Concentrations Of Markers Research Articles

MiR-652-3p Suppressed the Protective Effects of Isoflurane Against Myocardial Injury in Hypoxia/Reoxygenation by Targeting ISL1.

This research is concentrated on investigating the role and mechanism of miR-652-3p in the protective effects of isoflurane (ISO) against myocardial ischemia-reperfusion (I/R) injury. H9c2 cells underwent pretreatment with varying concentrations of ISO, and subsequently, a hypoxia/reoxygenation (H/R) model was constructed. The levels of miR-652-3p, ISL LIM homeobox 1 (ISL1), and inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor-alpha (TNF-α) were evaluated through reverse transcription polymerase chain reaction (RT-qPCR). Enzyme-linked immunosorbent assay was employed to investigate concentrations of myocardial injury markers, such as creatine kinase-MB (CK-MB) and cardiac troponin I (cTnI). Cell counting kit-8 was used to evaluate cell viability, while flow cytometry was utilized to measure apoptosis. Additionally, a dual luciferase reporter assay was conducted to validate the targeting relationship between ISL1 and miR-652-3p. Herein, we confirmed thatthe level of miR-652-3p wasgradually increased with prolonged hypoxia; nevertheless, this increase was suppressed by ISO pretreatment (P < 0.05). Additionally, ISO pretreatment prevented the decrease in cell viability, increase in apoptosis, and overproduction of IL-6, TNF-α, CK-MB, and cTnI induced by H/R (P < 0.05). However, the inhibitory effects of ISO were counteracted by the increased levels of miR-652-3p (P < 0.05). ISL1 is a potential target of miR-652-3p. H/R induction suppressed ISL1 levels compared to the control, but ISO treatment increased its expression (P < 0.05). Overexpression of ISL1 inhibited the elimination of the protective effect of ISO on myocardial damage induced by the elevation of miR-652-3p (P < 0.05). The findings of this research confirm that miR-652-3p attenuated the protective effect of ISO on cardiomyocytes in myocardial ischemia by targeting ISL1.

Timing of Assessment of Humoral and Cell-Mediated Immunity after Influenza Vaccination.

Assessment of the immune response to influenza vaccines should include an assessment of both humoral and cell-mediated immunity. However, there is a lack of consensus regarding the timing of immunological assessment of humoral and cell-mediated immunity after vaccination. Therefore, we investigated the timing of immunological assessments after vaccination using markers of humoral and cell-mediated immunity. In the 2018/2019 influenza season, blood was collected from 29 healthy adults before and after vaccination with a quadrivalent inactivated influenza vaccine, and we performed serial measurements of humoral immunity (hemagglutination inhibition [HAI] and neutralizing antibody [NT]) and cell-mediated immunity (interferon-gamma [IFN-γ]). The HAI and NT titers before and after vaccination were strongly correlated, but no correlation was observed between the markers of cell-mediated and humoral immunity. The geometric mean titer and geometric mean concentration of humoral and cellular immune markers increased within 2 weeks after vaccination and had already declined by 8 weeks. This study suggests that the optimal time to assess the immune response is 2 weeks after vaccination. Appropriately timed immunological assessments can help ensure that vaccination is effective.

Effect of milk stasis on mammary gland involution and the microRNA profile

The presence of an autocrine factor in milk that can trigger mammary gland involution was proposed more than 50 yr ago. To provide evidence for the existence of one or more autocrine factors, 10 multiparous cows in late lactation were quarter-milked for 7 d. Following this baseline period, the right front quarter of each cow was left unmilked, and the other quarters were milked for 7 d. Before the last milking of that period, milk (mammary secretions) was collected aseptically from both front quarters. After that milking, 250 mL of the collected samples were infused in the cows' respective rear quarters. No quarters were milked for the following 7 d (milk stasis period), and quarter milking was then resumed in all quarters for the last 7 d of the experiment (remilking period). Quarter milk samples were collected during the baseline period, before the milk stasis period, and during the remilking period. These samples were used for measuring milk components and the concentration of involution markers (SCC, BSA, and lactoferrin). Samples of mammary secretions were collected manually from the quarters during the milk stasis period for involution marker determination. We extracted RNA from samples collected from front quarters before the last milking before the milk stasis period for microRNA (miRNA) determination. As anticipated, the longer milk stasis period implemented for the right front quarter resulted in a more advanced involution than in the left front quarter, based on the concentration of involution markers in the mammary secretions, lower milk production recovery, and changes in milk composition during the remilking period. All 3 involution marker concentrations in the mammary secretions increased in both rear quarters, but were greater in the right quarter secretions than in the left quarter secretions. Resuming milking reinitiated milk production in all quarters, but milk production recovery in the right rear quarters was less robust than that in the left rear quarters (54.3 ± 1.4% vs. 61.6 ± 1.4%, respectively). Milk from the quarters infused with mammary secretions (right rear) had a lower lactose content, but a higher milk protein content and higher SCC than the quarters infused with milk. We detected a total of 359 miRNAs, 76 of which were differentially expressed in milk and mammary secretions. Expression of bta-miR-221 and bta-miR-223 was upregulated in mammary secretions 34- and 40-fold, respectively. The results of the present experiment support the contention that milk stasis leads to the accumulation of one or more factors that trigger involution. The results also indicate that milk stasis leads to changes in the miRNA profile of the milk, but whether such changes are a cause or a consequence of the involution process remains to be established.

#3127 Bedside cycling modulated the bone-resorption markers in patients with end-stage renal disease

Abstract Background and Aims Exercise provided multifactorial benefits in improving the health of the CKD patients. The exercise itself improved the bone remodeling. For CKD patients, chronic kidney disease mineral bone disorder (CKD-MBD) is an important complication and the treatment for renal osteodystrophy is unsatisfactory. The aim of study is to validate the role of bedside cycling in bone remodeling markers for patients with end-stage renal disease (ESRD). Method Participants were grouped into 4 groups: health control, chronic kidney disease (GFR 15∼60 ml/min) and ESRD (GFR&amp;lt;15 ml/min with maintenance dialysis). For ESRD patients, 12-week bedside cycling ((30 minutes per session; 3 times per week) were given for subjects. The pre-and post-exercise bone remodeling markers and clinical outcome were compared in ESRD with and without exercise in ESRD subjects. Results Bone formation markers (Bone specific alkaline phosphatase (15.57 ± 1.59 pg/dL) and Procollagen type 1 amino-terminal propeptide (1269.12 ± 116.34 pg/dL) were higher in ESRD patients with correlation with indoxyl sulfate and intact parathyroid hormone concentration(p &amp;lt; 0.05) respectively. In ESRD patients, the post-exercise concentration of Tartrate-resistant acid phosphatase 5b (3.64 ± 0.25 pg/dL to 4.23 ± 0.31 pg/mL, p = 0.003) and N-terminal telopeptide-1 (113.67 ± 20.53 pg/mL to 260.61 ± 30.9 pg/mL, p = 0.001)increased after 12 weeks of bed-side cycling. The hospitalization and mortality were similar between ESRD with or without exercise. Conclusion The bone formation marker concentration increased along with the severity of CKD. Bedside cycling increased the concentration of the bone resorption markers in ESRD patients.

#1329 Assessing eGFR performance against measured GFR stratifying for ethnicity and albumin: development of the AIM CKD UK study

Abstract Background and Aims Accurate measurement of kidney function using estimated glomerular filtration rate (eGFR) is essential for appropriate risk stratification of cardiovascular events, kidney failure and prescribing medications. The AIM CKD UK study (IRAS 320215) will assess the accuracy of current and novel creatinine-based eGFR equations compared to exogenous filtration marker derived measured GFR (mGFR) for people of different ethnicities in large national cohort. We performed a preliminary analysis from a single centre in AIM CKD UK to inform future data collection and assessed: Method All adults (≥18-years) who had paired mGFR and eGFR tests within 30 days between 2009-2022 were included. Patients with mGFR ≥150 ml/min/1.73 m2, incomplete datasets and previous amputations were excluded. Age, reported ethnicity, biological sex, referral specialty, height, weight, exogenous filtration marker, creatinine and albumin concentration were recorded. eGFR was calculated from creatinine, age and sex using five equations (Fig. 1). Each eGFR equation was compared to mGFR using median bias, precision, 30% accuracy (P30). Agreement was demonstrated with Bland-Altman plots. Results were stratified for time between eGFR-mGFR (≤7 days vs 8-30 days) and albumin concentration; low (&amp;lt;35 g/L) vs normal / high (≥35 g/L). Assessment of equation performance in different ethnicity was also stratified for the three largest ethnicity groups (White, Black and South Asian ethnicity). Data were analysed using software R V4.2.2. Results There were 2623 patients who met the inclusion criteria. Mean age was 54.8 ± 13.1 years, with 57.8% male (N = 1515). The majority of patients were White ethnicity (58.7%; N = 1539). Median mGFR was 77 mL/min/1.73 m2 (IQR 62–91 mL/min/1.73 m2). Median serum creatinine concentration was 74 umol/L (IQR 61–90 umol/L). The CKD-EPI 2009 equation, compared to mGFR, demonstrated a median bias of +12.8 mL/min/1.73 m2, percentage bias of 16.9% and P30 of 68.6%. The Lund-Malmo revised equation performed best with median bias of 2.1 mL/min/1.73 m2 (percentage bias 2.9%) and a P30 of 82.7%. Agreement, with ethnicity stratification is summarised in Table 1. The majority of patients (N = 2439; 93.0%) had paired mGFR-eGFR tests within 7-days. There was no significant difference between the median bias of each group (P = .749). 2610 (99.5%) patients had concurrent albumin tests performed; 183 (7.0%) of these patients had low albumin concentrations. Median bias was significantly higher in this group compared to normal/high albumin for each equation (e.g. Low vs Normal/high albumin median bias for CKD-EPI equation = 20.3 vs 12.5 mL/min/1.73 m2; P &amp;lt;. 001). Conclusion The LM revised and European Kidney Function Consortium equations perform best in this cohort. Limitations include selection bias and retrospective data. Future data analysis for the AIM CKD study can include paired samples within 30 days as there is no significant difference between 0-7 and 8-30 days. The impact of serum albumin on mGFR-eGFR association in a larger cohort stratified for confounding variables, such as liver disease, malignancy and body mass index is required.

Multi-element analysis of metals in human pathological and unchanged thyroid glands - pilot study.

Disturbances in the homeostasis of the elemental composition of thyroid tissue may have serious metabolic and health consequences. It is believed that the accumulation of some metals or the deficiency of others may even cause lethal tumours. Due to the fact that metallomics most often uses human serum to analyse macro and microelements as well as trace elements, it was decided to use material that is more difficult to obtain, but also adds credibility to the research - thyroid tissue samples biopsy. The experiments were conducted on 17 patients diagnosed with: nodular (10) and colloidal goitre (2), chronic thyroiditis (2), follicular adenoma (2) and papillary carcinoma (1). They were recruited by collecting a tumour fragment, control fragment and serum from each of them. The content of Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Zn was examined using ICP-OES (Inductively Coupled Plasma - Optical Emission Spectrometers). Simultaneously, biochemical methods were used to determine the markers of inflammation, glycation and peroxidation: malondialdehyde, pentosidine, reactive free amine content, compounds with thiol groups and galectin 3 in the sera of the examined patients. Three statistically significant correlations were identified: Ca-Mg and Cu-Zn in control tissues (p < 0.05) and Cr-Mn in pathological tissues (p < 0.05). A comparison of individual groups of patients shows that there are some potentail tendencies to increase or decrease in the concentration of certain elements or markers of inflammation and glycation, therefore we discuss potential relationships between a given parameter and a thyroid disorder. The pilot study is an introduction to a deeper analysis aimed at tracing the pathomechanism of the development of thyroid diseases, so that the risk of developing these diseases can be effectively minimized.

THE INFLUENCE OF AGE, PHYSICAL ACTIVITY, SMOKING AND THE PRESENCE OF MYOCARDIAL INFARCTION AND THYROID DISEASES IN THE FAMILY ON THE LEVEL OF ALDEHYDIC AND KETONIC DERIVATIVES OF OXIDATIVE MODIFICATION OF PROTEINS IN THE BLOOD OF WOMEN AND MEN WITH MYOCARDIAL INFARCTS AND HYPOTHYROIDISM

The importance of thyroid hormones in maintaining homeostasis of the cardiovascular system can be inferred from clinical and experimental data showing that even subtle changes in thyroid hormone concentrations - such as those observed in subclinical hypothyroidism or hyperthyroidism and low triiodothyronine syndrome - adversely affect the cardiovascular system. Some potential mechanisms linking the two conditions are dyslipidemia, endothelial dysfunction, changes in blood pressure, and the direct effects of thyroid hormones on the myocardium. Purpose: analysis of changes in the concentration of markers of oxidative stress, e.g. oxidation of protein amino acid residues [concentration of aldehydic and ketonic derivatives of oxidatively modified proteins (OMP)] in the blood of individuals with hypothyroidism and/or myocardial infarcts, living in central Pomeranian region, depending on age, physical activity, smoking and the presence of myocardial infarcts and thyroid diseases in the family. Methodology. The level of oxidative stress markers was assessed among 225 individuals, i.e. 132 males (58.67%) and 93 females (41.33%) aged 35-71 years residing in central Pomeranian region. In the obtained blood, an assessment of levels of aldehydic and ketonic derivatives of oxidatively modified protein analyses was carried out. Scientific novelty. According to our research, the level of aldehydic derivatives of oxidative modification of proteins was significantly higher in the elderly (more than 55 years old), and the level of ketonic derivatives was higher in younger individuals (less than 55 years old) with myocardial infarction and with both myocardial infarction and hypothyroidism. This proves the intensification of oxidative stress in both presented diseases, both among the elderly and younger individuals. The highest level of aldehydic and ketonic derivatives of oxidative modification of proteins was noted among individuals with myocardial infarctions and hypothyroidism with low physical activity and among non-smokers who suffer from hypothyroidism and additionally suffered myocardial infarctions, which may indicate an increase in oxidative stress at these diseases, regardless of physical activity and smoking. The highest level of aldehydic derivatives was noted in the group of individuals with myocardial infarction and hypothyroidism with myocardial infarction in family history, which may indicate an increase in oxidative stress in both diseases, especially in this studied group. The highest level of oxidative modification of proteins was noted among individuals with myocardial infarction and hypothyroidism, individuals with thyroid diseases in their family history, as well as those with no thyroid diseases in their family history. This proves the intensification of oxidative stress in both presented diseases, regardless of the factor of thyroid diseases in the family history. Conclusions. In the course of myocardial infarction, the level of aldehydic derivatives of oxidatively modified proteins is affected by gender, while the level of ketonic derivatives of oxidatively modified proteins is affected by the presence of a myocardial infarction in the family history. The level of aldehydic derivatives of oxidative modification of proteins in individuals with hypothyroidism is influenced by low physical activity and age over 55, while the level of ketonic derivatives of oxidative modification of proteins is affected by the gender of the individuals. In the course of myocardial infarction and hypothyroidism, the level of aldehydic derivatives of protein oxidative modification is affected by age and physical activity, while the level of ketonic derivatives of protein oxidative modification is affected by the age of individuals.

Changes in the concentration of bone turnover markers in men after maximum intensity exercise.

Physical activity is an important factor in modelling the remodelling and metabolism of bone tissue. The aim of the study was to evaluate the changes in indices demonstrating bone turnover in men under the influence of maximum-intensity exercise. The study involved 33 men aged 20-25, divided into two groups: experimental (n=15) and control (n=18). People training medium- and long-distance running were assigned to the experimental group, and non-training individuals to the control. Selected somatic, physiological and biochemical indices were measured. The level of aerobic fitness was determined using a progressively increasing graded test (treadmill test for subjective fatigue). Blood samples for determinations were taken before the test and 60 minutes after its completion. The concentration of selected bone turnover markers was assessed: bone fraction of alkaline phosphatase (b-ALP), osteoclacin (OC), N-terminal cross-linked telopeptide of the alpha chain of type I collagen (NTx1), N-terminal propeptide of type I progolagen (PINP), osteoprotegerin (OPG). In addition, the concentration of 25(OH)D3 prior to the stress test was determined. Additionally, pre and post exercise, the concentration of lactates in the capillary blood was determined. When comparing the two groups, significant statistical differences were found for the mean level of: 25(OH)D3 (p=0.025), b-ALP (p<0.001), OC (p=0.004) and PINP (p=0.029) prior to the test. On the other hand, within individual groups, between the values pre and post the stress test, there were statistically significant differences for the average level of: b-ALP (p<0.001), NTx1 (p<0.001), OPG (p=0.001) and PINP (p=0.002). A single-session maximum physical effort can become an effective tool to initiate positive changes in bone turnover markers.

P19 Defining the mechanism of skin barrier restoration by Lactobacillus rhamnosus

Abstract Introduction and aims We aim to investigate the lysate of Lactobacillus rhamnosus GG (LGG), a bacterium previously established as a gut probiotic, and to examine its postulated ability to enhance and restore human skin barrier function. Compromised skin barriers are implicated in various skin conditions including atopic dermatitis. Furthermore, skin barrier function decreases with age leading to dryness and sensitivity. Previous research has shown promising effects of LGG lysates on keratinocytes, including increasing transepidermal electrical resistance, increasing tight junction protein expression, cell migration promotion and mitigation of Staphylococcus aureus toxicity. This study aims to take a closer look at the molecular mechanisms behind the effect of LGG lysates on the skin, including terminal differentiation, lamin dispersal and nuclear degradation in keratinocytes. Methods Normal human epithelial keratinocytes and rat epithelial keratinocytes were grown and treated with the LGG lysate. Western blot analysis was used to determine the relative concentrations of tight junction proteins and other differentiation markers. A phosphoproteomic study was performed to identify key pathways activated by the LGG lysate. A Staphylococcus epidermidis lysate control was used to filter out any nonspecific bacterial effects. Immunostaining of nuclear proteins was used to visualize the effects of LGG lysate on lamin dispersal and nuclear degradation. short hairpin RNA filaggrin knockdown normal human keratinocytes were developed to create a ‘barrier impaired’ model to assess ability of the lysate to ‘rescue’ the skin barrier. Results The LGG lysate was shown to increase tight junction protein expression in normal human keratinocytes and increase keratinocyte terminal differentiation markers. We present Results that show decreased tight junction protein expression in filaggrin knockdown cells. Conclusions These Results show the potential for LGG lysate to enhance and restore skin barrier function and begin to suggest potential mechanisms for this effect.

Comparison of the pleiotropic effect of atorvastatin and rosuvastatin on postmenopausal changes in bone turnover: A randomized comparative study.

Statins are the first-line treatment for dyslipidemia, which is a major modifiable risk factor for atherosclerotic cardiovascular disease. Studies have shown that in addition to the beneficial lipid-lowering effect, statins also exhibit a number of pleiotropic effects that may find application in other diseases, including osteoporosis. This study aimed to assess the effect of statins on bone turnover, as measured by the concentration of bone turnover markers, and to compare the effect of atorvastatin as a lipophilic statin and rosuvastatin as a hydrophilic statin. This study included 34 postmenopausal women aged < 65 years with newly diagnosed dyslipidemia requiring statin therapy. Patients were randomly assigned to receive a statin drug. Statins were initiated at standard doses of 5 to 10 mg of rosuvastatin and 20 mg of atorvastatin. The levels of C-terminal telopeptide of type I collagen as a bone resorption marker and N-terminal propeptide of procollagen type I as a marker of bone formation, lipid concentrations and other biochemical parameters were assessed at baseline and after 6 and twelve months of treatment. There were no statistically significant differences between the levels of bone turnover markers before and 6 months after statin implementation (P > .05) - for all patients or subgroups according to statin use. Analysis of the results showed that after 12 months, there was a statistically significant decrease in N-terminal propeptide of procollagen type I concentration in all subjects (P = .004). By statin subgroup, a statistically significant decrease in N-terminal propeptide of procollagen type I was observed only in patients receiving rosuvastatin (P = .012) and not in those receiving atorvastatin (P = .25). Moreover, changes in bone turnover markers did not correlate with changes in lipid concentrations. These results may indicate the superiority of atorvastatin over rosuvastatin in inhibiting adverse changes in bone turnover in postmenopausal women. Confirmed by studies involving a larger population, the observed differences might find particular applications in clinical practice, and the choice of atorvastatin over rosuvastatin for women could be considered in the early postmenopausal period to reduce the risk of osteoporosis and subsequent osteoporotic fractures.

Increased TMEM166 Level in Patients with Postoperative Stroke after Carotid Endarterectomy

Postoperative stroke is a challenging and potentially devastating complication after elective carotid endarterectomy (CEA). We previously demonstrated that transmembrane protein 166 (TMEM166) levels were directly related to neuronal damage after cerebral ischemia–reperfusion injury in rats. In this subsequent clinical study, we aimed to evaluate the prognostic value of TMEM166 in patients suffering from post-CEA strokes. Thirty-five patients undergoing uncomplicated elective CEA and 8 patients who suffered ischemic strokes after CEA were recruited. We evaluated the protein level and expression of TMEM166 in patients diagnosed with postoperative strokes and compared it to those in patients who underwent uncomplicated elective CEA. Blood samples and carotid artery plaques were collected and analyzed. High expressions of TMEM166 were detected by immunofluorescence staining and Western Blot in carotid artery plaques of all patients who underwent CEA. Furthermore, circulating TMEM166 concentrations were statistically higher in post-CEA stroke patients than in patients allocated to the control group. Mean plasma concentrations of inflammatory markers, including interleukin 6 (IL-6) and C-reactive protein (CRP), were also elevated in patients with postoperative strokes. Therefore, based on these findings, we hypothesize that elevated TMEM166 levels, accompanied by a strong inflammatory response, serve as a useful biomarker for risk assessment of postoperative stroke following CEA.

The Impact of a 6-Week Nordic Walking Training Cycle and a 14-Hour Intermittent Fasting on Disease Activity Markers and Serum Levels of Wnt Pathway-Associated Proteins in Patients with Multiple Myeloma.

Background: Multiple myeloma (MM) accounts for about 10-15% of all diagnosed hematologic malignancies and about 1-2% of all cancer cases. Approximately 80-90% of MM patients develop bone disease and the changes rarely regress. It is only possible to stop or slow their progression. A major role in bone destruction in MM is attributed to the Wnt signaling pathway, and its action can be modified by various types of interventions including training and diet. Therefore, the aim of this project was to evaluate the effects of a Nordic Walking (NW) training cycle and intermittent fasting (IF) on the levels of selected bone turnover markers associated with the Wnt pathway in patients with MM. Materials and methods: Results from 35 patients divided into training (NW and IF NW) and non-training (IF and control) groups were included in the analysis. A 6-week training cycle involving 60 min workouts 3 times a week was conducted. Body mass and composition as well as the levels of vitamin D, calcium and phosphorus, beta2-microglobulin, and albumin were examined before and after the completion of the training cycle. Markers of bone turnover were also determined: sclerostin (SOST), Dickkopf-related protein 1 (DKK-1), osteoprotegrin (OPG), osteopontin (OPN), and Tartrate-resistant acid phosphatase 5b (TRACP 5b). Results: There was no negative effect of IF or combined training and fasting on the nutritional status of the patients (the level of albumins was unchanged). Both training groups showed an increase in serum concentrations of the active metabolite of vitamin D (IF NW and NW: p = 0.001 and p = 0.022, respectively). The change in the concentration of this vitamin negatively correlated with the concentration of TRACP 5b (r = -0.413, p = 0.014). Evaluating the concentrations of markers related to bone turnover, a reduction in the concentrations of SOST (time: p = 0.026, time vs. group: p = 0.033) and TRACP 5b (time: p < 0.001, time vs. group p < 0.001) was indicated. Conclusions: The obtained results allow one to indicate the training with the poles as a safe and beneficial form of physical activity that should be recommended to patients suffering from MM. However, the results obtained in the present study are not sufficient to show the beneficial effect of IF applied without trainings.

Association between vaginal microbiota and vaginal inflammatory immune markers in postmenopausal women.

In premenopausal individuals, vaginal microbiota diversity and lack of Lactobacillus dominance are associated with greater mucosal inflammation, which is linked to a higher risk of cervical dysplasia and infections. It is not known if the association between the vaginal microbiota and inflammation is present after menopause, when the vaginal microbiota is generally higher-diversity and fewer people have Lactobacillus dominance. This is a post hoc analysis of a subset of postmenopausal individuals enrolled in a randomized trial for treatment of moderate-severe vulvovaginal discomfort that compared vaginal moisturizer, estradiol, or placebo. Vaginal fluid samples from 0, 4, and 12 weeks were characterized using 16S rRNA gene sequencing (microbiota) and MesoScale Discovery (vaginal fluid immune markers: IL-1b, IL-1a, IL-2, IL-6, IL-18, IL-10, IL-9, IL-13, IL-8, IP10, MIP1a, MIP1b, MIP3a). Global associations between cytokines and microbiota (assessed by relative abundance of individual taxa and Shannon index for alpha, or community, diversity) were explored, adjusting for treatment arm, using linear mixed models, principal component analysis, and Generalized Linear Mixed Model + Microbiome Regression-based Kernel Association Test (GLMM-MiRKAT). A total of 119 individuals with mean age of 61 years were included. At baseline, 29.5% of participants had a Lactobacillus -dominant vaginal microbiota. Across all timepoints, alpha diversity (Shannon index, P = 0.003) was highly associated with immune markers. Individual markers that were associated with Lactobacillus dominance were similar to those observed in premenopausal people: IL-10, IL-1b, IL-6, IL-8 (false discovery rate [FDR] < 0.01), IL-13 (FDR = 0.02), and IL-2 (FDR = 0.09). Over 12 weeks, change in alpha diversity was associated with change in cytokine concentration (Shannon, P = 0.018), with decreased proinflammatory cytokine concentrations observed with decreasing alpha diversity. In this cohort of postmenopausal individuals, Lactobacillus dominance and lower alpha diversity were associated with lower concentrations of inflammatory immune markers, as has been reported in premenopausal people. This suggests that after menopause lactobacilli continue to have beneficial effects on vaginal immune homeostasis, despite lower prevalence.

Leveraging wastewater: Validating the national-scale SARS-CoV-2 surveillance system in Cyprus for elevated public health surveillance and enhanced epidemiological insight

The validation of a cross-country, national-scale SARS-CoV-2 wastewater surveillance system for the monitoring of public health in the Republic of Cyprus, took place during October 2021-September 2022. The chronicle of SARS-CoV-2 genetic marker concentrations and prevailing Variants of Concern (VoCs) in all large Wastewater Treatment Plants (WWTPs) of Cyprus for a continuous period of 1 year, was recorded through an integrated, multi-disciplinary sewage-based surveillance system set up to track, visualize and make the collected information and statistical analysis outcomes, publicly available. Composite sampling and physicochemical parameter normalization of Population Equivalents (PE) and SARS-CoV-2 genetic marker concentration loads were utilized.Chemical Oxygen Demand (COD) was selected for normalization of genetic fragment loads. The selection of a normalization method should be tailored according to the characteristics of each individual city/WWTP population structure, flows and wastewater origin. Metagenomic sequencing showed the transition of the SARS-CoV-2 genome from the Delta to the Omicron variant prevalence, in agreement with changes in wastewater gene concentrations. Differences in the early detection of SARS-CoV-2 genetic markers were found among cities. The established system was able to track and monitor the trend of SARS-CoV-2, even in times of absence of other public health measures such as tracing of confirmed COVID-19 cases, showing the capacity of the wastewater system to quickly capture shifts in infections.The work done herein, confirmed the ability of the national-scale system to timely observe pandemic trends, evaluate effects of control measures implemented and monitor the burden of infected patients that shed SARS-CoV-2 fragments.

Optimizing Echinodorus macrophyllus extracts: Assessing extraction conditions for chemical marker enrichment and evaluating inhibitory effects on cytokines release by THP-1 cells

This study investigated various extractive methods and solvents with the goal of producing standardized extracts of Echinodorus macrophyllus for future industrial applications. Extracts were prepared through percolation, dynamic maceration, and static maceration, utilizing 96°GL ethanol (EtOH) and hydroethanolic solutions (90% EtOH, 70% EtOH, and 50% EtOH) as solvents. The highest extraction yields were achieved with 70% EtOH and 50% EtOH across all extractive methods. Both percolation and static maceration produced 50% and 70% EtOH extracts with elevated concentrations of chemical markers (cis- and trans-aconitic acids, homoorientin, chicoric acid, swertisin, caffeoyl-feruloyl-tartaric acid, and di-feruloyl-tartaric acid) and demonstrated superior inhibition rates of TNF and IL-1β release by LPS-stimulated THP-1 cells. Notably, these extracts were obtained in the form of fine powders, distinguishing them from extracts derived from 96°GL EtOH and 90% EtOH solutions. While the 50% EtOH and 70% EtOH extracts prepared by percolation or static maceration exhibited similar chemical composition and biological effects, percolation emerged as the preferred technique due to its ability to generate larger extract volumes. Principal component analysis (PCA) revealed robust correlations between these extracts and the contents of chemical markers but did not unveil significant correlations between higher marker contents and the inhibition of TNF and IL-1β release by THP-1 cells. These findings provide a basis for the future production of high-quality herbal preparations, emphasizing the critical role of extraction methods and solvent selection in pharmaceutical applications.

Are shortened aPTT values always to be attributed only to preanalytical problems?

It has been recognized that shortened activated partial thromboplastin time (aPTT) may be caused by various preanalytical conditions. As coagulation Factor VIII is included in the invitro intrinsic coagulation cascade measured by aPTT, we hypothesized that the shortened aPTT could be a result of elevated FVIII activity. We aimed to inspect the connection of elevated FVIII with shortened aPTT, and the possible effect inflammation has on routine laboratory parameters. 40 patients from various hospital departments with aPTT measurement below the lower limit of the reference interval (<23.0 s) were included in the study. To compare the obtained results with aPTT measurements in the non-inflammatory state, samples from 25 volunteers (laboratory personnel) were collected. White blood cell count, C-reactive protein, aPTT, and FVIII values were measured in the control group. Only two samples among 40 patients with shortened aPTT (5 %) were clotted. Out of the remaining 38, 26 had FVIII activity above 150 % (upper limit of a reference interval), median value of 194 % (IQR: 143-243 %). Seven samples in the control group had shortened aPTT results (36 %). However, all coagulation samples were clot and hemolysis-free. Multiple regression identified only FVIII activity as an independent variable in predicting aPTT values (p=0.001). Our results support the thesis that shortened aPTT is rarely a consequence of preanalytical problems. Elevated FVIII activity causes shortened aPTT, not only in theinflammatory state but also in individuals with concentration of inflammatory markers within reference intervals.

Click evoked otoacoustic emissions in occupational exposure to lead, concentrations of selected essential elements and markers of oxidative stress

PurposeThis study focused on the selected markers of oxidative stress, impact of elevated lead levels on long-term hearing quality. We investigated whether the presence of certain essential minerals might provide protection to the auditory system against the effects of lead (and cadmium) compounds.MethodsThe research group included 280 male employees of the zinc and lead smelter, which was divided into: L-Pb—low blood lead concentration (PbB) subgroup, H-Pb—high PbB subgroup. Hearing tests were performed using the click evoked otoacoustic emission (CEOAE).ResultsZinc protoporphyrin level was significantly higher in the H-Pb subgroup by 68%. Cd concentration was significantly higher in H-Pb by 33%. The Ca concentration was significantly lower in the H-Pb by − 2%. Selected oxidative stress markers concentration were significantly higher in the H-Pb group: malondialdehyde (MDA) by 4%, and lipofuscin (LPS) by 9%. In the CEOAE results showed statistically significant differences between the L-Pb and H-Pb subgroups. Larger negative changes in otoemission amplitude were observed in H-Pb subgroup. All otoemission results showed a statistically significant negative correlation with age, time of work, MDA concentration, and with PbB. Selected CEOAE parameters showed a significant negative correlation with cadmium blood concentration (CdB), and a positive correlation with Ca and Zn.ConclusionElevated blood lead content in occupational exposure is associated with an increase in MDA and LPS concentration, which negatively correlates with CEOAE parameters. This suggests an important role of oxidative stress in the long-term deterioration of hearing.

Passive heat therapy to improve cardiometabolic health in persons with a spinal cord injury — a pilot study

Objective: To test the effect of repeated heat therapy on markers of inflammation, glucose metabolism and vascular health in persons with a spinal cord injury (SCI). It was hypothesized that the heat therapy intervention would lead to a reduction in the fasting concentration of inflammatory markers as well as an improvement in glucose tolerance and microvascular function.Methods: Five adults with thoracic SCI ((1 female, age: 43 (27 – 60) yrs, BMI: 29 (23 to 30) kg/m2, years since injury: 7 (1 – 11) yrs)) enrolled in an 8-week passive heat therapy intervention consisting of 24 session during which the oral temperature was elevated by 1°C by means of a water-perfused suit and heating blankets. The number of completed sessions, time to reach the desired oral temperature and adverse events were reported. Thermoregulatory and perceptual responses to a single passive heat therapy session were assessed during the first session of the intervention. Fasting interleukin-6 (IL-6), C-reactive protein (CRP) and glucose concentration were measured in plasma before and after the chronic intervention. Additionally, thermal hyperemia in sensate (i.e., arm) and insensate (i.e., leg) skin through laser Doppler flowmetry were determined at 39°C and 45°C pre- and post-intervention. Descriptive data are expressed as median (range).Results: The median time to reach a 1°C increase in oral temperature during passive heating was 53 (49 – 75) min, which was accompanied by a final mean skin temperature of 38.6 (36.5 – 38.9)°C, a thermal comfort of 2 (0 – 4) on a 0 to 5 ratings scale and a basic affect of +3 (+1 - +4) on a -5 to +5 rating scale. Four out of the five participants completed all prescribed passive heating sessions. However, in two participants a number of sessions (N = 10 and 2 out of 24 sessions) were stopped at 0.8°C due to thermal discomfort. There were no skin burns or signs of autonomic dysreflexia. All participants remained hemodynamically stable throughout the sessions. Resting IL-6 and CRP changed from 3.2 (2.7 – 10.5) pg/ml to 8.1 (7.6 – 11.1) pg/ml and 0.40 (0.14 – 1.16) to 1.11 (0.51 – 1.26) pg/ml, respectively. Fasting glucose concentration changed from 90.0 (85.5 – 206.0) mg/dl to 94.5 (69 – 266) mg/dl. Cutaneous vascular conductance at 39°C as a percentage of maximal vasodilation at the arm and leg changed from 23.2 (11.2 – 36.0)% to 30.4 (13.8 – 46.1)% and 34.3 (30.4 – 49.4)% to 47.4 (6.7 – 52.8)%, respectively. Conclusion: These preliminary findings suggest that it is feasible, well-tolerated and safe for persons with SCI to engage in repeated heat therapy. A full data set of this ongoing study is needed to provide more conclusive insight into its effects on cardiometabolic health outcomes. VA Rehabilitation R&amp;D (I21RX003724-01A1). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

ASSOCIATION OF SUBCLINICAL INFLAMMATION MARKERS WITH PRIMARY HYPERTENSION IN CHILDREN - A SYSTEMATIC REVIEW WITH META-ANALYSIS

Objective: Primary hypertension (PH) is a growing threat to children's health. The pathogenesis of this phenomenon is not fully known, and subclinical inflammation and activation of the immune system are potential explanations. This systematic review and meta-analysis aimed to systematically determine whether there is an association between low–grade inflammation markers and PH in children. Design and method: The MEDLINE, EMBASE, and Cochrane databases were searched up to March 2023, with no language restrictions, for cohort, cross-sectional, and case-control studies; additional references were obtained from reviewed articles. The included studies needed to investigate an association between any inflammation markers and PH defined by the authors. Participants of the study were children (&lt;18 years old) with PH and healthy controls. The main outcome measured was the concentration of any subclinical inflammation markers in children with PH and in healthy controls. This meta-analysis included 12 studies published between 2005 and 2022, enrolling 1220 patients (689 with PH and 531 healthy controls). The data were analyzed using Review Manager. Pooled mean difference (MD) with 95% confidence interval (95% CI) was used to assess the differences in inflammation markers levels. All analyses were based on the random-effect model. The risk of bias was assessed using the Newcastle-Ottawa Scale. Results: There was a significant difference between hypertensive and control groups in high-sensitivity C-reactive protein (hsRCP) concentration (MD: 0.07 95%CI [0.04,0.09]), intercellular adhesion molecule 1 (ICAM-1) (MD: 85.28 95%CI: [50.57-119.99]), vascular cell adhesion molecule 1 (VCAM-1) (MD: 259.78 95%CI: [22.65-496.91]), neutrophil count (MD: 0.80 95%CI [0.53-1.07]), monocyte count (MD: 0.07 95%CI: [0.02-0.12]), and neutrophil-to-lymphocyte ratio (NLR) (MD: 0.49 95%CI: [0.32-0.65]). There was no difference in terms of interleukin 6 (IL-6), lymphocyte count, platelet count, mean platelet volume (MPV), as well as in platelet-to-lymphocyte (PLR) and monocyte-to-lymphocyte (MLR) ratios. Conclusions: Some easily accessible markers of low-grade inflammation might be used as an additional tool for diagnosis and screening for hypertension in pediatric patients. These promising results should be validated in large and well-conducted studies.

Glutathione peroxidase 4 restrains temporomandibular joint osteoarthritis progression by inhibiting ferroptosis.

There are few effective therapeutic strategies for temporomandibular joint osteoarthritis (TMJOA) due to the unclear pathology and mechanisms. We aimed to confirm the roles of GPX4 and ferroptosis in TMJOA progression. ELISA assay was hired to evaluate concentrations of ferroptosis-related markers. The qRT-PCR assay was hired to assess gene mRNA level. Western blot assay and immunohistochemistry were hired to verify the protein level. CCK-8 assay was hired to detect cell viability. Human fibroblast-like synoviocytes (FLSs) were cultured to confirm the effects of GPX4 and indicated inhibitors, and further verified the effects of GPX4 and ferroptosis inhibitors in TMJOA model rats. Markers of ferroptosis including 8-hidroxy-2-deoxyguanosine (8-OHdG) and iron were notably increased in TMJOA tissues and primary OA-FLSs. However, the activity of the antioxidant system including the glutathione peroxidase activity, glutathione (GSH) contents, and glutathione/oxidized glutathione (GSH/GSSG) ratio was notably inhibited in TMJOA tissues, and the primary OA-FLSs. Furthermore, the glutathione peroxidase 4 (GPX4) expression was down-regulated in TMJOA tissues and primary OA-FLSs. Animal and cell experiments have shown that ferroptosis inhibitors notably inhibited ferroptosis and promoted HLS survival as well as up-regulated GPX4 expression. Also, GPX4 knockdown promoted ferroptosis and GPX4 overexpression inhibited ferroptosis. GPX4 also positively regulated cell survival which was the opposite with ferroptosis. In conclusion, GPX4 and ferroptosis regulated the progression of TMJOA. Targeting ferroptosis might be an effective therapeutic strategy for TMJOA patients in the clinic.