Stress Biomarkers Research Articles

Changes in Immunoglobulins G and A in the Saliva and Serum of Horses with Equine Gastric Ulcer Syndrome (EGUS) and Their Relationship with Other Immune and Redox Status Biomarkers

Equine Gastric Ulcer Syndrome (EGUS) is a widespread disease with a very high prevalence and importance in horses. It includes two conditions: Equine squamous gastric disease (ESGD), linked to acid disturbances, and Equine Glandular Gastric Disease (EGGD), associated with inflammation and immune system issues. In saliva, increased levels of immunological markers have been observed in this disease, indicating immune system involvement. In addition, changes in biomarkers of oxidative stress have been described. In horses, IgG and IgA are proteins that appear in saliva and serum and are considered to be the major proteins in specific immunity; however, their levels in horses with EGUS have not been previously studied. This report aims to evaluate IgG and IgA concentrations in the saliva and serum of horses with EGUS, measured by automated assays, and compare them with other biomarkers of the immune and oxidative stress system. This work validated the IgG and IgA assays in saliva and showed that IgA significantly increased in the saliva of horses with EGUS. When the correlation between IgG and IgA and other biomarkers of immune system issues and oxidative stress such as ADA, S100 A8-A9 (calprotectin), S100 A12 (calgranulin), uric acid, FRAS, and AOPP was studied, IgA in saliva showed a significant moderate correlation with adenosine deaminase, indicating its possible involvement in the immune reaction occurring in EGUS.

Advances of Oxidative Stress Impact in Periodontitis: Biomarkers and Effective Targeting Options.

Periodontitis is the most common inflammatory oral disease that affects around 15% of adults and contributes to severe periodontal tissue destruction with subsequent tooth loosening and loss. Among the main pathogenic mechanisms underlying periodontitis, excessive reactive oxygen species production and oxidative stress play a predominant role in inducing both local and systemic damage. Current therapeutic approaches have expanded the conventional methods combined with herbal antioxidant compounds to free radical-scavenging nanomaterials and infrared laser therapy, offering promising pre-clinical evidence in periodontitis management. Herein, we review the pathogenic mechanisms of reactive oxygen species tissue damage, along with recent advances in oxidative stress biomarkers and novel targeting options.

A growing-finishing diet formulated to reduce the soybean meal does not compromise the growth performance, health, behaviour and gut health of Italian heavy pigs

Soy contributes to the environmental impact of Italian pork production. This study aimed to evaluate the effect of a reduced soybean meal (SBM) and crude protein (CP) finishing diet on growth performance, health, behaviour and gut health of Italian heavy pigs. 1920 pigs (35.6 kg body weight; (BW)) balanced by sex and BW were assigned to the control diet (CO), or the treated diet (TRT) formulated by reducing SBM by 31%, 67% and 69% (replaced by pea and sunflower meal) in 3 feeding phases, respectively, and 1.2% CP in the third phase. 251 pigs were individually weighed at d11, d94 and d181. Feed intake (FI), behavioural indices and air gases at pen-level were monitored monthly. Faecal samples (20/pigs/group) for microbiota, ammonia and volatile fatty acids (VFAs) and hair for stress biomarkers were collected. Diet did not affect final BW, faecal ammonia, cortisol and dehydroepiandrosterone. From d11–d94, the CO group had higher gain to feed (G:F) (p = .007), favourable faecal VFAs profile and a lower environmental ammonia (p < .0001). From d10–d181, the TRT diet increased the ADG (p = .04) and G:F (p = .01), reduced FI (p < .0001) and the lesions score index at d102 (p = .03) and promoted Methanobrevibacter (d94; padj. = 0.013) and Clostridium sensu stricto (d181; padj. = 0.001). Overall, the TRT diet combined with the stress of the transport and acclimatisation to the farm may limit the growth of pigs in the initial period, but it can increase their growth in the long term. Concluding, replacing 56% of SBM with sustainable alternatives seems promising for heavy pig.

Are Oxidative Stress Biomarkers Reliable Part of Multimarker Panel in Female Patients with Type 2 Diabetes Mellitus?

Background: Oxidative stress and inflammation are the key features of metabolic diseases, including type 2 diabetes mellitus (T2D). However, studies that explored redox homeostasis parameters in relation to T2D show discrepant results. Accordingly, we aimed to examine the potential reliability of oxidative stress biomarkers [i.e., determined by malondialdehyde (MDA), advanced oxidation protein products (AOPP) and catalase (CAT)] in addition to traditional cardiometabolic parameters in relation to T2D in female cohort. Methods: A total of 214 women (of them 40.6% T2D) were consecutively recruited in the study. Principal component analysis with varimax rotation was performed to determine the adequate number of factors consisting of anthropometric, traditional cardiometabolic and redox status markers. Results: MDA and AOPP concentrations were lower, but CAT activity was higher in T2D group as compared with controls (P < 0.001, P = 0.002, P < 0.001). Traditional markers related factor (i.e., with positive loading of waist circumference, triglycerides, uric acid, high sensitivity C-reactive protein and negative loadings of high-density lipoprotein cholesterol) was found to be independently related with T2D in multivariate binary regression analysis, whereas oxidative stress related factor (i.e., with positive loading of MDA and AOPP) lost its independent prediction after adjustment for confounding factors (i.e., age, menopausal status, antihypertensive, and hypolipemic therapies). Increased Traditional markers related factor was associated with more than three times higher probability for T2D onset (OR = 3.319, p < 0.001). Conclusion: Oxidative stress biomarkers, i.e., MDA, AOPP, and CAT are not superior over traditional cardiometabolic markers in relation to T2D in female population. Future studies with both gender included are needed to confirm such results.

Effects of intensive and conventional farming on oxidative stress and meat quality biomarkers in holstein and simmental cattle

This study investigates the intricate factors influencing meat quality, including breed, rearing conditions, and processing, with a primary focus on oxidative stress in Holstein Friesian and Simmental cattle within conventional and intensive production systems. A notable difference in oxidative stress was found between animals subjected to intensive-farming versus conventional practices, with Holstein cattle showing a more pronounced antioxidant gene response than Simmental. The analysis revealed that intensive rearing conditions resulted in increased DNA repair activity and expression of stress-response proteins like heat shock proteins, suggestive of greater cellular damage and an adaptive stress response. Muscle tissue analyses, revealed a clear distinction in gene expression associated with meat quality between the breeds and the type of farming system. A negative correlation emerged between oxidative stress levels and genes related to muscle development, which affects meat quality. Intensive farming conditions altered the expressions of apoptotic proteins, impacting meat quality at the molecular level. These results underscore the profound effect rearing conditions have on meat quality, driven by stress-related molecular responses. This highlights the need for further research into the influence of husbandry practices on animal welfare and meat quality, with the intention of developing strategies to mitigate the negative consequences of intensive-farming.

Honey Enriched with Additives Alleviates Behavioral, Oxidative Stress, and Brain Alterations Induced by Heavy Metals and Imidacloprid in Zebrafish

Environmental concerns have consistently been a focal point for the scientific community. Pollution is a critical ecological issue that poses significant threats to human health and agricultural production. Contamination with heavy metals and pesticides is a considerable concern, a threat to the environment, and warrants special attention. In this study, we investigated the significant issues arising from sub-chronic exposure to imidacloprid (IMI), mercury (Hg), and cadmium (Cd), either alone or in combination, using zebrafish (Danio rerio) as an animal model. Additionally, we assessed the potential protective effects of polyfloral honey enriched with natural ingredients, also called honey formulation (HF), against the combined sub-chronic toxic effects of the three contaminants. The effects of IMI (0.5 mg·L−1), Hg (15 μg·L−1), and Cd (5 μg·L−1), both individually and in combination with HF (500 mg·L−1), on zebrafish were evaluated by quantifying acetylcholinesterase (AChE) activity, lipid peroxidation (MDA), various antioxidant enzyme activities like superoxide dismutase and glutathione peroxidase (SOD and GPx), 2D locomotor activity, social behavior, histological and immunohistochemical factors, and changes in body element concentrations. Our findings revealed that all concentrations of pollutants may disrupt social behavior, diminish swimming performances (measured by total distance traveled, inactivity, and swimming speed), and elevate oxidative stress (OS) biomarkers of SOD, GPx, and MDA in zebrafish over the 21-day administration period. Fish exposed to IMI and Hg + Cd + IMI displayed severe lesions and increased GFAP (Glial fibrillary acidic protein) and S100B (S100 calcium-binding protein B) protein expression in the optic tectum and cerebellum, conclusively indicating astrocyte activation and neurotoxic effects. Furthermore, PCNA (Proliferating cell nuclear antigen) staining revealed reduced cell proliferation in the IMI-exposed group, contrasting with intensified proliferation in the Hg + Cd group. The nervous system exhibited significant damage across all studied concentrations, confirming the observed behavioral changes. Moreover, HF supplementation significantly mitigated the toxicity induced by contaminants and reduced OS. Therefore, the exposure to chemical mixtures offers a more complete picture of adverse impacts on aquatic ecosystems and the supplementation with bioactive compounds can help to reduce the toxicity induced by exposure to environmental pollutants.

ВОЗМОЖНОСТИ ПРИМЕНЕНИЯ НЕИНВАЗИВНЫХ ОФТАЛЬМОЛОГИЧЕСКИХ МЕТОДОВ ДИАГНОСТИКИ ПАТОЛОГИЧЕСКИХ СОСТОЯНИЙ ПРОЦЕССА РАННЕГО СТАРЕНИЯ НА МОДЕЛИ ПРИМАТОВ

Currently, there is an urgent need for early diagnostics, searching for means of predicting the development of cognitive deficit, and assessing the dynamics of changes during treatment. Given the common origin of the brain and eye, which undergo the same onto- and phylogenetic development, some researchers have suggested that the eye can act as a “window” for easily accessible and non-invasive methods for studying brain pathology [3]. The popularity of fundamental research is growing, and interest in creating disease models in primates is becoming more relevant. The creation of such models will significantly expand the possibilities of obtaining the most reliable information, especially in the field of gerontology and brain pathology (cataract, senile ring, age-related changes in the fundus and other signs of early aging). In this part of the work, approaches to studying the fundus, physiological parameters of the structures and functions of the visual organ in rhesus macaques and hamadryas baboons of different ages are developed; potential ophthalmological biomarkers for diagnosing early aging processes are considered. A combined approach using ophthalmological markers, including non-invasive visualization of the retina and detection of pathological proteins in the intraocular fluid, lens or retina, in combination with biomarkers of oxidative stress (malondialdehyde, lipofuscin, protein carbonyls, 8-OH-deoxyguanosine) will allow us to develop a methodology for diagnosing the development and correction of early aging in humans on a primate model.

ВОЗМОЖНОСТИ ПРИМЕНЕНИЯ НЕИНВАЗИВНЫХ ОФТАЛЬМОЛОГИЧЕСКИХ МЕТОДОВ ДИАГНОСТИКИ ПАТОЛОГИЧЕСКИХ СОСТОЯНИЙ ПРОЦЕССА РАННЕГО СТАРЕНИЯ НА МОДЕЛИ ПРИМАТОВ

Currently, there is an urgent need for early diagnostics, searching for means of predicting the development of cognitive deficit, and assessing the dynamics of changes during treatment. Given the common origin of the brain and eye, which undergo the same onto- and phylogenetic development, some researchers have suggested that the eye can act as a “window” for easily accessible and non-invasive methods for studying brain pathology [3]. The popularity of fundamental research is growing, and interest in creating disease models in primates is becoming more relevant. The creation of such models will significantly expand the possibilities of obtaining the most reliable information, especially in the field of gerontology and brain pathology (cataract, senile ring, age-related changes in the fundus and other signs of early aging). In this part of the work, approaches to studying the fundus, physiological parameters of the structures and functions of the visual organ in rhesus macaques and hamadryas baboons of different ages are developed; potential ophthalmological biomarkers for diagnosing early aging processes are considered. A combined approach using ophthalmological markers, including non-invasive visualization of the retina and detection of pathological proteins in the intraocular fluid, lens or retina, in combination with biomarkers of oxidative stress (malondialdehyde, lipofuscin, protein carbonyls, 8-OH-deoxyguanosine) will allow us to develop a methodology for diagnosing the development and correction of early aging in humans on a primate model.

Protective effects of Sphaeranthus indicus floral extract against BPS-induced testicular damage in rats occurs through downregulation of RIPK1/3-MLK-driven necroptosis and Fas-FasL-mediated apoptosis

AbstractBisphenol S (BPS) is one of the monomers preferred in the manufacturing of polycarbonate plastics. Unfortunately, its estrogenic and genotoxic effects are similar to those of bisphenol A. The protective effects of Sphaeranthus indicus floral extract (SFE) against reprotoxic effects of BPS (50 µg/kg per body weight) in rats exposed to it via drinking water was investigated. Different SFE doses (25, 50, and 100 mg/kg) were administered via oral gavage for 10 weeks. High-performance liquid chromatography (HPLC) results indicated that SFE was rich in polyphenols, with rutin and quercetin being important bioactive molecules modulating BPS-induced necroptosis and apoptosis. Biochemical analyses unveiled that rats administered BPS only exhibited considerable elevation of biomarkers of nitro-oxidative stress, necroptotic (RIPK1/RIPK3 and MLKL), and apoptotic mediators (Fas/FasL and caspase 3/caspase-8). These events caused changes in sperm characteristics (sperm motility, sperm head, and sperm viability), sperm count, and hormonal profile (thyroid stimulating hormone, luteinizing hormone, and follicle-stimulating hormone) of the rats. Histological analysis suggested that there was pronounced sloughing of Sertoli cells, reduced spermatogenic cell density, increased levels of seminiferous tubules, and disorganized morphometric parameters related to seminiferous tubules. The SFE supplementation in rats with BPS-containing water restored nitro-oxidative stress biomarkers, which led to the reduction of necroptosis and apoptosis. Reinstatement of all the biomarkers of oxidative stress, inflammation, necroptosis, and apoptosis after SFE supplementations restored the hormonal profile and normal histoarchitecture of the testes. Virtual screening elucidated that the key regulators of the necroptosis are RIPK3-rutin and RIPK1-quercetin complexes. Further studies are needed to assess its pharmacodynamics, kinetics, and effective concentration for daily use in humans.

Evaluation of a probiotic blend on psychosocial health and biomarkers of inflammatory, immune and stress response in adults with subthreshold depression: a double-blind, randomised, placebo-controlled trial.

This study examined the efficacy of a probiotic in reducing depressive symptom severity in people with subthreshold depression. In a double-blind, randomised, placebo-controlled trial, a probiotic (1 × 10^9 live cells per strain: Limosilactobacillus fermentum LF16 (DSM26956), Lacticaseibacillus rhamnosus LR06 (DSM21981), Lactiplantibacillus plantarum LP01 (LMG P-21021) and Bifidobacterium longum 04 (DSM23233)) or placebo was taken daily for 12 weeks. Data were collected at baseline, 6 and 12 weeks including psychological symptom severity (Beck Depression Inventory, BDI; Patient Health Questionnaire, PHQ; Hospital Anxiety Depression Scale, HADS; and Depression Anxiety and Stress Scale, DASS). Biomarkers of glycaemia, inflammation (high-sensitivity C-reactive protein, hs-CRP), antioxidant status (total glutathione (GSH)) and stress (cortisol awakening response, CAR) were also measured. Thirty-nine participants (nineteen probiotic;twenty placebo) were enrolled. There were no significant between-group differences in the examined psychological symptom severity scores, despite certain significant within-group changes observed in both groups from baseline to 6 and/or 12 weeks of follow-up. Regarding biomarkers, the probiotic group showed reduced hs-CRP (-1520; 95 % CI -273·7, -2766·2 ng/dl) and CAR (-0·28; 95 % CI -0·05, -0·51 μg/dl) at 12 weeks, but increased total GSH (3·9; 95 % CI 0·1, 7·5 ng/dl) at 6 weeks, compared with the placebo. The current study reported favourable decreases in depressive symptoms in both groups. Although the within-group changes observed in the probiotic group were supported by favourable inflammatory, antioxidant status and stress biomarker changes compared with the placebo, further research is required to shed more light on the role of gut microbiota modulation on emotional regulation.

Hair cortisol concentration, a biomarker of stress, is gradually increased in patients with acute myocardial infarction before the acute episode

Abstract Background/Introduction Chronic stress is considered to be an important element of residual cardiovascular risk and modern guidelines recommend its systematic assessment and treatment. Its pathophysiologic contribution, however, in acute myocardial infarction (AMI) was not, until recently, easy to elucidate. This arises from difficulties in objectively quantifying stress levels using conventional research methods, like questionnaires. Purpose The purpose of our study was to measure stress levels at the days preceding an acute myocardial infarction using a solid biomarker, Hair Cortisol Concentration (HCC). Methods 102 consecutive patients with AMI (71 patients with ST-elevation Myocardial Infarction [STEMI] and 31 with Non ST-elevation Myocardial Infarction) from our institution were recruited for this study. We took hair samples from them to precisely measure HCC, which reflects cumulative cortisol levels for the time period preceding the acute event. We also did full clinical, biochemical and psychological evaluation. HCC levels were then compared with 50 age-matched healthy controls. Results To assess the temporal trend of HCC in hair segments, corresponding to specific time periods, we fitted linear mixed models. The mixed effects linear model including the main effects of time and group (AMI versus controls) and their interaction showed that cortisol concentration in hair increased over time with higher rate of increase in AMI patients versus controls (beta of interaction = 3.57; 95%CI: 1.74, 5.40; p &amp;lt; 0.001) The model explained a high percentage of the variance (conditional R2=0.940). No other baseline variable had a significant effect when added to the basic model. There was no significant difference between STEMI and NSTEMI (beta of interaction = 1.15; 95%CI: -1.330, 3.59; p = 0.358) Conclusions Hair cortisol levels, presumably reflecting stress levels, were gradually increasing before an acute myocardial infarction with higher rate of increase in patients compared to healthy controls. There was no significant difference between STEMI and NSTEMI patients. Picture 1. Predicted hair concentrations of cortisol over time prior to the event, based on linear mixed models.

Myocardial SGLT2 expression and cardiac remodeling in patients with severe aortic stenosis: the BIO-AS Study

Abstract Background Cardiac remodeling plays a major role in the prognosis of patients with aortic stenosis (AS) and could impact the benefits of aortic valve replacement. Blood and tissue biomarkers have a differential pattern and expression level in patients with AS, which may retain a pathophysiological role in cardiac remodeling and metabolism. However, the data available are limited and contradictory. Aim Our study aimed to evaluate the expression of sodium-glucose cotransporter 2 (SGLT2) gene and protein in patients with severe AS stratified in high gradient (HG) and low flow-low gradient (LF-LG) AS and its association with cardiac functional impairments. Methods Gene expression and protein levels of main biomarkers of cardiac fibrosis (Galectin-3, sST2, Serpin-4, PINP, PICP, Collagen, TGF-b), inflammation (GDF-15, IL-6, NF-kB), oxidative stress (SOD1, SOD2), and cardiac metabolism (NHE, PPAR-a, PPAR-g, GLUT1, and GLUT4) were evaluated in blood samples and heart biopsies of 45 patients with AS. Results Our study showed SGLT2 gene and protein hyper-expression in patients with LF-LG AS, compared to controls and HG AS (p&amp;lt;0.05). These differences remained significant even after adjusting for age, gender, body mass index, history of diabetes mellitus, arterial hypertension, and coronary artery disease. SGLT2 gene expression was positively correlated with: i) TGF-b (r=0.72, p&amp;lt;0.001) and collagen (r=0.73, p&amp;lt;0.001) as markers of fibrosis; ii) NF-kB (r=0.36, p&amp;lt;0.01) and myocardial IL-6 (r=0.68, p&amp;lt;0.001) as markers of inflammation: iii) SOD2 (r=-0.38, p&amp;lt;0.006) as a marker of oxidative stress; iv) GLUT4 (r= 0.33, p&amp;lt;0.02) and PPAR-a (r=0.36, p&amp;lt;0.01) as markers of cardiac metabolism (Figure 1). Conclusion In patients with LF-LG AS, SGLT2 gene and protein were hyper-expressed in cardiomyocytes and associated with myocardial fibrosis, inflammation, and oxidative stress. The hyper-expression of the SGLT2 gene and protein in patients with LF-LG might pave the way for using SGLT2-Is in this subset population, with the potential to improve the outcomes of AVR and reduce the likelihood of re-hospitalization within one year.Figure 1

Lipoprotein(a) regulates vascular redox state and inflammatory response in patients with coronary artery disease

Abstract Background Circulating Lipoprotein(a) (Lp(a)) is associated with cardiovascular disease, and over 90% of its variation in plasma is genetically determined. However, neither the causal nature of this association nor the underlying mechanisms are fully documented. Purpose We explored the hypothesis that Lp(a) triggers atherogenesis by dysregulating vascular redox-sensitive inflammatory state. Methods 1027 patients with advanced coronary artery disease (CAD) undergoing cardiac surgery were genotyped and a genetic signature (LPA) determining Lp(a) levels was generated. RNA-sequencing and vascular superoxide (O2.-) measurements were performed in internal mammary arteries (IMA), and the contribution of NADPH-oxidases and uncoupled endothelial nitric oxide synthase (eNOS) were determined. Patients were followed up for a median of 5.07 years. Results A genetic screening identified 7 SNPs (LPA) that were associated with increased Lp(a) plasma levels (A). Patients in the alternative LPA variant group, as well as patients with higher plasma Lp(a) levels, had significantly higher basal arterial O2.-. Patients with diabetes had significantly lower plasma Lp(a) compared to non-diabetics and the association between plasma Lp(a) and arterial O2.- was driven by non-diabetic patients. The effect of Lp(a) on vascular redox state was primarily due to eNOS uncoupling, resulting from reduced vascular tetrahydrobiopterin (BH4) bioavailability. Indeed, patients in the alternative LPA variant group or with higher plasma Lp(a) levels had a greater arterial eNOS-derived (LNAME-inhibitable) O2.- production (B,C). Levels of eNOS essential cofactor BH4 (defined by the ratio of plasma BH4 to its oxidation product BH2) were lower in patients with higher plasma Lp(a) levels (D). There was no significant impact of Lp(a) variability on vascular NADPH oxidase-derived O2.-. RNA sequencing of arterial tissue revealed dysregulation of nitrosative and inflammatory signalling in patients with high Lp(a), although there was no association with systemic biomarkers of inflammation (i.e. hsCRP) or oxidative stress (i.e. malondialdehyde). Finally, both LPA and high plasma Lp(a) were associated with elevated risk for cardiovascular mortality (E,F). When the cox regression models were additionally corrected for arterial O2.- production, both the models for the LPA genetic signature (HR[95% CI]=2.053[0.180-23.428], p=0.563) and plasma Lp(a) (HR[95% CI]=1.700[0.325-8.885], p=0.529) lost significance. This implies that redox-sensitive inflammatory signalling may be a link between Lp(a) and cardiovascular risk. All above associations were independent from plasma ApoB. Conclusions This study demonstrates for the first time that a genetically determined increase in plasma Lp(a) results in dysregulated vascular redox/nitrosative signalling in patients with atherosclerosis, revealing new therapeutic opportunities in cardiovascular prevention.

Chronic stress is gradually accumulated and peaks prior the occurrence of Takotsubo Syndrome: preliminary results based on hair cortisol concentration

Abstract Background Takotsubo syndrome (TTS) is frequently triggered by stressful events suggesting that increased sympathetic activity has a role in its pathophysiology. Hair cortisol concentration (HCC) is a promising biomarker of excess chronic stress by measuring cortisol secreted into hairs over several weeks. Purpose We explored whether premorbid ongoing stress over several weeks assessed by HCC at 2 weeks intervals is associated with the occurrence of TTS. Methods A case-control prospective study was performed by recruiting 18 consecutive TTS patients and 36 age- and sex-matched individuals free from cardiovascular or psychiatric disease. Hair samples were collected from participants vertex. Provided that 1 cm of hair represents cortisol levels over ~1 month period, 2.5 cm of hair samples, with many strands each, were cut in equal parts of 0.5 cm, representing cortisol levels in time intervals of 0-15, 15-30, 30-45, 45-60 and 60-75 days prior to hair collection. Results All TTS patients were women with a mean age of 71.8±11.2 years. HCC was significantly higher in TTS group versus controls starting from 45 days prior to the event (Figure 1A). Based on the linear mixed effects model, HCC was higher in the TTS group compared to controls at any time point and increased over time starting from 75 days prior to the event, while the rate of increase was significantly higher in TTS versus controls (beta of interaction = 0.48; 95%CI: 0.36 – 0.60; p &amp;lt; 0.001). The model explained a high percentage of the variance (conditional R2 = 0.885). The plot of the predicted HCC over time by diagnosis group is shown in Figure 1B. Conclusions Our preliminary data suggest that accumulation of chronic stressful events may cause hypothalamic-pituitary-adrenal axis dysregulation and hyperactivity leading to HCC increase, and this might be implicated in the pathophysiology of TTS.Figure 1.Plot of observed and predicted HCC

Chromatic covalent organic frameworks enabling in-vivo chemical tomography

Covalent organic frameworks designed as chromatic sensors offer opportunities to probe biological interfaces, particularly when combined with biocompatible matrices. Particularly compelling is the prospect of chemical tomography – or the 3D spatial mapping of chemical detail within the complex environment of living systems. Herein, we demonstrate a chromic Covalent Organic Framework (COF) integrated within silk fibroin (SF) microneedles that probe plant vasculature, sense the alkalization of vascular fluid as a biomarker for drought stress, and provide a 3D in-vivo mapping of chemical gradients using smartphone technology. A series of Schiff base COFs with tunable pKa ranging from 5.6 to 7.6 enable conical, optically transparent SF microneedles with COF coatings of 120 to 950 nm to probe vascular fluid and the surrounding tissues of tobacco and tomato plants. The conical design allows for 3D mapping of the chemical environment (such as pH) at standoff distances from the plant, enabling in-vivo chemical tomography. Chromatic COF sensors of this type will enable multidimensional chemical mapping of previously inaccessible and complex environments.

Exposome, oxidative stress and inflammation in multiple sclerosis: the EXPOSITION study protocol

Abstract Background The functional exposome approach examines the relationship between biological effects (internal exposome) and the sum of environmental exposures (external exposome). Multiple sclerosis (MS) is characterized by selective demyelination and acute inflammation; miRNAs and microbiota are emerging in the pathogenesis as inflammation and oxidative stress biomarkers. The EXPOSITION study protocol has been designed to study such biomarkers variation in response to the external exposome (air pollution, urbanization, lifestyles) in people with MS (pwMS). Methods EXPOSITION is a cross-sectional observational study collecting a sample of 200 pwMS residing in Lombardy, Northern Italy. Sample size is based on the prediction of differential expression of candidate miRNAs through bioinformatic analyses. All participants will be investigated for social, demographic and clinical data as well as general and specific external exposome factors. The internal exposome factors will be collected for each participant from blood and stool samples, as well as from nasal swabs. Exposome clustering and correlation statistical techniques will be applied and variation of miRNA and microbiota will be tested in association with the variation of the external exposome in urban and extra-urban environments. Results and conclusions The expected results will help pwMS and researchers understanding which biomarkers and which components of the external exposome are associated to oxidative stress and inflammation, to improve precision medicine and contrast disease progression. The relationship between urban exposome and biodiversity is an important dimension in the study of environmental impact on human health. This study may provide preliminary data to assess which nature-based solutions are suitable for enhancing the biodiversity of urban areas. Key messages • The functional exposome approach could be used to link environmental risk factors and disease biomarkers using the right methology. • EXPOSITION will quantify the variation of miRNA and microbiota in association with the variation of external exposome, in urban and extra-urban environments.

Emphasizing the Communal Demands of a Leader Role Makes Job Interviews Less Stressful for Women But Not More Successful

AbstractThe cultural construal of leadership as masculine impedes women’s attainment of leader roles. This research examined whether adding feminine demands to a leader role relieved the greater stress experienced by women than men in a job interview for a leadership position and considered the processes that mediated women’s less favourable interview outcomes. In a hiring simulation, management students (N = 209; 112 women, 97 men) interviewed for a leader role framed by either stereotypically feminine or masculine role requirements. As shown by the stress biomarker salivary cortisol, the feminine role framing alleviated women’s, but not men’s, physiological stress response during the interview. However, under both masculine and feminine role framing, women, compared with men, reported lesser fit, expected poorer interview performance, appraised greater threat relative to challenge, and evaluated their performance less favourably, as did external raters. An additional vignette study (N = 305; 189 women, 111 men, 5 diverse) found that the feminine role framing increased the leader role’s communal demands but still conveyed strong agentic demands not different from those of the masculine role. In conclusion, although a feminine role framing alleviated women’s physiological stress response, it did not change their less favourable outcomes, as indicated by participants’ self-reports and others’ reports.

Cardioprotective potential of oleuropein, hydroxytyrosol, oleocanthal and their combination: unravelling complementary effects on acute myocardial infarction and metabolic syndrome

Abstract Background/Introduction Clinical studies have established the role of olive products in primary and secondary prevention of cardiovascular (CV) disease, but the single compounds or compound combinations responsible for the protective effects are still under investigation. Purpose We sought to explore and compare the cardioprotective potential of Oleuropein (OL), Hydroxytyrosol (HT), Oleocanthal (OC) and Oleanolic Acid (OA) and three combinational treatments after chronic oral administration in mice subjected to Ischemia/Reperfusion Injury (IRI) in the presence of metabolic syndrome (MS) and verify the effect of combination in a proof-of-concept clinical study in Chronic Coronary Artery Syndrome (CCAS) patients. Methods First, we explored the IRI-limiting potential of 6-weeks daily oral treatment with OL (20.6 mg/kg), HT (5.9 mg/kg), OC (11.6 mg/kg), OA (17.4 mg/kg) or vehicle (DMSO 5%) in healthy C57Bl6 mice. At week 6, myocardial ischemia/reperfusion (30 minutes/2 hours) was induced and infarct size was measured. Next, MS was established by 14 weeks of Western diet and the isolated compounds were administered for the last 6 weeks. At week 14, IRI was surgically induced, and infarct size was determined. Basic MS parameters (fasting blood glucose, plasma cholesterol, body weight and blood pressure) were recorded at baseline, weeks 8 and 14. According to the exerted benefits, three combinational treatments were designed and tested on the MS model of IRI. The underlying cardioprotective mechanism was investigated on the mixture with preponderant CV benefits (OL-HT-OC, Combo 2). ApoE-/- mice fed with Western diet for 12 weeks were treated with Combo 2 orally for the last 4 weeks and aortic atherosclerotic lesion extent was determined to detect possible antiatherosclerotic effects. After multi-organ toxicity evaluation, a Combo 2-based supplement was administered in CCAS patients for evaluation of cardiac function, inflammation and oxidative stress biomarkers. Results OL, OC and OA, but not HT, significantly reduced infarct size in vivo both in healthy and MS mice. OL was the only compound to reduce hyperglycemia, while OA significantly reduced plasma cholesterol. All mixtures (Combo 1: OL-HT-OA, Combo 2: OL-HT-OC, Combo 3: OL-HT-OC-OA) were cardioprotective, but only Combo 2 prevented MS progression by limiting hyperglycemia. Combo 2 protection against IRI was attributed to apoptosis suppression (reduced Bax/BcL-xL ratio) and increased expression of the antioxidant enzymes Superoxide Dismutase and Catalase in the ischemic myocardium. Combo 2 treatment also reduced atherosclerotic lesion extent. In our proof-of-concept clinical study, Combo 2 treatment ameliorated cardiac, vascular and endothelial function in CCAS patients. Conclusions Combinational treatment with OL-HT-OC exerts potent cardioprotective, antihyperglycemic and antiatherosclerotic properties in vivo, with remarkable and clinically translatable CV benefits in high-risk patients.

In-vitro and in-vivo assessment of the bactericidal potential of peracetic acid and hydrogen peroxide disinfectants against A. hydrophila infection in Nile tilapia and their effect on water quality indices and fish stress biomarkers

This study aimed to assess the in vitro and in vivo disinfectant potential of peracetic acid (PAA) (1 mg/L) and hydrogen peroxide (H2O2) (20 mg/L) on the physicochemical and microbiological water quality parameters of fish aquaria, the microbial density of Nile tilapia muscular tissue, fish hepatic cortisol levels, and antioxidant biomarkers. In vitro, PAA and H2O2 reduced A. hydrophila colony viability by 5 log units after 30 and 5 min of contact time, respectively. PAA and H2O2 were added to aquaria water twice a week for the three-week experiment. Increased fish escape reflexes were observed only in the PAA group, which returned to normal within 10 min. No mortalities were reported in either the PAA or H2O2 groups. An in vivo experimental challenge with a pathogenic strain of A. hydrophila revealed a 20% reduction in mortality in the PAA group, with no mortalities in the H2O2 group. Cortisol levels and antioxidant markers were measured to assess the impact of PAA and H2O2 on fish health. Cortisol levels in the PAA and H2O2 groups were significantly higher than in the control group after disinfectant exposure, but they progressively returned to normal. A significant reduction in superoxide dismutase (SOD) and catalase (CAT) activity, along with considerably higher glutathione peroxidase (GPx) and malondialdehyde (MDA) enzymatic activity, was observed in the PAA and H2O2 groups compared to the control group. A substantial increase in total antioxidant capacity (TAC) was recorded in the PAA group. Physicochemical analyses revealed reduced pH and increased dissolved oxygen levels in the PAA and H2O2 groups. Microbiological analyses showed a significant reduction in bacterial density in water by 64% and 76% after 30 min of exposure to PAA and H2O2, respectively, with a non-significant increase in microbial count after bacterial challenge. Additionally, aerobic bacterial count, Aeromonas spp., and psychotropic bacterial count in fish muscle showed a significant reduction in the H2O2 group compared to the PAA and control groups before and after infection. The study concludes that regular application of PAA and H2O2 can temporarily reduce bacterial load in aquaria and fish muscle, regulate stress responses, and improve fish health by reducing A. hydrophila-induced infections and improving survival.

The gut microbiota metabolite trimethylamine-N-oxide in children with β-thalassemia: potential implication for iron-induced renal tubular dysfunction.

Renal tubular dysfunction is common in transfusion-dependent β thalassemia (β-TM). Iron overload, chronic anemia, and hypoxia are precipitating factors for renal insult. However, gut microbiota engagement in the renal insult has not been explored. Our work aimed to assess the potential link between iron overload, gut leakage/dysbiosis, and kidney dysfunction in these children. We enrolled 40 children with β-TM and 40 healthy controls. Gut leakage/dysbiosis biomarkers (trimethylamine-N-oxide [TMAO] and fecal short-chain fatty acids [SCFAs]), oxidative stress and inflammatory biomarkers, TMAO-regulated proteins such as serum sirtuin 1 (S.SIRT1) and serum high mobility box group-1 (S.HMGB1), and tubular dysfunction biomarkers were assessed. Correlations and regression analysis were performed to assess the relation between different parameters. Iron overload, redox imbalance, and generalized inflammation were evident in children with β-TM. Renal tubular dysfunction biomarkers and S.TMAO were significantly elevated in the patient group. Furthermore, fecal SCFAs were significantly lower with upregulation of the investigated genes in the patient group. The correlation studies affirmed the close relationship between circulating ferritin, TMAO, and renal dysfunction and strongly implicated SIRT1/HMGB1 axis in TMAO action. Gut dysbiosis may have a role in the pathogenesis of renal injury in children with β-TM. Renal tubular dysfunction is a prominent health issue in β thalassemia major (β-TM). Iron overload, chronic anemia, and hypoxia are known precipitating factors. However, gut microbiota engagement in renal insult in these patients has not yet been explored. We aimed to assess potential link between iron overload, gut leakage/dysbiosis, and kidney dysfunction in β-TM children and to highlight the SIRT1/HMGB1 axis, a signal motivated by the gut microbiota-dependent metabolite trimethylamine-N-oxide (TMAO), involvement in such insults. We found that gut leakage/dysbiosis may have a role in kidney dysfunction in β-TM children by exacerbating the iron-motivated oxidative stress, inflammation, ferroptosis, and modulating SIRT1/HMGB1 axis.