Activation Markers Research Articles

Exosomes activate hippocampal microglia in atrial fibrillation through long-distance heart–brain communication

BackgroundThere is growing evidence that atrial fibrillation (AF) is a risk factor for cognitive impairment (CI) and dementia in the presence or absence of stroke. The purpose of this study was to explore the mechanism of CI caused by AF.MethodsEighteen male canines were randomly divided into a sham group, a pacing group, and a pacing + GW4869 group. An experimental model of AF was established by rapid atrial pacing (450 beats/min) for 2 weeks, and the sham group received pacemaker implantation without atrial pacing. The GW4869 group received an intravenous GW4869 injection (0.3 mg/kg, once a day) during pacing. All canines were locally injected with Ad-CD63-RFP in epicardial adipose tissue (EAT) to trace the exosomes. Ultracentrifugation was employed to isolate EAT-derived exosomes, followed by RNA sequencing and quantitative real-time PCR (qRT-PCR) to assess RNA in both exosomes and hippocampal tissue. The miRanda database was used to predict the targeting relationships between miRNA and mRNA, which were further validated by luciferase reporter assays. Western blot analysis was conducted to detect exosomal markers (CD63, CD81, TSG101) in EAT exosomes, while immunofluorescence was used to detect Ad-CD63-RFP signals in both EAT and hippocampal tissues, as well as microglial activation marker IBA-1. To further explore the effects of exosomes on microglial cells, in vitro experiments using brain microvascular endothelial cells (bEnd3) and microglial cells (BV2) were conducted. IBA-1 expression and RNA levels in BV2 cells were analyzed by immunofluorescence and qRT-PCR, respectively.ResultsAfter 14 days of pacing of the canine atrium, compared to the sham group, both the pacing and GW4869 groups exhibited an increased number of AF inductions, along with prolonged AF duration. The fluorescence intensity of Ad-CD63-RFP and the microglial activation marker IBA-1 were markedly greater in the hippocampus. RNA sequencing showed that the differentially expressed gene cfa-miR-22e in EAT exosomes was upregulated, and its target gene IL33 was downregulated in the hippocampus. qRT-PCR showed that the levels of cfa-miR-22e were increased in both EAT exosomes and the hippocampus, while the expression of IL-33, a target of cfa-miR-22e, was decreased in the hippocampus. The administration of GW4869 abolished these effects. The in vitro results from bEnd3 and BV2 cell experiments were consistent with the conclusions drawn from the in vivo studies.ConclusionOur study indicated that the exosomes secreted by EAT in canines with AF can penetrate the BBB and activate microglia in the hippocampus through the cfa-miR-22e/IL33 signalling pathway.

Observational Analyses of Ex Vivo Native American Platelet Responses

Platelet activation plays an essential role in clot formation to prevent blood loss following vascular damage. In pathologic conditions, platelet activation can lead to obstructive clots, disrupting blood flow and resulting in thrombosis. Native Americans suffer disproportionately from arterial disease and previous research has shown that Blacks are enriched in genetic polymorphisms that correlate with higher platelet reactivity contributing to an increased risk for thrombosis. Therefore, the current study sought to determine phenotypic variations in Native American platelet responses following stimulation with agonists, simulating vascular damage. Several donors from a small cohort of Native Americans showed atypical robust platelet aggregation when stimulated with submaximal concentrations of agonists. Further, when comparing α-granule secretion, a specific marker of platelet activation, Native Americans were more likely to have elevated responses to multiple agonist conditions of stimulation compared to Whites. Interestingly, there were no noticeable differences in integrin activation between Native Americans and Whites. Our study is the first to observe elevated Native American platelet responses compared to Whites, supporting further mechanistic studies and investigation of treatment approaches for the prevention of thrombosis.

Quantifying choroidal and retinal thicknesses variations via optical coherence tomography in different stages of pediatric keratoconus

PurposeKeratoconus (KCN) is characterized by corneal thinning and bulging, leading to vision impairment. Assessing choroidal thickness (CT) in pediatric KCN ( pKCN) patients can provide insights for better understanding and managing the disease. CT may serve as a potential indicator of disease activity in KCN patients. This study aims to evaluate CT and retinal thickness (RT) in different stages of pKCN patients and compare the findings with an age-matched control group.Methods and patientsThis cross-sectional study included patients under 18 years old who met specific criteria. CT and RT was measured in all subjects using an optical coherence tomography device (Spectralis OCT, version 6.0, Heidelberg Engineering, Germany) with enhanced depth imaging mode, without pupil dilation. Mean CT and RT in pKCN was compared with healthy subjects and assessed among different disease grades using the ABCD grading method.ResultsThe study included 125 eyes (66 patients) in the pKCN group and 42 control eyes (21 individuals). Grade 2 KCN showed the highest prevalence at 26.4% (N = 33), while grades 3 and 1 had prevalence rates of 24% (N = 30). CT in all specified areas (all P-values < 0.001), as well as RT in the subfoveal area (P-value < 0.001) and 1500 μm nasal to the fovea (P-value = 0.025), were significantly greater in the pKCN group compared to controls. Furthermore, CT and RT differed significantly among the pKCN grades (P-values < 0.001).ConclusionCT was found to be elevated in pKCN patients, similar to adult KCN cases. CT could potentially serve as a clinical marker for disease activity in pKCN; however, further studies are needed.

EFFECT OF EXERCISE ON BURST FREQUENCY A MARKER OF SYMPATHETIC ACTIVITY IN HYPERTENSIVE INDIVIDUALS: A SYSTEMATIC REVIEW.

Background: Burst frequency is a factor that is used to know the relationship between sympathetic activity and blood pressure in individuals with hypertension. Various forms of exercise reduce the burst frequency among individuals with hypertension. This systematic review provides suggestion of exercise prescription for reducing blood pressure. Objective: To determine the effects of exercise training and its regulatory mechanisms on burst frequency (BF) a marker of muscle sympathetic nerve activity (MSNA) in hypertensive individuals. Methods: A systematic literature search of electronic databases such as PubMed, Web of Science and Scopus was conducted from inception to July 2024 included randomized controlled trials (RCTs), non-randomized experimental studies (non-RCTs) or pre-post experimental trials on hypertensive patients without associated metabolic disorders, with or without pharmacological treatment, aged between 18 and 70 years, both genders. Patients underwent at least 4 weeks of aerobic, resistance, or breathing exercises with BF as an outcome measure were included. The Revised Cochrane Risk of Bias tool (RoB 2) and Risk of Bias in Non-randomized Studies of Interventions tool (ROBINS-I) was used to evaluate the risk of bias. Results: Five trials were included. Four studies demonstrated a significant reduction in BF among exercise-trained hypertensive individuals, while one study reported no change in BF. Overall, two studies had "some concerns" about performance and attrition bias, one had "high RoB" for selection bias, one had “moderate RoB” due to expected confounding, and one had “serious RoB” due to uncontrolled confounding factors. Conclusion: These findings suggests that HIIT significantly reduces BF, while aerobic and breathing exercises offers lesser benefits, further research should focus on training dosages. Implications: It provides evidence to utilize tailored exercise programme that may reduce sympathetic overactivity in hypertensive patients for better blood pressure control and reduce cardiovascular risk.

TMED inhibition suppresses cell surface PD-1 expression and overcomes T cell dysfunction

BackgroundBlockade of the programmed cell death protein 1 (PD-1) immune checkpoint (ICB) is revolutionizing cancer therapy, but little is known about the mechanisms governing its expression on CD8 T cells....

Shifting GnRH neuron ensembles underlie successive preovulatory luteinizing hormone surges.

The gonadotropin-releasing hormone (GnRH) neurons operate as a neuronal ensemble exhibiting coordinated activity once every reproductive cycle to generate the preovulatory GnRH surge. Using GCaMP fibre photometry at the GnRH neuron distal dendrons to measure the output of this widely scattered population in female mice, we find that the onset, amplitude, and profile of GnRH neuron surge activity exhibits substantial variability from cycle to cycle both between and within individual mice. This was also evident when measuring successive proestrous luteinizing hormone surges. Studies combining short (c-Fos and c-Jun) and long (genetic Robust Activity Marking) term indices of immediate early gene activation revealed that, while ∼50% of GnRH neurons were activated at the time of each surge, only half of these neurons had been active during the previous proestrous surge. These observations reveal marked inter- and intra-individual variability in the GnRH surge mechanism. Remarkably, different sub-populations of overlapping GnRH neurons are recruited to the ensemble each estrous cycle to generate the GnRH surge. While engendering variability in the surge mechanism itself, this likely provides substantial robustness to a key event underlying mammalian reproduction.Significance Statement The mid-cycle luteinizing hormone (LH) surge driven by the gonadotropin-releasing hormone (GnRH) neurons represents the key event triggering ovulation in all mammals. Using GCaMP fibre photometry and genetic activation markers, we unexpectedly find that different sub-populations of GnRH neurons are responsible for driving consecutive LH surges every 4-5 days in cycling female mice. This remarkable oscillatory pattern of network plasticity within the ensemble occurs under normal physiological conditions and likely contributes to the variable timing of the onset of LH surge both within and between individuals. The ability of individual GnRH neurons to take turns within the ensemble in driving the LH surge likely provides a robust fail-safe mechanism for ovulation and contributes to the robustness of mammalian fertility.

Interleukin 9 in Oral Lichen Planus: an immunohistochemical study before and after treatment by intralesional steroid injection.

Oral Lichen Planus (OLP) is a chronic inflammatory mucocutaneous disorder of the oral mucosa. Th9 cells secrete IL9, which induces elevated levels of MMP9, exacerbating OLP disease severity. IL9 also increases Th17 levels in OLP lesions. This study aimed to detect IL9 tissue expression in patients with OLP compared to normal controls and to correlate its expression with disease severity and response to intralesional steroid therapy. This study involved 18 patients with OLP and 18 healthy, age- and sex-matched volunteers. The REU scoring system was used to monitor OLP lesions before and after treatment with 20mg/mL intralesional triamcinolone acetonide every 2 weeks for four sessions. Biopsies for H&E and IL-9 expression were taken from patients and controls, with repeat biopsies after the fourth session for patients. A highly statistically significant increase in IL9 expression was observed in the patient group compared with the control group. A highly statistically significant decrease in both the REU score and post-treatment IL-9 expression was detected in the patient group. We can conclude that IL9 is a tissue marker of OLP activity. Future studies on therapies targeting IL-9 in OLP are needed.

Zolmitriptan niosomal transdermal patches: combating migraine via epigenetic and endocannabinoid pathways and reversal of migraine hypercoagulability.

Conventional zolmitriptan (ZOL) has limited oral bioavailability, many adverse effects, and poor membrane penetrability that negatively influences its accessibility to its 5-HT1B/1D receptor binding pocket, located transmemberanous. This work aimed at preparing transdermal ZOL-nanoformulation (niosomes) to surpass these limitations and to explore novel antimigraine mechanisms for ZOL via modulation of the epigenetically-altered chronification genes (RAMP-1, NPTX-2) or microRNAs and affecting the endocannabinoid CB-1/MAPK pathway. The prepared ZOL niosomes (Fsp60/6-1:1) exhibited %EE of 57.28%, PS of 472.3nm, PDI of 0.366, and ZP of -26 mV were cast into patch with content uniformity of 93.12%, maintained endurance after 200-times folding, no interaction between its components (FT-IR), a biphasic release pattern and good stability after storage at 4°C for 6 months. In-vivo ZOL-patch application in rats with nitroglycerin-induced migraine showed significant management of migraine pain symptoms and photophobia assessed behaviorally, decreased brain levels of the trigeminal neuronal activation marker (c-fos), the migraine pain neurotransmitter (CGRP) and the serum levels of different migraine pain markers (substance P, nitric-oxide, and TNF-α). It also significantly decreased RAMP-1, NPTX-2, miR-382-5p, and CB-1/MAPK gene expression reflecting improved efficacy and brain receptors delivery to a much greater extent than conventional ZOL has done. Additionally, this nanoformulation significantly opposed migraine-induced platelet activation and hypercoagulable status in both central and peripheral circulations as evidenced by the significant decrease in adenosine diphosphate, thrombin, factor X, CD41, and Von-Willebrand factor levels assessed peripherally and centrally. TPFsp60/6-1:1 significantly improved ZOL efficacy and accessibility to brain-receptors to a much greater extent than conventional ZOL-solution.KeywordsEndocannabinoid receptors; Epigenetically-altered genes; Hemostatic pathways; Niosomal patch; Transdermal; Zolmitriptan.

Circulating Adipokines and Response to Treatment in Patients With Early Rheumatoid Arthritis.

The objective of this study was to determine if baseline adiponectin, leptin, and resistin levels are associated with response to antirheumatic treatment in early rheumatoid arthritis (RA). This study included 341 participants of the Nordic Rheumatic Diseases Strategy Trials and Registries trial with untreated early RA, randomized at baseline into four treatment arms: methotrexate combined with (1) prednisolone, (2) certolizumab, (3) abatacept, or (4) tocilizumab. Follow-up was up to 48 weeks. Adipokines were measured in plasma at baseline with enzyme-linked immunosorbent assay. The primary outcome for this report was the difference in remission (Clinical Disease Activity Index [CDAI] ≤2.8) over 48 weeks stratified by median adipokine levels. At baseline, levels of adiponectin and leptin were not associated with markers of RA activity, whereas participants with higher resistin levels had higher C-reactive protein (CRP) levels, swollen joint count, and Disease Activity Score in 28 joints based on CRP compared to participants with lower resistin. Overall, participants with baseline adipokine levels above the median and those with adipokine levels below the median had similar mean CDAI and changes in CDAI throughout follow-up for up to 48 weeks. Adjusted Cox proportional hazards models did not show any effect of baseline adiponectin, leptin, and resistin levels on the likelihood of achieving CDAI remission (adiponectin: hazard ratio [HR] 1.08, 95% confidence interval [CI] 0.80-1.45, P = 0.62; leptin: HR 0.89, 95% CI 0.64-1.26, P = 0.52; resistin: HR 0.86, 95% CI 0.65-1.13, P = 0.26). Baseline adiponectin, leptin, and resistin levels are not associated with the likelihood of achieving CDAI remission over 48 weeks of treatment in a large cohort of people with untreated early RA.

Automated and closed clinical-grade manufacturing protocol produces potent NK cells against neuroblastoma cells and AML blasts

Natural killer (NK) cells are a promising allogeneic immunotherapy option due to their natural ability to kill tumor cells, and due to their apparent safety. This study describes the development of a GMP-compliant manufacturing protocol for the local production of functionally potent NK cells tailored for high-risk acute myeloid leukemia (AML) and neuroblastoma (NBL) patients. Moreover, the quality control strategy and considerations for product batch specifications in early clinical development are described. The protocol is based on the CliniMACS Prodigy platform and Natural Killer Cell Transduction (NKCT) (Miltenyi Biotec). NK cells are isolated from leukapheresis through CD3 depletion and CD56 enrichment, followed by a 12-hour activation with IL-2 and IL-15 cytokines. Three CliniMACS Prodigy processes demonstrated the feasibility and consistency of the modified NKCT process. A three-step process without expansion, however, compromised the NK cell yield. T cells were depleted effectively, indicating excellent safety of the product. Characterization of the NK cells before and after cytokine activation revealed a notable increase in the expression of activation markers, particularly CD69, consistent with enhanced functionality. Intriguingly, the NK cells exhibited increased killing efficacy against patient-derived CD33 + AML blasts and NBL cells in vitro, suggesting a potential therapeutic benefit in AML and NBL.

Bone Mineral Density, C-Terminal Telopeptide of Type I Collagen, and Osteocalcin as Monitoring Parameters of Bone Remodeling in CML Patients Undergoing Imatinib Therapy: A Basic Science and Clinical Review

Background: Chronic myeloid leukemia (CML) is one of the most commonly found types of myeloproliferative neoplasms, characterized by increased proliferation of granulocytic cells without losing their differentiation ability. Imatinib, a tyrosine kinase inhibitor (TKI), can be effectively used as therapy for CML. However, Imatinib can affect bone turnover thus having clinical implications on the bones of CML patients undergoing long-term Imatinib therapy. However, parameters that can accurately describe the bone condition in CML patients receiving Imatinib still need further study. A combination of imaging techniques such as bone mineral density (BMD) and bone turnover activity markers such as C-terminal telopeptide of type I collagen (CTX-1) and osteocalcin has the potential to be used as monitoring parameters for bone density abnormalities in CML patients receiving Imatinib. Objectives: This article explains the rationale for using BMD, CTX-1, and osteocalcin as monitoring parameters of bone remodeling in CML patients receiving Imatinib. Results: First, the physiological process of bone turnover will be explained. Then, we describe the role of tyrosine kinase in bone metabolism. Next, the impact of Imatinib on BMD, CTX-1, and osteocalcin will be explained. Conclusion: The assessment of bone health of CML patients on Imatinib should include both BMD tests and bone turnover marker assays such as CTX-1 and osteocalcin.

Interferon-gamma driven elevation of CXCL9: a new sepsis endotype independently associated with mortality

Endotype classification becomes the cornerstone of understanding sepsis pathogenesis. Macrophage activation-like syndrome (MALS) and immunoparalysis are the best recognized major endotypes, so far. Interferon-gamma (IFNγ) action on tissue macrophages stimulates the release of the cytotoxic chemokine CXCL9. It was investigated if this mechanism may be an independent sepsis endotype. In this cohort study, 14 patient cohorts from Greece, Germany and Italy were studied. The cohorts were 2:1 randomly split into discovery and validation sets. Sepsis was defined by the Sepsis-3 definitions and blood was sampled the first 24h from meeting the Sepsis-3 definitions. Concentrations of IFNγ, CXCL9, IP-10 (IFNγ induced protein-10), soluble CD163 and ferritin were measured. The endotype of IFNγ-driven sepsis (IDS) was defined in the discovery set as the combination of a) blood IFNγ above a specified cut-off associated with the minimal risk for immunoparalysis (defined as ≥8000 HLA-DR receptors on CD45/CD14-monoytes); and b) increase of CXCL9. Results were compared to the validation set. 5503 patients were studied; 3670 in the discovery set and 1833 in the validation set. IDS was defined as IFNγ more than 3pg/ml and CXCL9 more than 2200pg/ml. The frequency of IDS in the discovery set was 19.9% (732 patients; 95% confidence intervals-CIs 18.7-21.3%) and in the validation set 20.0% (366 patients; 95% CIs 18.2-21.9%). Soluble CD163, a marker of macrophage activation, was greater in IDS and IDS had features distinct from MALS. The mortality in IDS patients was 43.0% (315 patients; 95% CIs 39.5-46.6%) in the discovery set and 40.4% in the validation set (148 patients; 95% CIs 35.5-45.5%) (p=0.44 compared to patients of the discovery set). IDS was an independent risk factor for death in the presence of other endotypes, severity scores and organ dysfunctions of the multivariate model [hazard ratio 1.71 (95% CIs 1.45-2.01) in the discovery set and 1.70 (95% CIs 1.34-2.16) in the validation set]. Decreases of IFNγ and CXCL9 blood levels within the first 72h were associated with better outcome. IDS is a new sepsis endotype independently associated with unfavorable outcome. Hellenic Institute for the Study of Sepsis; Horizon 2020 project ImmunoSep; Swedish Orphan BioVitrum AB (publ) and German Federal Ministry of Education and Research.

Detection of HBV pgRNA by catalytic hairpin assembly combined with two Kinds of immunochromatographic strips

Serum HBV pregenomic RNA (pgRNA), as an important marker of HBV replication activity, has significant potential in virological monitoring and efficacy evaluation. In this study, two highly sensitive HBV pgRNA detection platforms were developed based on catalytic hairpin assembly technology, combined with flow lateral immunochromatography and colloidal gold immunochromatography. The signal amplification achieved by CHA technology avoids the dependence of RT-qPCR on thermal cycling equipment. The experimental results showed that the minimum detection limit of CHA-LFIA was 1 pM, and the minimum detection limit of CHA-GICA was 10 pM. Both CHA-LFIA and CHA-GICA had good specificity and could effectively distinguish between target RNA and non-target sequences. The results of CHA-LFIA and CHA-GICA were highly consistent with those of RT-qPCR, and had the potential for clinical application. This study provides a simple, rapid and innovative solution for the detection of HBV pgRNA, which does not rely on expensive equipment, and can be used for the early diagnosis and treatment monitoring of HBV infection.

Skin conductance response and habituation to emotional facial expressions and words.

Skin conductance response (SCR) serves as a dependable marker of sympathetic activation used to measure emotional arousal. This study investigates the impact of presentation modality (face or word) on the degree of emotional discrimination elicited by SCR. Facial expressions or words associated with six basic emotions-anger, happiness, disgust, fear, sadness, and surprise-were studied among 102 participants. The amplitude of SCR was accurately predicted by subjective arousal ratings of these stimuli, but not by valence ratings. The habituation process to emotional and neutral stimuli across six successive presentations was characterized by an exponential decay function, capturing the rate at which SCR response diminishes in relation to the preceding trial of the same stimulus. Through the subtraction of the response to neutral stimuli from the emotion-evoked SCR, it was demonstrated that the initial presentation of each emotion elicits a substantial response, particularly attributable to the emotional content. Notably, the initial emotional response to faces expressing happiness, disgust, and sadness surpassed that of words conveying the same emotions. The results indicate that different emotional responses can be quantified using a simple electrical instrument.

Restricted Feeding Of Weight Control Diets Induces Weight Loss And Affects Body Composition, Voluntary Physical Activity, Blood Metabolites, Hormones, And Oxidative Stress Markers, And Fecal Metabolites And Microbiota Of Obese Cats.

Feline obesity puts many cats at risk for comorbidities such as hepatic lipidosis, diabetes mellitus, urinary tract diseases, and others. Restricted feeding of specially formulated diets may improve feline health and safely support weight loss while maintaining lean mass. The objective of this study was to determine the effects of restricted intake of weight control diets on weight loss, body composition, voluntary physical activity, serum metabolic and inflammatory markers, and fecal metabolites and microbiota of obese cats. Twenty-four obese adult domestic shorthair cats [body weight (BW) = 5.51 ± 0.92kg; body condition score (BCS) = 8.44 ± 0.53] were used. A leading grocery brand diet was fed during a 4-wk baseline to identify intake needed to maintain BW. After baseline (wk 0), cats were allotted to one of two weight control diets (DRY or CAN) and fed to lose 1.5% BW per wk for 18wk. At baseline and 6, 12, 18wk after weight loss, dual-energy x-ray absorptiometry scans were performed, blood and fecal samples were collected, and voluntary physical activity was measured. Change from baseline data were analyzed statistically using the Mixed Models procedure of SAS, with P<0.05 being significant and P<0.10 being trends. BW was reduced by 1.54 ± 0.51% per wk. Restricted feeding of both diets led to BW (P<0.01) and fat mass loss (P<0.01), reduced BCS (P<0.01), reduced leptin (P<0.01) and insulin (P<0.01) concentrations, and increased superoxide dismutase (P<0.01) and active ghrelin (P<0.01) concentrations. Change from baseline fecal scores were reduced (P<0.01) with restricted feeding and weight loss, while total short-chain fatty acid, acetate, and propionate concentration reductions were greater (P<0.05) in cats fed CAN than those fed DRY. Fecal bacterial alpha diversity measures increased (P<0.01) with restricted feeding and weight loss. Fecal bacterial beta diversity was altered by time in all cats, with wk 0 being different (P<0.05) than wk 6, 12, and 18. Change from baseline relative abundances of 3 fecal bacterial phyla and over 30 fecal bacterial genera were impacted (P<0.05) or tended to be impacted (P<0.10) by dietary treatment. Our data demonstrate that restricted feeding of both weight control diets was an effective means for weight loss in obese adult domestic cats. Some changes were also impacted by diet, highlighting the importance of diet formulation and format, and nutrient composition in weight control diets.

Analysis of vitamin D metabolism, thyroid and autoimmune markers in the vitiligo pathways and their possible interaction with sleep.

Vitiligo is an autoimmune skin disease that can be influenced by stress, including that resulting from sleep deprivation and sleep disturbances. Sleep is essential in the regulation of several hormonal, metabolic and autoimmune pathways that may have important roles in vitiligo. This study aimed to investigate the potential interplay between hormonal, metabolic, and autoimmune markers in vitiligo patients, and the possible influence of sleep quality in these vitiligo pathways. A cohort of 30 vitiligo patients and 26 healthy controls were assessed for various laboratory markers, including thyroid-stimulating hormone (TSH), parathyroid hormone (PTH), serum calcium, 1.25(OH)2D, 25(OH)D, anti-thyroid peroxidase (anti-TPO), anti-thyroglobulin (anti-TG), and antinuclear antibodies (ANA). The study evaluated sleep quality using the Pittsburgh Sleep Quality Index (PSQI). Positive anti-TPO were found in the vitiligo group, but did not in the control group. Vitamin D 25(OH)D mean levels were clinically insufficient in both groups (< 30mg/dL). Reactive ANA was analyzed with 2 variables related to vitiligo: phototherapy and skin activity. No statistical correlation was found in the chi-square test on this relationship. Descriptive findings have shown that the positivity to anti-TPO and anti-TG, associated or not with reactive ANA, was higher in vitiligo group. Great part (85.7%) of vitiligo group were "poor sleepers" (PSQI > 5), which has increased (88.2%) when considering only individuals with signs of vitiligo activity. Autoimmune hypothyroidism and positive anti-TPO are expected in vitiligo, although this marker is not usually measured in the first laboratory screening to this disease. Adequate vitamin D levels may be a key adjuvant in skin pigmentation, and be related to sleep quality and immune regulation, as this vitamin can be related to better sleep and immunomodulation in autoimmune diseases. Evaluating ANA before phototherapy can be controversial, but it should be considered in cases with a poor response to this treatment, or when there is a higher risk of other autoimmune diseases. Poor sleep predominated in the vitiligo group, based on PSQI scores that reported worse subjective sleep in these patients. Worse sleep predominated in individuals with signs of skin activity and reactive autoimmune markers. Screening these components could be important in the management of vitiligo, as maintaining body homeostasis can help to improve the disease course. Sleep should be considered as a potential modulator of several multidirectional vitiligo pathways.

Lymphocyte Antigen 6 Family Member D (LY6D) Affects Stem Cell Phenotype and Progression of Pancreatic Adenocarcinoma.

The membrane-bound protein lymphocyte antigen 6 family member D (LY6D), a marker of early B cell lineage is reportedly expressed in several human malignancies and has been implicated in cancer stemness. However, its expression and role in cancer stemness remain largely unexplored in pancreatic ductal adenocarcinoma (PDAC). The aim of this study was to clarify the role of LY6D in PDAC. We conducted functional analysis of LY6D to evaluate its impact on the malignant features of PDAC cells in vitro. Using our in-house developed stem cell separation technique, which isolates cells with low proteasome activity and CD44 v9 cell surface marker for cancer stem cells, we performed sphere formation and chemosensitivity tests and tumor formation assay in mice, through knockdown of LY6D expression. Immuno-histopathological analysis was also conducted to reveal the clinical significance of LY6D in PDAC. In vitro functional assays demonstrated that LY6D was critically involved in promoting the cancer malignant phenotype, including increased invasive ability, drug resistance, migration capacity, and cancer stemness. Immunohistopathological analysis revealed that high LY6D expression levels were associated with high recurrence rates and poorer prognosis in PDAC. Our study showed that LY6D is a novel prognostic indicator and plays a key role in regulation of cancer stemness in PDAC.

Neuroplasticity and neuroimmune interactions in fatal asthma.

Alteration of airway neuronal function and density and bidirectional interaction between immune cells and sensory peripheral nerves have been proposed to trigger and perpetuate inflammation that contribute to asthma severity. To date, few studies analysed neuroplasticity and neuroinflammation in tissue of asthmatic individuals. We hypothesized that the presence of these phenomena would be a pathological feature in fatal asthma. We have quantified the expression of the pan-neuronal marker PGP9.5 and the neuronal sensory-derived neuropeptide calcitonin gene-related peptide (CGRP) in the large airways of 12 individuals deceased due to an asthma attack and compared to 10 control lung samples. The proximity between nerve bundles to eosinophils, mast cells and CADM1+ cells was also quantified. We have additionally developed a hPSC-derived sensory neuron/mast cell co-culture model, from where mast cells were purified and differences in gene expression profile assessed. Fatal asthma patients presented a higher PGP9.5 and CGRP positive area in the airways, indicating sensory neuroplasticity. Eosinophils, mast cells and CADM1+ cells were observed in close contact or touching the airway nerve bundles, and this was found to be statistically higher in fatal asthma samples. Invitro co-culture model showed that human mast cells adhere to sensory neurons and develop a distinct gene expression profile characterized by upregulated expression of genes related to heterophilic adhesion, activation and differentiation markers, such as CADM4, PTGS2, C-KIT, GATA2, HDC, CPA3, ATXN1 and VCAM1. Our results support a significant role for neuroplasticity and neuroimmune interactions in fatal asthma, that could be implicated in the severity of the fatal attack. Accordingly, the presence of physical neuron and mast cell interaction leads to differential gene expression profile in the later cell type.

Immune Activation Is Associated With Neurocognitive Performance in Ugandan Adolescents Living With HIV.

We examined relationships between neurocognition and immune activation in Ugandan adolescents with perinatally acquired HIV (PHIV). Eighty-nine adolescents in Kampala, Uganda (32 virally suppressed [<400 copies/mL] PHIV and 57 sociodemographically matched HIV-negative controls), completed a tablet-based neurocognitive test battery. Control-derived z-scores for 12 individual tests and a global/overall z-score were calculated. We measured plasma (soluble CD14 and CD163), monocyte (proportions of monocyte subsets), and T-cell (expression of CD38 and HLA-DR on CD4 + and CD8 + ) activation and gut markers. Spearman rank correlations and median regressions examined associations between test performance and immune activation. The median [IQR] age was 15 [13-16] years, and 40% were girls. The median time on antiretroviral therapy was 10 years [7-11] for PHIV; 87% had viral load <50 copies/mL. Compared with controls, global z-scores were lower among PHIV ( P = 0.05) and significantly worse on tests of executive functioning and delayed recall ( P 's ≤ 0.05). Overall, monocyte activation significantly correlated with worse test performance on global z-score (r = 0.21, P = 0.04), attention, processing speed, and motor speed (r = 0.2-0.3, P ≤ 0.01). T-cell activation was significantly correlated with worse performance on tests of learning, executive functioning, and working memory (r = 0.2-0.4, P ≤ 0.04). In PHIV, after adjusting for age, sex, and antiretroviral therapy duration, activated CD4 T cells remained associated with worse memory (β-0.3, 95% CI: -0.55 to -0.07, P = 0.01). PHIV with virologic suppression on antiretroviral therapy shows evidence of worse neurocognitive test performance compared with controls. Monocyte and T-cell activation is correlated with worse neurocognition in Ugandan youth with and without HIV, which has not been previously investigated in this setting.

Resting-State Functional Magnetic Resonance Imaging Reveals the Effects of rTMS on Neural Activity and Brain Connectivity After Experimental Stroke.

Limited understanding of neurobiological mechanisms of repetitive transcranial magnetic stimulation (rTMS) prevents us from choosing optimal therapeutic regimen for patients to improve therapeutic efficiency. Resting-state functional magnetic resonance imaging (rs-fMRI) has been demonstrated to obtain comparable functional readouts across species. Intermittent and continuous theta burst stimulation were used to stimulate ipsilesional and contralesional hemisphere, respectively, during the subacute phase after stroke. We used a rat middle cerebral artery occlusion stroke model. The amplitude of low-frequency fluctuations and functional connectivity analyses of rs-fMRI were chosen to detect neuron activity and functional connectivity. The expression of neuron activation marker c-Fos and axonal plasticity marker GAP43 was examined by an immunochemistry method to corroborate the results of rs-fMRI. iTBS altered the long-term neuronal activity in bilateral sensorimotor cortex, whereas cTBS influenced immediate neuronal activity of bilateral sensorimotor cortex. In addition, cTBS enhanced interhemispheric and intrahemisheric functional connectivity in contralesional hemisphere, accompanied by axonal and dendritic remodeling in the perilesional cortical areas and contralesional homologous areas after large stroke. rTMS exerted complex effects on brain structural and functional connectivity in addition to affecting cortical excitability. cTBS promoted the compensatory effect of contralesional hemisphere after stroke with large lesions.