Assay Technique Research Articles

Evaluation of Tetracycline and Enrofloxacin Residues in Bovine Milk in Tehran Utilizing ELISA and HPLC Methods

Background: Milk is regarded as one of the most sources of nutrition in the world and has a high value for individual's health. Milk is consumed by sensitive groups including pregnant women, older adults, and children. Therefore, the significance of antibiotic on human health makes it crucial to monitor their existence in food. This investigation aims to evaluate enrofloxacin and tetracycline in the raw and pasteurized milk in Tehran.
 Methods: In this cross-sectional study, 112 raw and 112 pasteurized milks were accumulated in spring and winter from six reputable brands and traditional dairy stores in Tehran from December 2021 until May 2022 for six months, and antibiotic residues were examined by Immunosorbent Assay and High-Performance Liquid Chromatography techniques.
 Results: The findings indicated that the prevalence of tetracycline in raw and pasteurized milk was 41.07 and 26.78% in spring and 85.71 and 35.71% in winter, respectively. The median concentration of tetracycline was 33.27 and 22.65 ppb in spring and 55.81 and 21.91 ppb in winter, respectively. The prevalence of enrofloxacin in raw milk and pasteurized milk samples were 33.92 and 14.28% in spring and 64.28 and 32.14% in winter, respectively. The median concentrations of enrofloxacin were 9.13 and 9.38 ppb in spring and 10.57 and 11.23 ppb in winter, respectively. The raw and pasteurized milk samples collected in winter had higher percentage of antibiotic residue in terms of enrofloxacin than samples collected in spring (p<0.05). Furthermore, the quantity of tetracycline antibiotic in raw milk was significantly higher in winter than in spring (p=0.002). However, the pasteurized milk fails to have significant difference between two seasons. The finding showed the tetracycline and enrofloxacin in all samples were less than 100 ppb (standard limit), and there is no significant difference with the standard limit.
 Conclusion: Based on the obtained results, monitoring the antibiotic residues in milk, controlling and minimizing these residues for human health are regarded crucial.

Interrelation of Hypoxia-Inducible Factor-1 Alpha (HIF-1 α) and the Ratio between the Mean Corpuscular Volume/Lymphocytes (MCVL) and the Cumulative Inflammatory Index (IIC) in Ulcerative Colitis.

We intended to investigate the presence and medical application of serum hypoxia-inducible factor-1 alpha (HIF-1α) along with the already known systemic inflammatory markers and the new one's inflammatory indices, the proportion of mean corpuscular volume and lymphocytes (MCVL) and the cumulative inflammatory index (IIC), for patients with ulcerative colitis (UC). We sought to establish correlations that may be present between the serum levels of HIF-1α and these inflammatory indices, as well as their relationship with disease activity and the extent of UC, which can provide us with a more precise understanding of the evolution, prognosis, and future well-being of patients. Serum samples were collected from 46 patients diagnosed with UC and 23 controls. For our assessment of the serum levels of HIF-1α, we used the Enzyme-Linked Immunosorbent Assay (ELISA) technique. Thus, for HIF-1α we detected significantly higher values in more severe and more extensive UC. When it came to MCVL and IIC, we observed statistically significant differences between the three groups being compared (Severe, Moderate, and Mild). Our study highlighted that HIF-1α correlated much better with a disease activity score, MCVL, and IIC. With MCVL and IIC, a strong and very strong correlation had formed between them and well-known inflammation indices. By examining the ROC curves of the analyzed parameters, we recognized that TWI (accuracy of 83.70%) provides the best discrimination of patients with early forms of UC, followed by HIF-1α (73.90% accuracy), MCVL (70.90% accuracy), and PLR (70.40%). In our study, we observed that HIF-1α, MCVL, and PLR had the same sensitivity (73.33%) but HIF-1α had a much better specificity (60.87% vs. 58.70%, and 54.35%). Also, in addition to the PLR, HIF-1α and MCVL can be used as independent predictor factors in the discrimination of patients with early forms of UC.

Association between Serum 8-Iso-Prostaglandin F2α as an Oxidative Stress Marker and Immunological Markers in a Cohort of Preeclampsia Patients.

We aimed to analyze the presence and clinical use of serum 8-iso-prostaglandin F2-alpha (8-iso-PGF2α) as an oxidative stress marker and some inflammatory status biomarkers (tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), IL-10, high-sensitivity C-reactive protein (hs-CRP), and pentraxin-3 (PTX3)) for patients with preeclampsia (PE). Sixty pregnant women, including thirty diagnosed with PE and thirty who were healthy (NP), were included in this study. For the assessment of serum levels of biomarkers, we used the Enzyme-Linked Immunosorbent Assay (ELISA) technique. Our preliminary study showed that the expression level of serum 8-iso-PGF2α in the PE group was higher than in the PE after delivery (PE-AD) group (742.00 vs. 324.00 pg/mL, p < 0.0001). Groups of preeclamptic patients (PE + PE-AD) expressed significantly elevated levels for all of the assessed inflammatory mediators as compared to NP. Significant strong positive correlations with 8-iso-PGF2α levels were found for systolic blood pressure (SBP), and TNF-α (Spearman's rho = 0.622, p-value = 0.020 and rho = 0.645, p-value = 0.002, respectively). Our study demonstrates that 8-iso-PGF2α and PTX3 have the greatest diagnostic value for pregnant women with PE. 8-iso-PGF2α and PTX3 can be used as independent predictor factors, along with already-known cytokines, that could represent a prophylactic way to help clinicians identify or predict which pregnant women will develop PE.

Corydalis yanhusuo Polysaccharides Ameliorate Chronic Stress-Induced Depression in Mice through Gut Microbiota-Derived Short-Chain Fatty Acid Activation of 5-Hydroxytryptamine Signaling.

Depression, a prevalent psychiatric disorder, presents a serious health risk to humans. Increasing evidence suggested that the gut microbiota and the 5-hydroxytryptamine (5-HT) pathway both contribute significantly to depression. This research aimed to investigate how Corydalis yanhusuo polysaccharides (CYP) could potentially alleviate depression induced by chronic unpredictable mild stress in mice, as well as its underlying mechanism. The sucrose preference test, tail suspension test, and forced swimming test were employed to evaluate the behavior of mice. Enzyme-linked immunosorbent assay and PCR techniques were utilized to measure depression-related factors (dopamine [DA], 5-HT, norepinephrine [NE], brain-derived neurotrophic factor [BDNF], tryptophan hydroxylase 2 [TPH-2], 5-hydroxytryptophan [5-HTP], and tryptophan hydroxylase [TPH-1] levels). Hematoxylin and eosin staining and Nissl staining were conducted to observe histopathological changes in the hippocampus, the differences in the diversity of gut flora between groups were analyzed using 16S rRNA sequencing, and gas chromatography-mass spectrometry metabolomics was utilized to evaluate short-chain fatty acid (SCFA) concentrations. The findings indicated that CYP treatment increased the sucrose preference index, decreased the immobility time, and improved neuropathological injury. In depressed mice, CYP improved the dysregulation of the gut microbiota, and increased the SCFA levels. In addition, CYP enhanced the DA, 5-HT, NE, BDNF, and TPH-2 levels in the brain and the expression of 5-HTP and TPH-1 in the colon, while SCFAs were positively correlated with these levels. In summary, our study suggested that CYP may mitigate depression by ameliorating gut microbiota dysregulation, promoting the generation of SCFAs, and activation of 5-HT signaling expression.

First Serological Evidence of West Nile Virus among Individuals with Febrile Illness in a Tertiary Hospital in Port Harcourt, Nigeria

Aims: West Nile Virus (WNV) infection can cause severe illness. Very little is known about the seroepidemiology of WNV infection in individuals with febrile illness in Nigeria and many other developing countries. This study was carried out to determine the seroprevalence of WNV in individuals with febrile illness attending a tertiary hospital in Port Harcourt, Nigeria and to determine if there was an association between WNV infection with age and sex.&#x0D; Study Design: Cross-Sectional Study&#x0D; Place and Duration of Study: Port Harcourt in Rivers State, Nigeria from September 2019 to December 2019.&#x0D; Methods: Human sera were obtained and WNV IgG was determined using the Enzyme-Linked Immunosorbent Assay (ELISA) technique.&#x0D; Results: Of the 90 study subjects tested, WNV IgG antibodies were present in 27 (30.0%) study participants while 63 (70.0%) study participants were seronegative for WNV IgG antibody. With age, a higher prevalence of WNV occurred among 61-70-year-olds (31.3%, n= 5) compared to 41-60 (30.8%, n= 12) and 20-40 (28.6%, n= 10). A higher prevalence of WNV IgG antibodies occurred in males (34.3%, n=12) than their female counterparts (30.9%, n=17). This study indicated that there is no association between WNV infection with age and sex.&#x0D; Conclusion and Recommendations: These results show that WNV is circulating in Rivers State and has accounted for malaria-like infection in the region. It is recommended that WNV serological testing for malaria-infected individuals should be included as a routine test since they are most likely to present similar symptoms of WNV fever. Also, proper hygiene which includes eliminating mosquito breeding sites is recommended to mitigate the spread of West Nile Virus infection.

Perspectives on antimicrobial properties of Paulownia tomentosa Steud. fruit products in the control of Staphylococcus aureus infections

Ethnopharmacological relevancePaulownia tomentosa Steud. (P. tomentosa) is a medium-sized tree traditionally used in Chinese folk medicine for the treatment of infectious diseases. It is a rich source of prenylated phenolic compounds that have been extensively studied for their promising biological activities. Aim of the studyDue to the increasing development of antibiotic resistance, our study investigated plant-derived natural products from the fruits of P. tomentosa that could control Staphylococcus aureus infections with novel targets/modes of action and reduce antimicrobial resistance. Materials and methodsThe ethanolic extract was fractionated and detected by liquid chromatography. The antistaphylococcal effects of the plant formulations were studied in detail in vitro by various biological methods, including microdilution methods for minimum inhibitory concentration (MIC), the checkerboard titration technique for synergy assay, fluorescence measurements for membrane disruption experiments, autoinducer-2-mediated bioassay for quorum sensing inhibition, and counting of colony-forming units for relative adhesion. Morphology was examined by transmission electron microscopy. ResultsTotal ethanolic extract and chloroform fraction showed MICs of 128 and 32 μg/mL, respectively. Diplacol, diplacone, and 3′-O-methyl-5′-hydroxydiplacone inhibited S. aureus growth in the range of 8–16 μg/mL. Synergistic potential was shown in combination with mupirocin and fusidic acid. The ethanolic extract and the chloroform fraction destroyed the cell membranes by 91.61% and 79.46%, respectively, while the pure compounds were less active. The ethanolic extract and the pure compounds reduced the number of adhered cells to 47.33–10.26% compared to the untreated control. All tested plant formulations, except diplacone, inhibited quorum sensing of S. aureus. Transmission electron microscopy showed deformation of S. aureus cells. ConclusionsThe products from the fruit of P. tomentosa showed antimicrobial properties against S. aureus alone and in combination with antibiotics. By affecting intracellular targets, geranylated flavonoids proposed novel approaches in the control of staphylococcal infections.

Influence of Storage Time on the DNA Integrity and Viability of Spermatozoa of the Spider Crab Maja brachydactyla.

Natural populations of the spider crab Maja brachydactyla constitute a fishery resource of great economic importance in many countries. As in the rest of eubrachyurans, the females of this species have ventral-type seminal receptacles where they store sperm from copulations. Sperm can be stored in these structures for months and even years before egg fertilisation, with the consequent degradation of the sperm cells during the time. In this work, we analyse the viability and the possible genetic damage in sperm accumulated in the seminal receptacles of M. brachydactyla females as a function of the storage time (from 0 to 14 months) using the comet assay technique. On one hand, we developed an algorithm for comet image analysis that improves the comet segmentation compared with the free software Open comet v1.3.1 (97% vs. 76% of detection). In addition, our software allows the manual modification of the contours wrongly delimited via the automatic tool. On the other hand, our data show a sharp decline in sperm viability and DNA integrity in the first four months of storage, which could lead to a decrease in the fecundity rate and/or viability of the embryos or larvae from the second and third clutches of the annual cycle if the repair capacity in these gametic cells is low.

Seminal calbindin 2 level in azoospermia and oligoasthenoteratozoospermia and its correlation with seminal and hormonal parameters.

We aimed to assess seminal calbindin 2 (CALB 2) expression in men with different semen parameters as well as its correlation with reproductive hormones in azoospermic patients and different semen parameters in oligoasthenoteratozoospermic patients. CALB 2 is also known as calretinin and 29 kDa calbindin. This prospective study was performed on 96 cases from the andrology outpatient clinic divided into 3 groups as follows: group 1 including 32 non obstructive azoospermic (NOA) patients, group 2 including 32 patients with oligoasthenoteratozoospermia (OAT), and Group 3 including normozoospermic individuals as controls. Semen analysis and estimation of seminal CALB 2 concentrations by enzyme linked immunosorbent assay (ELISA) technique were performed for all participants. Reproductive hormones were measured in nonobstructive NOA patients. The mean seminal CALB 2 level was higher in OAT patients compared to NOA patients and controls (7.8 ± 1.30 ng/ml, 7.3 ± 0.80 and 7.4 ± 1.0, respectively). Furthermore, the study had shown strong positive correlations between CALB 2 and sperm normal forms in controls and OAT patients. In contrast, there was no significant correlation between seminal CALB 2 and any of the reproductive hormones measured in NOA patients. Seminal CALB 2 may play a role in increasing the abnormal forms in OAT patients.

A Perusal of Phenolics Within Polar Peel Fractions of Malus domestica var Maharaji via Hyphenated Techniques Cum Anti-microbial with REMA and Tyrosinase Inhibitory Potential thereof

This study aims to scrutinize the phenolic secondary metabolites in the polar peel decocture of Malus domestica var Maharaji via hyphenated techniques along with the study of the antibacterial, anti-candida, and tyrosinase inhibitory potential of bioactive compound-rich fractions. Preliminary phenolics go over was performed together with thin layer chromatography before the polar decocture was subjected to hyphenated techniques. FTIR investigation revealed the C-O bonds as in phenols, O-H bond stretch, and vibrations of alcohols and carboxylic acids as well as portrayed the C-H and &gt;C=O stretches among other functional groups all of which are representative of phenolic and polyphenolic compounds. GC-MS perusal demonstrated the presence of bioactive compounds like Quercetin (13.04%), Ascorbic acid (6.48%), p-Coumaric acid (6.17%), Caffeic acid (5.69 %), Mallic acid (5.44%), Apigenin (5.28%), Citric acid (5.15%), Gallic acid (4.38%), Cyanidin (3.52%), and Ferulic acid (3.51%). Kirby-Bauer method followed by the resazurin microtiter assay technique (REMA) for MIC/MBC against six MTCC bacterial strains and one yeast, all producing stubborn opportunistic infection in humans, was used to assess the antibacterial property of all the bioactive rich fractions. Some fractions comparatively revealed a good activity index (AI) against tested microbes. MIC concentrations for bacteria ranged from 15-24 mg/ml while a lower MBC value recorded was 18 mg/ml. Methanol fraction revealed significant tyrosinase inhibitory activity by revealing IC50 of 980.98 µg/ml when L-Tyrosine was substrate and IC50 of 830.68 µg/ml when L-DOPA was substrate when juxtaposed to standard kojic acid that revealed IC50 of 128.822 µg/ml when L-Tyrosine was substrate and IC50 of 149.43 µg/ml when L-DOPA was substrate. The bioactive compounds possessed by the fractions, may be synergistically, turned out to be more effective in the diphenolase reaction and kojic acid acts more effectively in the monophenolase one. It was inferred that peel phenolics of this malus variety have a lot of therapeutic potential in the context of bacterial infections and pigmentation disorders.

Antimicrobial Potency and E. coli β-Carbonic Anhydrase Inhibition Efficacy of Phenazone-Based Molecules.

In this investigation, 4-antipyrinecarboxaldhyde was reacted with methyl hydrazinecarbodithioate to afford the carbodithioate derivative 3. The as-prepared carbodithioate derivative 3 is considered to be a key molecule for the preparation of new antipyrine-1,3,4-thiadiazole-based molecules (4-9) through its reaction with the appropriate hydrazonoyl halides. Furthermore, a typical Biginelli three-component cyclocondensation reaction involving ethyl acetoacetate, 4-antipyrinecarboxaldhyde, and thiourea under the standard conditions is carried out in the presence of sulfuric acid to afford the corresponding antipyrine-pyrimidine hybrid molecule (10). The latter was submitted to react with hydrazine monohydrate to provide the corresponding hydrazide derivative (11) which, under reaction with ethyl acetoacetate in refluxing ethanol containing catalytic amount of acetic acid, afforded the corresponding derivative (12). The structure of the newly synthesized compounds was affirmed by their spectral and microanalytical data. We also screened for their antimicrobial potential (ZOI and MIC) and conducted a kinetic study. Additionally, the mechanism of biological action was assessed by a membrane leakage assay and SEM imaging technique. Moreover, the biological activities and the binding modes of these compounds were further supplemented by an in silico docking study against E. coli β-carbonic anhydrase. The amount of cellular protein released by E. coli is directly correlated to the concentration of compound 9, which was found to be 177.99 µg/mL following treatment with 1.0 mg/mL of compound 9. This finding supports compound 9's antibacterial properties and explains how the formation of holes in the E. coli cell membrane results in the release of proteins from the cytoplasm. The newly synthesized compounds represent acceptable antimicrobial activities with potential action against E. coli β-carbonic anhydrase. The docking studies and antimicrobial activity test proved that compound (9) declared a greater activity than the other synthesized compounds.

Effects of probiotic saccharomyces bourlardii on cytokine levels and outcomes of childhood severe malaria: Study protocol for a randomized controlled trial

Background: Severe malaria is acute malaria with signs of organ dysfunction and/or hyperparasitaemia. About 90% of the world’s severe and fatal malaria is estimated to affect young children in sub-Sahara Africa. Systemic complications and neuropathology in severe malaria include ‘cytokine storm hypothesis’. Probiotic immunomodulation may show therapeutic benefits in severe malaria. Aim: This study aims to evaluate the effects of probiotic saccharomyces bourladii on serum cytokine levels (IL-6 and TNF), clinical course and outcome of children with severe malaria. Methods: Participants shall be recruited based on WHO diagnostic criteria for severe malaria and their clinical-demographic data shall be obtained with a pretested questionnaire. The study design is randomized controlled trial (RCT); participants shall be randomized into two study sub-groups (intervention vs. control). All participants shall receive standard anti-malarial therapies. In addition, probiotic saccharomyces bourladii 250mg twice a day shall be administered to those in the intervention sub-group for 3 days. There after, their serum cytokines (IL-6 and TNF) shall be measured quantitatively by a Sandwich enzyme-linked immunosorbent assay (ELISA) technique. Primary outcomes shall be cytokine levels and length of stay (LOS) while secondary measures shall be coma scores, seizure, neurological deficits and mortality. The data shall be analyzed using SPSS version 26.0 statistical software for Windows. Both ‘intention-to-treat analysis’ and ‘analysis as per protocol’ shall be done. P-value&lt; 0.05 shall be considered significant in all tests. Discussion: At the end of this clinical trial, it is expected that the potential benefits of probiotic saccharomyces bourladii in modulating pro-inflammatory cytokines, averting systemic complications and reducing mortality in childhood severe malaria would have been verified. Protocol No: ADM/E22/A/ VOL.VII/148312145; University of Benin Teaching Hospital’s Health Research Ethics Committee Approval; August 5th, 2022.

Role of Human β-Defensin-3 in Rheumatoid Arthritis: An Observational Single-Center Study

Background: It's believed that HBD-3 is involved in the tissue remodeling process of articular cartilage. Also, HBD-3 has anti-inflammatory properties. Objectives: The purpose of this study is to assay human beta-defensine-3 (HBD-3) in serum from rheumatoid arthritis (RA) patients and investigate its correlation with proinflammatory cytokines. Methods: In this case-control study, fifty-eight RA patients were aged 20–65 years, and 29 age-matched healthy subjects (HS) had no inflammatory rheumatic diseases. The disease activity score-28 joint erythrocyte sedimentation rate (DAS28-ESR) was used to measure RA activity. CRP, ACPA, HBD-3, TNF-α, and IL-1β were assessed using the enzyme-linked immunosorbent assay technique (ELISA). Results: There was a significant increase in RF, ACPA, CRP, proinflammatory cytokines, and HBD-3 in the RA group compared with the HS group. There was no significant difference in HBD-3 levels according to the activity of diseases. The results of the correlation between HBD-3 and proinflammatory cytokines showed a significant positive relationship in the RA group. Conclusions: Inflammatory markers and S.HBD-3 demonstrated fair diagnostic performance to differentiate RA from HS. The current study supports the hypothesis that there is a correlation between HBD-3 and the immunoregulatory response.

Parental stress parameter difference after caring for congenital adrenal hyperplasia children with genital ambiguity

Abstract Introduction: Congenital adrenal hyperplasia (CAH) disrupts cortisol biosynthesis, which may cause genital ambiguity in females. Caring for children with genital ambiguity creates parental distress. Common stress parameters include biological (e.g., serotonin cortisol) and psychological (e.g., Depression, Anxiety, and Stress Scale [DASS] scores). This study elaborated parental stress parameter differences among parents of CAH children with and without genital ambiguity. Methods: This cross-sectional study included 34 fathers and 44 mothers of children with CAH. All participants agreed to fill in the DASS-42 and socioeconomic questionnaire. Morning serum serotonin and cortisol samples were examined using the enzyme-linked immune-absorbent assay technique. Results: The median (minimum–maximum) value of serotonin concentration was lower in fathers compared to mothers (281.18 [78.44–122.91] ng/ml vs. 399.78 [122.91–1050.0] ng/ml, P = 0.11); in parents of children with genital ambiguity (292.22 [78.44–1050.00] ng/ml vs. 426.71 [150.78–1050.00] ng/ml, P = 0.01); and in parents of children with a diagnosis time &lt;1 year (225.00 [93.41–570.33] ng/ml vs. 371.815 [78.44–1050.00] ng/ml, P = 0.046). Higher cortisol concentration was observed in parents of children with diagnosis time &lt;1 year (78.65 [50.89–126.89] ng/ml vs. 63.33 [6.14–283.67] ng/ml, P = 0.046). The proportion of depression was 24.4%, anxiety 45.1%, and stress 21.8%. Conclusion: Fathers, parents of children with genital ambiguity, and being recently diagnosed had lower serum serotonin levels. Parents of recently diagnosed children had higher cortisol levels. Anxiety was more common than depression and stress.

Immunogenicity of Recombinant Erwinia asparaginase (JZP458) for Acute Lymphoblastic Leukemia (ALL) or Lymphoblastic Lymphoma (LBL)

Introduction: L-asparaginase is important in multi-agent treatment regimens for pediatric and adult patients (pts) with ALL/LBL. However, hypersensitivity reactions (HSRs) to E. coli-derived asparaginases (ASP) often lead to treatment delay or discontinuation. JZP458, a recombinant Erwinia ASP derived from a Pseudomonas fluorescens expression platform, is indicated as part of multi-agent chemotherapy regimens for ALL/LBL in adult and pediatric pts ≥1 month old who developed HSRs to E. coli-derived ASP. Here we report the immunogenicity profile of JZP458 based on study AALL1931. Methods: AALL1931 was an open-label, 2-part, multicenter, phase 2/3 trial conducted with the Children's Oncology Group to investigate efficacy, safety, and pharmacokinetics (PK) of JZP458 in ALL/LBL (NCT04145531). Eligible pediatric and adult pts with ALL/LBL who developed HSRs (grade ≥3 allergic reaction or silent inactivation) to E. coli-derived ASP received JZP458 as part of their multi-agent treatment regimen. Each dose of E. coli-derived pegaspargase was substituted with 6 doses of JZP458 either intramuscularly (IM, Part A) or intravenously (IV, Part B) on Mon/Wed/Fri over 2 weeks, defined as 1 course. Efficacy endpoints were the proportion of pts maintaining therapeutic nadir serum ASP activity (SAA) ≥0.1 IU/mL at the last 72-hour (primary endpoint) or 48-hour (key secondary endpoint) time point during course 1. Other endpoints included safety and tolerability, PK, and antidrug antibody (ADA) formation. Immunogenicity samples were collected prior to dose 1 in each course, prior to dose 6 in course 1, and at end-of-study safety follow-up visit (30±3 days after last dose). For pts experiencing HSRs, additional samples were tested for ADAs. A multi-tiered approach was used to measure ADAs and neutralizing antibodies (NAbs) to evaluate immunogenicity of JZP458. First, samples were screened for ADAs that bind JZP458. Samples with potentially ADA-positive (+) results were retested for binding specificity confirmation. Samples confirmed as ADA+ were tested further for NAbs. For pts with ADA+ samples obtained prior to study end, follow-up ADA samples up to approximately 6 months after the last dose of the last course were collected. The ADA assay was a validated binding assay based on an electro-chemiluminescence immunosorbent assay technique and the NAb assay was a validated enzymatic activity assay. Results: The median (range) age of pts was 10 (1-25) years. Analysis of pts receiving IM JZP458 (n=167) showed that 82 (49%) pts had confirmed ADA toward JZP458; of these, 7 (4%) were ADA+ at pre-dose 1. Twenty-two pts (15% of 146 pts with samples) remained ADA+ at the end of study. Overall, 18 (11%) pts developed treatment-related HSRs after receiving IM JZP458. Among ADA+ pts (n=82), 11 (13%) experienced treatment-related HSR and 4 (5%) developed positive NAbs. Of ADA-negative (-) pts, 7/85 (8%) experienced treatment-related HSRs. Among pts receiving IV JZP458 (n=61), 34 (56%) had confirmed ADA toward JZP458; of these, 1 (2%) was ADA+ at pre-dose 1. Seventeen pts (34% of 50 pts with samples) remained ADA+ at the end of study. Overall, 16 (26%) pts developed treatment-related HSRs after receiving IV JZP458. Among ADA+ pts (n=34), 12 (35%) experienced treatment-related HSRs and 2 (6%) developed positive NAbs. Of ADA- pts, 4/27 (15%) experienced treatment-related HSRs. Anti-JZP458 antibody status was evaluated as a potential covariate for PK of IM or IV JZP458 within the population PK analysis and it was not found to be a significant covariate. SAA levels were similar between ADA+ and ADA- pts. Overall treatment-related adverse events (TRAEs) in ADA+ and ADA- pts were similar and consistent with the most common TRAEs expected with ASP-containing chemotherapy for ALL (Table). TRAEs led to JZP458 discontinuation for 14 pts with ADA+ status in Part A (8%) and 13 in Part B (21%). Conclusions: The majority of pts who developed ADAs did not develop HSRs or NAbs. The overall rate of HSRs was similar to literature-reported rates for ASP and appeared to be higher with IV vs IM JZP458. ADA status did not appear to impact PK and other AEs for JZP458. Based on these results, no specific immunological concerns were identified with JZP458 beyond known immunogenicity risks associated with ASP and no specific clinical decisions are recommended based on immunogenicity testing.

Relationship between Levels of Interleukin-6 and Tumour Necrosis Factor Alpha and Khorana Scores of Newly Diagnosed Ambulatory Cancer Patients

Abstract Background: In cancer microenvironment, there is increased production of inflammatory cytokines, including tumour necrosis factor (TNF)-α, interleukin (IL)-1a, and IL-6. These cytokines are capable of causing thrombus formation through the activation of tissue factors. Then, the Khorana risk score can be Utilised to stratify the risk of venous thromboembolism in cancer patients. Aim: The objective of this study was to determine the relationship between levels of IL 6, TNF-α, and Khorana scores of chemotherapy-naïve ambulatory cancer patients (CNACPs). Materials and Methods: This was a cross-sectional study. The case group (n = 100) consisted of newly diagnosed CNACP recruited from adult oncology clinics of University of Nigeria Teaching Hospital, Ituku-Ozalla Enugu, Nigeria, whereas the control group (n = 100) consisted of healthy individuals. The participants’ blood samples were assayed by enzyme-linked immunosorbent assay (ELISA) technique for IL-6 and TNF-α. Demographic variables were analysed using descriptive statistics in the form of frequencies and percentages and P &lt; 0.05 was considered statistically significant. Results: There were significant differences between the mean levels of IL-6 (P = 0.036) and TNF-α (P = 0.001) in three Khorana score groups of the case group. Further, comparisons of mean IL-6 levels between three Khorana score groups showed that differences were between high-risk versus low-risk groups (P = 0.026) and between intermediate-risk versus low-risk groups (P = 0.014). Comparison of mean TNF-α levels in three Khorana score groups of CNACP showed that differences were between high-risk versus low-risk groups (P = 0.026) and between intermediate-risk versus low-risk groups (P = 0.014). Furthermore, there was a significant positive correlation between the Khorana scores and IL-6 (r = 0.28, P = 0.031) and TNF-α (r = 0.254, P = 0.011). The mean serum levels of IL-6 and TNF-α were significantly higher in CNACP than in healthy control (8.98 [8–12] pg/ml vs. 8.43 [2–10] pg/ml, P &lt; 0.001) and (P &lt; 0.001 and &lt; 0.01), respectively. Conclusion: Inflammatory biomarkers are elevated in ambulatory CNACP and their values are significantly related with Khorana scores. There is a need for more studies on the possible benefit of prophylactic anticoagulation for newly diagnosed CNACP.

Multiplicity counting using organic scintillators to distinguish neutron sources: An advanced teaching laboratory

In this advanced instructional laboratory, students explore complex detection systems and nondestructive assay techniques used in the field of nuclear physics. After setting up and calibrating a neutron detection system, students carry out timing and energy deposition analyses of radiation signals. Through the timing of prompt fission neutron signals, multiplicity counting is used to carry out a special nuclear material (SNM) nondestructive assay. Our experimental setup is comprised of eight trans-stilbene organic scintillation detectors in a well-counter configuration, and measurements are taken on a spontaneous fission source as well as two (α,n) sources. By comparing each source's measured multiplicity distribution, the resulting measurements of the (α,n) sources can be distinguished from that of the spontaneous fission source. Such comparisons prevent the spoofing, i.e., intentional imitation, of a fission source by an (α,n) neutron source. This instructional laboratory is designed for nuclear engineering and physics students interested in organic scintillators, neutron sources, and nonproliferation radiation measurement techniques.

Nanotechnology-based Assays for the Detection of Cancer through Sputum

Abstract: Cancer remains a formidable global health challenge, underscoring the critical need for early and accurate diagnostic tools. In recent years, nanotechnology has emerged as a promising frontier in cancer detection, offering innovative approaches to enhance sensitivity, specificity, and convenience in diagnostics. This comprehensive review delves into the intersection of nanotechnology and sputum-based assays for cancer detection. Sputum, a readily accessible biomarker source, has gained increasing attention due to its potential to house early cancer biomarkers. By harnessing the unique properties of nanoparticles, this review explores how nanotechnology is revolutionizing sputum- based cancer detection. We examine the intrinsic attributes of sputum as a diagnostic fluid, the diverse applications of nanoparticles, and the evolving landscape of cancer-specific biomarkers. Moreover, we provide insights into various nanotechnology assay techniques and their clinical implications while highlighting the challenges and future prospects in this dynamic field. This review aims to elucidate the transformative impact of nanotechnology on cancer diagnostics through sputum, offering a roadmap towards earlier and more accurate cancer detection.

Tracking tumor alteration in glioma through serum fibroblast activation protein combined with image

PurposeDetecting tumor progression of glioma continues to pose a formidable challenge. The role of fibroblast activation protein (FAP) in gliomas has been demonstrated to facilitate tumor progression. Glioma-circulating biomarkers have not yet been used in clinical practice. This study seeks to evaluate the feasibility of glioma detection through the utilization of a serum FAP marker.MethodsWe adopted enzyme-linked immunosorbent assay (ELISA) technique to quantify the relative FAP level of serum autoantibodies in a cohort of 87 gliomas. The correlation between preoperative serum autoantibody relative FAP levels and postoperative pathology, including molecular pathology was investigated. A series of FAP tests were conducted on 33 cases of malignant gliomas in order to ascertain their efficacy in monitoring the progression of the disease in relation to imaging observations. To validate the presence of FAP expression in tumors, immunohistochemistry was conducted on four gliomas employing a FAP-specific antibody. Additionally, the investigation encompassed the correlation between postoperative tumor burden, as assessed through volumetric analysis, and the relative FAP level of serum autoantibodies.ResultsA considerable proportion of gliomas exhibited a significantly increased level of serum autoantibody relative FAP level. This elevation was closely associated with both histopathology and molecular pathology, and demonstrated longitudinal fluctuations and variations corresponding to the progression of the disease The correlation between the rise in serum autoantibody relative FAP level and tumor progression and/or exacerbation of symptoms was observed.ConclusionsThe measurement of serum autoantibody relative FAP level can be used to detect the disease as a valuable biomarker. The combined utilization of its detection alongside MR imaging has the potential to facilitate a more accurate and prompt diagnosis.

Seasonal Study of Aflatoxin M1 Contamination in Cow Milk on the Retail Dairy Market in Gorgan, Iran

Milk and milk products are the main nutritional foods for all age groups, especially for infants and children. Milk may be dangerous to consume due to the presence of a harmful substance called Aflatoxin M1 (AFM1). The objective of this study was to assess the levels of AFM1 in milk, particularly those that exceed the standards set by the European Union (50 ng/L), the Food and Drug Administration (500 ng/L), and the Iranian National Standards Organization (100 ng/L). The study included one hundred and eighty samples of raw cow’s milk from various retail dairy markets in Gorgan, with 45 samples collected during each season. The level of Aflatoxin M1 in the samples was measured using the enzyme-linked immunosorbent assay (ELISA) technique. AFM1 was detected in 139 (72.2%) raw cow milk samples with a range of 3.5–357 ng/L. All of the samples collected had Aflatoxin M1 concentration levels that were below the maximum limit of 500 ng/L set by the FDA. However, 41 samples (22.7%) exceeded the EU’s limit of 50 ng/L, and 26 samples (14.4%) exceeded the INSO’s limit of 100 ng/L for Aflatoxin M1 in raw cow’s milk. The lowest and highest AFM1 levels of contamination were detected in the summer and winter seasons, which constituted 32 (71.1%) and 38 (84.4%) samples, respectively. The consumption of raw cow milk can lead to health risks for individuals from various age groups because regulatory limits are not being followed.

Performance of IS6110-LAMP assay for detection of Mycobacterium tuberculosis complex in blood and urine samples from patients with extrapulmonary tuberculosis

BackgroundThe diagnosis of extrapulmonary tuberculosis (EPTB) shows numerous difficulties because of non-specific symptomatology and low sensitivity of conventional methods. Loop-mediated isothermal amplification (LAMP) is a fast and low-cost technique, which can amplify under isothermal conditions an amount of target DNA copies into approximately a billion copies. ObjectiveThe present study aimed to evaluate a IS6110-LAMP system for Mycobacterium tuberculosis detection in blood and urine samples from patients with EPTB. MethodsThe collected samples (n = 122) were stratified in two groups: Group EPTB – patient samples with confirmed EPTB (n = 61); Group non-TB – patient samples without TB (n = 61). The urine samples underwent decontamination, and the components of blood samples were separated (plasma and PBMC). DNA extractions were performed in all biological samples followed by IS6110-LAMP assay technique. The detection limit was evaluated through dilution curves (1:10) using Mtb reference strain (H37Rv) genomic DNA. FindingsThe detection limit of IS6110-LAMP was 10 fg/μL (∼10–20 bacilli/μL). The IS6110-LAMP technique sensitivity and specificity were 95.65 % and 79.25 %, respectively, with a general kappa agreement index of 0.762. Main conclusionsBased on these results, IS6110-LAMP test showed considerable diagnostic parameters, being able to aid in the speed and accuracy of the final EPTB diagnosis.