Journal of immunoassay & immunochemistry最新文献

The incidence of multiple sclerosis (MS) has increased in recent years. Its pathogenesis involves the interaction between various elements, with interleukin 27 (IL-27) playing a key role in autoimmunity. The presence of the IL-27 receptor on astrocytes emphasizes its involvement in the disease's progression.

Purpose: The study aims to investigate possible associations between IL27 rs181206, serum level of IL27, and the development of MS.

Methods: The study comprised 70 MS patients and 70 seemingly healthy controls. They were genotyped for IL27 rs181206 using the Taqman allelic discrimination approach, and their serum IL27 levels were estimated using ELISA.

Results: The frequency of TT genotype, T allele, and IL27 serum level were significantly higher among MS patients compared to controls. There was no significant difference between IL27 serum levels among different genotypes in both MS patients and controls; however, individuals with TT genotype showed higher levels of IL27 than those with CC genotype.

Conclusion: TT genotype and T allele can increase the risk of developing MS. On the other hand, carrying the C allele may be associated with a lower risk of MS development. Understanding IL27 genetics and epistatic interactions can help clarify IL27's role in MS pathogenesis and utilize it as a therapeutic target.

Enzyme-linked immunosorbent assay (ELISA) is a rapid, sensitive, and economical tool for detecting hormones in diverse biological matrices, making it valuable in forensic endocrinology. This review summarizes ELISA's principles, applications, advantages, and limitations in medico-legal practice. ELISA enables quantification of hormones such as cortisol, testosterone, estrogen, melatonin, thyroid hormones, progesterone, and human chorionic gonadotropin (hCG). It supports postmortem toxicology, sexual assault investigations, doping control, and forensic psychiatry. Cortisol may help reconstruct perimortem stress, while hCG detection assists in pregnancy confirmation in assault or maternal death cases. In sports, ELISA screens for anabolic steroids, erythropoietin, and growth hormone, with LC-MS/MS required for confirmation. Its compatibility with blood, saliva, urine, and hair enhances versatility. Key challenges include antibody cross-reactivity, matrix interference, degradation, variability among commercial kits, and limited multiplexing. False positives, hook effects, and inconsistent validation affect admissibility, necessitating strict quality assurance, ISO/IEC 17025 compliance, and confirmatory testing. ELISA is unsuitable for paternity determination; DNA profiling remains the legal standard. Emerging advances - digital ELISA, nanotechnology, AI, and biosensors - promise greater sensitivity and automation but face regulatory, cost, and training barriers. ELISA should be regarded as a complementary, high-throughput screening method integrated into validated, multimodal forensic workflows.

Tuberculosis (TB) is caused by an infection with Mycobacterium tuberculosis. The Bacille Calmette-Guérin (BCG) vaccine is used to treat TB. However, its efficacy varies in different countries. A new TB vaccine is urgently required to inhibit the spread of TB infection. DNA vaccines are promising for providing an immune response against the disease. The resuscitation-promoting factors B and D (rpfB/rpfD) genes are promising vaccine candidates. In this study, the vaccine candidates pcDNA3.1-rpfB and pcDNA3.1-rpfD were evaluated for their ability to inhibit M. tuberculosis infection in BALB/c mice. Epitope analysis indicates that the recombinant protein sequences of RpfB and RpfD used in this study possess epitopes recognized by T and B lymphocytes. Although the presence of M. tuberculosis cells in lung tissue was not detected, histopathological analysis revealed the absence of lymphoid aggregates in mice vaccinated with pcDNA3.1-rpfB or pcDNA3.1-rpfD, in contrast to those administered with phosphate-buffered saline or pcDNA3.1. In addition, analysis of the humoral immune response showed the highest IgG2a antibody titer in mice immunized with both vaccine candidates. These results support our previous findings, which indicate that pcDNA3.1-rpfB and pcDNA3.1-rpfD have considerable potential as TB vaccine candidates.

Psoriasis is characterized by increased levels of pro-inflammatory cytokines, including TNF-α, IL-1β, IL-17A, and NF-κB, as well as keratinocyte hyperproliferation and epidermal thickening. Its pathophysiology is significantly influenced by oxidative stress and abnormal activation of redox-sensitive signaling pathways, such as NF-κB and MAPK. The present study examined the potential of Syringaldehyde (SYD) in the imiquimod (IMQ) induced psoriasis model. Psoriasis Area Severity Index (PASI) scores, back skin thickness, and spleen hypertrophy decreased at dose levels of SYD 25, 50, and 100 mg/kg. Furthermore, a significant reduction in PASI scores following SYD administration indicated a marked improvement in the disease severity and lesion morphology. High-affinity binding of SYD to NF-κB (PDB ID: 4KIK; -34.76 kcal/mol) and MAPK (PDB ID: 1A9U; -32.87 kcal/mol) was found by molecular docking, indicating interference with nuclear translocation and phosphorylation processes. The treatment groups 50 mg/kg and 100 mg/kg indicated restoration of normal histological features. The biochemical evaluation showed decrease in NF-κB, IL-1β, TNF-α, and IL-17 on treatment with SYD. Thus, SYD appears to be a potential therapeutic option for psoriasis, but additional research is needed to confirm its efficacy and safety.

Human β Defensin-2 (HBD2) is an Antimicrobial peptide (AMP) that serves a dual function in host defense and fertility. On the other hand, a plethora of microbial infectious agents have been associated with an increased risk of spontaneous abortion (SA). This study examined the impact of HBD-2 on women with SA with a particular focus on its relationship with clue cells and other microbial infections. 100 women aged 17-50 with a history of SA participated. We utilized two vaginal swabs. They were cultivated first. The Vitek2 technique was used after biochemical tests identified the isolates. vaginosis diagnosed by Amsel criteria. Serum HBD-2 levels were measured using Enzyme-linked immunosorbent assay (ELISA). An association was observed between bacterial vaginosis and SA in the second trimester. HBD-2 levels fell with age. Women with five or more abortions had lower HBD-2 levels (p = 0.034). In cases containing clue cells, HBD-2 averaged 586.0 pg/mL, whereas in cases without clue cells, it decreased (p = 0.007). Gram-negative bacterial infections decreased HBD-2. During fungal infection, HBD-2 levels dropped to 176.1 pg/ml. A significant relationship was observed between HBD2 levels and vaginitis, suggesting its potential as a biomarker and diagnostic tool for reproductive health.

Filgrastim, a therapeutic protein for the treatment of neutropenia, exerts its biological activity by interacting with the granulocyte colony-stimulating factor receptor. However, the affinity of filgrastim to its receptor is usually overlooked during characterization and comparability tests, since its biological activity is assessed only by in vitro proliferation assays. In this work, we propose the use of a cell-based colorimetric method to determine the relative affinity of filgrastim to its receptor expressed in murine myeloid leukemia cells. After performing a validation exercise, the method proved to be accurate, specific, and linear within an affinity interval of 75 to 130%; precise with a confidence interval of 93.6 to 114.2% and a coefficient of variation of 12.6%. The validation exercise proved that the proposed cell-based method is a viable alternative to evaluate the biological activity of filgrastim via the affinity for its receptor and it is suitable to be included as part of its biological characterization exercises as well as in comparability exercises for products coming from distinct manufacturing processes.

Background: The rising global burden of breast cancer demands early detection and effective treatment, with a focus on prognostic and predictive markers. The eighth edition of the American Joint Committee on Cancer staging manual introduced a new prognostic staging system to increase the predictive power of the existing anatomical staging system of breast cancer. The current study aimed to establish the correlation between Ki67 expression with molecular subtypes and with the pathological prognostic stage of invasive ductal carcinoma.

Materials and methods: A total of 40 patients were included in the study with samples from 32 modified radical mastectomies and 8 biopsies. Hematoxylin and Eosin staining, histopathological analysis and Ki67 immunostaining were conducted. Descriptive and inferential statistical analyses were performed.

Results: Bloom Richardson Grade II was the predominant histological grade. In Grade II cases, 15 of 24 had a Ki67 labeling index of 26-45%, while 6 exceeded 45% (p = 0.001). Pathological prognostic staging reclassified 27 cases, with 24 (75%) downstaged, 3 (9.38%) upstaged, and 5 (15.63%) retaining their clinical stage.

Conclusions: Ki67 immunohistochemistry is an effective tool for assessing proliferative activity of invasive ductal carcinoma, aiding in pathological prognostic stage stratification and offering insights into tumor biology.

Background: Application of antibodies in therapeutics and diagnostics are growing Continually. Herein, we aimed to find the most qualified immunoglobulin (Ig) chemical preparation method.

Methods: A rabbit was immunized against recombinant SARS-CoV-2 nucleocapsid (NP) and reactive polyclonal antibodies were prepared using the ammonium sulfate (AS), caprylic acid (CA), polyethylene glycol (PEG), and caprylic acid/ammonium sulfate (CA/AS) methods. Different antibody solutions were analyzed by SDS-PAGE and subsequently quantified by ImageJ software for further analysis in terms of Ig purity, Ig recovery, and albumin impurity. Ultimately, the prepared antibodies were assessed via Western blotting and ELISA to evaluate their ability to bind NP.

Results: Prepared Ig solutions via the CA/AS method had the highest Ig purity (followed by CA, PEG, and AS) and lowest albumin impurity (followed by CA, AS, and PEG). The PEG method had the highest recovery followed by AS, CA, and CA/AS methods. Moreover, antibodies prepared via different methods demonstrated comparable binding capacities to NP in ELISA and Western blotting.

Conclusions: CA/AS, closely followed by CA, proved to be the most qualified method for the preparation of Ig yielding the highest Ig purity while the PEG method resulted in the highest Ig recovery rate.

Background: Noncoding RNAs have recently proven their contributing role in the pathogenesis of numerous diseases. The etiology of psoriasis is still not well defined, and genetic and environmental factors are blamed. We meant to explore the association of circRNAs/microRNAs in psoriasis and inspect their correlation with the clinical features of the disease.

Design and methods: This study included 166 subjects: 83 cases with plaque psoriasis and 83 age- and sex-matched healthy persons as the control group. The expression levels of circRNA000102, circRNA0045272, circRNA0084764, and microRNAs -93,181, 16a, and 21 were estimated by quantitative real-time PCR.

Results: CircRNA0045272 was ten-fold up-regulated, whereas circRNA0084764 and circRNA000102 were about two-fold downregulated in patients vs. control. MicroRNAs -93 and 181 were significantly downregulated in patients vs. control, whereas microRNA -16a and 21 were up-regulated in patients vs. control. CircRNA0045272 was positively correlated with microRNA 181, and circRNA0084764 was positively correlated with microRNA 93 and microRNA 181. CircRNA0045272 had the highest sensitivity (98.8%) and specificity (93.98%), and microRNA-16a had sensitivity (98.80%) with specificity (93.98%).

Conclusion: We assume that circRNAs can predict psoriasis occurrence and progression by sponging a set of microRNAs and can modulate several genes that disturb immune cell balance and inflammatory elements in psoriasis.