Enzyme-linked Immunosorbent Assay Method Research Articles

Novel solid-phase extraction promotes simultaneous colloidal gold immunochromatographic assay of malachite green, leuco-malachite green, chloramphenicol, and semi-carbazone metabolites in aquatic products.

Solid phase extraction (SPE) using traditional column materials offers several advantages, but its ability to simultaneously extract and purify commonly used veterinary drugs, such as chloramphenicol, malachite green, leuco-malachite green and semi-carbazone, remains unproven.Therefore, in the present study, a rapid analytical method based on ultrasound-assisted SPE combined with colloidal gold test strips was developed and applied herein to determine the residues of these veterinary drugs in common aquatic products. Under optimized conditions, the method achieved a limit of detection in the range of 0.01-0.5μg/kg, inter-day RSDs of the spiked samples in the ranges of 0.99-5.58% and 1.6-7.2%, and absolute recoveries in the range of 84.2-112.9% using the colloidal gold immunoassay and enzyme-linked immunosorbent assay methods, respectively. The SPE column efficiently enriched the four veterinary drugs simultaneously, showing considerable potential for determining common veterinary drug residues in aquatic products.

Paralytic Shellfish Toxin Concentrations Measured in Alaskan Arctic Clams Using ELISA and HPLC Methods

Clams are efficient vectors of potent algal neurotoxins, a suite of saxitoxin (STX) congeners collectively called paralytic shellfish toxins (PSTs), to higher trophic levels. The Alaskan Arctic is a region facing an expanding threat from PSTs due to ocean warming, yet little is known about PSTs in clams from this region. Quantifying total toxicity in bivalves requires analytical techniques, such as high-performance liquid chromatography (HPLC). Enzyme-linked immunosorbent assays (ELISAs) are an efficient but only semi-quantitative method for measuring clam toxicity. PSTs (STX eq.) were measured in split clam samples (n = 16) from the Alaskan Arctic using ELISA and HPLC methods to develop a preliminary linear model for conservatively estimating total toxicity in clams from ELISA toxin values (R2adj = 0.95, p < 0.001). Profiles of PST congeners and total toxicity using HPLC were also assessed in additional clams (n = 36 additional, n = 52 total). Clams contained mostly potent PST congeners, and over half of the clams had PST concentrations above the seafood regulatory limit. These data will help assess the exposure risks of PSTs in Arctic marine food webs, as harmful algal bloom activity is predicted to increase as the Arctic continues to warm.

Relationship Between Systemic Inflammatory Response Exponents, Levels of ADAM10, ADAM17 Proteins and Selected Clinical Parameters in Patients with Colorectal Cancer: Original Research Study

Chronic inflammation is a confirmed risk factor for colorectal cancer (CRC). Indicators of systemic inflammatory response (SIR), such as neutrophil-to-lymphocyte ratio (NLR) or platelet-to-lymphocyte ratio (PLR), are easily accessible indicators of the generalized inflammatory response. At the molecular level, inflammation-related carcinogenesis involves proteins from the adamalysin family: ADAM10 and ADAM17. The aim of the study was to assess NLR and PLR and their relationship with selected clinical parameters in CRC patients, as well as the correlation between ADAM10 and ADAM17 in tumor tissue and matched surgical margins with NLR and PLR values. Tumor tissue material matched surgical margins, and blood was collected from 66 patients who underwent surgery because of CRC. The concentrations of ADAM10 and ADAM17 in the collected material were tested using the enzyme-linked immunosorbent assay (ELISA) method. SIR parameters (NLR, PLR) were also determined. The results were statistically analyzed and compared with selected clinical parameters. Results: The study showed that PLR was lower in patients with comorbid cardiovascular diseases (CVD). In patients who underwent preoperative treatment, both the NLR and PLR values were higher than in patients who underwent primary surgery. There was also a negative correlation between ADAM17 concentrations in the surgical margin and PLR values. In conclusion, the presence of additional diseases such as CVD or diabetes mellitus type 2 (DMT2) or the use of preoperative treatment should be taken into account when assessing SIR parameters in CRC patients. Moreover, no clear correlations have been found between ADAM10 and ADAM17 and SIR parameters.

High-affinity VNARs targeting human hemoglobin: Screening, stability and binding analysis.

Hemoglobin, composed of α- and β-chains, is essential for oxygen transport and is key in diagnosing and treating gastrointestinal and blood disorders. It also aids in detecting blood contamination and estimating transfusion volumes. Immunological methods, based on antigen-antibody interactions, are distinguished by their high sensitivity and accuracy. Consequently, it is necessary to develop hemoglobin-specific antibodies characterized by high specificity and affinity to enhance detection accuracy. The variable domain of the new antigen receptor (VNAR) from sharks, the smallest antigen-binding unit, is ideal for disease diagnosis and treatment due to its small size, stability, and high affinity. In this study, Chiloscyllium plagiosum was immunized with human hemoglobin protein. Nine VNAR immune libraries with sizes ranging from 1 × 108 to 1.82 × 109 colony-forming units (CFU) were constructed and biopanned using phage display, resulting in three hemoglobin-specific VNAR sequences (5-10C, 7-11 A, T-12-4D). These sequences were inserted into pTT5-TEV-Fc vectors and transfected into HEK 293F cells. The resulting VNAR-Fc fusion proteins were purified from the cell culture supernatants. Binding activity, cross-reactivity, physicochemical stability, and epitope competition were evaluated using non-competitive enzyme-linked immunosorbent assay (ELISA) and biolayer interferometry (BLI). T-12-4D-Fc exhibited the highest affinity with a KD value of 7.59 nM and superior physicochemical stability. It maintained over 80 % binding activity at 90 °C, over 51 % in extreme pH conditions (pH 2 and 12), and above 65 % in urea concentrations up to 8 mol/L. Its binding activity remained largely unaffected after 6 h of incubation in human plasma-like medium (HPLM). The binding epitope competition results showed that 5-10C-Fc and T-12-4D-Fc targeted the same hemoglobin epitope. Molecular dynamics simulations revealed hydrogen bonds as the primary interaction force between VNARs and hemoglobin. Furthermore, a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) method was established for the detection of human hemoglobin, utilizing T-12-4D-Fc as the coating antibody. This technique demonstrated high accuracy, reproducibility and specificity when applied to human whole blood samples. Hence, the identified VNARs, particularly T-12-4D, demonstrated good stability, specificity, and high affinity, filling the gap of hemoglobin-targeting shark-derived single-domain antibodies and offering a foundation for diagnosing and monitoring hemoglobin-related diseases.

P0523 Comparative analysis of two ELISA methods for measuring serum ustekinumab levels in Inflammatory Bowel Disease ¿Is interchangeability feasible?

Abstract Background Ustekinumab (UTK) is a recently introduced biologic therapy for inflammatory bowel disease (IBD), and consequently, the clinical use of UTK trough concentrations is less standardized compared to other biologic therapies. Therefore, accurate measurement of serum UTK levels is essential for adjusting treatment efficacy. The objective of the present study is to compare two enzyme-linked immunosorbent assay (ELISA) methods to evaluate their correlation, concordance, and systematic bias, given the variability between methods. Methods The study assessed the correlation and the agreement between two different methods available for UTK measurement. Thirty-six blood samples from different patients, which had a previous diagnosis of IBD, were analyzed. All patients were recruited from September to November 2024 from the autoimmunity laboratory of the Hospital Universitario 12 de Octubre, Madrid. Serum UTK levels were measured using the two ELISA techniques: IDKmonitor® ELISA assay (Immundiagnostik AG; Bensheim, Germany) (T1) and the alternative Promonitor® ELISA assay (Grifols; Barcelona, Spain) (T2). Measurements were further categorized into therapeutic ranges for response classification: non-responders (0–1 µg/mL), clinical response (1–4.5 µg/mL), and endoscopic response (>4.5 µg/mL). Results A strong linear correlation was observed between the two methods (Pearson’s r = 0.9693, p < 0.0001, R² = 0.9395), confirming a proportional relationship despite systematic differences. Lin’s concordance coefficient was moderate-to-good (0.7025), indicating agreement despite systematic differences. Bland-Altman analysis revealed a mean difference of 2.79, with increasing variability at higher concentrations, consistent with proportional error. Passing-Bablok regression revealed a systematic bias, indicating an overestimation by the Promonitor-Grifols ELISA: the slope was 1.4342 (95% CI: 1.2530–1.6387) and the intercept 1.2591 (95% CI: 0.7591–1.8309). Considering these results, a linear regression adjustment was conducted. Categorization into therapeutic ranges revealed poor concordance between the methods (Kappa index = 0.28), suggesting that systematic errors substantially affect clinical interpretation. T2 consistently assigned higher therapeutic ranges, potentially impacting treatment decisions. Conclusion While the two ELISA methods show strong correlation, moderate concordance and significant systematic biases limit their interchangeability. Adjustment strategies should be adopted to harmonize results and ensure reliable therapeutic decisions in IBD management. References - Kwon Y, Kang B, Kim ES, Choe YH, Kim MJ. Comparison of Ustekinumab Trough Concentrations Measured by 2 ELISA Kits and Evaluation of Clinical Response in Crohn’s Disease. Ther Drug Monit. 2022;44(4):535-542. doi:10.1097/FTD.0000000000000976 - Rodríguez-Moranta F, Argüelles-Arias F, Hinojosa Del Val J, et al. Therapeutic drug monitoring in inflammatory bowel diseases. Position statement of the Spanish Working Group on Crohn’s Disease and Ulcerative Colitis (GETECCU). Gastroenterol Hepatol. 2024;47(5):522-552. doi:10.1016/j.gastrohep.2024.01.007

C-reactive protein as a diagnostic marker for ovarian carcinoma

Ovarian carcinoma is a leading cause of death in gynecological cancers, making early detection crucial for improving survival rates. C-reactive protein (CRP) has shown promise as a cost-effective biomarker to distinguish ovarian carcinoma from benign ovarian masses. Elevated CRP levels are associated with an increased risk of ovarian cancer. This cross-sectional study included 87 patients: 59 with ovarian carcinoma and 28 with ovarian cysts. The aim was to evaluate CRP as a diagnostic marker to improve early detection and clinical management of ovarian carcinoma. CRP levels were measured using the enzyme-linked immunosorbent assay method. Statistical analysis was conducted to assess the differences in CRP levels between the ovarian carcinoma group and the ovarian cyst group. All statistical analyses were performed using the Statistical Program for Social Sciences (IBM SPSS 24, IL, USA). Most subjects in the study were 50 years old or younger (69%) and had ovarian carcinoma (67.8%). Age over 50 [odds ratio (OR) 5.71, p=0.01] and menopausal status (OR 4.72, p=0.01) were significant risk factors for ovarian carcinoma. No significant difference in CRP levels was found between ovarian carcinoma and ovarian cyst patients (p=0.23). Based on the results, CRP cannot be used as an effective predictor to differentiate ovarian carcinoma from ovarian cysts.

Increased SCUBE-1 levels in the aqueous humour of patients with pseudoexfoliation syndrome

ABSTRACT Clinical relevance Pseudoexfoliation syndrome (PXS) is a common age-related disorder associated with glaucoma and cataract. Despite its clinical importance, the pathogenesis of PXS is not yet fully understood. Background To evaluate levels of SCUBE-1 (signal peptide, CUB domain, and epidermal growth factor-like domain containing protein 1) in the serum and aqueous humour of patients with PXS in comparison with non-PXS controls. Methods This prospective study included patients with and without PXS undergoing phacoemulsification surgery for cataract. Patients with pseudoexfoliative glaucoma were excluded. Aqueous humour samples collected at the beginning of phacoemulsification surgery were analysed by using the sandwich enzyme-linked immunosorbent assay method (human SCUBE-1 enzyme-linked immunosorbent assay kit; Elabscience). Results There was a significant difference in mean aqueous humour SCUBE-1 levels between PXS patients (1.69 ± 0.41 ng/µL) and non-PXS controls (0.89 ± 0.33 ng/µL) (p = 0.012). However, the difference in mean serum SCUBE-1 levels was not significant (0.87 ± 0.36 ng/µL and 0.81 ± 0.35 ng/µL, respectively; p = 0.560). There was no significant correlation between aqueous humour and serum SCUBE-1 levels in either group (p > 0.05). Conclusion SCUBE-1 is an important marker of ischaemia and oxidative stress. Increased SCUBE-1 levels in the aqueous humour of patients with PXS may indicate a local effect of SCUBE-1 in the pathogenesis of PXS.

Evaluating Methods for Aflatoxin B1 Monitoring in Selected Food Crops Within Decentralized Agricultural Systems.

Aflatoxin B1 (AFB1) contamination of food crops pose severe public health risks, particularly in decentralized agricultural systems common in low-resource settings. Effective monitoring tools are critical for mitigating exposure, but their adoption is limited by barriers such as cost, infrastructure, and technical expertise. The objectives of this study were: (1) to evaluate common AFB1 detection methods, including enzyme-linked immunosorbent assays (ELISA) and lateral-flow assays (LFA), validated via high-performance liquid chromatography (HPLC), focusing on their suitability for possible applications in decentralized, low-resource settings; and (2) to conduct a barriers-to-use assessment for commonly available AFB1 detection methods and their applicability in low-resource settings. Among four ELISA kits, the AgraQuant Aflatoxin B1 2/50 ELISA Kit demonstrated the highest accuracy and precision, reliably quantifying AFB1 in maize and tortillas across 5-150 ppb with minimal cross-reactivity. For LFA, a smartphone-based algorithm achieved a high presence/absence accuracy rate of 84% but struggled with concentration prediction. The barriers-to-use analysis highlighted the practicality of low-cost tools like moisture readers for field screening but underscored their qualitative limitations. Advanced methods like HPLC and LC-MS offer greater precision but remain impractical due to their high costs and infrastructure requirements, suggesting a potential role for adapted ELISA or LFA methods as confirmatory approaches. These findings support the development of multi-tiered frameworks integrating affordable field tools with regional or centralized confirmatory testing. Addressing systemic barriers through capacity building, partnerships, and improved logistics will enhance AFB1 monitoring in decentralized systems, protecting public health in vulnerable communities.

CXCL13 levels in cerebrospinal fluid in relapsing-remitting multiple sclerosis: The role of Borrelia in neuroinfections.

This study was compared the Borrelia antibodies and chemokine ligand 13 (CXCL13) levels in cerebrospinal fluid (CSF) samples from cases diagnosed with relapsing-remitting multiple sclerosis (RRMS), radiologically isolated syndrome (RIS), and pseudotumour cerebri (PTC). A total of 43 CSF samples were collected from patients diagnosed with RRMS, RIS and PTC. We prospectively investigated Borrelia IgG and IgM antibodies in the CSF samples of the cases by enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) method, and CXCL13 levels by ELISA. Data were statistically analysed using the the Spearman rank correlation test. Five antigens (protein 19, 20, 21, 58, and outer surface protein C (OspC)) were positive due to confirmation of the positive samples for Borrelia antibodies by the WB method. There were no significant differences in CSF CXCL13 levels between the three groups. The CXCL13 level was found to be statistically higher in the demyelinating group compared to the PTC group (p=0.001). The Borrelia antibodies were found positive in CSF samples of RRMS patients. The coexistence of high CXCL13 (may be a potential biomarker) suggests that LNB may also play a role in the etiopathogenesis of RRMS. In addition, the positive detection of OspC and p58 WB bands in most cases suggests that these protein bands can be used as in the differential diagnosis of neurodegenerative diseases and Lyme disease.

Relationship between BDNF content in cord blood and early neurobehavior in newborns with subclinical hypothyroidism during pregnancy: a preliminary study.

Research on neurobehavioral abnormalities in neonates of mothers with subclinical hypothyroidism (SCH) is limited. The link between umbilical cord blood brain-derived neurotrophic factor (BDNF) levels and neurobehavioral outcomes in neonates has not been explored. This study investigates the correlation between alterations in umbilical cord blood BDNF levels and early neurobehavioral abnormalities in neonates born to pregnant women with SCH. This study recruited 72 pregnant women with SCH and 76 healthy controls (HC). The study collected general information for all subjects, including body mass index, parity, thyroid function assessed during early to late pregnancy, and neonatal birth weight. Neonatal behavioral and neural abilities were evaluated using the Neonatal Behavioral Neurological Assessment (NBNA). BDNF levels in umbilical cord blood were measured using the Enzyme-Linked Immunosorbent Assay method. The results indicated that neonates with SCH during pregnancy had lower total NBNA scores, behavioral ability, passive muscle tone, active muscle tone, primitive reflexes, general assessment, and lower levels of cord blood BDNF compared to healthy controls. The cord blood BDNF of newborns with SCH during pregnancy was positively correlated with total NBNA score, behavioral ability, active muscle tone, and general assessment. Moreover, multiple linear regression analysis demonstrated an association between cord blood BDNF levels in pregnant patients with SCH and multiple measures of newborn health, including total NBNA score, behavioral ability, active muscle tone, and general assessment. Infants born to pregnant women with SCH exhibit reduced behavioral and neural abilities linked to BDNF levels in umbilical cord blood.

Pharmacokinetic and Pharmacodynamic Equivalence of Biosimilar and Reference Ultra-Rapid Lispro: A Comparative Clamp Study in Healthy Volunteers.

Ultra-rapid insulin lispro is an innovative insulin analogue designed to achieve rapid onset and short duration of action, aimed at optimizing glycemic control in patients with diabetes. This was a double-blind, randomized, 2-period, crossover clamp study to evaluate the pharmacokinetics (PK) and pharmacodynamics (PD), along with safety profiles, of a potential biosimilar ultra-rapid insulin lispro compared to the reference product in healthy White men. A total of 35 healthy volunteers completed hyperinsulinemic euglycemic clamp procedures across both study periods. Blood samples were collected at predefined intervals up to 8hours to assess PK parameters. Plasma glucose levels were monitored every 5minutes during the 8-hour clamps, with adjustments to the glucose infusion rate to maintain the target range. Insulin quantification in plasma was conducted using a validated enzyme-linked immunosorbent assay method. PD assessment was based on glucose infusion rate profiles during both clamps. Geometric mean ratios for maximum plasma concentration and area under the concentration-time curve from insulin administration to the last measurable concentration for the test and reference drugs fell within the bioequivalence range of 80%-125%. Furthermore, the investigational drugs demonstrated comparable PK/PD profiles of insulin lispro. Both formulations exhibited similar safety profiles primarily characterized by mild injection site reactions.

Analysis of Glucocorticoids as Potential Adulterants in Cosmetic Products: A Dual Approach for Qualitative and Quantitative Evaluation Based on ELISA and HPLC-MS Methods

The analysis of cosmetic products represents an important field of analytical chemistry, since the demand for new formulations is continuously increasing. Regulations about prohibited/regulated compounds are applied in each country. Among the substances that are banned in cosmetics, corticosteroids represent a potential harm for consumers since the prolonged exposure to these compounds can affect health status. However, corticosteroids can be found in cosmetics as an illegal addition since they are able to alleviate the symptoms of inflammatory skin problems. In this work, two different approaches for detecting corticosteroids as potential adulterants in cosmetic products were compared. First, a reversed-phase HPLC-MS method was optimized and fully validated in order to identify and quantify eight corticosteroids (methylprednisolone, beclomethasone, flunisolide, budesonide, betamethasone 17-valerate, beclomethasone dipropionate, flumethasone, and dexamethasone). This reference method was then compared with an enzyme-linked immunosorbent assay (ELISA). Indeed, immunological techniques allow for rapid, low-cost, and sensitive detection of target analytes even in complex matrices, and they can be performed with simple instrumentation and by non-skilled personnel. The application of these methods on spiked cosmetic products was compared in terms of performance and advantages in order to evaluate the possibility of exploiting a complementary approach for optimizing the time for and costs of the analysis.

Postoperative evaluation of interleukin-8 and C1q/TNF-related protein-12 in patients undergoing coronary artery bypass graft surgery.

Coronary artery bypass graft surgery is one of the most frequently performed surgeries worldwide. Coronary artery bypass graft surgery induces an inflammatory response. Interleukin-8 is a pro-inflammatory cytokine that plays a role in the pathogenesis of cardiovascular diseases. C1q/TNF-related protein-12 is implicated in mitigating inflammation and cardiomyocyte damage. This study aimed to compare interleukin-8 and C1q/TNF-related protein-12 levels before and 45 days after coronary artery bypass graft surgery. A total of 43 patients who underwent coronary artery bypass graft surgery were studied. Serum concentrations of interleukin-8 and C1q/TNF-related protein-12 were evaluated using the enzyme-linked immunosorbent assay method before and 45 days after the surgery. No significant differences were observed in interleukin-8 levels between pre- and post-coronary artery bypass graft surgery (p=0.077). However, serum levels of C1q/TNF-related protein-12 were found to be lower 45 days after coronary artery bypass graft surgery compared to pre-surgery levels (p<0.001). Moreover, changes in C1q/TNF-related protein-12 were not associated with diabetes, hypertension, and body mass index (p>0.05), but C1q/TNF-related protein-12 alterations were found to be associated with opium addiction. These findings suggest that the evaluation of C1q/TNF-related protein-12 can be beneficial for the late assessment of post-coronary artery bypass graft surgery inflammation. The reduction of C1q/TNF-related protein-12 levels might indicate increased levels of inflammation after surgery at this time point, which requires the assessment of further inflammatory factors to confirm this finding.

Assessing aflatoxin exposure in the United Arab Emirates (UAE): Biomonitoring AFM1 levels in urine samples and their association with dietary habits.

Aflatoxins, known for their carcinoginc properties and produced by Aspergillus fungi, pose a substantial threat to public health, particularly in regions with hot and humid climates, where individuals are exposed to these toxins through contaminated food. The primary objective of this study was to assess the extent of aflatoxin exposure in the Emirate of Sharjah employing Aflatoxin M1 (AFM1) as a biomarker in urine samples from adult participants. Furthermore, this study aimed to explore the relationship between dietary habits and AFM1 levels in order to establish a potential link. In a cross-sectional study design, a total of 144 adults (73 females and 71 males) were recruited for participation. The urine samples obtained from participants were subjected to analysis for AFM1 concentrations utilizing the enzyme-linked immunosorbent assay (ELISA) method. Additionally, structured questionnaires were administered to collect information on the dietary and lifestyle habits of the participants. To explore the relationship between dietary factors and AFM1 levels, various statistical analyses, including linear regression and the Mann-Whitney U test, were performed. AFM1 was detected in 69 % of the samples under invstigation, wherein males exhibited a higher mean level (0.912 ng/mg creatinine) in comparison to females (0.676 ng/mg creatinine). The overall mean concentration of AFM1 was determined to be 0.792 ng/mg creatinine. It is worth noting that there was a significant correlation between rice consumption and heightened AFM1 exposure among males, while no such correlation was observed among females. This study conducted in the UAE provides novel perspectives on aflatoxin exposure, shedding light on the gender-specific correlation between rice consumption and aflatoxin levels among males. These findings hold significant implications for guiding public health interventions and underscore the pivotal role of ongoing surveillance and stringent food safety regulations in mitigating the hazards associated with aflatoxin contamination.

Prussian blue-doped CaCO3 nanoparticle-labeled secondary antibodies for electrochemical immunoassay of interleukin-6 with migraine patients.

The sensitive and accurate identification of interleukin-6 (IL-6) in biological fluids is essential for assessing migraine due to its role in different physiological and pathological processes. In this study, we designed a simple and feasible electrochemical immunosensing method for the voltammetric measurement of IL-6. The electrochemical immunosensor was fabricated through covalent conjugation of anti-IL-6 capture antibodies on the glassy carbon electrode with a typical carbodiimide coupling method. Anti-IL-6 secondary antibodies were labeled on the surface of Prussian blue-doped CaCO3 nanoparticles (PBCaNP) via the epoxy-amino reaction. The assay was carried out with a sandwich-type immunoreaction. In the presence of target IL-6, the analyte was sandwiched between the capture antibody and detection antibody. Thereafter, the carried PBCaNP accompanying the secondary antibody could be detected by using square wave voltammetry (SWV). The voltammetric peak current was dependent on the concentration of target IL-6. Under optimum conditions, the electrochemical immunosensor exhibited good analytical properties, and allowed detection of IL-6 within a wide linear range from 0.1 to 1000 pg mL-1. The limit of detection was estimated to be 0.078 pg mL-1 of IL-6 at the 3sB criterion. An intermediate reproducibility of ≤10.59% was accomplished with batch-to-batch identification, and good anti-interference capacity against other biomolecules was achieved. Importantly, clinical human serum samples obtained from 15 migraine patients were analyzed with the developed electrochemical immunosensors, giving results well-matched with those obtained from the referenced enzyme-linked immunosorbent assay (ELISA) method.

Role of Dardarin and Isthmin-1 in the Protective Effect of Hydroxytyrosol Against Corn Syrup-Induced Liver Damage.

In this study, it was investigated whether dardarin (LRRK2) and isthmin-1 (ISM1) play a role in the protective effect of hydroxytyrosol (HT) used to prevent liver damage caused by corn syrup in rats. Rats were divided into four groups with six in each group: 1) control, 2) HT, 3) corn syrup, and 4) corn syrup + HT. Rats were given water containing 30% corn syrup for six weeks. At the same time, HT-containing liquid was given orally at 4 ml/kg/day, alone and together with corn syrup for six weeks. The weights of the rats were measured every week. LRRK2 and ISM1 molecules in liver tissue were evaluated by histopathological methods. Biochemical parameters were also examined with the enzyme-linked immunosorbent assay (ELISA) method. It was found that weight gain was less in rats receiving HT than in those consuming corn syrup. The increase in cholesterol, triglyceride, and liver enzyme levels because of corn syrup consumption decreased with HT consumption. As a result of histopathological analysis, it was observed that the increase in LRRK2 and ISM1 levels observed in the liver tissue in the corn syrup-administered group decreased when HT was administered together with corn syrup. In addition, it was determined that the increase in sinusoidal expansion and hepatocyte necrosis observed in the liver tissue as a result of corn syrup application decreased as a result of the application of HT together with corn syrup. The protective effect of HT against the damage caused by corn syrup in the liver has been demonstrated once again; however, LRRK2 and ISM1 are thought to contribute to this issue.

Serum Vitamin D Level Correlates Significantly With Leptin and Tumor Necrosis Factor-Alpha in Overweight Postmenopausal Women With Hypertension.

Association of serum vitamin D (vitD) with leptin (Lep) and tumor necrosis factor-alpha (TNF-α) is not precisely known in overweight hypertensive (OW-HT) postmenopausal (PMP) women. Hence, the present study was carried out to investigate the body mass index (BMI)-based correlation of serum vitD with Lep and TNF-α in OW-HT PMP women. Women subjects in their early PMP (n = 346, age: 51 - 60 years) categorized into three groups had main inclusion criteria of specified range of age, BMI and blood pressure (BP). Enzyme-linked immunosorbent assay (ELISA) and other kit methods were employed to investigate the role of various variables in three subject groups (normal weight normotensive (NW-NT, n = 116, BMI (kg/m2): 22 - 24.9), normal weight hypertensive (NW-HT, n = 115, BMI: 22 - 24.9) and OW-HT (n = 115, BMI: 25 - 29.9) PMP women). A significant negative linear correlation of vitD with serum Lep and TNF-α, and a significant positive linear correlation of BMI with Lep and TNF-α in OW-HT PMP women were obtained. Significantly higher levels of serum Lep, TNF-α and interleukin-6 (IL-6) were found in OW-HT PMP women, as compared to NW-HT PMP women. The present study suggests that decreased serum vitD levels correlate with the Lep and TNF-α in OW-HT PMP women. However, further studies may help understand the impact of vitD in cardiovascular events and the influencing factors in OW-HT PMP women.

Do Salivary Cullin7 Gene Expression and Protein Levels Provide Advantages over Plasma Levels in Diagnosing Breast Cancer?

In addition to the tumor suppressor role of Cullin 7 (Cul7), one of the proteins belonging to the Cullin (Cul) family, studies have also suggested that Cul7 may act as an oncogene under certain conditions. The role of the Cul7 molecule in breast cancer is still unclear, and understanding its function could have significant implications for identifying novel therapeutic targets or improving diagnostic strategies in breast cancer management. In this study, the levels of the Cul7 molecule in plasma and noninvasive material saliva were investigated, and its possibility as a marker for breast cancer was discussed. Protein levels of blood and saliva samples taken from breast cancer patients and a healthy control group were measured by the ELISA (Enzyme-Linked Immunosorbent Assay) method. Gene expression levels between the two groups were analyzed by the qPCR (quantitative Polymerase Chain Reaction) method. In our study, Cul7 mRNA and protein expression levels were examined in 60 breast cancer patients and 20 healthy female controls, and a statistically insignificant difference was found between the patient and control groups in both plasma and saliva samples (p > 0.05). No correlation was found between the clinical characteristics of the patients and plasma and saliva Cul7 gene expression and protein levels (p > 0.05). Considering the possibility of Cul7 being a biomarker at the protein and mRNA levels, plasma is thought to be a better study material for Cul7. Our findings suggest that in the context of a study on salivary material, the expression of Cul7 at the mRNA level may have better potential utility as a biomarker.

Dynamics of cortisol indicators in patients with mild arterial hypertension I-II stages in the conditions of a stress test against the background of excessive anxiety and hyperventilation syndrome

Annotation. The aim of the study was to investigate the dynamics of cortisol levels in 105 patients with mild (grade 1) arterial hypertension (AH) of stage I-II (st.) against the hyperventilation syndrome (HVS) and excessive anxiety, in the conditions of a 6-minute walk test (6MWT). The control group of patients included 28 patients with mild hypertension, I-II stages, without signs of vegetosomatic disorders. The state of anxiety and HVS was determined by the Spielberger-Hanin scale and the Nijmegen questionnaire. The level of cortisol in saliva was determined by enzyme-linked immunosorbent assay method on the “Stat Fax 303 Plus” analyzer. Statistical analysis was performed using the Statistica v.10 program (USA). It was found that normal baseline cortisol levels in patients of the main group with AH I stage were found in 36.96% of patients and amounted to 5.63±0.33 nmol/l, in AH II stage - in 41.46% of patients. The conditional norm was the result obtained in the group of practically healthy individuals. In the control group with I-II stage of hypertension, normal baseline values were observed in 65.5% and 58.33% of patients, respectively. Increased baseline cortisol levels (6.58±0.32) nmol/l were detected in 47.83% of patients in the main group with AH I stage, and in 46.34% of patients with AH II stage (6.72±0.26) nmol/l (p˂0.05); in the control group, respectively, in 37.5% and 41.67% of patients. The rest of the patients, reduced baseline cortisol levels were found. After 6 MWT, in the main group with stage I hypertension, the number of cases with elevated cortisol levels increased by 8.70%, and the number of patients with low levels decreased by 10.87%. In stage II hypertension, after 6 MWT, the number of patients with elevated cortisol levels increased by 12.20%, while the number of cases with low levels decreased by 7.32% and the number of normal levels decreased by 4.88%. In the control group with stage I hypertension after 6 MWT, a decrease in normal cortisol levels was noted in 6.25% of patients and a similar increase in the number of patients with elevated levels. In stage II hypertension after 6 MWT, cases with normal cortisol levels decreased by 16.33%; at the same time, the number of patients with elevated levels increased by 8.33%. The dynamics of cortisol levels after exercise in mild hypertension with excessive anxiety and hypertension indicates the tension of neuroendocrine regulation mechanisms, the risk of destabilization of the underlying disease under stress.

The Association of Serum Lactoferrin Level with Psychological Symptoms, Cognition, and Executive Function in Schizophrenia Patients

Objective: Lactoferrin, a glycoprotein, has known neuroprotective effects, yet its role in the pathophysiology of neuropsychiatric disorders, particularly schizophrenia, remains unclear. This study aims to assess changes in lactoferrin levels during different phases of schizophrenia and explore its relationship with cognitive symptoms and performance. Method: This before/after interventional study involved 30 patients diagnosed with schizophrenia. Participants were evaluated at two time points: upon hospital admission and after the resolution of acute symptoms. The Positive and Negative Syndrome Scale (PANSS) was utilized to measure symptom severity, while the Brief Assessment of Cognition in Schizophrenia (BACS) assessed the neurocognitive function. Serum lactoferrin levels were quantified using the Enzyme-Linked Immunosorbent Assay (ELISA) method. Results: Serum lactoferrin levels significantly decreased from 130.63 ± 52.49 ng/mL at admission to 85.42 ± 29.03 ng/mL at discharge (P &lt; 0.001). No significant correlation was found between lactoferrin levels and PANSS scores (r = 0.011, P = 0.975). However, an inverse correlation was observed between changes in lactoferrin levels and the executive function subscale of the BACS (r = -0.360, P = 0.050). Cognitive assessments indicated significant improvements in verbal memory (P = 0.033), working memory (P = 0.002), and executive function (P = 0.039) post-treatment. Conclusion: The study demonstrates a significant reduction in serum lactoferrin levels during the acute phase of schizophrenia, suggesting its potential role in modulating cognitive functions, particularly the executive function, rather than influencing positive or negative symptoms.