Structured illumination microscopy for cancer identification in diagnostic breast biopsies.

Breast cancer is the second most common cancer diagnosed in women in the US with almost 280,000 new cases anticipated in 2023. Currently, on-site pathology for location guidance is not available during the collection of breast biopsies or during surgical intervention procedures. This shortcoming contributes to repeat biopsy and re-excision procedures, increasing the cost and patient discomfort during the cancer management process. Both procedures could benefit from on-site feedback, but current clinical on-site evaluation techniques are not commonly used on breast tissue because they are destructive and inaccurate. Ex-vivo microscopy is an emerging field aimed at creating histology-analogous images from non- or minimally-processed tissues, and is a promising tool for addressing this pain point in clinical cancer management. We investigated the ability structured illumination microscopy (SIM) to generate images from freshly-obtained breast tissues for structure identification and cancer identification at a speed compatible with potential on-site clinical implementation. We imaged 47 biopsies from patients undergoing a guided breast biopsy procedure using a customized SIM system and a dual-color fluorescent hematoxylin & eosin (H&E) analog. These biopsies had an average size of 0.92 cm2 (minimum 0.1, maximum 4.2) and had an average imaging time of 7:29 (minimum 0:22, maximum 37:44). After imaging, breast biopsies were submitted for standard histopathological processing and review. A board-certified pathologist returned a binary diagnostic accuracy of 96% when compared to diagnoses from gold-standard histology slides, and key tissue features including stroma, vessels, ducts, and lobules were identified from the resulting images.

The nonvesicular sterol transporter Aster-C plays a minor role in whole body cholesterol balance.

Introduction: The Aster-C protein (encoded by the Gramd1c gene) is an endoplasmic reticulum (ER) resident protein that has been reported to transport cholesterol from the plasma membrane to the ER. Although there is a clear role for the closely-related Aster-B protein in cholesterol transport and downstream esterification in the adrenal gland, the specific role for Aster-C in cholesterol homeostasis is not well understood. Here, we have examined whole body cholesterol balance in mice globally lacking Aster-C under low or high dietary cholesterol conditions. Method: Age-matched Gramd1c +/+ and Gramd1c -/- mice were fed either low (0.02%, wt/wt) or high (0.2%, wt/wt) dietarycholesterol and levels of sterol-derived metabolites were assessed in the feces, liver, and plasma. Results: Compared to wild type controls (Gramd1c +/+) mice, mice lackingGramd1c (Gramd1c -/-) have no significant alterations in fecal, liver, or plasma cholesterol. Given the potential role for Aster C in modulating cholesterol metabolism in diverse tissues, we quantified levels of cholesterol metabolites such as bile acids, oxysterols, and steroid hormones. Compared to Gramd1c +/+ controls, Gramd1c -/- mice had modestly reduced levels of select bile acid species and elevated cortisol levels, only under low dietary cholesterol conditions. However, the vast majority of bile acids, oxysterols, and steroid hormones were unaltered in Gramd1c -/- mice. Bulk RNA sequencing in the liver showed that Gramd1c -/- mice did not exhibit alterations in sterol-sensitive genes, but instead showed altered expression of genes in major urinary protein and cytochrome P450 (CYP) families only under low dietary cholesterol conditions. Discussion: Collectively, these data indicate nominal effects of Aster-C on whole body cholesterol transport and metabolism under divergent dietary cholesterol conditions. These results strongly suggest that Aster-C alone is not sufficient to control whole body cholesterol balance, but can modestly impact circulating cortisol and bile acid levels when dietary cholesterol is limited.

Systematic characterization of multi-omics landscape between gut microbial metabolites and GPCRome in Alzheimer's disease.

Shifts in the magnitude and nature of gut microbial metabolites have been implicated in Alzheimer's disease (AD), but the host receptors that sense and respond to these metabolites are largely unknown. Here, we develop a systems biology framework that integrates machine learning and multi-omics to identify molecular relationships of gut microbial metabolites with non-olfactory G-protein-coupled receptors (termed the "GPCRome"). We evaluate 1.09 million metabolite-protein pairs connecting 408 human GPCRs and 335 gut microbial metabolites. Using genetics-derived Mendelian randomization and integrative analyses of human brain transcriptomic and proteomic profiles, we identify orphan GPCRs (i.e., GPR84) as potential drug targets in AD and that triacanthine experimentally activates GPR84. We demonstrate that phenethylamine and agmatine significantly reduce tau hyperphosphorylation (p-tau181 and p-tau205) in AD patient induced pluripotent stem cell-derived neurons. This study demonstrates a systems biology framework to uncover the GPCR targets of human gut microbiota in AD and other complex diseases if broadly applied.

Do flickering lights impact turkey hen behavior, stress, and fear?

Little is known about the effect of light-flicker frequency on poultry, particularly on turkeys. This experiment examined the impact of light-flicker frequency on the behavior, stress, and fear response of Nicholas Select turkey hens reared to 11 wk. The experiment was a randomized complete block design (2 trials), with a one-way factorial analysis evaluating 3 light-flicker frequencies (30, 90, or 195 Hertz; Hz). Birds (n = 3,276 per trial) were housed in 9 individual environmentally controlled rooms (3 replicates per treatment per trial). Data collected included: behavior (4, 8, and 10 wk), incidence of aggressive damage, heterophil-to-lymphocyte ratio, and novel object test (daily d 1-7 and at 4, 8, and 11 wk). Data were analyzed using Proc Mixed (SAS 9.4), with significance declared at P ≤ 0.05. Behavior data are presented as the percentage of time spent performing the behavior. At 4 wk, gentle feather pecking and exploratory behaviors were higher under 195 Hz compared to 30 Hz (P = 0.04 and P = 0.05, respectively). Preening was higher under 90 Hz compared to 30 Hz (P = 0.05). At 8 wk, wing flapping was lowest under 195 Hz (P < 0.01). Gentle feather pecking was higher under 90 and 195 Hz compared to 30 Hz (P = 0.02). Fighting (P = 0.05), aggressive pecking (P = 0.02), and aggressive behaviors (P = 0.01) were lower under 30 Hz compared to 90 Hz. At 10 wk, preening was decreased under 30 Hz (P = 0.03). Incidences of aggressive damage were reduced under 30 Hz compared to 90 Hz (0 d-4 wk; P = 0.01) and under 30 compared to both 90 and 195 Hz (4-8 wk; P = 0.01). At 11 wk, heterophil-to-lymphocyte ratios were lowest under 30 Hz (P = 0.04). The novel object test was unaffected by flicker treatment. In conclusion, many behaviors and the stress and fear responses were unaffected by either visible or non-visible flicker. However, visible flicker (30 Hz) reduced some comfort and exploratory behaviors early in life, and the impact on preening continued to older ages, suggesting minor negative impacts of flicker, particularly early in life.

Membrane-bound O-acyltransferase 7 (MBOAT7) shapes lysosomal lipid homeostasis and function to control alcohol-associated liver injury.

Recent genome-wide association studies (GWAS) have identified a link between single-nucleotide polymorphisms (SNPs) near the MBOAT7 gene and advanced liver diseases. Specifically, the common MBOAT7 variant (rs641738) associated with reduced MBOAT7 expression is implicated in non-alcoholic fatty liver disease (NAFLD), alcohol-associated liver disease (ALD), and liver fibrosis. However, the precise mechanism underlying MBOAT7-driven liver disease progression remains elusive. Previously, we identified MBOAT7-driven acylation of lysophosphatidylinositol lipids as key mechanism suppressing the progression of NAFLD (Gwag et al., 2019). Here, we show that MBOAT7 loss of function promotes ALD via reorganization of lysosomal lipid homeostasis. Circulating levels of MBOAT7 metabolic products are significantly reduced in heavy drinkers compared to healthy controls. Hepatocyte- (Mboat7-HSKO), but not myeloid-specific (Mboat7-MSKO), deletion of Mboat7 exacerbates ethanol-induced liver injury. Lipidomic profiling reveals a reorganization of the hepatic lipidome in Mboat7-HSKO mice, characterized by increased endosomal/lysosomal lipids. Ethanol-exposed Mboat7-HSKO mice exhibit dysregulated autophagic flux and lysosomal biogenesis, associated with impaired transcription factor EB-mediated lysosomal biogenesis and autophagosome accumulation. This study provides mechanistic insights into how MBOAT7 influences ALD progression through dysregulation of lysosomal biogenesis and autophagic flux, highlighting hepatocyte-specific MBOAT7 loss as a key driver of ethanol-induced liver injury.

Polycomb protein binding and looping in the ON transcriptional state.

Polycomb group (PcG) proteins mediate epigenetic silencing of important developmental genes by modifying histones and compacting chromatin through two major protein complexes, PRC1 and PRC2. These complexes are recruited to DNA by CpG islands (CGIs) in mammals and Polycomb response elements (PREs) in Drosophila. When PcG target genes are turned OFF, PcG proteins bind to PREs or CGIs, and PREs serve as anchors that loop together and stabilize gene silencing. Here, we address which PcG proteins bind to PREs and whether PREs mediate looping when their targets are in the ON transcriptional state. While the binding of most PcG proteins decreases at PREs in the ON state, one PRC1 component, Ph, remains bound. Further, PREs can loop to each other and with presumptive enhancers in the ON state and, like CGIs, may act as tethering elements between promoters and enhancers. Overall, our data suggest that PREs are important looping elements for developmental loci in both the ON and OFF states.

TLR2 regulates hair follicle cycle and regeneration via BMP signaling.

The etiology of hair loss remains enigmatic, and current remedies remain inadequate. Transcriptome analysis of aging hair follicles uncovered changes in immune pathways, including Toll-like receptors (TLRs). Our findings demonstrate that the maintenance of hair follicle homeostasis and the regeneration capacity after damage depend on TLR2 in hair follicle stem cells (HFSCs). In healthy hair follicles, TLR2 is expressed in a cycle-dependent manner and governs HFSCs activation by countering inhibitory BMP signaling. Hair follicles in aging and obesity exhibit a decrease in both TLR2 and its endogenous ligand carboxyethylpyrrole (CEP), a metabolite of polyunsaturated fatty acids. Administration of CEP stimulates hair regeneration through a TLR2-dependent mechanism. These results establish a novel connection between TLR2-mediated innate immunity and HFSC activation, which is pivotal to hair follicle health and the prevention of hair loss and provide new avenues for therapeutic intervention.

Preventing fibrosis in IBD: update on immune pathways and clinical strategies.

INTRODUCTION: Intestinal fibrosis is a common and serious complication of inflammatory bowel diseases (IBD) driving stricture formation in Crohn's disease patients and leading to submucosal damage in ulcerative colitis. Recent studies provided novel insights into the role of immune and nonimmune components in the pathogenesis of intestinal fibrosis. Those new findings may accelerate the development of anti-fibrotic treatment in IBD patients. AREAS COVERED: This review is designed to cover the recent progress in mechanistic research and therapeutic developments on intestinal fibrosis in IBD patients, including new cell clusters, cytokines, proteins, microbiota, creeping fat, and anti-fibrotic therapies. EXPERT OPINION: Due to the previously existing major obstacle of missing consensus on stricture definitions and the absence of clinical trial endpoints, testing of drugs with an anti-fibrotic mechanism is just starting in stricturing Crohn's disease (CD). A biomarker to stratify CD patients at diagnosis without any complications into at-risk populations for future strictures would be highly desirable. Further investigations are needed to identify novel mechanisms of fibrogenesis in the intestine that are targetable and ideally gut specific.

Bexarotene leads to durable improvements in visual evoked potential latency: A follow-up study of the Cambridge Centre for Myelin Repair One trial.

The Cambridge Centre for Myelin Repair One (CCMR-One) trial showed that 6 months of bexarotene reduces visual evoked potential (VEP) latency in people with relapsing-remitting multiple sclerosis (MS). In a single-centre follow-up study of these participants, we re-examined full-field VEP and clinical assessments. Twenty participants (12 bexarotene and 8 placebo) were seen on average 27 months after their trial involvement. In an analysis of all eyes with recordable signal (24 bexarotene and 14 placebo), the adjusted bexarotene-placebo treatment difference in P100 latency was -7.79 (95% confidence interval (CI) = -14.76, -0.82) ms, p = 0.044. We conclude that there were durable improvements in VEP latency, suggesting long-term benefits from exposure to a remyelinating drug.

Marital Quality and Depressive Symptoms Among Older Hispanic Adults in the United States.

OBJECTIVES: Despite the cultural importance of marriage as a social support system and its well-established link to mental health, older Hispanic adult populations, which are the largest racial and ethnic minoritized groups, remain understudied. The current study examined how positive and negative dimensions of marital quality are associated with depressive symptoms. METHODS: Data from Hispanic adults aged 51 years and older (n = 1,012) were obtained from the 2016 and 2018 Health and Retirement Study waves. The Center for Epidemiological Studies-Depression scale (0-8 symptoms) was modeled as a function of positive and negative marital quality measures (1-4), as well as the relevant covariates. RESULTS: Results from a negative binomial regression model showed that a 1-unit change in positive and negative marital quality was associated with a 23.61% reduction and a 23.74% increase, respectively, in depressive symptoms. The interaction terms with marital quality and gender, as well as marital quality and religion, were not statistically significant. DISCUSSION: In the United States, a large percentage of older Hispanic adults are immigrants, and their extended family tends to reside in their countries of origin. As such, older Hispanic adults may have smaller social networks, and marital quality most likely represents a culturally important social support network in later life. Significant associations between depressive symptoms and marital quality among older Hispanic adults should receive more attention in family and public health policy discussions, particularly given the increasing diversity in U.S. society.

Visual outcome measures in clinical trials of remyelinating drugs.

One of the most promising approaches to delay, prevent or reverse disability progression in multiple sclerosis (MS) is to enhance endogenous remyelination and limit axonal degeneration. In clinical trials of remyelinating drugs, there is a need for reliable, sensitive and clinically relevant outcome measures. The visual pathway, which is frequently affected by MS, provides a unique model system to evaluate remyelination of acute and chronic MS lesions in vivo and non-invasively. In this review, we discuss the different measures that have been used and scrutinise visual outcome measure selection in current and future remyelination trials.

Vivaria housing conditions expose sex differences in brain oxidation, microglial activation, and immune system states in aged hAPOE4 mice.

Apolipoprotein E ε4 allele (APOE4) is the predominant genetic risk factor for late-onset Alzheimer's disease (AD). APOE4 mouse models have provided advances in the understanding of disease pathogenesis, but unaccounted variables like rodent housing status may hinder translational outcomes. Non-sterile aspects like food and bedding can be major sources of changes in rodent microflora. Alterations in intestinal microbial ecology can cause mucosal barrier impairment and increase pro-inflammatory signals. The present study examined the role of sterile and non-sterile food and housing on redox indicators and the immune status of humanized-APOE4 knock-in mice (hAPOe4). hAPOE4 mice were housed under sterile conditions until 22 months of age, followed by the transfer of a cohort of mice to non-sterile housing for 2 months. At 24 months of age, the redox/immunologic status was evaluated by flow cytometry/ELISA. hAPOE4 females housed under non-sterile conditions exhibited: (1) higher neuronal and microglial oxygen radical production and (2) lower CD68+ microglia (brain) and CD8+ T cells (periphery) compared to sterile-housed mice. In contrast, hAPOE4 males in non-sterile housing exhibited: (1) higher MHCII+ microglia and CD11b+CD4+ T cells (brain) and (2) higher CD11b+CD4+ T cells and levels of lipopolysaccharide-binding protein and inflammatory cytokines in the periphery relative to sterile-housed mice. This study demonstrated that sterile vs. non-sterile housing conditions are associated with the activation of redox and immune responses in the brain and periphery in a sex-dependent manner. Therefore, housing status may contribute to variable outcomes in both the brain and periphery.

Do flickering LED lights reduce productivity of layer pullets and hens?

Most characteristics of artificial light sources are well studied, however light-flicker frequency (F) has been overlooked. The purpose of this study was to determine the effect of F on performance of Lohmann LSL-Lite (LW) pullets and Lohmann Brown-Lite (LB) pullets. In addition, pullets were followed through to the laying phase to evaluate long-term effects of F during rearing on productivity. Two trials were conducted with 3 F (30, 90, or 250 Hz) treatments. LW and LB pullets (n = 2,688 per strain [S]) were randomly assigned to floor pens within 8 light-tight rooms (15 pen replicates per F × S for 30 and 250 Hz; 18 pen replicates per F × S for 90 Hz). At 16 wk, pullets were transferred to conventional layer cages, with no flicker treatment applied. Pullet data collected included BW, feed disappearance, flock uniformity, and overall mortality. Hen data collected included BW, feed intake (feed efficiency calculated), mortality, egg production, and egg quality. Data were analyzed using Proc Mixed (SAS 9.4) and differences were considered significant when P ≤ 0.05. Frequency did not affect pullet uniformity or feed disappearance (0-8 wk and 0-16 wk). Pullets reared under 30 Hz had higher mortality (caused by "other") than those reared under 250 Hz. Lohmann Brown-Lite pullets reared under 30 Hz had the highest feed disappearance. Overall mortality was higher for LW pullets reared under 30 Hz compared to LB reared under 30 Hz or 250 Hz. Lohmann Brown-Lite hens reared under 30 Hz were heavier at the beginning of the hen phase (17 wk), however differences related to F were not seen at 40 or 48 wk. Hen day production (%) was higher for hens reared under 30 compared to 90 Hz (P = 0.03), however no other egg parameters were affected by F. Hen feed efficiency and mortality were unaffected by F. These results indicate minor effects of F, during either the pullet or hen phases. The data also suggest that S (LW vs. LB) may affect response to F.

The relative efficacy of multiple syringe tip disinfection techniques against virulent staphylococcus contamination.

BACKGROUND: A recent study confirmed significant contamination of syringe tips following routine anaesthesia practice of at least 6 h in duration. AIM: We assessed the relative efficacy of clinically relevant syringe tip disinfection techniques following contamination with the hyper transmissible and more pathogenic Staphylococcus aureus sequence type 5 (S. aureus ST5) strain characteristic associated with increased strength of biofilm formation and greater desiccation tolerance. METHODS: Syringe tips (N=40) contaminated with S. aureus ST5 were randomized to 70% isopropyl pads with 10 or 60 s of drying time, scrubbing alcohol disinfection caps with 10 or 60 s of dwell time, or to non-scrubbing alcohol disinfection caps with 60 s of dwell time. The primary outcome was residual 24-h colony forming units (cfu) >10. RESULTS: Scrubbing disinfection caps were more effective than alcohol pads (25% (12/48) <10 cfu for scrubbing caps (10- or 60-s dwell times) vs 0% (0/48) <10 cfu for alcohol pads (10 or 60 s of drying time), Holm-Sidak adjusted P=0.0016). Scrubbing disinfection caps were more effective than non-scrubbing alcohol disinfection caps (25% (12/48) <10 cfu for scrubbing alcohol caps (10- or 60-s dwell times) vs 2% (1/48) for non-scrubbing alcohol caps (60-s dwell time), adjusted P=0.0087). CONCLUSIONS: Scrubbing alcohol caps are more effective than alcohol pads or non-scrubbing disinfecting caps for microbial reduction of syringe tips contaminated with the more pathogenic S. aureus ST5.

Lymphatic endothelial-like cells promote glioblastoma stem cell growth through cytokine-driven cholesterol metabolism.

Glioblastoma is the most lethal primary brain tumor with glioblastoma stem cells (GSCs) atop a cellular hierarchy. GSCs often reside in a perivascular niche, where they receive maintenance cues from endothelial cells, but the role of heterogeneous endothelial cell populations remains unresolved. Here, we show that lymphatic endothelial-like cells (LECs), while previously unrecognized in brain parenchyma, are present in glioblastomas and promote growth of CCR7-positive GSCs through CCL21 secretion. Disruption of CCL21-CCR7 paracrine communication between LECs and GSCs inhibited GSC proliferation and growth. LEC-derived CCL21 induced KAT5-mediated acetylation of HMGCS1 on K273 in GSCs to enhance HMGCS1 protein stability. HMGCS1 promoted cholesterol synthesis in GSCs, favorable for tumor growth. Expression of the CCL21-CCR7 axis correlated with KAT5 expression and HMGCS1K273 acetylation in glioblastoma specimens, informing patient outcome. Collectively, glioblastomas contain previously unrecognized LECs that promote the molecular crosstalk between endothelial and tumor cells, offering potentially alternative therapeutic strategies.

Determination of the absolute intensity of the 1205 keV γ-ray emission from 91Y.

A radiochemically pure solution of 91Y was produced by the thermal neutron fission of 235U followed by successive chemical separations to remove fission product impurities. The gamma emission rate of the 91Y 1205 keV gamma was measured using multiple high purity germanium gamma spectrometers previously calibrated for counting efficiency using a certificated mixed nuclide gamma standard. The activity concentration of the 91Y was subsequently standardised by liquid scintillation counting. From the combination the activity concentration and gamma emission intensity, the absolute intensity of the 1205 keV gamma emission was derived as 0.2297(39)%. This data agrees within the quoted uncertainties with the absolute intensity of 0.26(4)% published in nuclear data sheets A=91 (Baglin, 2013), but reduces the uncertainty by an order of magnitude.

Quantifying the Representation of Black Adolescents in Suicide Intervention Research.

This systematic review aimed to quantify the representation of Black youth in U.S. suicide intervention research. Specifically, we sought to evaluate Black youth representation in terms of (a) equity of inclusion (i.e., the inclusion of Black youth in research study samples at a rate consistent with the overall national rate of Black adolescents in the US) and (b) equity of intervention efficacy (i.e., evaluating the presence of racial disparities in intervention efficacy/effect sizes). In addition, we aimed to evaluate whether an association existed between funding status of research and representation of Black youth in studies, and to provide recommendations for future research in this area. To this end, the present study extracted and analyzed demographic information of studies included in recent meta-analyses conducted by Robinson and colleagues (2018), which were not previously analyzed, in addition to new literature published between September 2017 and January 2021. Results showed that the prevalence of Black youth included in studies was representative (14.67%; ntotal = 4451, nBlack = 664), with a median inclusion rate of 13%; however, absolute sample and group sizes were so small that it precluded comparison of differential treatment outcomes for Black youth. Thus, out of 22 studies identified, only one was able to investigate treatment outcomes for suicide in Black youth specifically. This study points to the conclusion that without adequately powered studies, disparities in treatment efficacy for Black youth cannot be compared or addressed, and the existing disparity in suicidal outcomes for Black youth will grow even larger.