A Potential Role of Interleukin-5 in the Pathogenesis and Progression of Amyotrophic Lateral Sclerosis: A New Molecular Perspective.

Cumulative data suggest that neuroinflammation plays a prominent role in amyotrophic lateral sclerosis (ALS) pathogenesis. The purpose of this work was to assess if patients with ALS present a specific peripheral cytokine profile and if it correlates with neurological disability assessed by ALSFRS-R, the rate of disease progression, and the pattern of disease progression (horizontal spreading [HSP] versus vertical spreading [VSP]). We determined the levels of 15 cytokines in the blood of 59 patients with ALS and 40 controls. We identified a positive correlation between levels of pro-inflammatory cytokines (interleukin [IL]-17F, IL-33, IL-31) and the age of ALS patients, as well as a positive correlation between IL-12p/70 and survival from ALS onset and ALS diagnosis. Additionally, there was a positive correlation between the ALSFRS-R score in the upper limb and respiratory domain and IL-5 levels. In our ALS cohort, the spreading pattern was 42% horizontal and 58% vertical, with patients with VSP showing a faster rate of ALS progression. Furthermore, we identified a negative correlation between IL-5 levels and the rate of disease progression, as well as a positive correlation between IL-5 and HSP of ALS. To the best of our knowledge, this is the first study reporting a "protective" role of IL-5 in ALS.

The Prognostic Utility of Cytokines in Hospitalized COVID-19 Patients.

Introduction: The severity of COVID-19 relies on several factors, but the overproduction of pro-inflammatory cytokines remains a central mechanism. The aim of this study was to investigate the predictive utility of interleukin (IL)-6, IL-8, IL-10, IL-12, tumor necrosis factor alpha (TNF-α), and interferon gamma (IFN-γ) measurement in patients with COVID-19. Material and Methods: We prospectively enrolled 181 adult patients with COVID-19 admitted to the 1st Infectious Disease County Hospital Târgu Mureș from December 2020 to September 2021. Serum cytokine levels were measured and correlated with disease severity, need for oxygen therapy, intensive care unit (ICU) transfer, and outcome. Results: We found significantly higher serum levels of IL-6, IL-8, and IL-10 in patients with severe COVID-19 and in those with a fatal outcome. The logistic regression analysis showed a significant predictive value for IL-8 regarding disease severity, and for IL6 and IL-10 regarding ICU transfer and fatal outcome. Conclusions: Serum levels of IL-6, IL-8, and IL-10 were significantly increased in patients with COVID-19, but their predictive value regarding disease severity and the need for oxygen therapy was poor. We found IL-6 and IL-10 to have a good predictive performance regarding ICU transfer and fatal outcome.

The Relationship between Psychological Suffering, Value of Maternal Cortisol during Third Trimester of Pregnancy and Breastfeeding Initiation.

Background and Objectives: Cortisol, the stress hormone, is an important factor in initiating and maintaining lactation. Maternal suffering during pregnancy is predictive for the initiation and shorter duration of breastfeeding and can also lead to its termination. The aim of this study is to evaluate the relationship between the level of salivary cortisol in the third trimester of pregnancy and the initiation of breastfeeding in the postpartum period in a cohort of young pregnant women who wanted to exclusively breastfeed their newborns during hospitalization. Materials and Methods: For the study, full-term pregnant women were recruited between January and May 2022 in the Obstetrics and Gynecology Clinic of the Mureș County Clinical Hospital. Socio-demographic, clinical obstetric and neonatal variables were collected. Breastfeeding efficiency was assessed using the LATCH Breastfeeding Assessment Tool at 24 and 48 h after birth. The mean value of the LATCH score assessed at 24 and 48 h of age was higher among mothers who had a higher mean value of salivary cortisol measured in the third trimester of pregnancy (p < 0.05). A multivariate logistic regression model was used to detect risk factors for the success of early breastfeeding initiation. Results: A quarter of pregnant women had a salivary cortisol level above normal limits during the third trimester of pregnancy. There is a statistically significant association between maternal smoking, alcohol consumption during pregnancy and the level of anxiety or depression. Conclusions: The most important finding of this study was that increased salivary cortisol in the last trimester of pregnancy was not associated with delayed initiation/absence of breastfeeding.

Dynamic Evaluation of Natural Killer Cells Subpopulations in COVID-19 Patients.

The aim of the study was to evaluate the dynamic changes of the total Natural Killer (NK) cells and different NK subpopulations according to their differentiated expression of CD16/CD56 in COVID-19 patients. Blood samples with EDTA were analyzed on day 1 (admission moment), day 5, and day 10 for the NK subtypes. At least 30,000 singlets were collected for each sample and white blood cells were gated in CD45/SSC and CD16/CD56 dot plots of fresh human blood. From the lymphocyte singlets, the NK cells subpopulations were analyzed based on the differentiated expression of surface markers and classified as follows: CD16-CD56+/++/CD16+CD56++/CD16+CD56+/CD16++CD56-. By examining the CD56 versus CD16 flow cytometry dot plots, we found four distinct NK sub-populations. These NK subtypes correspond to different NK phenotypes from secretory to cytolytic ones. There was no difference between total NK percentage of different disease forms. However, the total numbers decreased significantly both in survivors and non-survivors. Additionally, for the CD16-CD56+/++ phenotype, we observed different patterns, gradually decreasing in survivors and gradually increasing in those with fatal outcomes. Despite no difference in the proportion of the CD16-CD56++ NK cells in survivors vs. non-survivors, the main cytokine producers gradually decline during the study period in the survival group, underling the importance of adequate IFN production during the early stage of SARS-CoV-2 infection. Persistency in the circulation of CD56++ NK cells may have prognostic value in patients, with a fatal outcome. Total NK cells and the CD16+CD56+ NK subtypes exhibit significant decreasing trends across the moments for both survivors and non-survivors.

Inflammatory Biomarkers Associated with In-Hospital Mortality in Critical COVID-19 Patients.

The COVID-19 pandemic poses global healthcare challenges due to its unpredictable clinical course. The aim of this study is to identify inflammatory biomarkers and other routine laboratory parameters associated with in-hospital mortality in critical COVID-19 patients. We performed a retrospective observational study on 117 critical COVID-19 patients. Following descriptive statistical analysis of the survivor and non-survivor groups, optimal cut-off levels for the statistically significant parameters were determined using the ROC method, and the corresponding Kaplan-Meier survival curves were calculated. The inflammatory parameters that present statistically significant differences between survivors and non-survivors are IL-6 (p = 0.0004, cut-off = 27.68 pg/mL), CRP (p = 0.027, cut-off = 68.15 mg/L) and IL-6/Ly ratio (p = 0.0003, cut-off = 50.39). Additionally, other statistically significant markers are creatinine (p = 0.031, cut-off = 0.83 mg/dL), urea (p = 0.0002, cut-off = 55.85 mg/dL), AST (p = 0.0209, cut-off = 44.15 U/L), INR (p = 0.0055, cut-off = 1.075), WBC (p = 0.0223, cut-off = 11.68 × 109/L) and pH (p = 0.0055, cut-off = 7.455). A survival analysis demonstrated significantly higher in-hospital mortality rates of patients with values of IL-6, IL-6/Ly, AST, INR, and pH exceeding previously mentioned thresholds. In our study, IL-6 and IL-6/Ly have a predictive value for the mortality of critically-ill patients diagnosed with COVID-19. The integration of these parameters with AST, INR and pH could contribute to a prognostic score for the risk stratification of critical patients, reducing healthcare costs and facilitating clinical decision-making.

Reply to Chambers, P. Comment on "Huțanu et al. Low Serum Vitamin D in COVID-19 Patients Is Not Related to Inflammatory Markers and Patients' Outcomes-A Single-Center Experience and a Brief Review of the Literature. Nutrients 2022, 14, 1998".

We thank Patrick Chambers for his interest [...].

Low Serum Vitamin D in COVID-19 Patients Is Not Related to Inflammatory Markers and Patients' Outcomes-A Single-Center Experience and a Brief Review of the Literature.

The aim of the study was to evaluate the vitamin D status in hospitalized COVID-19 patients and the correlation with C reactive protein (CRP), ferritin, fibrinogen, and peripheral blood leukocytes, as well as inflammatory derived indices. A prospective study was performed on 203 COVID-19 hospitalized patients, classified by disease severity. Blood was collected after admission, and inflammatory biomarkers and vitamin D status were assessed using routine laboratory procedures. No significant correlation was found between vitamin D serum levels and disease severity stratified by different age groups. However, the highest vitamin D levels were found in patients with mild disease: median 29.39 (IQR 12.12-44.02) ng/mL, while for moderate and severe forms the serum levels were significantly lower: median 15.10 (IQR 9.56-24.11) ng/mL for moderate, and 18.86 (IQR 12.50-27.88) ng/mL for severe; p = 0.009. Patients with no comorbidities showed a significantly higher level of vitamin D median 24.72 (IQR 16.05-31.52) ng/mL compared to subjects with at least one comorbidity: median 16.02 (IQR 9.81-25.22) ng/mL, p = 0.004. We did not find an association between vitamin D levels and inflammatory biomarkers except for significantly lower vitamin D levels in moderate and severe COVID-19 compared to mild disease forms.

Tribulus terrestris Efficacy and Safety Concerns in Diabetes and Erectile Dysfunction, Assessed in an Experimental Model.

The present project aims to evaluate Tribulus terrestris (TT) extracts by addressing various possible mechanisms of action in order to see whether the use of TT supplements in diabetes and diabetes complications is justified. Diabetic rats were divided into three groups: diabetic control group, TT extract with low protodioscin content group (TT-LPC) and TT extract with high protodioscin content group (TT-HPC). After twelve weeks of treatment, fasting blood glucose, insulin, LH, FSH and testosterone levels were measured. Both TT preparations reduced elevated blood glucose level. Insulin and luteinizing hormone levels were not significantly different compared with the control group; however, the FSH and testosterone levels were significantly higher in the TT-HPC group compared with the diabetic control group. The testosterone level is correlated in part with the protodioscin concentration in extracts and is probably mediated through an FSH-linked pathway.

Plasma Biomarkers as Potential Predictors of Functional Dependence in Daily Life Activities after Ischemic Stroke: A Single Center Study.

OBJECTIVE: Despite advances made in the treatment of ischemic stroke, it still remains one of the leading causes of mortality and disability worldwide. The main objective of this study was to identify from a panel of 10 inflammatory markers and chemokines those biomarkers that have a potential predictive role in the evolution of disability and functional dependence in daily activities after an ischemic stroke. METHODS: The study included 116 patients with ischemic stroke and 40 healthy volunteers matched for gender and age. Stroke severity was assessed by the National Institute of Health Stroke Scale (NIHSS) on admission and during hospitalization and functional mobility in daily activities by Barthel index (BI). Multiplex panel with 10 biomarkers [brain-derived neurotrophic factor (BDNF), platelet-derived growth factor (PDGF)-AA, PDGF-AB/BB, neural cell adhesion molecule (NCAM), cathepsin D, soluble vascular cell adhesion molecule (sVCAM), soluble intercellular cell adhesion molecule (sICAM), myeloperoxidase (MPO), regulated on activation, normal T cell expressed and secreted (RANTES), plasminogen activator inhibitor (PAI)-1] was analyzed on days 1 and 5 after admission using the xMAP technology. RESULTS: Plasma concentrations of RANTES and NCAM were significantly lower in patients with ischemic stroke compared with healthy controls, while MPO and sICAM were significantly higher in patients versus controls. Plasma concentrations of sICAM, sVCAM, and RANTES significantly decreased during the analyzed period. For the first-day measurement, the bivariate analysis revealed the association of NIHSS on admission with sVCAM, and on discharge negative association with PDGF-AA, PDGR-AB/BB, BDNF, and RANTES. Plasma levels of PDGF-AA, PDGF-AB/BB, BDNF, and RANTES were found to be significantly lower in patients with BI ≤ 80, on day 5 after disease onset. PDGF-AA, PDGF-AB/BB, and BDNF were univariate and multivariate predictors for functional dependence in daily life activity (BI ≤ 80), having a protective effect (odds ratio < 1). CONCLUSION: Plasma levels of BDNF, PDGF-AA, and PDGF-AB/BB are independent predictors for functional dependency in daily life activities and may be useful prognostic markers in the evaluation of ischemic stroke patients.

The adipokines and inflammatory status in the era of pediatric obesity.

INTRODUCTION: Obesity is associated with a chronic inflammation due to the deficiency in immune activity related to adipose tissue. A wide-spectrum of pro-inflammatory cytokines secreted by adipocytes play an important role in the assessment of obesity-associated inflammatory status. The aim of this study was to assess the relationship between IL and 1ß, IL-6, TNF α, leptin, and inflammatory status in children with overweight/obesity. MATERIAL AND METHODS: We performed a cross-sectional study on 193 children, admitted to a Pediatric Tertiary Hospital in Romania. The children were divided according to BMI into: the study group-91 children with overweight/obesity, and the control group-102 children with normal BMI. Demographic, anthropometric, and laboratory parameters including the serum levels of several adipokines (leptin, IL-1ß, IL-6, and TNF α) were assessed in both groups. RESULTS: Our findings revealed significantly higher values of leukocytes, lymphocytes, platelets, AST, and ALT, as well as for the lipid metabolism parameters including cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides, and CRP, in children with overweight/obesity. We found significantly higher levels of adipokines in the serum of children with overweight and obesity assessed for leptin, IL6, and TNF α (p = 0.0145/p < 0.0001/p = 0.004/), except for IL-1ß serum levels. CONCLUSIONS: Childhood overweight and/or obesity is associated with a systemic inflammatory status demonstrated by increased levels of CBC parameters. Pro-inflammatory adipokines are also an essential factor in obesity-related inflammatory status according to our findings that underlined the importance of increased serum levels of IL-6, TNF α, and leptin in pediatric patients with overweight/obesity. Clinically, CBC parameters along with adipokines might represent useful diagnostic tools for low-grade systemic inflammation in children with overweight or obesity.

Vascular protease-activated receptor 4 upregulation, increased platelet aggregation, and coronary lipid deposits induced by long-term dabigatran administration - results from a diabetes animal model.

Essentials The impact of long-term thrombin inhibition outside the coagulation cascade is far from clear. We aimed to assess the impact of dabigatran etexilate (DE) in diabetic and control rats. In diabetic rats, DE increased platelet aggregation and lead to coronary lipid deposits. Long-term thrombin inhibition may increase atherosclerotic and atherothrombotic risk. SUMMARY: Background Besides its role in the coagulation cascade, thrombin contributes to platelet aggregation and to a plethora of non-hemostatic functions. Objectives To assess the impact of long-term thrombin inhibition with dabigatran etexilate (DE) on platelet aggregation and on extrahemostatic thrombin-related functions in diabetic and control rats. Methods Markers of inflammation, endothelial dysfunction, oxidative stress, angiogenesis and cell adhesion molecules were quantified in control rats (Control; n = 6), DE-treated control rats (Control-Dabi; n = 8), diabetic rats (Diabetes; n = 5), and DE-treated diabetic rats (Diabetes-Dabi; n = 8). Agonist-induced platelet aggregation, aortic and coronary lipid deposits and aortic protease-activated receptor 4 (PAR4) expression were also assessed. Results Control-Dabi rats showed significantly higher high-sensitivity C-reactive protein, von Willebrand factor (VWF), vascular endothelial growth factor (VEGF) and fibronectin levels, and significantly lower PAR4 agonist-induced aggregation, than Control rats. Control-Dabi rats also showed mild aortic lipid deposits, whereas no such changes were observed in Control rats. Diabetes-Dabi rats showed significantly higher VWF, VEGF and fibronectin levels than Diabetes rats, and similar PAR4 agonist-induced aggregation as Diabetes rats, and significantly higher ADP-induced aggregation than Diabetes rats. Coronary lipid deposits were observed in 75% of Diabetes-Dabi rats and in none of the Diabetes rats. PAR4 expression was 20.4% higher in Control-Dabi rats and 27.4% higher in Diabetes-Dabi rats than in their non-treated peers. Conclusions This study indicates that long-term thrombin inhibition increases vascular PAR4 expression, promotes atherosclerosis-related mechanisms, and, in diabetic rats, increases platelet aggregation and favors the occurrence of coronary lipid deposits. These experimental data suggest that long-term thrombin inhibition may increase atherosclerotic and atherothrombotic risk, particularly in the presence of diabetes.

Ischemic damage and early inflammatory infiltration are different in the core and penumbra lesions of rat brain after transient focal cerebral ischemia.

Clinical and experimental observations emphasize that inflammation is a direct risk factor for stroke. We performed a detailed histological and immunohistochemical analysis, assisted by digital morphometry, to compare the representative brain lesions in the ischemic core and penumbra in a rat model. Focal neuronal necrosis and degeneration were significantly more intense in the core, whereas inflammatory infiltration, MPO, CD68, CD3, FXIII, Cox-2, iNOS2, Arg-1 expressions were stronger in the penumbra. Our findings indicate that neuroinflammation affects the penumbra more than the core and suggest that targeted modulation of the cellular infiltrate could be exploited to save brain volume.

Placental Growth Factor, Soluble fms-Like Tyrosine Kinase 1, Soluble Endoglin, IL-6, and IL-16 as Biomarkers in Preeclampsia.

Preeclampsia (PE), an important cause of maternal and perinatal morbidity and mortality worldwide, is a pregnancy-related disease characterized by hypertension and proteinuria after 20 weeks of gestation. The aim of our study was to find a practical panel of biomarkers useful in early diagnosis of PE. This study was carried out at the Obstetrics and Gynecology Department in Tîrgu Mureș University Hospital, Romania, between January 2014 and July 2015 and included 68 pregnant women (47 preeclamptic women and 21 controls) with gestational age between 16 and 20 weeks at enrollment. The biomarkers PlGF, sFlt-1, sEng, IL-6, and IL-16 were determined by ELISA test. We compared the serum levels of soluble markers analysed in preeclamptic women and controls during the second and third trimesters of pregnancy and we found that the best predictor for PE was PlGF with a sensitivity of 100% at a concentration threshold of 120.16 pg/mL, a diagnosis accuracy of 70.8%, and AUC of 0.684 (p = 0.005). We also estimated the risk for PE according to BMI and we found that pregnant women with weight >90 kg had 7 times higher risk for PE. Second-trimester PlGF serum level may serve as an early biomarker for the diagnosis of PE.

The Role of Angiopoietine-2 in the Diagnosis and Prognosis of Sepsis.

INTRODUCTION: Angiopoietin-2 (ANG-2) is a new biomarker whose blood-serum values increase in sepsis and its expression is elevated in line with the severity of the degree of inflammation. The aim of this study was to identify the diagnostic role of ANG-2 in patients with non-surgical sepsis addmitted to an intensive care unit. MATERIAL AND METHODS: This was a prospective randomized study including 74 patients admitted in the Clinic of Intensive Care of the County Clinical Emergency Hospital Tirgu Mureș, divided into two groups: Group S: patients with sepsis (n=40, 54%) and Group C:control, without sepsis (n=34, 46%). ANG-2 levels were determined in both groups. RESULTS: From the Group S, 14 patients (35%) had positive haemocultures. ANG-2 values varied between 1 and 43 ng/mL, with an average of 6.0 ng/mL in patients without sepsis and 10.38 ng/mL in patients with sepsis (p=0.021). A positive correlation between ANG-2 and SAPS II, SOFA and APACHE II severity scores was demonstrated, as was a positive correlation between serum levels of ANG-2 and procalcitonine. ANG-2 had a 5.71% specificity and 74.36% sensitivity for diagnosis of sepsis. CONCLUSIONS: ANG-2 serum levels were elevated in sepsis, being well correlated with PCT values and prognostic scores. ANG-2 should be considered as a useful biomarker for the diagnosis and the prognosis of this pathology.

An Evaluation of Serum Procalcitonin and C-Reactive Protein Levels as Diagnostic and Prognostic Biomarkers of Severe Sepsis.

BACKGROUND: Recommendations have been made, following the multicenter Surviving Sepsis Campaign study, to standardize the definition of severe sepsis with reference to several parameters such as haemodynamic stability, acid-base balance, bilirubin, creatinine, International Normalized Ratio (INR), urine output and pulmonary functional value of the ratio between arterial oxigen partial pressure and inspiratory oxigen concentration. Procalcitonin (PCT) is considered to be a gold standard biomarker for the inflammatory response, and recent studies have shown that it may help to discover whether a seriously ill person is developing sepsis. C-reactive protein (CRP) is also used as a marker of inflammation in the body, as its blood levels increase if there is any inflammation in the body. The aim of this study was to evaluate serum procalcitonin and C-reactive protein levels as diagnostic and prognostic biomarkers of severe sepsis. MATERIAL AND METHOD: Sixty patients, diagnosed as being "septic", were admitted to the intensive care unit (ICU). Based on laboratory results and clinical findings a diagnosis of "severe sepsis" was made, and correlated with PCT and CRP values. The APACHE II, SAPS II and SOFA severity scores were calculated, analyzed and correlated with PCT and CRP. RESULTS: Fifty two patients (86.67%) presented with criteria for severe sepsis. Multivariate correlation analysis indicated a significant positive association between procalcitonin and all severity scores (APACHEII p<0.0001, SOFA p<0.0001, SAPS II p<0.0001). CRP proved to be significantly correlated only with the SAPS II score (p=0.0145). Mortality rate was high, with 48 patients (80%) dying. There was no significant correlation between the levels of the PCT and CRP biomarkers and severe sepsis (p=0.2059 for PCT, p=0.6059 for CRP). CONCLUSIONS: The procalcitonin levels are highly correlated with the severity scores (APACHE II, SAPS II, SOFA) regularly used in ICUs and therefore can be used for determining the severity of the septic process. Quantitive procalcitonin and C-reactive protein analysis was not shown to be useful in diagnosing severe sepsis. However, PCT and CRP can be used to predict the fatal progression of the septic patient.