMP was robustly related, in the expected direction, to individual differences in self–reported personality questionnaires and managed to predict both self–reported and objectively measured behaviours. The main advantage of SMP over classical explicit measures of personality is its higher resistance to social desirability tendencies, although its psychometric properties are somewhat lower than those pertaining to explicit measures of personality. Finally, comparisons of our results with studies that used other implicit measures of personality self–concept indicate that the SMP has higher criterion validity. Copyright © 2012 John Wiley & Sons, Ltd.

To estimate the prevalence of refractive errors in adult Chinese Americans, and to evaluate factors associated with myopia and high myopia.A population-based, cross-sectional study.Chinese Americans 50 years and older residing in Monterey Park, California, were recruited. Noncycloplegic automated refraction with supplemental subjective refraction was performed. Myopia, high myopia, hyperopia, and high hyperopia were defined as a spherical equivalent of-0.5 diopter (D),-5.0 D,+0.5 D, and ≥+3.0 D, respectively. Astigmatism and high astigmatism were defined as a cylinder of0.5 D and2.25 D, respectively. Risk factor assessment was guided by a conceptual model.Data from 4144 participants were analyzed. The overall prevalence of myopia, high myopia, hyperopia, high hyperopia, astigmatism, and high astigmatism in the right eye was 35.1% (95% confidence interval, 33.6%-36.6%), 7.4% (6.6%-8.3%), 40.2% (38.7%-41.8%), 2.7% (2.2%-3.3%), 45.6% (44.1%-47.2%), and 3.7% (3.1%-4.3%), respectively. The prevalence of myopia and high myopia was lower among older individuals (P.05). Reversed age trends were observed for the other refractive errors (P.05). There was no sex difference in the prevalence of refractive errors, except for a higher prevalence of hyperopia among female subjects (P = .010). Age, acculturation, education, income, marital status, birth country, history of ocular disease, nonocular comorbidities, and recent eye examination were associated with prevalence of myopia. All of these factors, except for acculturation, were also associated with high myopia.Our data present the first population-based estimates of the prevalence of refractive errors among adult Chinese Americans. Compared with whites, Hispanics, and blacks, Chinese Americans have a higher burden of myopia, high myopia, and astigmatism.

Tissue clearing methods promise to provide exquisite three-dimensional imaging information; however, there is a need for simplified methods for lower resource settings and for non-fluorescence based phenotyping to enable light microscopic imaging modalities. Here we describe the simplified CLARITY method (SCM) for tissue clearing that preserves epitopes of interest. We imaged the resulting tissues using light sheet microscopy to generate rapid 3D reconstructions of entire tissues and organs. In addition, to enable clearing and 3D tissue imaging with light microscopy methods, we developed a colorimetric, non-fluorescent method for specifically labeling cleared tissues based on horseradish peroxidase conversion of diaminobenzidine to a colored insoluble product. The methods we describe here are portable and can be accomplished at low cost and can allow light microscopic imaging of cleared tissues, thus enabling tissue clearing and imaging in a wide variety of settings.

Objective: Functional recovery after stroke has been observed in both human and animal studies. Post-stroke brain stimulations are promising neurorestorative techniques as they allow direct manipulation of the target area’s excitability. Previously we have demonstrated that optogenetic neuronal stimulation of the ipsilesional primary motor cortex (iM1) promotes functional recovery. To determine an optimal brain stimulation target, we test whether optogenetic neuronal stimulation of the contralesional cerebellar dentate nucleus (cLCN) can promote recovery. We hypothesize that stimulation of cLCN may be more effective, as it sends excitatory outputs to multiple motor and premotor areas. Methods: Thy-1-ChR2-YFP line-18 transgenic male mice were used. Mice underwent stereotaxic surgery to implant an optical fiber in cLCN or iM1, followed by an intraluminal middle cerebral artery suture occlusion. Three groups of mice were used: control non-stimulated stroke mice, short stimulated stroke mice (short-stim, day5-14 post-stroke) and long stimulated stroke mice (long-stim, day5-28 post-stroke). Sensorimotor behavior tests were used to assess their recovery. Results: Our data showed that cLCN-stimulated stroke mice recovered quickly, with significant improvement in distance traveled as early as day7 (p Conclusion: Our data suggest that cLCN stimulations post-stroke can promote persistent functional recovery. Furthermore, cLCN-stimulated mice recovered faster than iM1-stimulated mice, indicate that cLCN may be a more effective brain stimulation target. Current studies examine the brain activation patterns of cLCN-stimulated mice, as well as the mechanisms of cLCN-induced recovery.

Objective: Recovery can occur after stroke in both human and animals, and this is attributed in part by rewiring of neural connections in areas adjacent to or remotely connected to the infarct. As stroke can cause brain-wide network changes, it is important to interrogate regenerative processes in the whole brain after stroke. In this study we use a high resolution 3D whole brain imaging technique called CLARITY to visualize the cellular and structural changes in stroke mice during recovery. Hypothesis: We hypothesize that the CLARITY procedure will provide a more detailed and complete visualization of post-stroke regenerative processes in the whole brain. Material/Methods: We used C57Bl6 WT mice with 10-12 weeks of age. Ischemic stroke was induced by transient middle cerebral artery occlusion using an intraluminal suture. Mice were sacrificed at different time points (Post-stroke day 1, 7, 14 and 28) and brains were processed with the CLARITY protocol. Brains were immunostained repeatedly with antibodies for neurons, glia, oligodendrocytes, microglia/macrophages and blood vessels. Results: Stroke brains collected at different post-stroke timepoints became optically transparent after the CLARITY process. Interestingly, the ischemic area turned transparent for all timepoints except post-stroke day 7 and 28. At this time point the ischemic area remains opaque after CLARITY process, suggesting that the cellular composition at this timepoint is resistant to the CLARITY treatment. Immunostaining with MAP2 demonstrates labeling of neuronal processes in the whole brain. Current studies investigate other cell types and cellular processes, such as glial scarring (GFAP), microglia/macrophage accumulation (CD68) and angiogenesis (Collagen IV) to map the reorganization at the cellular level with high resolution in 3D whole brain. Conclusion: Stroke brains can be made optically transparent after the CLARITY process. This allows high resolution whole brain 3D imaging to probe for the cellular and molecular mechanisms during stroke recovery.