Solid-phase Enzyme-linked Immunosorbent Assay Research Articles

CHARACTERISTICS OF FOLLICULAR FLUID MORPHOLOGY AND LEVELS OF MATRIX METALLOPROTEINASE-1 AND ITS INHIBITOR-1 IN PATIENTS WITH POLYCYSTIC OVARIES IN ASSISTED REPRODUCTIVE TECHNOLOGY

Assisted reproductive technologies in patients with polycystic ovary syndrome (PCOS) is an effective way to realize reproductive function with a global pregnancy rate of 25–30 %. The purpose of the study is to establish the correlation between the levels of matrix metalloproteinase-1 (MMP-1) and its inhibitor-1 (TIMP-1) with follicular fluid morphology in PCOS patients undergoing assisted reproductive technology. Materials and Methods. The authors examined 26 PCOS patients aged 24–40. All patients were diagnosed with primary and secondary infertility lasting from 1 to 10 years. A comparative analysis of patients was carried out depending on the presence (n=12) or absence (n=14) of concomitant pathology, a history of endometrial hyperplasia. Clinical, anamnestic, instrumental and laboratory methods were used for analysis. Concentrations of MMP-1 and TIMP-1 in follicular fluid was determined by quantitative solid-phase ELISA. Follicular fluid morphology was studied according to wedge-shaped dehydration method. Statistica 10.0 wasnused for statistical data processing. Results. TIMP-1 level in PCOS patients was 2550±57.8 ng/ml, in PCOS patients with endometrial hyperplasia – 2694.4±87.8 ng/ml (p<0.01). At the same time, the average MMP-1 level was low – 1.5±0.1 ng/ml. In PCOS patients, the 3rd (reactive) type of facies and markers indicating impaired blood supply were detected in the follicular fluid. In PCOS patients with endometrial hyperplasia, the 4th (chaotic) type of facies and markers indicating impaired elasticity, vascular sclerosis, and vasospasm was observed. It was found that the higher the TIMP-1 level, the more markers indicating destructive changes in the vessels of ovarian tissue (r=0.7) and the lower the chances of getting pregnant in assisted reproducrtive technology programs. Conclusions. In PCOS patients over 32 years of age, infertility duration of more than 4 years, having endometrial hyperplasia and surgical interventions in the past medical history (tubectomies for sactohydrosalpinx), TIMP-1 level significantly (p<0.05) increases (≥2694 ng/ml) in the follicular fluid. Moreover, markers indicating destructive changes (vascular sclerosis, vasospasm and endothelial dysfunction) are identified. Thus, the chances of getting pregnant in assisted reproducrtive technology programs become lower.

The development of anticyclic citrullinated peptide (anti-CCP) antibody following severe COVID-19.

The dysregulated immune response is one of the cardinal features of severe coronavirus disease 2019 (COVID-19). This study was conducted to clarify the occurrence of autoantibodies (AABs) associated with systemic autoimmune rheumatic diseases (SARDs) in hospitalized patients with a moderate, severe, and critical form of COVID-19. The serum samples obtained from 176 hospitalized COVID-19 patients were investigated in this study, including patients with moderate (N = 90), severe (N = 50), and critical (N = 36) forms of COVID-19. Also, the serum samples collected from healthy subjects before the COVID-19 pandemic were used as controls (N = 176). The antinuclear antibodies (ANAs), antidouble-stranded DNA (anti-dsDNA), cytoplasmic-anti neutrophil cytoplasmic antibody (c-ANCA), perinuclear ANCA (p-ANCA), antiphospholipid antibodies (aPLs), and anticyclic citrullinated peptide (anti-CCP) occurrence was evaluated using a solid-phase enzyme-linked immunosorbent assay (ELISA). The results showed that the occurrence of ANAs, anti-dsDNA, anti-CCP, c-ANCA, and p-ANCA was significantly higher in the COVID-19 patients compared to serum obtained from healthy subjects (p < .0001, p < .0001, p < .0001, p < .05, and p < .001, respectively). The positive number of anti-CCP tests increased significantly in severe COVID-19 compared to the moderate group (p < .01). Our study further supports the development of autoantibodies related to systemic autoimmune rheumatologic diseases. To the best of our knowledge, this is the first study with a large sample size that reported the occurrence of anti-CCP in a severe form of COVID-19.

THE INFLUENCE OF ADJUVANTS ON THE EXPRESSION OF PRO-INFLAMMATORY CYTOKINES DURING SPINAL ANESTHESIA

Objective. To investigate the influence of adjuvants (fentanyl and dexmedetomidine) on TNF-α and IL-6 expression during spinal anesthesia based on bupivacaine.&#x0D; Materials and Methods. A total of 150 patients with an average age of 41.3±8.2 years undergoing endoscopic biportal discectomy were examined. Patients were divided into groups: Group 1 received intrathecally 2.5 ml hyperbaric bupivacaine; Group 2 received intrathecally 2.5 ml hyperbaric bupivacaine and 25 μg fentanyl; Group 3 received intrathecally 2.5 ml hyperbaric bupivacaine and 5 μg dexmedetomidine. Peripheral venous blood was collected 1 hour before surgery, at 12 and 24 hours postoperatively, to measure TNF-α and IL-6 levels through solid-phase enzyme-linked immunosorbent assay. Results. The expression of pro-inflammatory cytokines (tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6)) may play a crucial role in modulating postoperative inflammatory reactions. However, contemporary scientific literature presents conflicting data on the influence of anesthetics on cytokine release.&#x0D; Groups 1 and 2 showed a significant increase in TNF-α levels at all observation stages. In Group 3, a significant elevation in TNF-α was observed only 12 hours postoperatively, with no statistically significant difference in the subsequent 24 hours. Inter-group analysis of TNF-α dynamics revealed the smallest increase in Group 3, while Group 1 had significantly higher values compared to Group 3 and Group 2. In all study groups, there was a progressive, statistically significant increase in IL-6 levels within 24 hours postoperatively. However, Group 1 exhibited consistently higher values, with statistically significant differences compared to Group 2 and Group 3. In Groups 2 and 3, statistically significant inter-group differences in IL-6 were noted 12 hours postoperatively, but there was no statistical significance in IL-6 dynamics 24 hours postoperatively.&#x0D; Conclusion. Intrathecal use of bupivacaine with dexmedetomidine had a lesser impact on postoperative TNF-α and IL-6 expression during endoscopic biportal discectomy.

Comparative Assessment of the Effectiveness of HSP70 / HIF-1α System Modulators after Prenatal Hypoxia

Prenatal hypoxia (PH) poses a significant threat to fetal development and may be responsible for neonatal mortality or neurodevelopmental abnormalities. The proteins HSP70 and HIF-1, which hold a distinct significance in the cellular reaction to PH, can be regarded as potential targets for pharmaceutical interventions aimed at mitigating the repercussions of chronic PH. This study aimed to identify a possible correlation between offspring survival and stages of expression of endogenous neuroprotective factors (HSP70 and HIF-1) after chronic prenatal hypoxia with course administration of potential HSP70 modulators (angiolin, piracetam, thiotriazoline, nicomex, cerebrocurin, tamoxifen, L-arginine, glutoredoxin, HSF-1, and mildronate). In the rat offspring after PH we determined the plasma concentrations of HSP70 and HIF-1 by solid-phase ELISA immunoassay, and the expression of HIF-1 mRNA and HSP70 mRNA by real-time PCR. For the first time, we found a positive correlation between offspring survival after PH and the expression of HIF-1 and HSP70, both in groups without experimental therapy and in groups receiving pharmacological agents. The course administration of HSP70/HIF-1α modulators, especially angiolin (50 mg/kg), cerebrocurin (150 mg/kg), and HSF-1 (50 mg/kg), to rats that underwent PH reduces postnatal lethality, increases blood plasma concentrations of HSP70 and HIF-1α, and positively affects the expression level of HIF-1α mRNA in the rat brain. These drugs can be considered as the most promising drug candidates for new therapeutic strategies of pharmacological correction of the consequences of chronic PH.

Determination of Mycotoxigenic Fungi and Total Aflatoxins in Stored Corn from Sites of Puebla and Tlaxcala, Mexico

This paper aimed to evaluate the contamination with mycotoxigenic fungi and total aflatoxins in stored corn from different sites in Puebla and Tlaxcala, Mexico. Methodology. The study was conducted at two sites in Puebla (San Salvador El Seco and Junta Auxiliar La Resurrección) and two sites in Tlaxcala (Tlaltepango and Nativitas). A total of 80 samples of stored corn were collected. Identification of Aspergillus flavus was performed by microculture techniques and specific taxonomic keys (macromorphological and micromorphological). Then, samples of contaminated corn were selected, and aflatoxin production was confirmed using a direct solid-phase ELISA kit. A total of 25 A. flavus strains were identified. Other possible mycotoxin-producing fungi were Penicillium (n=52) and Fusarium (n=19). Regarding total aflatoxin contamination, all samples were contaminated within a range of 1.589 to 11.854 μg/kg, and the average concentration was 6.3 μg/kg corn. Implications. The detection of mycotoxigenic fungi in the samples tested and of aflatoxins in corn highlights the importance of monitoring these fungi. Since food safety is at risk, it shows the need for methods to control these fungi and their metabolites.

Correlation of Serum Galactose-Deficient IgA1 and Oxford Class in Cases of IgA Nephropathy.

Galactose-deficient immunoglobulin A1 (Gd-IgA1) deposition in the renal mesangium plays a role in the pathogenesis of IgA nephropathy. To assess the serum Gd-IgA1 level in biopsy-proven IgA nephropathy cases on diagnosis and 3 months post treatment and its relation with histologic Oxford classification. In this hospital-based prospective cohort study, 40 cases and 20 controls were enrolled. Serum samples of biopsy-proven IgA nephropathy cases collected on the day of biopsy and 3 months post treatment were evaluated. Solid-phase ELISA (enzyme-linked immunosorbent assay) was performed for assessment of Gd-IgA1 level. All renal biopsies were scored by using Oxford Classification (C-MEST score). The association of serum Gd-IgA1 levels with other established prognostic parameters was assessed. To estimate the prognostic value of markers, logistic regression analysis and Kruskal-Wallis ANOVA (analysis of variance) were used. Significant difference was observed in the serum Gd-IgA1 level values in the IgA nephropathy cases and healthy controls (P = .001) at baseline. However, no significant correlation between serum Gd-IgA1 levels at baseline and 3 months of follow-up (P = .31) or between baseline levels and age, proteinuria, hematuria, or estimated glomerular filtration rate was noted. There was no significant correlation between C-MEST score and serum Gd-IgA1 levels at baseline (P > .05); however, the distribution of Gd-IgA1 at 3 months was found to differ significantly between different grades of S score (P = .008). Serum Gd-IgA1 levels may be of utility in predicting disease progression in IgA nephropathy cases. Measurement of serum Gd-IgA1 levels for the diagnosis and prognosis of IgA nephropathy may preclude the need for invasive renal biopsies.

Utility of insulin-like growth factor-1 (IGF-1) in growth hormone-treated short children – An experience from a tertiary level center

Introduction: Very few studies have been conducted in low-middle-income countries to assess the response to growth hormone (GH) therapy by measuring the improvement in insulin-like growth factor-1 (IGF-1) concentrations. Objectives: The objective is to assess IGF-1 concentrations in patients receiving GH therapy to correlate the height increment with respect to increase in the IGF-1 concentrations, including IGF-1 Z-scores as a measure of safety profile. Methods: Clinical and anthropometric data were extracted retrospectively from hospital records of children aged 0–18 who received GH and had IGF-1 measured during the study period. Serum IGF-1 concentrations were analyzed by a solid-phase enzyme-linked immunosorbent assay. Patients with GH deficiency (GHD) and multiple pituitary hormone deficiency (MPHD) were grouped as group A, while participants receiving GH therapy for non-GHD/MPHD were clubbed as group B. Results: We report a significant positive correlation between an increase in IGF-1 values and increase in height. The improvement in height Z-scores was significantly higher in participants of group A as compared to B, with no significant difference in increase in IGF-1 Z-scores between the two groups. A total of 18.75% of subjects had IGF-1 Z-scores between +2 and +3, and these were largely subjects belonging to group B. Conclusion: The increment in height and height velocity Z-scores in the patients on GH correlated with increase in the IGF-1 concentrations, particularly in GHD/MPHD group of patients.

COT-TT vaccine attenuates cocaine-seeking and cocaine-conditioned place preference in rats

ABSTRACT Vaccination active, promising alternative immunological strategy to treat of CUD. Various models of cocaine vaccines have been evaluated in animals and humans with relative success. In this sense, it is necessary to improve or optimize the cocaine vaccines already evaluated. Our laboratory previously reported the efficacy of the tetanus toxoid-conjugated morphine vaccine (M6-TT). The M6-TT vaccine can generate high titers of antibodies and reduce heroin-induced behavioral effects in rodents. So, it would be plausible to assume that if we modify the M6-TT vaccine by changing the hapten and maintaining the rest of the structural elements of the vaccine, we will maintain the properties of the M6-TT vaccine (high antibody titers). The objective of this study was to determine whether the antibodies generated by a tetanus toxoid-conjugated cocaine vaccine (COC-TT) can recognize and capture cocaine and decrease the cocaine-induced reinforcing effects. Male Wistar rats were immunized with the COC-TT. A solid-phase antibody-capture ELISA was used to monitor antibody titer responses after each booster dose in vaccinated animals. The study used cocaine self-administration and place-preference testing to evaluate the cocaine-reinforcing effects. The COC-TT vaccine could generate high levels of anti-cocaine antibodies. The antibodies reduced the cocaine self-administration and cocaine place preference. In addition, they decreased the cocaine-induced Fos protein expression. These findings suggest that the COC-TT vaccine generates a robust immunogenic response capable of reducing the reinforcing effects of cocaine, which supports its possible future use in clinical trials in patients with CUD.

Serum neurofilament light chains in assessing the course of multiple sclerosis

To study a role of serum neurofilament light chains (sNFL) in assessment of course and progression of multiple sclerosis (MS) in the population of patients with MS in the Tomsk region. The study involved 93 patients with relapsing-remitting MS (RRMS) and secondary progressive MS (SPMS) (nRRMS=75, nSPMS=18). The study was carried out in a two-stage design: the first stage was a cross-sectional study for the entire sample; the second stage was a prospective observation with two visits for patients with relapse. sNFL concentration was determined by solid-phase ELISA. There was no statistically significant difference between RRMS and SPMS, and relapse and remission groups in terms of sNFL levels. Patients with a MS duration exceeding 14 years had higher rates of sNFL than those with a shorter duration (p=0.02). The subjects of the second study stage showed a decrease in sNFL from 2.05 (1.86; 2.19) pg/ml to 1.92 (1.87; 2.04) pg/ml (p=0.005), and slowdown in sNFL reduction correlated with the severity of cognitive impairment (k=0.52; p<0.05). Dynamic monitoring of sNFL allows the evaluation of the activity of the disease, as well as making an assumption about the compensatory possibilities of subsequent recovery.

Cellular localization and potential ligands of a novel scavenger receptor class B/CD36 protein homolog (Pt-SRB2) identified in the marine crab, Portunustrituberculatus

The scavenger receptor class B family proteins (SRB) are multiligand membrane receptor proteins. Herein, a novel SRB homolog (Pt-SRB2) was identified in Portunus trituberculatus. The open reading frame of Pt-SRB2 was predicted to encode 520 amino acid residues comprising a typical CD36 domain. Phylogenetic analysis showed that Pt-SRB2 distinctly clustered with the SRB homologs of most crustaceans and Drosophila but was separate from all vertebrate CD36/SRB. Semi-quantitative and Real-time quantitative PCR revealed that the abundance of Pt-SRB2 transcripts was the highest in hepatopancreas than in other tested tissues. Overexpressed Pt-SRB2 was distributed primarily in the cell membrane and cytoplasm of HEK293T or Drosophila Schneider 2 cells. In crab hemocytes, Pt-SRB2 was distributed primarily in the cell membrane by immunofluorescence staining. In addition, the immunofluorescence staining showed that green fluorescence signals were mainly located in the inner lumen membrane of the hepatopancreatic tubules. Moreover, solid-phase enzyme-linked immunosorbent assay revealed that rPt-SRB2-L exhibited relative high affinity with lipopolysaccharides, and relative moderate binding affinity with lipoteichoic acid or peptidoglycan. Of note, rPt-SRB2-L showed high binding affinity with eicosapentaenoic acid among a series of long-chain polyunsaturated fatty acids. Taken together, this study provided valuable data for understanding the functions of the crab CD36/SRB.

A Comparative Analysis of NT-proBNP Levels in Pregnant Women and the Impact of SARS-CoV-2 Infection: Influence on Birth Outcome.

The cardiac biomarker NT-proBNP is released by the ventricles in response to increased cardiac wall tension, showing cardiac activity in heart failure. The primary objective of this comparative study was to analyze the variations of NT-proBNP levels among pregnant patients and to determine the potential influence of SARS-CoV-2 infection on these values. Secondly, the study focused on NT-proBNP levels and their influence on the type of birth. Blood samples were taken from 160 pregnant mothers in order to determine, through the solid-phase enzyme-linked immunosorbent assay (ELISA) method, the NT-proBNP concentrations from the plasma. The cohort was separated into two distinct groups based on SARS-CoV-2 diagnostic results: negative to the infection, and positive to the infection. The SARS-CoV-2-positive group of patients presented with higher levels of NT-proBNP and had higher rates of cesarean sections. (4) Conclusions: Our research highlights the crucial relationship between elevated NT-proBNP values and the mode of giving birth, natural delivery or cesarean section, and also the influence of SARS-CoV-2 viral infection and this biomarker.

Concordance between the In Vivo Content of Neurospecific Proteins (BDNF, NSE, VILIP-1, S100B) in the Hippocampus and Blood in Patients with Epilepsy.

The identification of reliable brain-specific biomarkers in periphery contributes to better understanding of normal neurophysiology and neuropsychiatric diseases. The neurospecific proteins BDNF, NSE, VILIP-1, and S100B play an important role in the pathogenesis of neuropsychiatric disorders, including epilepsy. This study aimed to assess the correspondence of the expression of BDNF, NSE, VILIP-1, and S100B in the blood (serum and peripheral blood mononuclear cells (PBMCs)) to the in vivo hippocampal levels of subjects with drug-resistant epilepsy who underwent neurosurgery (N = 44) using multiplex solid-phase analysis, ELISA, and immunohistochemical methods, as well as to analyze the correlations and associations of the blood and hippocampal levels of these proteins with clinical parameters. We first studied the concordance between in vivo brain and blood levels of BDNF, NSE, VILIP-1, and S100B in epileptic patients. A positive correlation for NSE between hippocampal and PBMC levels was revealed. NSE levels in PBMCs were also significantly correlated with average seizure duration. BDNF levels in PBMCs were associated with seizure frequency and hippocampal sclerosis. Thus, NSE and BDNF levels in PBMCs may have potential as clinically significant biomarkers. Significant correlations between the levels of the neurospecific proteins studied herein suggest interactions between BDNF, NSE, VILIP-1, and S100B in the pathophysiology of epilepsy.

Serum levels of cytokines in patients with retinal neurodegeneration in diabetic retinopathy on the background of type 2 diabetes mellitus

The aim of the present study was to determine the serum level of IL-1, TNF- IL-17A, INF-, IL-10 in patients with retinal neurodegeneration against the background of type 2 diabetes mellitus. The study included 80 patients with type 2 diabetes mellitus, which were divided into 4 groups depending on the presence of vascular symptoms of diabetic retinopathy and retinal neurodegeneration. The level of cytokines was determined using solid-phase ELISA. In groups of patients with retinal neurodegeneration, a significant (p0.05) increase in IL-1 and a decrease in IL-10 were detected. The presence of vascular changes in the retina did not affect the level of cytokines studied. The identified changes show the role of systemic imbalance between pro- and anti-inflammatory cytokines in the induction of retinal neurodegeneration in diabetic retinopathy. Further study of this problem is required in order to search for possible immune markers of retinal neurodegeneration in patients with type 2 diabetes.

Immunological indexes for assessing parameters of cardiovascular system in athletes of different qualification

Timely diagnosis and prevention of cardiovascular diseases is based on markers that detect changes in the athlete’s body at an early stage of disease. To implement this task, it is important to use novel laboratory techniques. We have carried out a comparative determination of immunological markers, specific antibodies to angiotensin, bradykinin, histamine, dopamine, serotonin and functional indicators of the cardiovascular system in athletes of various qualifications. The object of study included athletes of the Russian national teams who underwent an in-depth examination (IME) as part of medical survey. The participants were divided into groups depending on the sport arts and qualifications. Representatives of the group “Cyclic sports” included athletes without a category of 30 people and 29 qualified athletes (1st step, Candidate Master, Master of Sports, etc.). The “Combat sports” group consisted of 32 people without a category and 31 athletes with qualifications similar to those indicated above. Athletes of “Speed-strength” sports are represented by a group of 31 people without a category and 29 athletes with qualifications. The functional parameters of the cardiovascular system included analysis of heart rate, systolic blood pressure and diastolic blood pressure, which were compared with control values established for the athletes. The immunological indexes were determined in blood serum of the athletes and in subjects from control group by means of the solid-phase ELISA method using conjugated antigens of angiotensin, bradykinin, histamine, dopamine, serotonin for absorption on the plates. To compare the indexes, a control group of 30 people without cardiovascular symptoms was examined. Diastolic blood pressure for athletes of cyclic sports and combat sports was below the control values, and, for athletes of speed-strength sports, it was higher than in control group. A decrease in diastolic blood pressure for the above subgroup of athletes may indicate characteristic adaptive physiological changes in myocardium. The levels of immune-related indexes for serotonin, dopamine for the athletes of all groups corresponded to the control values. The only exception concerned qualified athletes of cyclic sports, which significantly differed from the normal values, compared to athletes without a sports category. Significantly high immunological parameters for histamine and angiotensin did not depend on the athlete’s qualification. In athletes involved in speed-strength sports, the levels of indexes for bradykinin did not differ from the normal values, and, for athletes in cyclic sports and combat sports, they significantly exceeded the norm.We have shown that different levels and types of sports activities affect the performance of cardiovascular system in their own way. The changes in immunological parameters reflect regulatory state of cardiovascular system. Accordingly, their simultaneous increase against the control may indicate participation in the development of cardiovascular diseases. The differences in their levels for athletes involved in combat sports, cyclic and speed-strength sporting activities show deeper changes in regulatory systems associated with duration and level of physical activities.

Physical exercise improves memory in sedentary middle-aged adults: Are these exercise-induced benefits associated with S-Klotho and 1,25-dihydroxivitamin D? The FIT-AGEING randomized controlled trial.

We aimed to investigate the effect of three types of exercise interventions on memory (i.e., immediate memory (IM), long-term memory (LTM), and recognition). We also investigated whether exercise-induced changes in circulating S-Klotho and 1,25-dihydroxivitamin D (1,25(OH)2 D) levels were related to those observed in memory in healthy middle-aged sedentary adults. A 12-week randomized controlled trial was performed with a parallel-group design. Seventy-four participants (45-65 years old: 53% women) were randomly assigned to (1) no exercise (control) group, (2) concurrent training based on the international physical activity recommendations (PAR) group, (3) high-intensity interval training (HIIT) group, or (4) HIIT plus whole-body electromyostimulation (HIIT-EMS) group. Memory outcomes were assessed using the Wechsler Memory Scale-third edition. S-Klotho plasma levels were determined according to a solid-phase sandwich enzyme-linked immunosorbent assay kit while 1,25(OH)2 D plasma levels were measured using a DiaSorin-Liaison immunochemiluminometric analyzer. IM-Verbal Paired Associates (IM-VPA) and IM-Logical Memory (IM-LM) were improved in both the HIIT and HIIT-EMS groups compared with the control group (all p ≤ 0.045). Exercise-induced changes in S-Klotho plasma levels were positively associated with those observed in IM, LTM, and recognition (all p ≤ 0.007), whereas exercise-induced changes in 1,25(OH)2 D plasma levels were directly related to changes in IM and LTM (all p ≤ 0.048). In conclusion, a 12-week HIIT intervention with or without WB-EMS seems to be the most effective exercise program to improve IM. The significant and positive associations between exercise-induced changes in S-Klotho and 1,25(OH)2 D levels with those observed in memory outcomes suggest that these factors may be potentially related to exercise-induced improvements of memory in middle-aged adults.

Value of Immunological Biomarkers in Early Prediction of Bacillus Calmette-Guerin Failure in High-Risk Non-muscle-invasive Bladder Cancer

Objectives To investigate the predictive value of different immunological markers on treatment outcomes after bacillus Calmette-Guerin (BCG) induction in high-risk non-muscle-invasive bladder cancer (NMIBC). Patients and Methods Patients who underwent transurethral resection of bladder tumors for NMIBC were assessed for study eligibility. Urine and blood samples were taken from patients at baseline (immediately before the first dose of induction). Urine samples were evaluated for interleukin (IL)-6, IL-8, IL-10, IL-11, and interferon- γ by solid-phase enzyme-linked immunosorbent assay (ELISA). Blood samples were evaluated for epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor-2 (HER2) using quantitative reverse transcriptase-polymerase chain reaction analysis. Each marker was assessed in relation to tumor recurrence. Results Between June 2016 and December 2019, 160 patients were included. Tumor recurrence occurred in 47 (29.38%) patients over a median (IQR) follow-up of 24 (12: 49) months. Using univariate analysis, the following urinary cytokines were associated with higher recurrence: urinary IL-6, 8, 10, 11, and interferon-γ. Also, serum EGFR and HER2 were associated with higher recurrence. On multivariate Cox regression analysis, significant variables include HER2 [HR (95%CI): 2.675 (1.367-5.233), p= 0.004], and IL-11 [HR (95%CI): 0.889 (0.825-0.957), p= 0.002]. Conclusions Serum HER2 and urinary IL-11 could be applied in clinical practice to predict BCG failure in patients with high-risk NMIBC, so those patients could be offered other modalities (radical cystectomy) early with better survival. Further studies are recommended to establish their exact role.

The effect of chronic stress on the immunogenicity and immunoprotection of the M6-TT vaccine in female mice

Active vaccination is an effective therapeutic option to reduce the reinforcing effects of opioids. Several studies showed that chronic stress affects the immune system decreasing the efficiency of some vaccines. Heroin withdrawal is a stressor and it is a stage in which the patient who abuses heroin is vulnerable to stress affects the immune response and consequently its immunoprotective capacity, then, the objective was to determine the effect of heroin-withdrawal and heroin-withdrawal plus immobilization, on the immune (immunogenicity) and protective response (behavioral response) of morphine-6-hemisuccinate-tetanus toxoid (M6-TT) vaccine in animals of two inbred mice strains with different sensitivity to drug-opioid and stress.Female BALB/c and C57Bl/6 inbred mice were immunized with the M6-TT. A solid-phase antibody-capture ELISA was used to monitor antibody titer responses after each booster dose in vaccinated animals. During the vaccination period, the animals were subjected to two different stress conditions: drug-withdrawal (DW) and immobilization (IMM). The study used tail-flick testing to evaluate the heroin-induced antinociceptive effects. Additionally, heroin-induced locomotor activity was evaluated. Stress decreased the heroin-specific antibody titer generated by the M6-TT vaccine in the two inbred mouse strains evaluated. In the two stress conditions, the antibody titer was not able to decrease the heroin-induced antinociceptive effects and locomotor activity. These findings suggest that stress decreases the production of antibodies and the immunoprotective capacity of the M6-TT vaccine. This observation is important to determine the efficacy of active vaccination as a potential therapy for patients with opioid drug use disorder, since these patients during drug-withdrawal present stress disorders, which could affect the efficacy of therapy such as active vaccination.

The 2023 ACR/EULAR Antiphospholipid Syndrome Classification Criteria.

To develop new antiphospholipid syndrome (APS) classification criteria with high specificity for use in observational studies and trials, jointly supported by the American College of Rheumatology (ACR) and EULAR. This international multidisciplinary initiative included 4 phases: 1) Phase I, criteria generation by surveys and literature review; 2) Phase II, criteria reduction by modified Delphi and nominal group technique exercises; 3) Phase III, criteria definition, further reduction with the guidance of real-world patient scenarios, and weighting via consensus-based multicriteria decision analysis, and threshold identification; and 4) Phase IV, validation using independent adjudicators' consensus as the gold standard. The 2023 ACR/EULAR APS classification criteria include an entry criterion of at least one positive antiphospholipid antibody (aPL) test within 3 years of identification of an aPL-associated clinical criterion, followed by additive weighted criteria (score range 1-7 points each) clustered into 6 clinical domains (macrovascular venous thromboembolism, macrovascular arterial thrombosis, microvascular, obstetric, cardiac valve, and hematologic) and 2 laboratory domains (lupus anticoagulant functional coagulation assays, and solid-phase enzyme-linked immunosorbent assays for IgG/IgM anticardiolipin and/or IgG/IgM anti-β2 -glycoprotein I antibodies). Patients accumulating at least 3 points each from the clinical and laboratory domains are classified as having APS. In the validation cohort, the new APS criteria versus the 2006 revised Sapporo classification criteria had a specificity of 99% versus 86%, and a sensitivity of 84% versus 99%. These new ACR/EULAR APS classification criteria were developed using rigorous methodology with multidisciplinary international input. Hierarchically clustered, weighted, and risk-stratified criteria reflect the current thinking about APS, providing high specificity and a strong foundation for future APS research.

Reduced levels of hepcidin associated with lower ferritin concentration and increased number of previous donations in periodic blood donors: a pilot study.

Hepcidin is one of the major negative regulators of iron balance. Periodic blood donors are highly susceptible to have iron deficiency. Our goal was to evaluate the possible association between serum hepcidin levels and iron homeostasis parameters in periodic blood donors. We enrolled a total of n=39 periodic healthy blood donors (n=24 M and n=15 F). A solid phase enzyme-linked immunosorbent assay (ELISA) was used to measure endogenous hepcidin-25 levels in serum biospecimens collected from each study participant. Statistical analysis was performed to evaluate possible associations between hepcidin levels and ferritin, transferrin, total iron binding capacity (TIBC), unsaturated iron binding capacity (UIBC), transferrin saturation (TSAT), and number of previous donations. Reduced serum hepcidin levels significantly correlated with reduced serum ferritin concentration (r = 0.56, IC95%: 0.51-0.60, p < 0.01). A multiple linear regression analysis showed that hepcidin levels were independently and negatively correlated with ferritin (p < 0.01). In addition, the number of previous blood donations was significantly associated with reduced hepcidin levels, independently of the other covariates (p < 0.01). Reduced serum hepcidin levels were significantly associated with reduced levels of ferritin and with increased number of previous donations suggesting its possible clinical role as non-invasive "point-of-care" in predicting iron deficiency among periodic blood donors.

Change in circulating klotho in response to weight loss, with and without exercise, in adults with overweight or obesity.

Introduction: Klotho is a protein associated with protection from aging-related diseases and health conditions. Obesity is associated with lower Klotho concentrations. Thus, this secondary analysis of adults with obesity examined 1) the change in serum Klotho concentration in response to a behavioral weight loss intervention by the magnitude of weight loss achieved; and 2) the association among serum Klotho concentration and weight, body composition, and cardiorespiratory fitness. Methods: Participants were randomized to either diet alone (DIET), diet plus 150min of physical activity per week (DIET + PA150), or diet plus 250min of physical activity per week (DIET + PA250). Participants [n = 152; age: 45.0 ± 7.9years; body mass index (BMI): 32.4 ± 3.8kg/m2] included in this secondary analysis provided blood samples at baseline, 6-, and 12months, and were classified by weight loss response (Responder: achieved ≥10% weight loss at 6 or 12months; Non-responder: achieved <5% weight loss at both 6 and 12months). Serum Klotho was measured using a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA). Analyses of covariance (ANCOVA's) were used to examine changes in weight, body composition, cardiorespiratory fitness, and Klotho concentration by weight loss response across the 12-month weight loss intervention. Results: Responders had a greater reduction in measures of weight and body composition, and a greater increase in cardiorespiratory fitness, compared to Non-Responders (p < 0.05). Change in Klotho concentration differed between Responders and Non-Responders (p < 0.05), with the increase in Klotho concentration from baseline to 6months for Responders being statistically significant. The 6-month change in Klotho concentration was inversely associated with the 6-month change in weight (r s = -0.195), BMI (r s = -0.196), fat mass (r s = -0.184), and waist circumference (r s = -0.218) (p-values <0.05). Discussion: Findings provide evidence within the context of a behavioral intervention, with and without exercise, that change in Klotho concentration is significantly different between adults with weight loss ≥10% compared to <5% across 12months. These findings suggest that weight loss and reduction in fat mass may be favorably associated with the change in Klotho concentration. This may reduce the risk of negative health consequences associated with accelerated aging in middle-aged adults.