Specific isolation and quantification of PD-L1 positive tumor derived exosomes for accurate breast cancer discrimination via aptamer-functionalized magnetic composites and SERS immunoassay.

PD-L1 positive tumor derived exosomes (TEXsPD-L1) play a significant role in disease progression, tumor metastasis and cancer immunotherapy. However, the overlap of PD-L1 between TEXs and non-tumor derived exosomes (non-TEXs) restricts the specific isolation and quantification of TEXPD-L1 from clinical samples. Herein, a new aptamer-functionalized and hydrophilic immunomagnetic substrate was designed by decorating generation 5 polyamidoamine dendrimers (G5 PAMAM), zwitterionic trimethylamine N-oxide (TMAO) and EpCAM (Epithelial cell adhesion molecule) aptamers on magnetic cores sequentially (Fe3O4@PAMAM@TMAO@Aptamer, named as FPTA) for rapid target and efficient capture of TEXs. The FPTA substrate gathered excellent characters of strong magnetic responsiveness of Fe3O4, abundant affinity sites of PAMAM, strong hydrophilicity of TMAO and enhanced affinity properties of EpCAM aptamers. Because of these advantages, FPTA can isolate TEXs quickly within 30min with high capture efficiency of 90.5 % ± 3.0 % and low nonspecific absorption of 8.2 % ± 2.0 % for non-TEXs. Furthermore, PD-L1 (Programmed cell death-ligand 1) positive TEXs (TEXsPD-L1) from the captured TEXs were recognized and quantitatively analyzed by utilizing SERS (surface-enhanced Raman spectroscopy) reporter molecules 4-NTP (4-Nitrothiophenol) on PD-L1 aptamers-functionalized gold immunoaffinity probe. The signal of TEXsPD-L1 was converted to SERS signal of 4-NTP at 1344 cm-1 which exhibited a linear correlation to concentration of TEXsPD-L1(R2 = 0.9905). With these merits, this strategy was further applied to clinical plasma samples from breast cancer (BC) patients and healthy controls (HC), exhibited an excellent diagnosis accuracy with area under curve (AUC) of receiver operating characteristic (ROC) curve reaching 0.988. All these results demonstrate that the FPTA immunomagnetic substrate combined with SERS immunoaffinity probe may become a generic tool for specific isolation and quantitative analysis of PD-L1 positive tumor-derived exosomes in clinics.

PCR GeneScan Analysis of Rearranged Immunoglobulin or T-Cell Receptor Genes for Clonality Diagnostics in Suspect Lymphoproliferations.

Assessment of the presence of clonal lymphoproliferations via polymerase chain reaction (PCR)-based analysis of rearranged immunoglobulin (IG) or T-cell receptor (TR) genes is a valuable method in the diagnosis of suspect lymphoproliferative disorders. Additionally, this methodology can be used for evaluating dissemination of lymphoma cells and for studying the clonal relationship between multiple (different locations) and consecutive (over time) lymphomas. Here we describe an integrated approach to assess clonality via analysis of Ig heavy chain (IGH), Ig kappa (IGK), TCR beta (TRB), and TCR gamma (TRG) gene rearrangements, based on the standardized multiplex PCRs as originally developed by the European BIOMED-2 consortium (currently named EuroClonality). The described protocol covers the pre-analytical phase of DNA isolation (from formalin-fixed paraffin-embedded and fresh tissues, body fluids, peripheral blood, and bone marrow), the analytical phase of PCR GeneScan analysis, and the post-analytical interpretation of the obtained profiles, following established guidelines.

Effects of combined postweaning social isolation and ketamine administration on schizophrenia-like behaviour in male Sprague Dawley rats.

The pathophysiology behind negative and cognitive symptoms of schizophrenia is not well understood, thus limiting the effectiveness of treatment on these symptoms. Developing reliable animal model of schizophrenia is vital to advance our understanding on the neurobiological basis of the disorder. Double hit is used to refer to the use of two schizophrenia inducing interventions viz ketamine exposure and social isolation. In this study we aim to investigate the robustness of double hit model of schizophrenia in inducing negative and cognitive symptoms of schizophrenia. On postnatal day (PND) 23, thirty-two male Sprague Dawley rats were randomly grouped into four equal groups as follows: group housed + saline (GH), group housed + ketamine (GHK), isolated + saline (SI), and isolated + ketamine (SIK). A single ketamine dose (16 mg/kg) was administered 3 times a week for four weeks. Isolated animals were housed singly throughout the study. The following behavioural tests were carried out: elevated plus maze, three chamber social interaction, resident intruder tests, and novel object recognition (NOR). The SIK group exhibited high anxiety levels, with increased ACTH, corticosterone and norepinephrine concentration when compared to the other groups. The SIK animals also presented with reduced social interaction and decreased oxytocin concentration. SIK rats were more aggressive towards a juvenile intruder but had low testosterone concentration. The SIK group or double hit model showed impaired visual learning and memory and increased expression of proinflammatory cytokines. This suggest that the double hit model is more robust in inducing negative and cognitive symptoms of schizophrenia than each treatment alone.

Management and associated outcomes of COVID-19 infection among Ghanaian autoimmune rheumatic disease patients.

Objective: This study assessed the prevalence of infection, management strategies and associated disease outcomes of COVID-19 among Autoimmune Rheumatic Disease (AIRD) patients in a teaching hospital in Ghana. Design: This was a retrospective cross-sectional study. Setting: Rheumatology Unit, Korle Bu Teaching Hospital. Participants: Autoimmune Rheumatic Disease patients. Results: Thirty-one (31) out of approximately 1700 AIRD patients in the unit tested positive for COVID-19, registering a COVID-19 prevalence of 1.82%. The majority, 25(80.6%), were females with a mean ± SD age of 41.7 ± 12.8 years. Systemic lupus erythematosus was the most affected autoimmune rheumatic condition, reporting fever as the commonest COVID-19-related symptom. Most participants, 22(71%), were managed by the "self-isolation"/home management" strategy. In comparison, 7(22.5%) were monitored at the hospital, with both strategies having resulted in complete recovery. The remaining 2(6.5%) patients who managed under "intensive care unit" strategy resulted in mortality. Conclusion: These findings highlight the relatively low frequency of COVID-19 infection among AIRD patients, the encouraging recovery, and the low severe disease rates observed within this cohort. Additionally, the outcome of self-isolation and home management strategies underscore the importance of personalised approaches to COVID-19 management in this population. Funding: None.

First Isolation and Genetic Characterization of Avian Nephritis Virus 4 from Commercial Poultry in India.

Avian nephritis virus (ANV), which belongs to the family Astroviridae, is associated with different clinical manifestations (enteric and kidney disorders) in poultry. Despite being a significant pathogen of the avian industry worldwide, information regarding genetic features of these viruses in India is scarce. In this study, 386 intestinal samples collected from 37 slaughterhouses in two north Indian states (Rajasthan and Haryana) were screened for ANV with reverse transcription PCR (RT-PCR) targeting the conserved ORF1b gene, followed by nucleotide sequencing of the amplified product. RT-PCR and sequencing confirmed the presence of ANV in 32 clinical samples (8.29%), with concurrent infections of infectious bronchitis virus, chicken astrovirus, and fowl adenoviruses observed in some clinical samples (n = 4). Virus isolations were successful from four out of 12 ANV-positive clinical samples passaged via the yolk-sac route in specific-pathogen-free embryonated chicken eggs. Additionally, the near-complete genomes of two viruses were determined through sequencing. Phylogenetic analysis based on full-length capsid protein sequences classified the viruses into ANV genotype 4 (ANV4), and to the best of our knowledge this is the first report of ANV4 from India. This study revealed the presence and circulation of new strains of ANV in Indian poultry. Genetic profiling and isolation of the viruses in this study will not only aid in the development of diagnostic tools and vaccines for ANV but also offer valuable insights into its epidemiology.

Primer aislamiento y caracterización genética del virus de la nefritis aviar 4 en la avicultura comercial de la India El virus de la nefritis aviar (ANV), que pertenece a la familia Astroviridae, se asocia con diferentes manifestaciones clínicas (trastornos entéricos y renales) en la avicultura. A pesar de ser un patógeno importante de la industria avícola en todo el mundo, la información sobre las características genéticas de estos virus en la India es escasa. En este estudio, se analizaron 386 muestras intestinales recolectadas en 37 plantas de procesamiento en dos estados del norte de la India (Rajasthan y Haryana) para detectar al virus de la nefritis aviar mediante un método de RT-PCR dirigido al gene conservado ORF1b, seguido de la secuenciación de nucleótidos del producto amplificado. El método de RT-PCR y la secuenciación confirmaron la presencia del virus de la nefritis aviar en 32 muestras clínicas (8.29%), observándose infecciones concurrentes por el virus de la bronquitis infecciosa, astrovirus del pollo y adenovirus de las aves en algunas muestras clínicas (n = 4). Los aislamientos del virus tuvieron éxito en cuatro de las 12 muestras clínicas positivas para el virus de la nefritis aviar inoculadas a través de la ruta del saco vitelino en huevos de gallina embrionados libres de patógenos específicos. Además, se determinaron los genomas casi completos de dos virus mediante secuenciación. El análisis filogenético basado en secuencias completas de proteínas de la cápside clasificó los virus en el genotipo 4 del virus de la nefritis aviar (ANV4) y hasta donde se sabe, este es el primer informe del virus de la nefritis aviar 4 en la India. Este estudio reveló la presencia y circulación de nuevas cepas del virus de la nefritis aviar en la avicultura comercial de la India. El perfil genético y el aislamiento de los virus en este estudio no solo ayudarán en el desarrollo de herramientas de diagnóstico y vacunas para el virus de la nefritis aviar, sino que también ofrecerán información valiosa sobre su epidemiología.

Effects of respiratory isolation for tuberculosis to reduce community-based transmission: a systematic review.

BACKGROUND: Respiratory isolation of people with pulmonary tuberculosis (TB), including after treatment initiation, is used to prevent community-based transmission; yet guidelines on duration are limited and implementation is heterogeneous. This systematic review synthesized evidence on respiratory isolation for TB to inform National TB Coalition of America guidelines. METHODS: After searching six databases, eight reviewers screened and extracted data in duplicate on effects of respiratory isolation compared to no isolation or masking. Studies were stratified by outcomes: TB infection or disease in contacts, mortality, hospitalization duration, patient and health system costs, and impact on mental health or stigma. We used a convergent integrated approach to synthesize quantitative and qualitative findings and assess limitations. RESULTS: Seventeen studies were included. There were limited data directly comparing isolation to non-isolation interventions, including effects after treatment initiation. One randomized controlled trial suggested treatment in a sanatorium versus at home did not affect TB incidence in contacts. Modelling studies suggest isolation may reduce transmission, but relied on various assumptions, and isolation was implemented alongside other interventions. Descriptive, mixed-methods, and qualitative studies described adverse impacts of isolation on employment, education, food/housing security, and mental health due to transmission fears, stigma and social isolation. Impacts were compounded in marginalized groups including indigenous and incarcerated persons. CONCLUSION: Data to support current isolation practices, particularly after effective treatment initiation, to reduce TB transmission in communities are limited. Public health guidance should weigh the negative impacts on people with TB against decreased community transmission to make evidence-based decisions about respiratory isolation.

"What Does 'Age-Friendly' Mean to You?" The Role of Microaggressions in a Retirement and Assisted Living Community.

BACKGROUND AND OBJECTIVES: In conversations about expanding age-friendly ecosystems, the concept of "age-friendliness" has not been explored in relation to residential settings. RESEARCH DESIGN AND METHODS: This multiple case study compared four residents' perspectives on the age-friendliness of a retirement and assisted living community, combining individual semi-structured interviews with observational data and organizational document analyses in a contextualist thematic examination. RESULTS: Three themes depict (A) existing experiences of the setting as "age-friendly" and the tension of the built design vs. identity; (B) resident-to-resident microaggressions, delineated into four sub-themes including identity-related, intergenerational, condition-related microaggressions and their influence on social isolation; and (C) desired experiences of the setting as "age-friendly" reflecting the social design. In cases of visibly perceptible diversity (white cane, darker skin tone), residents fared worse in experiencing microaggressions stemming from ableism, racism, and age differences. Conversely, in cases of visibly imperceptible diversity, residents had more positive or entirely positive experiences. Although the setting met many environmental and healthcare needs, it lacked design factors prioritizing meaningful social relationships between residents, impacting social isolation. DISCUSSION AND IMPLICATIONS: Resident-to-resident social relationships are key in the experience of a retirement and assisted living community as age-friendly. Resident-to-resident microaggressions undermine perceptions of the community as age-friendly, and influence social isolation. We reflect on the organizational role in mitigating against negative social relationships and social isolation to maximize dignity.

Spatially Controlled DNA Frameworks for Sensitive Detection and Specific Isolation of Tumor Cells.

High-affinity, specific, and sensitive probes are crucial for the specific recognition and identification of tumor cells from complex matrices. Multivalent binding is a powerful strategy, but the irrational spatial distribution of the functional moieties may reduce the probe performance. Here, we constructed a Janus DNA triangular prism nanostructure (3Zy1-JTP-3) for sensitive detection and specific isolation of tumor cells. Benefiting from spatial features of the triangular prism, the fluorescence intensity induced by 3Zy1-JTP-3 was almost 4 times that of the monovalent structure. Moreover, the DNA triangular prisms were connected to form hand-in-hand multivalent DNA triangular prism structures (Zy1-MTP), in which the fluorescence intensity and affinity were increased to 9-fold and 10-fold of 3Zy1-JTP-3, respectively. Furthermore, 3Zy1-JTP-3 and Zy1-MTP were combined with magnetic beads, and the latter showed higher capture efficiency (> 90%) in whole blood. This work provides a new strategy for the efficient capture of rare cells in complex biological samples.

Validation of ablation site classification accuracy and trends in the prediction of potential reconnection sites for atrial fibrillation using the CARTONET® R12.1 model.

Background: CARTONET® enables automatic ablation site classification and reconnection site prediction using machine learning. However, the accuracy of the site classification model and trends of the site prediction model for potential reconnection sites are uncertain. Methods: We studied a total of 396 cases. About 313 patients underwent pulmonary vein isolation (PVI), including a cavotricuspid isthmus (CTI) ablation (PVI group) and 83 underwent PVI and additional ablation (i.e., box isolation) (PVI+ group). We investigated the sensitivity and positive predictive value (PPV) for automatic site classification in the total cohort and compared these metrics for PV lesions versus non-PV lesions. The distribution of potential reconnection sites and confidence level for each site was also investigated. Results: A total of 29,422 points were analyzed (PV lesions [n = 22 418], non-PV lesions [n = 7004]). The sensitivity and PPV of the total cohort were 71.4% and 84.6%, respectively. The sensitivity and PPV of PV lesions were significantly higher than those of non-PV lesions (PV lesions vs. non-PV lesions, %; sensitivity, 75.3 vs. 67.5, p < .05; PPV, 91.2 vs. 67.9, p < .05). CTI and superior vena cava could not be recognized or analyzed. In the potential reconnection prediction model, the incidence of potential reconnections was highest in the posterior, while the confidence was the highest in the roof. Conclusion: The automatic site classification of the CARTONET®R12.1 model demonstrates relatively high accuracy in pulmonary veins excluding the carina. The prediction of potential reconnection sites feature tends to anticipate areas with poor catheter stability as reconnection sites.

A modified dual preparatory method for improved isolation of nucleic acids from laser microdissected fresh-frozen human cancer tissue specimens.

A central theme in cancer research is to increase our understanding of the cancer tissue microenvironment, which is comprised of a complex and spatially heterogeneous ecosystem of malignant and non-malignant cells, both of which actively contribute to an intervening extracellular matrix. Laser microdissection (LMD) enables histology selective harvest of cellular subpopulations from the tissue microenvironment for their independent molecular investigation, such as by high-throughput DNA and RNA sequencing. Although enabling, LMD often requires a labor-intensive investment to harvest enough cells to achieve the necessary DNA and/or RNA input requirements for conventional next-generation sequencing workflows. To increase efficiencies, we sought to use a commonplace dual preparatory (DP) procedure to isolate DNA and RNA from the same LMD harvested tissue samples. While the yield of DNA from the DP protocol was satisfactory, the RNA yield from the LMD harvested tissue samples was significantly poorer compared to a dedicated RNA preparation procedure. We determined that this low yield of RNA was due to incomplete partitioning of RNA in this widely used DP protocol. Here, we describe a modified DP protocol that more equally partitions nucleic acids and results in significantly improved RNA yields from LMD-harvested cells.

Exploring the association between multidimensional social isolation and heterogeneous cognitive trajectories among older adults: evidence from China.

Objective: This study aims to elucidate the heterogeneous cognitive trajectories among older adults in China through a comprehensive, nationally representative longitudinal study. Furthermore, it seeks to investigate the impact of multidimensional social isolation on heterogeneous cognitive trajectories among older adults in China. Methods: Utilizing data from three successive waves of the Chinese Longitudinal Aging Social Survey (CLASS) spanning 2016 to 2020, this investigation quantified baseline social isolation across three dimensions-family isolation, friend isolation, and subjective isolation-alongside cognitive function scores of older adults, measured across all three waves. Through latent class growth models, heterogeneous cognitive trajectories were delineated. The influence of family isolation, friend isolation, and subjective isolation on these cognitive trajectories was examined employing multinomial logistic regression analysis. Results: The study included 6,378 participants aged 60 and above, revealing three primary cognitive trajectories: High baseline stable group (68.8%), High baseline but declining group (21.7%), and Low baseline deteriorating group (9.5%). Adjusting for variables such as personal physical characteristics, social networks, living and working conditions, and the surrounding policy environment, the findings indicated that family isolation did not significantly affect cognitive function's high-level decline or low-level deterioration. Conversely, friend isolation markedly increased the risk of high-level cognitive decline (OR = 1.289) and low-level cognitive deterioration (OR = 1.592). Similarly, subjective isolation significantly heightened the risk for both high-level decline (OR = 1.254) and low-level deterioration (OR = 1.29) in cognitive function. Conclusion: Mitigating friend and subjective isolation among older adults appears to be a more effective strategy in preventing or delaying cognitive decline, potentially reducing the strain on healthcare and social welfare systems.

A Protocol for the Isolation of Oval Cells without Preconditioning.

Oval cells (OCs) is the name of hepatic progenitor cells (HPCs) in rodents. They are a small population of cells in the liver with the remarkable ability to proliferate and regenerate hepatocytes and cholangiocytes in response to acute liver damage. Isolating OCs generally requires a pretreatment with special diets, chemicals, and/or surgery to induce hepatic damage and OC proliferation in mice. Unfortunately, these pretreatments are not only painful for the mice but also increase the cost of the assays, and the effects on the different organs as well as on various liver cells are still unclear. Therefore, the search for a protocol to obtain OCs without prior liver damage is mandatory. In our study, we present a protocol to isolate murine OCs from healthy liver (HL-OCs) and compare them with OCs isolated from mice pretreated with 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC-OCs). Our results demonstrated that cells derived from untreated mice exhibited similar behavior to those from treated mice in terms of surface marker expression, proliferation, and differentiation capacity. Therefore, given the impracticability of isolating human cells with prior hepatotoxic treatment, our model holds promise for enabling the isolation of progenitor cells from human tissue in the future. This advancement could prove invaluable for translational medicine in the understanding and treatment of liver diseases.

Microfluidic Affinity Selection of B-Lineage Cells from Peripheral Blood for Minimal Residual Disease Monitoring in Pediatric B-Type Acute Lymphoblastic Leukemia Patients.

Assessment of minimal residual disease (MRD) is the most powerful predictor of outcome in B-type acute lymphoblastic leukemia (B-ALL). MRD, defined as the presence of leukemic cells in the blood or bone marrow, is used for the evaluation of therapy efficacy. We report on a microfluidic-based MRD (MF-MRD) assay that allows for frequent evaluation of blood for the presence of circulating leukemia cells (CLCs). The microfluidic chip affinity selects B-lineage cells, including CLCs using anti-CD19 antibodies poised on the wall of the microfluidic chip. Affinity-selected cells are released from the capture surface and can be subjected to immunophenotyping to enumerate the CLCs, perform fluorescence in situ hybridization (FISH), and/or molecular analysis of the CLCs' mRNA/gDNA. During longitudinal testing of 20 patients throughout induction and consolidation therapy, the MF-MRD performed 116 tests, while only 41 were completed with multiparameter flow cytometry (MFC-MRD) using a bone marrow aspirate, as standard-of-care. Overall, 57% MF-MRD tests were MRD(+) as defined by CLC numbers exceeding a threshold of 5 × 10-4%, which was determined to be the limit of quantitation. Above a threshold of 0.01%, MFC-MRD was positive in 34% of patients. The MF offered the advantage of the opportunity for efficiently processing small volumes of blood (2 mL), which is important in the care of pediatric patients, especially infants. The minimally invasive means of blood collection are of high value when treating patients whose MRD is typically tested using an invasive bone marrow biopsy. MF-MRD detection can be useful for stratification of patients into risk groups and monitoring of patient well-being after completion of treatment for early recognition of potential impending disease recurrence.

A Fresh Look at Islet Isolation from Rabbit Pancreases.

Islet transplantation represents a promising therapeutic approach for diabetes management, yet the isolation and evaluation of pancreatic islets remain challenging. This study focuses on the isolation of islets from rabbit pancreases, followed by a comprehensive assessment of their viability and functionality. We developed a novel method for isolating islet cells from the pancreas of adult rabbits. We successfully isolated viable islets, which were subsequently evaluated through a combination of viability assays, an insulin enzyme-linked immunosorbent assay (ELISA), and fluorescence lifetime imaging microscopy (FLIM). The viability assays indicated a high percentage of intact islets post-isolation, while the insulin ELISA demonstrated robust insulin secretion in response to glucose stimulation. FLIM provided insights into the metabolic state of the islets, revealing distinct fluorescence lifetime signatures correlating with functional viability. Our findings underscore the potential of rabbit islets as a model for studying islet biology and diabetes therapy, highlighting the efficacy of combining traditional assays with advanced imaging techniques for comprehensive functional assessments. This research contributes to the optimization of islet isolation protocols and enhances our understanding of islet functional activity dynamics in preclinical settings.

Damping Characteristics of a Novel Bellows Viscous Damper.

Micro-vibrations during the operation of space remote sensing equipment can significantly affect optical imaging quality. To address this issue, a bellows-type viscous damper serves as an effective passive damping and vibration isolation solution. This paper introduces a bellows-type viscous damper with adjustable damping capabilities, designed for mid- to high-frequency applications. We developed a system damping model based on hydraulic fluid dynamics to examine how different factors-such as viscous coefficients, damping hole lengths, hole diameters, chamber pressures, and volumes-influence the damping characteristics. To validate the theoretical model, we constructed an experimental platform. The experimental results show that the theoretical damping curves closely match the measured data. Moreover, increasing the chamber pressure effectively enhances the damper's damping coefficient, with the deviation from theoretical predictions being approximately 4%.

Advancements in Optical Resonator Stability: Principles, Technologies, and Applications.

This paper provides an overview of the study of optical resonant cavity stability, focusing on the relevant principles, key technological advances, and applications of optical resonant cavities in a variety of high-precision measurement techniques and modern science and technology. Firstly, the vibration characteristics, thermal noise, and temperature characteristics of the reference cavity are presented. Subsequently, the report extensively discusses the advances in key technologies such as mechanical vibration isolation, thermal noise control, and resistance to temperature fluctuations. These advances not only contribute to the development of theory but also provide innovative solutions for practical applications. Typical applications of optical cavities in areas such as laser gyroscopes, high-precision measurements, and gravitational wave detection are also discussed. Future research directions are envisioned, emphasising the importance of novel material applications, advanced vibration isolation technologies, intelligent temperature control systems, multifunctional integrated optical resonator design, and deepening theoretical models and numerical simulations.

Isolation and transcriptional profiling of antigen-presenting cells from mouse lung and MLNs following intranasal immunization.

Dendritic cells (DCs) play a central role in the initiation of the adaptive immune response. Here, we present a protocol for isolating and transcriptionally profiling antigen-presenting cells (APCs) from the mouse lung and mediastinal lymph nodes (MLNs) following intranasal immunization. We describe steps for preparing single-cell suspensions from the lung and MLN, along with the detection and RNA sequencing (RNA-seq) of antigen-presenting DCs. This protocol offers a broadly applicable approach for identifying variations in DC subpopulations under diverse experimental conditions. For complete details on the use and execution of this protocol, please refer to Youhui et al.1.

Protocol for mapping T cell activation using single-cell RNA-seq.

Stimulation of CD4 T cells with anti-CD3/CD28 is a commonly used model to study T cell activation. Here, we present a protocol for investigating T cell activation based on anti-CD3/CD28 bead stimulation and single-cell RNA sequencing (scRNA-seq). We describe the workflow from the isolation of human peripheral blood mononuclear cells (PMBCs) and CD4 T cell enrichment to anti-CD3/CD28 bead stimulation, scRNA-seq, and data analysis. For complete details on the use and execution of this protocol, please refer to Li et al.1.

Isolated during adolescence: long-term impact on social behavior, pain sensitivity, and the oxytocin system in male and female rats.

BACKGROUND: Adolescent social isolation (ASI) has profound long-term effects on behavioral and neural development. Despite this, the specific long-term impact of ASI during different adolescent stages and across sexes remain underexplored. METHODS: Our study addresses this gap by examining the effects of early- and late- adolescent social isolation on both male and female rats. Rats were either isolated (or group-housed) starting from PD 21 (early) or PD 42 (late) for three weeks and then rehoused into groups. In adulthood (PD 90), rats underwent a battery of tests: elevated plus-maze, open field, novel object recognition, social interaction and social recognition memory and hotplate tests. Finally, we analyzed oxytocin receptor binding in several regions in the brains of a second cohort of rats. RESULTS: Both, male and female rats from the late adolescent social isolation (LASI) groups spent significantly less time interacting in the social interaction test. Additionally, we observed a general decrease in social recognition memory regardless of sex. Both male ASI groups demonstrated heightened thermal pain sensitivity, while the opposite was observed in early adolescent social isolation (EASI) female rats. In the brain, we observed changes in oxytocin receptor (OTR) binding in the paraventricular nucleus of the hypothalamus (PVN) and paraventricular nucleus of the thalamus (PVT) and central amygdala (CeA) with the largest changes in EASI and LASI female rats. CONCLUSION: Our model demonstrates long-lasting alterations on behavior and oxytocin receptor binding levels following ASI providing insights into the long-term effects of ASI in a time- and sex-specific manner.

Social isolation, loneliness and low dietary micronutrient intake amongst older people in England.

BACKGROUND: Social isolation and loneliness are associated with increased risk of adverse health outcomes at older ages. This study evaluated whether isolation and loneliness are related to inadequate intake of micronutrients in the diet. METHODS: We tested associations between social isolation and loneliness and dietary micronutrient intake 2 years later in 3713 men and women (mean age 68.26, standard deviation 7.81 years) who completed two online 24-h dietary recalls. Associations of isolation and loneliness with intake of nine minerals and vitamins that fell below national recommendations were tested using logistic regressions, adjusting for age, gender, ethnicity, education, marital status, smoking and physical activity and total energy intake. RESULTS: The prevalence of low dietary intake varied markedly across micronutrients. Social isolation (1-point increase in a score ranging 0-5) was associated with increased odds (adjusted for covariates) of low intake of magnesium [odds ratio (OR) 1.153, 95% confidence interval (CI) 1.037-1.282, P = .009], potassium (OR 1.201, 95% CI 1.087-1.327, P < .001), vitamin B6 (OR 1.263, 95% CI 1.110-1.438, P < .001), folate (OR 1.211, 95% CI 1.093-1.341, P < .001) and vitamin C (OR 1.238, 95% CI 1.098-1.394, P < .001). These associations remained unchanged when food insecurity and impaired activities of daily living were taken into account. By contrast, loneliness was not related to the inadequate intake of any micronutrient. CONCLUSIONS: Low intake of micronutrients increases risk of age-related health problems. Attention to the dietary quality of older people with limited social contacts and little involvement in community activities might enhance health outcomes.