Enzyme-linked Immunosorbent Assay Kit Research Articles

Umbilical cord mesenchymal stem cells in ulcerative colitis treatment: efficacy and possible mechanisms

BackgroundMesenchymal stem cells (MSCs) possess powerful immunomodulatory ability. This study aimed to assess the efficacy and safety of human umbilical cord-derived mesenchymal stem cells (UMSCs) in patients with ulcerative colitis (UC) and to explore the potential mechanisms.MethodsThis prospective, self-controlled clinical study was conducted at Henan Provincial People’s Hospital. Patients with moderate-to-severe active UC, unresponsive to traditional drugs were continuously enrolled from September 2018 to March 2023. UMSCs were administered intravenously monthly for two months at a cell dosage of 1 × 106 per kg. The primary outcome was a clinical response at 2 months. The levels of cytokines and progerin in the plasma of the patients were analyzed using enzyme-linked immunosorbent assay kits, and longitudinal data was analyzed using generalized estimation equation.ResultsForty-one patients were enrolled and received UMSC therapy. At 2 months, 73.2% (30/41) of patients achieved a clinical response, and 41.5% (17/41) achieved a clinical remission. At 6 months, 2 patients were lost to follow-up; the corresponding figures were 70.0% (25/41) and 34.2% (14/41), respectively. After UMSC therapy, the Mayo score, Mayo endoscopy score, mean and maximum values of Ulcerative Colitis Endoscopic Index of Severity and Nancy index were significantly reduced compared with baseline values. Additionally, the levels of progerin and inflammatory markers, such as interleukin (IL)-1β, IL-6, IL-8, IL-12, and IL-17 A decreased, while hemoglobin, albumin, and IL-10/IL-17 A ratio increased, particularly in the response group. Multiple stepwise logistic regression analysis showed age was an independent risk factor affecting efficacy (odds ratio, 0.875 (95% confidence interval (0.787, 0.972)); the area under the receiver operating characteristic curve for age was 0.79. No serious adverse events were observed during or after UMSC therapy.ConclusionUMSCs are safe and effective for patients with UC, with age being an independent risk factor affecting efficacy. Mechanistically, UMSC treatment may ameliorate cell senescence and suppress the secretion of pro-inflammatory cytokines.Trial registrationThe study was retrospectively registered at www.chictr.org.cn/ (ChiCTR1900026035) on September 18, 2019.

Evaluation serum soluble interleukin 2 receptor with diagnosis and prognosis in canine solid tumour: 34 cases.

The soluble interleukin-2 receptor (sIL-2R) serve as a valuable biomarker for tumors in human patients, as its levels increase during the activation of T lymphocytes in clinical states such as inflammation, infection, and tumor. This study aimed to demonstrate that sIL-2R levels can be also elevated in dogs with tumors and evaluate its applicability as a diagnostic and prognostic factor in canine cancer patients. Serum was collected from 6 healthy dogs and 34 dogs with solid tumors. The concentration of sIL-2R was measured using a commercial enzyme-linked immunosorbent assay kit. The median sIL-2R concentration was significantly higher in dogs with solid masses than in healthy dogs (117.3 vs 68.33 pg/ml, p=0.016). The highest median sIL-2R concentration was found in dogs with malignant tumors, followed by those with benign tumors, and healthy dogs (119.6 vs 93.74 vs 68.33 pg/ml, respectively). In dogs with malignant tumors, the mortality rate was significantly higher in the group with high sIL-2R levels than in the group with low sIL-2R levels. Dogs with solid tumors, particularly those with malignant tumors, had higher concentrations of sIL-2R than healthy dogs. Among dogs with malignant tumors, a correlation between sIL-2R concentration and mortality rate was confirmed. Serum sIL-2R levels may be used to detect malignant tumors and serve as a prognostic factor in dogs with malignant tumors.

Sex-related differences in cognition and its correlation with lipid parameters and sex steroids among community-dwelling older persons in Nigeria

Abstract Introduction: Studies on sex-related differences in cognition and its correlation with serum lipids and sex steroids in older persons in sub-Saharan Africa (SSA) are few and inconsistent. Our objectives were to assess: (i) sex-related differences in cognition and (ii) the correlation of cognition with lipid parameters, age, years of formal education, and sex steroids in community-dwelling older persons in Kano state, Nigeria. Materials and Methods: About 143 eligible participants were randomly selected. We used the Montreal Cognitive Assessment-Basic (MoCA-B), verbal fluency test, 10-Item Word List Learning Test (10-IWLLT), delayed word list recall, and Stick Design Test (SDT) to assess cognitive function. Serum lipids were determined using chemical colorimetry, whereas sex steroids were determined using competitive enzyme-linked immunosorbent assay kits. Results: The results revealed that the participants had a mean age of 66 years. The mean MoCA-B score was 18.84 (males = 20.26, females = 17.73, P = 0.001). Males had better scores in orientation (P = 0.001), abstraction (P = 0.009), naming (P = 0.001), verbal fluency (P = 0.013), delayed word list recall score (P = 0.001), visuo-construction (P = 0.015), and visuo-perception (P = 0.003). Among whole participants, there was a positive correlation between total cholesterol and MoCA-B (r = 0.236, P = 0.005) and between low-density lipoprotein-cholesterol and MoCA-B (r = 0.207, P = 0.013). Serum testosterone had positive correlation with MoCA-B score (r = 0.212, P = 0.001) but negative correlation with delayed word list recall (r = −0.218, P = 0.009), and SDT (r = −0.181, P = 0.031); dihydrotestosterone had positive correlation with MoCA-B score (r = 0.210, P = 0.012) but negative correlation with delayed word list recall (r = −0.243, P = 0.003) and SDT (r = −0.183, P = 0.028). Estradiol had a negative correlation with the 10-IWLLT (r = −0.169, P = 0.044). Conclusion: This study concludes that males had better cognition than females. Higher levels of serum lipids and sex steroids are related to better cognition in community-dwelling older persons in Nigeria.

Magnoflorine attenuates Ang II-induced cardiac remodeling via promoting AMPK-regulated autophagy.

Heart failure (HF) remains one of the most common events in the progression of hypertension. Magnoflorine (MNF) has been shown beneficial effects on the cardiovascular system. However, the action of MNF on angiotensin (Ang) II-induced cardiac remodeling and its underlying mechanisms have not yet been characterised. Here, we assessed the action of MNF in the development of hypertension-related HF. C57BL/6 male mice were subjected to Ang II through a micro-osmotic pump infusion continuously for 4 weeks to induce hypertensive HF. MNF (10 and 20 mg/kg) was administered in the final 2 weeks. Ang II content was measured by enzyme-linked immunosorbent assay (ELISA) kit. Values of ejection fraction (EF) and fractional shortening (FS) were detected using an ultrasound diagnostic instrument. The mRNA levels of hypertrophic and fibrotic genes were determined by real-time quantitative polymerase chain reaction (RT-qPCR). Haematoxylin and eosin (H&E), wheat germ agglutinin (WGA), Masson trichrome, and Sirius Red staining were used to analyse pathologic changes in heart tissues. The expression levels of phosphorylated AMP-activated protein kinase (AMPK), light chain 3 microtubule associated protein II (LC3 II) to LC3 I, and p62 were detected by western blot assay. MNF significantly improved cardiac dysfunction and the content of creatine kinase-MB without altering blood pressure in Ang II-challenged mice. MNF obviously corrected the phenotypes of cardiac hypertrophy and fibrosis, including the high mRNA levels of atrial natriuretic peptide (Anp), brain natriuretic peptide (Bnp), collagen1a (Col1a1), transforming growth factor beta (Tgfb1), enlarged myocardial areas, and increased positive areas of Masson trichrome and Sirius Red staining. In addition, MNF alleviated oxidative injury, reflected by the upregulation of glutathione and the downregulation of reactive oxygen species and malondialdehyde. The activation of AMPK was elevated accompanied by an increased level of autophagy by MNF in hypertensive heart tissues. The therapeutic action of MNF was confirmed in Ang II-challenged H9c2 cells. Specifically, the AMPK inhibitor could eliminate the autophagy pathway in which MNF is involved. MNF has benefits in hypertension-induced cardiac remodeling, which was partially associated with the improvement of oxidative stress via the mediation of the AMPK/autophagy axis.

Acute Neuroinflammation Alters the Transport of a Model Therapeutic Protein from the Brain into Lymph and Blood.

The drainage of fluid and solutes along lymphatic pathways from the brain has been found to be impaired in mouse models of multiple sclerosis, Alzheimer's disease, and Parkinson's disease where neuroinflammation is present. We recently demonstrated that 3H-albumin, a model therapeutic protein (∼65 kDa), undergoes preferential lymphatic transport from the brain using a cervical lymph cannulation model in healthy rats. We thus hypothesized that neuroinflammation would impede the lymphatic transport of 3H-albumin from the brain. Our aim was to quantify the impact of acute neuroinflammation on drainage of the model therapeutic protein (3H-albumin) from the rat brain into blood and deep cervical lymph. To establish the required neuroinflammation model, male Sprague-Dawley rats were administered an intraperitoneal (IP) dose of 0.5-2 mg/kg lipopolysaccharide (LPS, Escherichia coli) or a saline control. After 12 or 24 h, brain samples were collected and analyzed for concentrations of interferon gamma (IFN-γ) using a commercial enzyme-linked immunosorbent assay (ELISA) kit. The impact of neuroinflammation on the drainage of 3H-albumin from the brain was determined via IP administration of 2 mg/kg LPS or saline followed by cannulation of the carotid artery for blood collection 24 h later with/without cannulation or ligation at the efferent deep cervical lymph trunk. Rats were then administered 3H-albumin via direct injection into the brain striatum or via intravenous (IV) injection (lymph-intact group only). Blood ± lymph samples were collected for up to 8 h following dosing. At the end of the study, brain and lymph node samples were harvested for biodistribution analysis, with samples analyzed for radioactivity levels via scintillation counting. Brain concentrations of the pro-inflammatory cytokine IFN-γ were only significantly elevated 24 h after IP administration of 2 mg/kg LPS compared to saline control. Therefore, this induction regimen was utilized for subsequent studies. The plasma concentrations of 3H-albumin over time were elevated in LPS-induced rats compared to saline-injected rats in the lymph-intact and lymph-ligated groups but not in the lymph-cannulated group. In the deep cervical lymph-cannulated animals, the lymph transport of 3H-albumin was not increased and appeared to be slower in the LPS-administered rats. Acute LPS-induced neuroinflammation therefore led to an enhanced overall transport of 3H-albumin from the brain into the systemic circulation. This appeared to be primarily due to increased transport of 3H-albumin from the brain directly into the blood circulation as 3H-albumin transport from the brain via the lymphatics was not increased in the LPS-induced neuroinflammation model. Such changes in the clearance of therapeutic proteins from the brain in the setting of neuroinflammation may impact the therapeutic efficacy and safety.

Evaluation of neutrophil gelatinase-associated lipocalin (NGAL), hypoxia-induced factor-1 alpha (HIF-1α) and apelin 13 levels as new potential biomarkers for pulmonary thromboembolism: A prospective clinical study

AimThe objective was to evaluate the serum levels of neutrophil gelatinase-associated lipocalin (NGAL), hypoxia-induced factor-1 alpha (HIF-1α), and apelin 13 in patients with acute pulmonary thromboembolism (PE) and to investigate their diagnostic and prognostic role in PE patients with different mortality risk groups. Material and methodsThis study was conducted in a tertiary referral center and included 124 subjects with 94 cases of PE and 30 cases of healthy control group. All subjects were 18 years of age or older. The diagnosis of PE was done with computed tomography angiography of the thorax. After the diagnosis of acute PE, the serum levels of neutrophil gelatinase-associated lipocalin (NGAL), hypoxia-induced factor-1 alpha (HIF-1α), and apelin 13 levels were measured with a commercial enzyme-linked immunosorbent assay (ELISA) kit. ResultsThe median and IQR (interquartile range) age of patients and control groups were 68 (56–76) and 61.5 (56–67) years, respectively. The majority of patients with PE had risk factors (97.88 %), and only two (2.12 %) had no known risk factors. HIF-1 alpha level was found to be higher in the patient group than in the control group (p = 0.03). At the same time, the HIF-1 alpha level was found to be higher in the high mortality risk group than in the control group, low mortality risk group and intermediate-low mortality risk group (p = 0.000, 0.011, 0.002, respectively). While there was no significant difference in NGAL level between the patient group and the control group, a significant difference was observed between the mortality groups. NGAL level was found to be higher in the high mortality risk group than the control group, low mortality risk group, and medium-low mortality risk group (p = 0.001, 0.000, 0.010, respectively). Apelin 13 levels did not differ significantly in all groups. ConclusionHIF-1 alpha is a promising biomarker in distinguishing between patients and control groups and in identifying those with high mortality risk in the patient group. At the same time, NGAL can be used as a successful biomarker in determining the group with high mortality risk in cases of PE.

Co-Occurrence of Cyanotoxins and Phycotoxins in One of the Largest Southeast Asian Brackish Waterbodies: A Preliminary Study at the Tam Giang—Cau Hai Lagoon (Vietnam)

The Tam Giang-Cau Hai lagoon (TGCH) in Thua Thien Hue province (Vietnam) is a marsh/lagoon system and ranks among the largest waterbodies in Southeast Asia. It plays a significant role in terms of both socio-economic and environmental resources. However, anthropogenic stress, as well as the discharge of untreated domestic and industrial sewage with agricultural runoff from its three major tributaries, dramatically damages the water quality of the lagoon. Especially after heavy rain and flash floods, the continuous degradation of its water quality, followed by harmful algal and cyanobacterial bloom patterns (HABs), is more perceptible. In this study, several physicochemical factors, cyanotoxins (anatoxins (ATXs), saxitoxins (STXs), microcystins (MCs)), phycotoxins (STXs, okadaic acid (OA), and dinophysistoxins (DTXs)) were analyzed in water and shellfish samples from 13 stations in June 2023 from 13 stations, using enzyme-linked immunosorbent assay (ELISA) kits for the ATXs and STXs, and the serine/threonine phosphatase type 2A (PP2A) inhibition assay kit for the MCs, OA, and DTXs. The results showed for the first time the co-occurrence of freshwater cyanotoxins and marine phycotoxins in water and shellfish samples in this lagoon. Traces of ATXs and STXs were detected in the shellfish and the orders of magnitude were below the seafood safety action levels. However, toxins inhibiting the PP2A enzyme, such as MCs and nodularin (NODs), as well as OA and DTXs, were detected at higher concentrations (maximum: 130.4 μg equiv. MC-LR/kg shellfish meat wet weight), approaching the actionable level proposed for this class of toxin in shellfish (160 μg of OA equivalent per kg of edible bivalve mollusk meat). It is very important to note that due to the possible false positives produced by the ELISA test in complex matrices such as a crude shellfish extract, this preliminary and pilot research will be repeated with a more sophisticated method, such as liquid chromatography coupled with mass spectroscopy (LC-MS), in the upcoming research plan.

Activation of NLRP3 inflammasome in patients with renal transplantation: relation to allograft dysfunction, inflammation, and renal fibrosis

ABSTRACT Background Chronic allograft dysfunction (CAD) represents a complex process of inflammatory response, fibrosis, and tissue restoration, mediated by a myriad of proinflammatory cytokines, enzymes, and growth factors. NOD-like receptor protein3 (NLRP3) inflammasome is one of the NLR protein families. It drives proteolysis of proinflammatory caspases which induces maturation of pro-interleukin (IL)-1β and pro-IL-18. The aim of this work is to study NLRP3 inflammasome activation in patients with renal transplantation in relation to allograft function, inflammation, and renal fibrosis. Methods Thirty renal transplant recipients (RTR): 15 patients with stable renal function (Group I), 15 patients with CAD (Group II), and 15 healthy subjects (Group III) were included. Serum and urinary NLRP3, caspase-1, and interleukin-1β were measured using an enzyme-linked immunosorbent assay (ELISA) kit. Serum high sensitivity C-reactive protein (hs-CRP) level was measured, and neutrophil-lymphocyte ratio (NLR) was calculated. Renal function was evaluated by serum creatinine (S.Cr) and estimated glomerular filtration rate (eGFR). Resistive index (RI) was calculated with Doppler ultrasonography. In patients with CAD, renal interstitial fibrosis (RIF) was graded in renal biopsies. Results Serum NLRP3, caspase-1, serum and urinary IL-1β, and hs-CRP showed a statistically significant increase in RTR, particularly in those with CAD compared to the controls with a positive correlation with each other, their urinary level, S. Cr, hs-CRP, NLR, RI, and RIF (p < 0.001). The receiver operating characteristic curve showed high diagnostic accuracy of serum NLRP3, caspase-1, and IL-1β in discriminating patients with CAD from patients with stable renal function. Conclusion The NLRP3-caspase1-IL-1β cascade is activated and upregulated in renal transplant recipients, particularly those with advanced renal impairment, and thus may play a role in developing CAD and renal fibrosis. They could be potential biomarkers for post-transplant outcome, follow-up, and early diagnosis. Moreover, their cascade could be a target for future anti-inflammatory therapeutic strategies for allograft dysfunction.

Sero-prevalence of Toxoplasma gondii before and during the COVID-19 pandemic in Northwestern Iran

BackgroundToxoplasma gondii (T. gondii) is a ubiquitous protozoan parasite on our planet that causes toxoplasmosis. This study evaluated the seroprevalence and related risk factors for T. gondii infection in a population referred to healthcare centers in Meshkin-Shahr, Northwest Iran.MethodsA total of 400 blood samples were randomly collected from the general population and assessed using the anti-Toxoplasma antibodies, Immunoglobulin G and M (IgG and IgM) Enzyme-linked immunosorbent assay (ELISA) Kits in two steps before and during the coronavirus disease 2019 (COVID-19) pandemic, 2019–2020. The results were analyzed through logistic regression via SPSS 26 software.ResultsBefore the COVID-19 pandemic, anti-toxoplasma antibodies were detected in 39% of individuals (IgG: 38%, IgM: 0.5%, and IgG-IgM: 0.5%). Among the eleven risk factors evaluated, contact with soil and people awareness were significantly associated with T. gondii infection (p < 0.05). However, factors such as females, 20–39 age groups, junior high schools, housewives, rural areas, raw meat or vegetable consumption, vegetable or fruits washed by water, not detergent, and cat owners did not show a significant relationship with seropositivity (p > 0.05). After the outbreak of the COVID-19 pandemic, the overall seroprevalence for anti-T. gondii antibody increased to 49.7% (IgG: 47.7%, IgM: 0.5%, and IgG and IgM: 1.5%). Among these patients, 26% were positive for COVID-19. Additionally, before the COVID-19 pandemic, 40 samples were negative for anti-T. gondii antibodies but later became positive. The crude and adjusted models suggested that toxoplasmosis may be a possible risk factor for increased susceptibility to COVID-19, with an odds ratio (OR) of 1.28 (95% confidence interval (CI), 0.82–1.99; P < 0.05). Conversely, a non-significant protective effect against latent toxoplasmosis was observed in COVID-19-positive individuals (OR = 0.99; 95% CI, 0.51–1.92; P > 0.05), and COVID-19 positivity did not increase the levels of anti-T. gondii IgG antibodies.ConclusionsThe general population in this region had a moderate seroprevalence of T. gondii. The increased number of COVID-19-positive patients with latent toxoplasmosis highlights the need to pay attention to the early diagnosis and proper treatment of toxoplasmosis in these patients and implement preventive programs in these areas for future possible viral infections.

Changes in Circulating Adipokine Levels in COVID-19 Patients.

Objective: The COVID-19 pandemic has posed significant global health challenges. Despite extensive research efforts, the inflammatory response triggered by SARS-CoV-2 remains to be further explored and understood. Our study aims to examine the changes in serum concentrations of pro-inflammatory adipokines-visfatin and leptin-in COVID-19 patients in relation to a healthy control group. Patients/Materials/Subjects and Methods: The study consisted of forty COVID-19 patients and twenty-four healthy patients in the control group. Two serum samples were collected: upon admission and on the seventh day of hospitalization. Concentrations of visfatin and leptin in the serum, alongside routine biochemical parameters, were measured using enzyme immunoassay or enzyme-linked immunosorbent assay kits. The Shapiro-Wilk test was used to assess normality. Differences between independent groups were compared using the Mann-Whitney U test and Kruskal-Wallis ANOVA. Correlations were evaluated with Spearman's rank correlation coefficient. Results: Our findings revealed significantly lower visfatin levels in COVID-19 patients compared to the control group upon admission (4.29 ng/mL, (3.0-6.88 ng/mL) vs. 37.16 ng/mL (24.74-50.12 ng/mL), p < 0.001 for visfatin 1 and 52.05 ng/mL, (31.2-69.66 ng/mL) vs. 37.16 ng/mL (24.74-50.12 ng/mL), p = 0.048 for visfatin 2). The visfatin level of COVID-19 patients returned to the normal levels, established in the control group. However, there was no significant difference in leptin levels between the two groups (p = 0.270 for leptin 1 and p = 0.129 for leptin 2). There was a positive correlation between BMI and leptin concentration (r = 0.66 and p = 0.00). Moreover, it was discovered that COVID-19 independently reduces visfatin levels during the first day of illness. Conclusions: The results of our research suggest that the onset of COVID-19 infection is correlated to visfatin levels. Association with leptin levels remains inconclusive. Further research is imperative to elucidate the intricate role of visfatin and leptin in SARS-CoV-2 infection and their potential as biomarkers for COVID-19 severity and prognosis.

Role of cyclophilin A in aggravation of myocardial ischemia reperfusion injury via regulation of apoptosis mediated by thioredoxin-interacting protein.

The progression and persistence of myocardial ischemia/reperfusion injury (MI/RI) are strongly linked to local inflammatory responses and oxidative stress. Cyclophilin A (CypA), a pro-inflammatory factor, is involved in various cardiovascular diseases. However, the role and mechanism of action of CypA in MI/RI are still not fully understood. We used the Gene Expression Omnibus (GEO) database for bioinformatic analysis. We collected blood samples from patients and controls for detecting the levels of serum CypA using enzyme-linked immunosorbent assay (ELISA) kits. We then developed a myocardial ischemia/reperfusion (I/R) injury model in wild-type (WT) mice and Ppia-/- mice. We utilized echocardiography, hemodynamic measurements, hematoxylin and eosin (H&E) staining, immunohistochemistry, enzyme-linked immunosorbent assay, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining to determine the role of CypA in myocardial I/R injury. Finally, we conducted an in vitrostudy, cell transfection, flow cytometry, RNA interference, and a co-immunoprecipitation assay to clarify the mechanism of CypA in aggravating cardiomyocyte apoptosis. We found that CypA inhibited TXNIP degradation to enhance oxidative stress-induced cardiomyocyte apoptosis during MI/RI. By comparing and analyzing CypA expression in patients with coronary atherosclerotic heart disease and in healthy controls, we found that CypA was upregulated in patients with Coronary Atmospheric Heart Disease, and its expression was positively correlated with Gensini scores. In addition, CypA deficiency decreased cytokine expression, oxidative stress, and cardiomyocyte apoptosis in I/R-treated mice, eventually alleviating cardiac dysfunction. CypA knockdown also reduced H2O2-induced apoptosis in H9c2 cells. Mechanistically, we found that CypA inhibited K48-linked ubiquitination mediated by atrophin-interacting protein 4 (AIP4) and proteasomal degradation of TXNIP, a thioredoxin-binding protein that mediates oxidative stress and induces apoptosis. These findings highlight the critical role CypA plays in myocardial injury caused by oxidative stress-induced apoptosis, indicating that CypA can be a viable biomarker and a therapeutic target candidate for MI/RI.

Next-Generation Electrochemical Biosensing Based on Ternary Nanocomposites of nMoS2@Rgo@MWCNT for Label-Free Cancer Biomarker Detection in Undiluted Human Serum

Sperm protein-17 (Sp17) is a cancer-testicular antigen that is expressed in various types of cancers and has been proposed as a potential biomarker for cancer. However, the detection of Sp17 in human serum is challenging due to its low concentration and the lack of specific and sensitive methods. Hence, there is an urgent need to develop fast, accurate, reliable, and cost-effective disposable biosensors for Sp17 detection. Herein, we report sensing of Sp17 with a disposable carbon cloth-based electrode (CC) modified with a ternary nanocomposite comprising molybdenum disulphide (MoS2), reduced graphene oxide (rGO), and multi-walled carbon nanotube (MWCNT). The nanocomposite was synthesized by a facile hydrothermal method and was well-characterized in terms of structural, morphological, and chemical attributes using X-ray diffraction, transmission electron microscopy, scanning electron microscopy, Raman spectroscopy spectral techniques, and electrochemical studies to prove the robust formation of the electroactive ternary nanocomposite and its suitability for Sp17 detection. The nanocomposite exhibited excellent electrocatalytic activity, biocompatibility, and stability. We immobilized a Sp17-specific antibody on the surface of the nanocomposite-modified electrode and used it as a biosensor for Sp17 detection. The biosensor showed a linear response range from 0.01 to 11 ng mL-1 and a limit of detection (LOD) and limit of quantification (LOQ) of 0.23 ng mL-1 and 0.79 ng mL-1, respectively having sensitivity of 9.97 mA ng mL-1 cm-2. The biosensor was successfully applied to detect Sp17 in human serum samples and showed good accuracy, reproducibility, and selectivity, making it helpful for use in translational medicine research and clinical applications. The biosensor also demonstrated a high correlation with a commercial enzyme-linked immunosorbent assay (ELISA) kit. Our results suggest that the proposed biosensor is a promising tool for Sp17 detection in human serum and could be useful for the diagnosis and prognosis of ovarian cancer and other Sp17-related cancers. Keywords: Carbon cloth; Biosensor; Electrochemistry; Sperm protein 17; Cancer biomarker; Nanocomposite

Estimating the Seroincidence of Scrub Typhus using Antibody Dynamics after Infection.

Scrub typhus, a vector-borne bacterial infection, is an important but neglected disease globally. Accurately characterizing the burden is challenging because of nonspecific symptoms and limited diagnostics. Prior seroepidemiology studies have struggled to find consensus cutoffs that permit comparisons of estimates across contexts and time. In this study, we present a novel approach that does not require a cutoff and instead uses information about antibody kinetics after infection to estimate seroincidence. We use data from three cohorts of scrub typhus patients in Chiang Rai, Thailand, and Vellore, India, to characterize antibody kinetics after infection and two population serosurveys in the Kathmandu Valley, Nepal, and Tamil Nadu, India, to estimate seroincidence. The samples were tested for IgM and IgG responses to Orientia tsutsugamushi-derived recombinant 56-kDa antigen using commercial enzyme-linked immunosorbent assay kits. We used Bayesian hierarchical models to characterize antibody responses after scrub typhus infection and used the joint distributions of the peak antibody titers and decay rates to estimate population-level incidence rates in the cross-sectional serosurveys. Median responses persisted above an optical density (OD) of 1.8 for 23.6 months for IgG and an OD of 1 for 4.5 months for IgM. Among 18- to 29-year-olds, the seroincidence was 10 per 1,000 person-years (95% CI, 5-19) in Tamil Nadu, India, and 14 per 1,000 person-years (95% CI: 10-20) in the Kathmandu Valley, Nepal. When seroincidence was calculated with antibody decay ignored, the disease burden was underestimated by more than 50%. The approach can be deployed prospectively, coupled with existing serosurveys, or leverage banked samples to efficiently generate scrub typhus seroincidence estimates.

P-Hydroxy benzaldehyde, a phenolic compound from Nostoc commune, ameliorates DSS-induced colitis against oxidative stress via the Nrf2/HO-1/NQO-1/NF-κB/AP-1 pathway

BackgroundUlcerative colitis (UC), a chronic idiopathic inflammatory bowel disease (IBD), presents with limited current drug treatment options. Consequently, the search for safe and effective drug for UC prevention and treatment is imperative. Our prior studies have demonstrated that the phenolic compound p-Hydroxybenzaldehyde (HD) from Nostoc commune, effectively mitigates intestinal inflammation. However, the mechanisms underlying HD's anti-inflammatory effects remain unclear. PurposeThis study delved into the pharmacodynamics of HD and its underlying anti-inflammation mechanisms. MethodsFor in vivo experiments, dextran sodium sulfate (DSS)-induced colitis mouse model was established. In vitro inflammation model was established using lipopolysaccharide (LPS)-induced RAW264.7 and bone marrow-derived macrophages (BMDMs). The protective effect of HD against colitis was determined by monitoring clinical symptoms and histological morphology in mice. The levels of inflammatory factors and oxidative stress markers were subsequently analyzed with enzyme-linked immunosorbent assay (ELISA) and biochemical kits. Furthermore, western blotting (WB), immunofluorescence (IF), luciferase reporter gene, drug affinity reaction target stability (DARTS) assay, molecular docking, and molecular dynamics (MD) simulation were used to determine the potential target and molecular mechanism of HD. ResultsOur findings indicate that HD significantly alleviated the clinical symptoms and histological morphology of colitis in mice, and curtailed the production of pro-inflammatory cytokines, including TNF-α, IL-6, IFN-γ, COX-2, and iNOS. Furthermore, HD stimulated the production of SOD, CAT, and GSH-px, enhanced total antioxidant capacity (T-AOC), and reduced MDA levels. Mechanically, HD augmented the expression of Nrf2, HO-1, and NQO-1, while concurrently downregulating the phosphorylation of p65, IκBα, c-Jun, and c-Fos. ML385 and siNrf2 largely attenuated the protective effect of HD in enteritis mice and RAW 264.7 cells, as well as the promotion of HO-1 expression levels. ZnPP-mediated HO-1 knockdown reversed HD-induced inhibition of colonic inflammation. Luciferase reporter assay and IF assay confirmed the transcriptional activation of Nrf2 by HD. DARTS analysis, molecular docking, and MD results showed high binding strength, interaction efficiency and remarkable stability between Nrf2 and HD. ConclusionThese outcomes extend our previous research results that HD can combat oxidative stress through the Nrf2/HO-1/NQO-1/NF-κB/AP-1 pathways, effectively alleviating colitis, and propose new targets for HD to protect against intestinal barrier damage.

Comparison of in-house enzyme-linked immunosorbent assay (ELISA) and six commercial ELISAs for Detection of antibodies to Bordetella pertussis

Enzyme-linked immunosorbent assays (ELISA) are currently the method of choice for the serodiagnosis of pertussis and play a key role in the diagnosis of pertussis in adolescents and adults, as well as in epidemiological studies. In the present study, the in-house developed indirect ELISA was comparatively evaluated with six commercial kits from various manufacturers. Antipertussis antibodies were measured in 40 serum samples from patients with clinical symptoms of respiratory tract infection, in two WHO standards, and in seven human ECDC control sera. IgA and IgG antibodies were detected at a diagnostically significant level by different ELISA kits of 5.0% to 27.0% and 12.0% to 70.0% of patients' sera, appropriately. The analysis of results carried out with six commercial kits showed only 17.5% consistent results in class IgG (either clearly positive or negative). The average percentage of errors in the level of antibodies determined in the control samples, reference serum samples, differed quite significantly and ranged from 9.5% to 35.4% depending on the kit. This poor correlation of the results obtained on various serological tests intended for the serodiagnosis of pertussis may cause very serious diagnostic problems, especially when examining a serum sample obtained once during the course of the disease.

Value of serum muscular creatinine phosphokinase levels in patients with adenomyosis as a non-invasive diagnostic marker.

In the present study, we aimed to compare serum CK-MM levels in patients with and without adenomyosis and to investigate whether CK-MM level can be a non-invasive marker for myometrial damage due to adenomyosis. Our study was a prospective case-control study in a tertiary center and consisted of 40 patients with a clinical/ultrasonographic diagnosis of adenomyosis and 40 patients without a clinical/ultrasonographic diagnosis of adenomyosis as the control group based on recently published morphological uterus sonographic assessment (MUSA) criteria. Individuals of similar age who signed a voluntary consent form were included in our study. Demographic, clinical, and laboratory findings of the patients in both groups were recorded. Blood serum samples were used for the determination of serum CK-MM levels of the participants in our study. The samples were analyzed by using the human CK-MM enzyme-linked immunosorbent assay (ELISA) kit. In our study, the mean serum CK-MM level was 16.2 ± 21.7 (ng/dL) in patients with adenomyosis and 2.6 ± 2.2 (ng/dL) in patients without adenomyosis. Serum CK-MM level was statistically significantly higher in the patient group with adenomyosis than in the control group (p < 0.001). The CK-MM threshold value of 3.43 ng/mL, with a sensitivity of 82.5% and specificity of 85%, has been found to be a valuable distinguishing level in patients with and without adenomyosis. In this study, we demonstrated that serum CK-MM can be used as a non-invasive diagnostic method in patients with adenomyosis. As the number of studies around this subject in the literature is insufficient, larger studies are needed to use CK-MM as a diagnostic marker in adenomyosis.

Differences in T cell-associated serum markers between ischemic cardiomyopathy and dilated cardiomyopathy.

Ischemic cardiomyopathy (ICM) and dilated cardiomyopathy (DCM) have similar clinical manifestations but differ in pathogenesis. We aimed to identify T cell-associated serum markers that can be used to distinguish between ICM and DCM. We identified differentially expressed genes (DEGs) with transcriptome sequencing data in GSE116250, and then conducted enrichment analysis of DEGs in the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Protein-protein interaction (PPI) networks were used to analyze the relationship between T cells-related genes and identify hub genes. Enzyme-linked immunosorbent assay (ELISA) kits were used to detect T cell-associated proteins in serum, and receiver operating characteristic (ROC) curves were used to evaluate the diagnostic efficacy of these serum markers. Using the limma package and Venn plots, we found that the non-failing donors (NFD) and DCM groups shared many of the same DEGs and DEGs-enriched functions compared to the ICM group, which were involved in T cell activation and differentiation, among other functions. Subsequently, the immune cell score showed no difference between NFD and DCM, but they were significantly different from ICM patients in CD8 T cells CD4 T cells memory resting and activated, T cells follicular helper, and M1 macrophage. After analyzing T cell-associated DEGs, it was found that 4 DEGs encoding secreted proteins were highly expressed in the ICM group compared with the NFD and DCM groups, namely chemokine (C-C motif) ligand 21 (CCL21), interleukin (IL)-1β, lymphocyte-activation gene 3 (LAG3), and vascular cell adhesion molecule-1 (VCAM-1). Importantly, the serum levels of CCL21, IL-1β, LAG3, and VCAM-1 in ICM patients were all significantly higher than those in DCM patients. The ROC curves showed that the area under the curve (AUC) values of serum CCL21, IL-1β, LAG3, and VCAM-1 were 0.775, 0.868, 0.934, and 0.903, respectively. We have identified four T cell-associated serum markers, CCL21, IL-1β, LAG3, and VCAM-1, as potential diagnostic serum markers that differentiate ICM from DCM.

Super-resolution Multispectral Imaging Nanoimmunosensor for Simultaneous Detection of Diverse Early Cancer Biomarkers.

In medical diagnosis, relying on only one type of biomarker is insufficient to accurately identify cancer. Blood-based multicancer early detection can help identify more than one type of cancer from a single blood sample. In this study, a super-resolution multispectral imaging nanoimmunosensor (srMINI) based on three quantum dots (QDs) of different color conjugated with streptavidin was developed for the simultaneous screening of various cancer biomarkers in blood at the single-molecule level. In the experiment, the srMINI chip was used to simultaneously detect three key cancer biomarkers: carcinoembryonic antigen (CEA), C-reactive protein (CRP), and alpha-fetoprotein (AFP). The srMINI chip exhibited 108 times higher detection sensitivity of 0.18-0.5 ag/mL (1.1-2.6 zM) for these cancer biomarkers than commercial enzyme-linked immunosorbent assay kits because of the absence of interfering signals from the substrate, establishing considerable potential for multiplex detection of cancer biomarkers in blood. Therefore, the simultaneous detection of various cancer biomarkers using the developed srMINI chip with high diagnostic precision and accuracy is expected to play a decisive role in early diagnosis or community screening as a single-molecule biosensor.

The Prevalence of Encephalitozoon cuniculi in Domestic Rabbits (Oryctolagus cuniculus) in the North-Western Region of Romania Using Serological Diagnosis: A Preliminary Study.

Encephalitozoon cuniculi is a microsporidian, domestic rabbits being the main host. The disease can be acute or subclinical, but treatment options are limited and usually with unrewarding results; therefore, diagnosis and prevention of encephalitozoonosis in rabbits are of the utmost importance. This study aims to obtain the first preliminary information of the prevalence of E. cuniculi in the north-western region of Romania. A total of 176 rabbits were clinically examined and 2 mL of blood was sampled from each. An enzyme-linked immunosorbent assay (ELISA) kit by Medicago (Medicago, Uppsala, Sweden) on the resulted blood serum was utilized. Statistical analysis of the results was conducted using the EpiInfo 2000 software (CDC, Atlanta, GA, USA). A total prevalence of 39.2% (69/176) was identified, with statistically significant differences in relation to the rabbits' clinical status, age, season of sampling, breeding system, body condition score and county of origin; the different family farms tested also had a statistically significant difference. This study gives the first preliminary information on this pathogen distribution on Romania's territory, but further studies need to be performed on larger regions to declare the prevalence in the country.

End products of glycation (AGEs) and inflammation in the clinic of cardiovascular complications and vascular calcification at different stages of chronic kidney disease (G1-G5D)

To clarify the role of advanced glycation end products (AGEs) and inflammation in the development of vascular calcification and cardiovascular complications at different stages of chronic kidney disease (CKD) G1-G5D. We examined 105 patients aged 19 to 75 years with stage C1-C5D CKD, 77 (74%) of whom were patients with diabetic nephropathy. The concentration of AGEs, interleukin (IL)-1, IL-6 and tumor necrosis factor α (TNF-α), troponin I, parathyroid hormone was determined by enzyme-linked immunosorbent assay (ELISA) using kits from BluGene biotech (Shanghai, China), Cloud-Clone Corp. (USA), ELISA Kit (Biomedica, Austria). A high content of AGEs, IL-1, IL-6, TNF-α was established, which directly correlated with the increase in renal failure and changes in the morpho-functional parameters of the left ventricle and aorta. An increase in serum concentrations of AGEs and inflammatory mediators, correlating with a decrease in renal function and changes in the morpho- functional parameters of the left ventricle and aorta, indicate their significant role in the processes of damage to the cardiovascular system in CKD.