Assay Method Research Articles

Assessment of serum levels of 8-hydroxy-2’ deoxyguanosine and f2-isoprostanes in newly diagnosed adult hypertensive with clinical depression in NAUTH, Nnewi, Nigeria

Despite the high prevalence of depression and hypertension, the relationship between the two diseases has received little attention. The potential roles of some biomarkes of oxidative stress in disease progression and management is very crucial. Aim: The present study therefore, evaluated the serum levels of 8-hydroxy-2’ deoxyguanosine (8-OHdG) and F-2 isoprostanes in newly diagnosed hypertensive individuals with and without depression at Nnamdi Azikiwe University Teaching Hospital, Nnewi. Method: This was a cross sectional study carried out on randomly selected 121 consented adult male and female participants within the ages of 18-65 years which comprises 30 newly diagnosed hypertensive individuals without depression (Hypertensive), 31 hypertensive individuals with depression selected from the department of internal medicine. 30 non hypertensive with depression (Depressive) and 30 apparently healthy non-hypertensive individuals without depression which served as control selected among the hospital staff. Their bio-data was obtained from their hospital records. 3 ml of venous blood were collected from each of the participants, dispensed into a plain container, centrifuged and serum separated into another container and stored at -20 0C for the assessment of serum 8-OHdG using Enzyme Linked Immunosorbent Assay (ELISA) Method. Result: The result showed that Hypertensive individuals with and without clinical depression (107.08±22.05, 129.16±82.49) had significantly higher serum level of 8-OHdG when compared with control group (53.95±28.25) (P<0.05 respectively). Hypertensive individuals (129.16±82.49) also had significantly higher 8-OHdG level when compared with non-hypertensive with depression (63.31±37.30) (P<0.05). Hypertensive individuals with depression (9.94±7.14) and depressive individuals (8.99±5.84) also had significantly higher F2-Isoprostanes level when compared with control (5.15±1.47) (P<0.05 respectively). There was a moderately strong positive correlation between 8-OHdG level and body mass index in both hypertensive and depressive individuals (r=0.682, P<0.05). Conclusion: The study observed higher 8-OHdG and F2-Isoprostanes in individuals with hypertension with and without clinical depression. In addition, there is evidence that oxidative stress is related to inflammation and endothelial activation in individuals with both conditions leading to disease severity.

Verification of analytical performance of Creatine Kinase isoenzyme MB (CK-MB) on the Alinity c® Experience from the Biochemistry Laboratory of Mohammed VI University Hospital in Oujda

The verification of analytical methods is a requirement of the NF EN ISO 15189 standard. It involves evaluating the performance of an analytical method according to a well-defined protocol and comparing it to pre-established analytical objectives. In this study, we present the results of the verification protocol for the measurement method of creatine kinase isoenzyme MB (CK-MB) on the Alinity c® analyzer, which is employed for functional exploration of the heart. Our study is conducted according to the criteria of Scope A detailed in the Verification/Validation Guide for Methods in Medical Biology, following the SH GTA 04 recommendations from COFRAC. The verification focused on the CK-MB assay on the Alinity c® analyzer using International Federation of Clinical Chemistry (IFCC) method based on immunoinhibition principle for an assessment of analytical performance in terms of repeatability and intermediate precision. these results were aligned with the manufacturer's specifications. The results obtained for various CK-MB assay verification criteria on the Alinity c® analyzer demonstrate satisfactory repeatability across one level with CV = 1.29%. Intra-laboratory reproducibility was satisfactory for one level with CV = 2.58%. The coefficient of variation (CV) values obtained in our study were compared with manufacturer's specifications, no CV reference values were established by French Society of Clinical Biology SFBC. The findings from this study have allowed us to verify the performance of the CK-MB assay method and to compare them with the objectives set in the accreditation process to which our laboratory is committed.

Verification of the analytical performance of the urinary creatinine assay on Abbott Architect ci8200

The urinary creatinine assay is a pivotal diagnostic tool for assessing kidney function, hydration status, and muscle metabolism. This study evaluated the analytical performance of the urinary creatinine assay method using an Abbott kit on the Architect ci8200® automated system in the biochemistry laboratory of CHU Mohammed VI, Oujda. Method verification was conducted in two phases: intermediate fidelity testing using internal quality controls and repeatability testing on patient urine samples across low and high creatinine concentration levels. The results demonstrated excellent analytical performance, with coefficients of variation (CV) for intermediate fidelity (CV1 = 1,95% and CV2 = 1.46%) and repeatability (CV1 = 0.78% and CV2 = 0.87%) well within the limits established by the French Society of Clinical Biology (SFBC). Levey-Jennings charts illustrated consistent and robust performance. This method also underwent a comparative analysis using the Bland-Altman diagram, which confirmed its accuracy and precision relative to established standards. These findings underscore the reliability of the urinary creatinine assay method, ensuring high-quality diagnostic outcomes.

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.

Occurrence of Alternaria Toxins, Ochratoxin A and Aflatoxins B1 in Green Arabic Coffee (Coffea arabica) in Saudi Arabia: Risk Assessment on HepG-2 Cell Line and Male Albino Rats

Arabica coffee (Coffee arabica L.) is among the most popular and widely consumed drinks globally today. The current study aimed to detect and quantify of potential mycotoxins in coffee beans and to investigate their potential risk on HepG-2 human liver cancer cells and male albino rats. Eighteen fully matured coffee beans samples were collected from different geographical locations in Saudi Arabia. Mycotoxins detected and quantified utilizing high-performance liquid chromatography (HPLC). Cytotoxicity of the detected toxins was investigated utilizing the MTT assay method toward HepG-2 cells and male albino rats. Cytotoxic properties of the detected mycotoxins were investigated at doses of 31.25, 62.50, 125, 250 and 500 µg/ml following exposure durations of 24 and 48 h in comparison with control. HPLC analysis revealed the detection of four Alternaria toxins (ALT-Ts) namely Altenuene (ALT), Tenuazonic acid (TA), Alternariol (AOH), and Altenuisol being AOH reported as the frequently detected mycotoxin in 18 coffee beans samples representing 75 % with quantity ranged 0.98 to 8.97 μg/g. OTA was frequently detected (10 samples, 55.5%) in concentrations varied from 0.75 to 9.87 μg/g. In addition, Aflatoxin B1 was also detected with low quantity ranged 0.89 to 3.22 μg/g. Treatment of HepG-2 cell line with combined Alternaria toxins (ALT, TeA, AOH, and Altenuisol), Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) as well as their combination triggered a decline in cell viability that was proportional to the dose administered. The elevated doses (62.50, 125, 250 and 500 µg/ml) notably reduced cell viability and induced cellular damages when contrasted with the lowest dose (31.25 µg/ml) and control. The IC50 for the tested mycotoxins registered for ALT-Ts, AFB1 and OTA as well as their combination were 115.95, 56.68, 59.86 and 31.05 µg/ml after 48 h of exposure. Secreting inflammatory markers (Interleukin-6 (IL-6), interleukin-1β (IL-1β) and COX-2), and oxidative markers (total antioxidant capacity (TAC), glutathione (GSH) and nitrite (NO) were downregulated by all mycotoxins with the highest decrease induced by their mixture. Malonaldehydes (MDA) and TAC were significantly increased by all toxins in treated rats. Hematological results revealed that AFB1, OTA and ALT-Ts-treated rats exhibited a notable decline in hemoglobin (Hb), red blood cells (RBCs), white blood cells (WBCs), and packed cell volume % (PCV). However, single and double dose-treated rats with OTA, Mix1 and Mix2 exhibited a marked increase in WBCs. The results demonstrated a notable decline in liver and kidney functions in all mycotoxins-treated rats. The studied organs (liver, kidney and spleen) exhibited significant damage, with the most severe alterations observed in the Mix1 and Mix2-treated group in comparison with the groups receiving individual treatment. This indicates that the mixture of AFB1, ALT-Ts and OTA had an enhanced toxic consequence against treated HepG-2 cell line and rats. Our results indicated the synergistic effect of the combined different mycotoxins against HepG-2 human liver cancer cells and male albino rats. This work could help fill in data gaps and enhance risk assessment for human health by addressing some of the possible risks associated with Alternaria toxins

Assessment of Inflammatory and Oxidative Stress Biomarkers for Predicting of Patients with Asymptomatic Carotid Artery Stenosis

Background/Objectives: Asymptomatic carotid artery stenosis is usually detected by physicians in patients, coincidentally, during an ultrasound examination of the neck. Therefore, measurable biomarkers in blood are needed to define the presence and severity of atherosclerotic plaque in patients to identify and manage it. We hypothesized that biomarkers that indicate pathways related to the pathogenesis of atherosclerosis could be used to identify the presence and severity of atherosclerotic plaque. For this purpose, the levels of participants’ inflammatory and oxidative stress biomarkers were determined. Kynurenine/tryptophan and neopterin levels were measured as relatively new biomarkers of inflammation in this study. Methods: Our study included 57 patients diagnosed with asymptomatic carotid artery stenosis and 28 healthy volunteers. Blood kynurenine and tryptophan levels were measured with LCMS/MS. Blood catalase, total superoxide dismutase (t-SOD), glutathione peroxidase (GPx), malondialdehyde, and neopterin levels were measured using the ELISA assay method. Result: The kynurenine/tryptophan ratio reflecting IDO activity was higher in patients than in healthy volunteers. Decreased tryptophan levels and increased kynurenine and neopterin levels were observed in patients who underwent carotid endarterectomy. In patients, catalase, t-SOD, and malondialdehyde levels were higher, while GPx activity was lower. These differences were found to be more significant in patients who underwent carotid endarterectomy. Conclusions: Increased kynurenine/tryptophan ratio and neopterin levels in patients with asymptomatic carotid artery stenosis were associated with the inflammatory status of the patients. Oxidative stress and inflammatory biomarkers can be considered effective diagnostic and severity indicators for asymptomatic carotid artery stenosis.

Comparative Analysis of 2 Commercially Available Assays for Therapeutic Drug Monitoring of Infliximab and Adalimumab.

Tumor necrosis factor is a crucial proinflammatory cytokine in immune-mediated diseases. Tumor necrosis factor inhibitors (TNFi), such as infliximab and adalimumab, effectively treat rheumatological and digestive disorders. However, challenges such as primary nonresponse, secondary treatment failure, or adverse reactions limit their efficacy. Monitoring TNFi levels is essential for optimizing treatment and improving outcomes. An enzyme-linked immunosorbent assay (ELISA; Promonitor) was compared with 2 commercially available methods for quantifying infliximab and adalimumab levels: the chemiluminescence assay (i-Track10) and fluorescence assay (Afias-10). Serum samples from 166 patients with inflammatory bowel disease were analyzed. Drug levels were measured using i-Track10, Afias-10, and Promonitor. Spearman's correlation analysis, Bland-Altman analysis, analysis of differences, Passing-Bablok regression, and Cohen kappa for agreement assessment were used for statistical analysis. Strong correlations were observed between Promonitor and Afias-10 for infliximab (rs = 0.982) and adalimumab (rs = 0.972), and with i-Track10 (rs = 0.935 for infliximab, rs = 0.947 for adalimumab). However, significant differences indicated noninterchangeability with ELISA. Passing-Bablok regression showed systematic and proportional biases. Cohen kappa exhibited higher concordance with Afias-10 for therapeutic ranges (κ = 0.962 for infliximab, κ = 0.849 for adalimumab) compared with i-Track10. Afias-10 and i-Track10 are suitable for TNFi monitoring but are not interchangeable with ELISA. Consistent assay methods should be used for patient monitoring to ensure accuracy and reliability.

Synthesis of chitosan and carboxymethyl cellulose connect flavonoid (CH-Fla-CMC) composite and their investigation of antioxidant, cytotoxicity activities.

This study successfully synthesised and characterised composites combining chitosan (CH), carboxymethyl cellulose (CMC), and various flavonoids (Fla). This innovative approach demonstrates the potential for developing functional materials with antioxidant and food preservation properties. The composites CH-Fla-CMC (1-5) was characterised using advanced techniques such as FT-IR, UV-Vis, XRD, SEM, TEM, and TGA, providing robust data on their structural, morphological, and thermal properties. CH-connected CMC has been used to prevent many diseases, based on the findings of this study. Therefore, dietary flavonoids (Fla = 1. 3-Hydroxyflavone; 2. rhamnetin; 3. natsudaidain; 4. isorhamnetin; 5. myricetin) was used to prepare the composites in this study. Dietary flavonoids play an important role in the prevention of degenerative diseases. In addition, oxygen permeability (OP), water solubility (WS), and moisture content (MS) were analysed. The synthesised composites were screened for antioxidant and cytotoxic activities. Multiple antioxidant assays (DPPH, H₂O₂, NO, ABTS•⁺, and AAPH) were conducted, confirming the superior radical scavenging activity of CH-Fla-CMC-5 compared to standards such as BHT and Trolox. The synthesised composite CH-Fla-CMC 5 was more active than the standard butylated hydroxytoluene (BHT) against 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2), and nitric oxide (NO) radical scavenging activity (DPPH: 10.30 vs. 33.88 μg/mL; H2O2: 13.26 vs. 27.16 μg/mL, and NO: 13.56 vs. 31.73 μg/mL), whereas 2,2-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid radical cation (ABTS•+) decolourisation assay and lipid peroxidation method (AAPH) CH-Fla-CMC 5 was more active than the standard Trolox (ABTS: 91.26 ± 0.59 % vs. 85.28 ± 0.97 %; AAPH: 91.02 ± 0.01 % vs. 62.39 ± 0.35 %). The synthesis and characterisation methods are laboratory-based. This study primarily focused on in vitro antioxidant and cytotoxicity assays. The performance of these composites in living organisms and real-life food packaging scenarios remains untested. Cytotoxicity against only check these cell lines such as MCF-7, HeLa, HepG2, and normal Vero cancer cell lines was assessed. Only five flavonoids were tested, potentially limiting the generalisability of the findings to other dietary flavonoids. In the future, we plan to compare more cell lines with flavonoids. These laboratory-based methods have been converted into industrial production. The CH-Fla-CMC-5 composite performed better than the other compounds in all tests. Based on our findings, the synthesised CH-Fla-CMC-5 composite could be used to pack dishes that are watery, acidic, or alcoholic, as well as to coat freshly cut fruits.

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.

Antitumor Activity of Whole-Plant Extracts from In Vitro Cultured and Wild-Growing Clinopodium vulgare Plants on a Panel of Human Tumor Cell Lines

Clinopodium vulgare L. is a valuable medicinal plant with various beneficial effects on health. In this study, water extracts from the aerial part of the wild and in vitro cultured C. vulgare plants were obtained. The polyphenol, flavonoid content and antioxidant activity of the extracts as well as their antitumor efficiency against a panel of cell lines were analyzed. The ability of C. vulgare to inhibit cancer cell migration and induce apoptosis in the tumor cells was examined by wound healing assay and fluorescence microscopic methods. The effect of the extracts on the cell cycle progression of the tumor cells was analyzed by flow cytometry. The presented results show that the antitumor activity of the extracts from in vitro cultured plants was similar to and even exceeded that of the wild plants. The cell viability and migration assays demonstrate the selective anticancer effect of the extract and significant inhibition of cancer cell proliferation and motility. The fluorescence microscopy and cell cycle analyses indicate that the antitumor activity of the in vitro plant extract was related to both antiproliferative and proapoptotic effects. These results show that C. vulgare plants obtained by in vitro micropropagation and cultivated ex vitro are promising candidates for anticancer drug therapy.

Synthesis and characterization of Zn(II) metalated chitosan based nano-hybrid: Antioxidant and antibacterial activity

An efficacious nano-hybrid has been developed comprising of a lacunary polyoxometalate (K8SiW11O39). The lacunary position has been utilized to incorporate alkyl-silane. In alkyl-silane the NH2 group is covalently attached to 2-hydroxy-1-naphthaldehyde through Schiff-base reaction and then metalated with zinc salt. After successful metalation it has been encapsulated into chitosan by ion gel technique to develop nano-hybrid. Each step of the reaction has been monitored by FT-IR spectroscopy. Insertion of alkyl-silane into lacunary polyoxometalate followed by Schiff-base reaction and metalation of zinc has been confirmed by UV-Vis spectroscopy. Further the characteristic bands of UV–Visible spectrum have confirmed the final encapsulation into chitosan to develop the nano-hybrid. EDX analysis reveals the successful metalation with zinc and confirms the presence of all elements in their expected stoichiometric ratio. The inductively coupled plasma (ICP) technique further supported and confirmed the EDX analysis. The antibacterial activity of initial moiety and nano-hybrid have been carried out against bacterial strains of E. Coli gram (-)ve and B. subtilis gram (+)ve. The nano-hybrid has shown better antibacterial activity as compared to the initial moieties L-POM@Sil-HN and L-POM@Sil-NH-Zn. We have also studied the antioxidant activity of nano-hybrid by DPPH assay method; it has shown excellent activity.

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.

The green combustion technique for synthesizing CuO nanoparticles with an extract from Azadirachta indica seeds: potential anticancer and photocatalytic studies using organic dye

The present study highlights the biosynthesis of CuO nanoparticles employing an A. indica seed extract and copper sulphate solution by combustion technique. The extract's phytocomponents facilitated the reduction process and the formation of copper oxide nanoparticles (CuONPs). TEM, SEM, FTIR, X-ray diffractometry, XPS, and ultravioletvisible spectroscopy were used to analyse the pure CuONPs. X-ray diffractometers characterized the CuONPs produced, demonstrating a 12 nm mean particle size. Cu-O stretching vibration bands were detected at 532 cm−1 in the FT-IR spectrum. In UV-vis, the CuO nanoparticles' optical band gaps were at 2.75 eV, with a maximum absorption wavelength of 232 nm. SEM and HRTEM were used to examine the CuONPs; they displayed spherical and undefined shapes with mean sizes of 17.4 nm. The pollution dye rhodamine B was used to test the CuONPs photocatalytic activity. In the presence of sunlight, a remarkable 85% degradation efficiency was attained in 60 minutes, and a degradation constant of k (0.9194 min-1) was observed. This suggests that Azardirachta indica seed extract-derived green-synthesized CuONPs have potential uses in photocatalysis. Furthermore, in the MTT assay method against human renal adenocarcinoma cancer cells, the CuO NPs showed strong anticancer activity, with 5 mg/mL as the lowest IC50 value. This novel green method has demonstrated copper oxide nanoparticle synthesis is a very successful and cost-effective pollutant adsorption technique for treating wastewater.

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.

In Silico Method for ssDNA Aptamer Binding with Aurora Kinase A Protein.

While traditional assay methods face challenges in detecting specific proteins, aptamers, known for their high specificity and affinity, are emerging as a valuable biomarker detection tool. Aurora kinase A (AURKA) plays a role in cell division and influences stem cell reprogramming. In this study, an in silico approach method was conducted for a random ssDNA aptamer sequence selection and its binding with AURKA. The aptamer was designed based on AURKA's structure and nucleic acid sequence, obtained from PDB RCSB. Using RNAfold and RNA composer, we predicted the aptamer's secondary and tertiary structures. Protein-aptamer binding was analyzed via HDOCK and HADDOCK, with 2D interactions visualized in LIGPLOT+ v1.4. Autodock 4 and NAMD 2.3 tools were used to conduct docking and MD simulation studies.

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.

Unlocking the mysteries of n-oxPTH: implications for CKD patients.

Parathyroid hormone (PTH) is a pivotal hormone that regulates serum calcium and phosphate and is closely associated with higher risk of cardiovascular disease and mortality in patients with chronic kidney disease (CKD). PTH can undergo oxidation at methionine 8 and methionine 18 of the molecule. This oxidation process leads to a lower binding affinity to the PTH receptor due to molecular refolding, particularly for PTH oxidized at methionine 8. Although, the oxidation of PTH has been reported for several decades, it is only recently that a method has been developed to detect non-oxidized PTH (n-oxPTH) levels. The utilization of this assay enables the precise detection of n-oxPTH levels and facilitates the evaluation of their correlation with poor prognosis in patients with CKD. However, the current available clinical research findings indicate that n-oxPTH does not demonstrate clinical superiority over iPTH. Here, we provide a comprehensive review on the mechanism of PTH oxidation, the n-oxPTH assay method, and its correlation with iPTH and clinical outcomes.