Enzyme-linked Immunoassay Kit Research Articles

Point-of-Care Detection of HER2 and CA 15-3 in Breast Cancer Patients: Dual-Channel Biosensor Implementation

Breast cancer remains a considerable health challenge, affecting numerous individuals annually. This research introduces an innovative method for detecting breast cancer utilizing dual-channel test strips capable of simultaneously assessing two key biomarkers—HER2 and CA 15-3. The test strip utilized in this study is not only cost-effective but also entirely non-invasive. The reusable device employs a printed circuit board with metal-oxide-semiconductor field-effect transistor amplification and Arduino-based control to convert voltage signals from test strips into digital readings efficiently. The device utilizes double-pulse measurement instead of direct current, effectively mitigating the screening effect. The detection limit for both biomarkers is exceptionally low at 10−15 g ml−1, surpassing commercial enzyme-linked immunoassay kits by four orders of magnitude. The sensor demonstrates remarkable sensitivity, with 78/dec for HER2 and 56/dec for CA 15-3. Human sample tests were conducted to validate the efficacy of the dual-channel strip, successfully distinguishing between healthy and cancerous groups. The results reveal significant p-values for both HER2 and CA 15-3 tests, underscoring the significance of this research. Note that this is a rapid testing process, completed in less than 2 s. These findings offer a promising avenue for swift and accurate breast cancer detection, furnishing crucial insights for early diagnosis and subsequent treatment.

Comparative Evaluation of Periodontal Ligament-associated Protein-1/Asporin Levels in Periodontal Tissue in Health and Disease

Background: Periodontal ligament-associated protein-1 (PLAP-1)/asporin is an extracellular matrix protein that plays a protective role in the pathogenesis of periodontitis. There is a paucity of information about the association between PLAP-1/asporin and periodontitis in human PDL. Thus, in this study, PLAP-1/asporin levels between participants with healthy periodontium and chronic periodontitis were compared and correlated with periodontal parameters. Materials and Methods: Fifty participants were recruited and divided into 25 in each group: Group 1 (control) and Group 2 (test). Probing pocket depth (PPD) and clinical attachment level (CAL) were recorded. Periodontal ligament (PDL) samples were collected from extracted teeth for estimating PLAP-1/asporin levels using the Human Asporin Enzyme-Linked Immunoassay Kit. Results: A statistically significant difference (P = 0.001) in the PLAP-1/asporin levels was observed between Group 1 and Group 2. A weak negative correlation was observed between PLAP-1/asporin levels and periodontal parameters (PPD and CAL) in both groups. Conclusion: In this study, higher PLAP-1/asporin levels in participants with healthy periodontium highlight the protective role of PLAP-1/asporin in maintaining periodontal homeostasis.

Comparative Evaluation of Periodontal Ligament-associated Protein-1/Asporin Levels in Periodontal Tissue in Health and Disease

Background: Periodontal ligament-associated protein-1 (PLAP-1)/asporin is an extracellular matrix protein that plays a protective role in the pathogenesis of periodontitis. There is a paucity of information about the association between PLAP-1/asporin and periodontitis in human PDL. Thus, in this study, PLAP-1/asporin levels between participants with healthy periodontium and chronic periodontitis were compared and correlated with periodontal parameters. Materials and Methods: Fifty participants were recruited and divided into 25 in each group: Group 1 (control) and Group 2 (test). Probing pocket depth (PPD) and clinical attachment level (CAL) were recorded. Periodontal ligament (PDL) samples were collected from extracted teeth for estimating PLAP-1/asporin levels using the Human Asporin Enzyme-Linked Immunoassay Kit. Results: A statistically significant difference (P = 0.001) in the PLAP-1/asporin levels was observed between Group 1 and Group 2. A weak negative correlation was observed between PLAP-1/asporin levels and periodontal parameters (PPD and CAL) in both groups. Conclusion: In this study, higher PLAP-1/asporin levels in participants with healthy periodontium highlight the protective role of PLAP-1/asporin in maintaining periodontal homeostasis.

Effect of butylphthalide on prevention and treatment of high altitude cerebral edema in rats

3-n-butylphthalide (NBP) contains one of the main active ingredients of celery seed. It has a series of pharmacological mechanisms, including reconstitution of microcirculation, protection of mitochondrial function, inhibition of oxidative stress, and inhibition of neuronal apoptosis. Based on the complex multi-targeting of NBP pharmacological mechanisms, the clinical applications of NBP are increasing, and more and more clinical studies and animal experiments have focused on NBP. In this study, we used male Sprague Dawley rats as an animal model to elucidate the intervention effect of butylphthalide on high altitude cerebral edema (HACE), and also compared the effect of butylphthalide and rhodiola rosea on HACE. Firstly, we measured the changes of body weight and brain water content and observed the pathological changes of brain tissues. In addition, the contents of inflammatory factors, oxidative stress and brain neurotransmitters were assessed by enzyme-linked immunoassay kits, and finally, the expression of apoptotic proteins in brain tissues was determined by western blotting. The results showed that NBP reduced brain water content, attenuated brain tissue damage, altered inflammatory factors, oxidative stress indicators, and brain neurotransmitter levels, and in addition NBP inhibited the expression of Caspase-related apoptotic proteins. Therefore, NBP has the potential to treat and prevent HACE.

3D-printed biosensors in biomedical applications exploiting plasmonic phenomena and antibody self-assembled monolayers.

In this work, a 3D-printed plasmonic chip based on a silver-gold bilayer was developed in order to enhance the optical response of the surface plasmon resonance (SPR) probe. More specifically, numerical and experimental results were obtained on the 3D-printed SPR platform based on a silver-gold bilayer. Then, the optimized probe's gold plasmonic interface was functionalized with a specific antibody directed against the p27Kip1 protein (p27), an important cell cycle regulator. The 3D-printed plasmonic biosensor was tested for p27 detection with good selectivity and a detection limit of 55 pM. The biosensor system demonstrated performance similar to commercially available ELISA (enzyme-linked immunoassay) kits, with several advantages, such as a wide detection range and a modular and simple-based architecture. The proposed biosensing technology offers flexible deployment options that are useful in disposable, low-cost, small-size, and simple-to-use biochips, envisaging future applications in experimental and biomedical research.

Interleukin-6 Concentration in Early Assessment of Severity and Prediction of Outcome among Patients with Acute Pancreatitis: A Prospective Cohort Study

Introduction: Various inflammatory markers, such as C-reactive Protein (CRP), Interleukin (IL), and Tumour Necrosis Factor (TNF), are used as indicators of illness severity in acute pancreatitis. Concentrations of inflammatory mediators are correlated to the severity of the disease. If detected before the occurrence of multiple organ dysfunction, early initiation of aggressive therapy might prevent its development. IL-6, released by macrophages in response to tissue injury, peaks 24 hours after the onset of inflammation and can be used to predict early disease severity. The Bedside Index of Severity in Acute Pancreatitis (BISAP) scoring system was used to assess the severity of acute pancreatitis. Aim: To study the association of IL-6 levels with the BISAP in predicting the outcome. Materials and Methods: A prospective cohort study was conducted in the Department of General Surgery at Government Medical College, Kottayam, Kerala, India. The duration of the study was 12 months, from January 1, 2019 to December 31, 2019. The study was done on patients of either gender, aged between 18 and 60 years, with clinical features suggestive of acute pancreatitis. After obtaining informed consent, a 4 mL venous blood sample was taken, and serum amylase and serum lipase levels were estimated. If serum amylase/lipase levels were three times above the normal range, a diagnosis of acute pancreatitis was made according to revised Atlanta criteria. Meanwhile, the BISAP score was calculated. After reaching a sample size of 77 for the present study, IL-6 estimation was performed using an Enzyme-linked Immunoassay (ELISA) kit. At the end of the study, IL-6 concentration was compared with the BISAP score to assess severe acute pancreatitis. The data entered in Microsoft Excel were analysed using the Chi-square test and Fisher’s-exact test. Results: The mean age of the study participants was 30 years, with 37 patients (48%) between the ages of 20-40 years and 40 patients (52%) between the ages of 40-60 years. Out of the 77 study subjects, 71 were males and six were females. There was a significant association between Intensive Care Unit (ICU) admission with persistent organ failure, which indicates severe acute pancreatitis. Conclusion: In the present study, no significant association was found between IL-6 and the BISAP score in the early assessment and prediction of the severity of acute pancreatitis.

Colorimetric and photothermal dual-mode aptasensor with redox cycling amplification for the detection of ochratoxin A in corn samples

Ochratoxin A (OTA) detection is critical for public health safety. This study proposes a G-quadruplex-Hemin/iodide (G4-Hemin/I−)-mediated non-enzyme redox cycling amplification (RCA) system for dual-modal (colorimetric and photothermal thermometer) OTA analysis. The proposed aptasensor platform for point-of-care testing employs a common thermometer for quantitative signal readouts. The OTA aptamer folds into a G4 structure, which significantly enhances the catalytic activity in the presence of I− after RCA reaction. Moreover, a notable temperature enhancement causes color changes, providing an ultrasensitive and label-free platform for OTA detection. Further, the designed sensor was applied to OTA content determination in corn samples and achieved satisfactory results compared to a commercial enzyme-linked immunoassay kit. The proposed dual-mode aptasensor is simple, highly sensitive (1 pg/mL for colorimetric method, 0.8 pg/mL for photothermal method), selective, and suitable for low-cost instrument-free bioanalysis in low-resource settings.

Determining the Seroprevalence and the Knowledge of Viral Hepatitis B Infection among Beauticians in Yenagoa LGA, Yenagoa, Bayelsa State, Nigeria

Cosmetology is a rapidly growing field, resulting in increasing numbers of beauty centers and beauticians. Ear piercing is a common practice in Nigeria and, in recent years piercing of other body parts has greatly increased in popularity. Beauty treatments, such as piercing, tattooing, manicuring, and barbing are used by many people. Individuals working in barber shops, hairdressing and beauty centers are likely to have contact with blood through applications such as shaving, manicure, pedicure and skin care. The aim of this study was to determine hepatitis B virus (HBV) seroprevalence in a sample of beauticians in Yenagoa local government area, Bayelsa State (Nigeria) and to assess the level of knowledge of these professionals regarding viral hepatitis. This was a descriptive cross-sectional study involving a total of 120 beauticians (hairdressers and manicurists/pedicurists) that were selected by a multistage sampling method. Data was collected by an interviewer-administered questionnaire for knowledge assessment and serum samples were tested for HBsAg positivity using commercial enzyme-linked immunoassay (ELISA) kits. Data collected were analyzed using the statistical package for social science (SPSS), version 25 software. Of the total 120 participants 16 (13.3%) were males and 104 (86.7%) were females. The prevalence of HBV infections among the respondents was 7.5%. The knowledge of beauticians on awareness of viral hepatitis B was 5.8%, and their knowledge on ways of transmission was 61.7%. Conclusions: the findings indicate that, due to their low level of awareness of viral hepatitis B existence, beauticians working in Yenagoa Local Government Area are in a risk of HBV infection even though the seroprevalence of HBV was low.

Analysis of Influence of Cigarette Smoke on Signaling Pathways of Innate Immune System in Monocytes of Peripheral Blood

INTRODUCTION: Tobacco smoking is an important medical problem since it has a significant impact on the development and progression of chronic obstructive pulmonary disease (COPD). The components of tobacco smoke can initiate and support local and systemic inflammation with participation of monocytes and macrophages.
 AIM: To study molecular mechanisms associated with the impact of cigarette smoke on signaling pathways of the innate immune system in monocytes of peripheral blood.
 MATERIALS AND METHODS: The methods of in silico analysis was used to identify genes associated with the impact of tobacco smoke. On the basis of the data obtained, a cellular model of inflammation was created in vitro using tobacco smoke extract and monocytes of peripheral blood isolated by immunomagnetic separation. An enzyme-linked immunoassay (ELISA) kit was used to measure the concentration of tumor necrosis factor- (TNF-), interleukin-1 (IL-1) in cell supernatants, and of Toll-like receptor 4 (TLR4), ATP-binding cassette A1 (ABCA1) in homogenates of cell membranes of native monocytes and monocytes exposed to 4% tobacco smoke extract. These data were compared with the levels of TNF-, IL-1, TLR4 and ABCA1 in monocytes of peripheral blood of patients with COPD with frequent exacerbation phenotype and with obliterating atherosclerosis of lower limb arteries (OALLA). For statistical processing and visualization of the data, MedCalc 20.1.4 and R (version 4.2.2) software was used.
 RESULTS: Tobacco smoke influences TLR4, TNF- signaling pathways and lipid metabolism. Cigarette smoke extract enhanced the expression of proinflammatory cytokines TNF- and IL-1 in cell supernatants, increased the level of TLR4 and decreased that of ABCA1 in plasmolemma of monocytes of peripheral blood. In patients with COPD with frequent exacerbation phenotype and with OALLA, there were shown increase in the levels of proinflammatory TNF- and IL-1 cytokines in cell supernatants, increase in the level of TLR4 and reduction of the level of ABCA1 in plasmolemma of monocytes of peripheral blood compared to native monocytes of healthy individuals.
 CONCLUSION: Cigarette smoke enhances the production of proinflammatory TNF- and IL-1 cytokines, increases the levels of TLR4 protein and reduces the amount of ABCA1 transporter in membranes of monocytes of peripheral blood. This may partially explain the cause of the influence of cigarette smoke on development of the pulmonary and cardiovascular diseases. COPD with frequent exacerbation phenotype and OALLA are characterized by enhancement of inflammation with participation of monocytes.

Jaceosidin inhibits NLRP3-mediated pyroptosis by activating SIRT1/NRF2 and ameliorating intestinal epithelial cell injury

Purpose: To investigate the effect of Jaceosidin in ulcerative colitis (UC).Methods: An ulcerative colitis cell model was established by stimulating normal human colon mucosal epithelial cell lines (NCM460 cells) with lipopolysaccharide (LPS). The cells were treated with 5, 10, 20 or 40 μM Jaceosidin. Cell viability was performed using cell counting kit 8 (CCK8) assay. Oxidative stress was measured with superoxide dismutase (SOD), lipid peroxidation MDA, reduced glutathione (GSH), oxidized glutathione (GSSG), and human myeloperoxidase enzyme-linked immunoassay (ELISA) kits. The mRNA levels were determined by quantitative polymerase chain reaction (qPCR) assay, while protein levels of SIRT1, NRF2, NLRP3, caspase-1, TNF-α, IL-1β, and IL-6 were determined by western blotting.Results: Jaceosidin significantly inhibited oxidative stress and accumulation of inflammatory cytokines in LPS-induced NCM460 cells, as well as NLRP3-mediated cell pyroptosis (p < 0.05). Jaceosidin also inhibited activation of NLRP3 inflammasome by activating SIRT1/NRF2 pathway, thereby preventing NCM460 cell pyroptosis.Conclusion: Jaceosidin inhibits NLRP3-mediated pyroptosis, thus suggesting that jaceosidin is a potential lead for UC secondary to NLRP3 inflammasome.

Serum Myonectin and Adropin Levels in Predicting Diabetes

Introduction: Myonectin is a myokine associated with type 2 diabetes mellitus and insulin resistance, and adropin is a peptide hormone that prevents IR and impaired glucose tolerance. Adropin is produced by the liver and regulates energy homeostasis. This research aimed to examine the serum levels of myonectin and adropin levels among prediabetes, newly-diagnosed type 2 diabetes mellitus, and healthy controls.
 Methods: For this cross-sectional study, a total of 167 individuals were divided into 3 subgroups using OGTT and HbA1c; 61 prediabetes (36.5%), 62 nT2DM (37.1%), and 44 healthy controls (26.3%). Serum levels of myonectin and adropin were assayed using enzyme-linked immunoassay kits. The anthropometric (age, gender, weight, height, waist/hip ratio -WHR-, waist and hip circumference, and body mass index -BMI-) and biochemistry findings of the participants were evaluated between the groups.
 Results: Our study showed that myonectin levels are associated with nT2DM and WHR (p = 0.028 and p = 0.015, respectively). The serum levels of myonectin are significantly correlated with systolic blood pressure values in the prediabetes group (p = 0.017). Linear regression analysis revealed that nT2DM affects the serum levels of myonectin, but not adropin values. The adropin levels are correlated with LDL-cholesterol, total cholesterol, and triglyceride levels in the nT2DM group (p = 0.002, p= 0.004, and p = 0.035, respectively).
 Conclusion: The findings of the previous studies are supported by those of this study and indicated that the serum levels of myonectin may be associated with newly-diagnosed T2DM patients. Serum myonectin levels could be a valuable marker for predicting diabetes mellitus.

Vitamin C supplementation promotes locomotor and exploratory behaviors in male Wistar rats exposed to varying stress models.

Constant exposure to environmental stress has negative behavioral outcomes. Considering the inverse relationship between stress and Vitamin C intake, this study was aimed at investigating variable stress techniques and Vitamin C supplementation on exploratory/locomotor behaviors in male Wistar rats. Twenty-eight male Sprague-Dawley rats (100g-120g) were allotted into four groups (n=7). Control received 10ml/kg distilled water, group two received 100 mg/kg vitamin C, group three was exposed to different models of stress while group four was stressed alongside 100 mg/kg vitamin C. Vitamin C treatments were given orally for 2 weeks. Animals in groups 3 and 4 were stressed every other day with models such as multiple cage changes, exposure to noise, overnight strange objects, overnight wetting of beddings, and immobility. Explorative and locomotor activities were assessed with the open field test, novel object recognition test, and Y maze test using a Logitech camera and ANY-maze software to track the movement of the rats. Cortisol was assayed in the serum using Enzyme-linked Immuno Assay (ELISA) kit. Superoxide Dismutase, catalase, and lipid peroxidase; malondialdehyde (MDA) were also assayed in the serum. The results show that locomotor activities such as distance traveled, average speed, and time spent in the center square was significantly reduced by stress. These activities were improved with the intake of vitamin C compared with stress. Explorative activities such as locomoting around the environment, orientating towards novelty, and touching or sniffing novel objects were significantly increased in the rats on Vitamin C supplements and reduced in the stressed group. In the serum, cortisol level was significantly increased in rats exposed to stress and decreased with Vitamin C intake. Stress also significantly increased MDA and decreased SOD and CAT while vitamin C supplement decreased MDA and increased SOD and CAT. In conclusion, oral intake of vitamin C enhanced explorative/locomotor behavior and increased oxidative stress in rats exposed to different models of stress.

The predictive values of soluble B7-DC in children with refractory mycoplasma pneumoniae pneumonia.

Refractory mycoplasma pneumoniae pneumonia (RMPP) is a serious mycoplasma pneumoniae infection and is difficult to diagnose early. The levels of serum soluble B7-dendritic cell (sB7-DC) in children with mycoplasma pneumoniae pneumonia (MPP) were assessed to explore the clinical significance of sB7-DC levels in RMPP. A total of 65 patients with mycoplasma pneumoniae pneumonia (MPP) were enrolled in this study between January 2017 and December 2018. The patients were divided into the general mycoplasma pneumoniae pneumonia (GMPP) (n=30) and RMPP groups (n=35); the data of 20 normal children served as a control group (n=20). An enzyme-linked immunoassay kit was used to detect the expression of soluble B7-dendritic cell (sB7-DC) and other inflammatory factors. Binary logistic regression was performed to identify the independent predictors of RMPP. Receiver operating characteristic (ROC) curves were drawn to evaluate the value of each independent risk factor in the early diagnosis of RMPP. The results showed that compared to the GMPP group, children in the RMPP group had a significantly longer hospital stay and had a significantly longer fever duration (P<0.05). The values of interferon-gamma (IFN-γ), interleukin 17 (IL-17), and sB7-DC in the RMPP group were significantly higher than those in the normal control and GMPP groups (all P<0.05). The results of the correlation analysis showed that sB7-DC was positively correlated with IFN-γ and IL-17 and these indicators could be used in combination to evaluate the severity of the disease. The binary logistic regression analysis identified IL-17 and sB7-DC as independent risk factors for RMPP (P<0.05). The ROC curve analysis showed that the cut-off values of IL-17 and sB7-DC were 309.6 pg/L and 1,109.7 pg/mL, respectively. The areas under the curve (AUCs) of IL-17 and sB7-DC were 0.741 and 0.794, respectively. The sensitivity of IL-17 to RMPP prediction was 83.3%, and the specificity was 62.9%. The sensitivity and specificity of sB7-DC to RMPP were 86.7% and 62.9%, indicating that sB7-DC had the highest predictive power for RMPP. The level of serum sB7-DC may play an important role in the early diagnosis of RMPP. Our research results provide a theoretical basis for the early diagnosis of RMPP.

Designing and developing a sensitive and specific SARS-CoV-2 RBD IgG detection kit for identifying positive human samples

More than 3 y into the coronavirus 2019 (COVID-19) pandemic, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to undergo mutations. In this context, the Receptor Binding Domain (RBD) is the most antigenic region among the SARS-CoV-2 Spike protein and has emerged as a promising candidate for immunological development. We designed an IgG-based indirect enzyme-linked immunoassay (ELISA) kit based on recombinant RBD, which was produced from the laboratory to 10L industry scales in Pichia pastoris. A recombinant-RBD comprising 283 residues (31kDa) was constructed after epitope analyses. The target gene was initially cloned into an Escherichia coli TOP10 genotype and transformed into Pichia pastoris CBS7435 muts for protein production. Production was scaled up in a 10L fermenter after a 1L shake-flask cultivation. The product was ultrafiltered and purified using ion-exchange chromatography. IgG-positive human sera for SARS-CoV-2 were employed by an ELISA test to evaluate the antigenicity and specific binding of the produced protein. Bioreactor cultivation yielded 4g/l of the target protein after 160h of fermentation, and ion-exchange chromatography indicated a purity>95%. A human serum ELISA test was performed in 4 parts, and the ROC area under the curve (AUC) was>0.96 for each part. The mean specificity and sensitivity of each part was 100% and 91.5%, respectively. A highly specific and sensitive IgG-based serologic kit was developed for improved diagnostic purposes in patients with COVID-19 after generating an RBD antigen in Pichia pastoris at laboratory and 10L fermentation scales.

Berberine inhibits RA-FLS cell proliferation and adhesion by regulating RAS/MAPK/FOXO/HIF-1 signal pathway in the treatment of rheumatoid arthritis.

Rheumatoid arthritis (RA) is a common chronic immune disease. Berberine, as its main active ingredient, was also contained in a variety of medicinal plants such as Berberaceae, Buttercup, and Rutaceae, which are widely used in digestive system diseases in traditional Chinese medicine with anti-inflammatory and antibacterial effects. The aims of this article were to explore the therapeutic effect and mechanism of berberine on rheumatoid arthritis. Cell Counting Kit-8 was used to evaluate the effect of berberine on the proliferation of RA fibroblast-like synoviocyte (RA-FLS) cells. The effect of berberine on matrix metalloproteinase (MMP)-1, MMP-3, receptor activator of nuclear factor kappa-Β ligand (RANKL), tumour necrosis factor alpha (TNF-α), and other factors was determined by enzyme-linked immunoassay (ELISA) kit. Transcriptome technology was used to screen related pathways and the potential targets after berberine treatment, which were verified by reverse transcription-polymerase chain reaction (RT-qPCR) and Western blot (WB) technology. Berberine inhibited proliferation and adhesion of RA-FLS cells, and significantly reduced the expression of MMP-1, MMP-3, RANKL, and TNF-α. Transcriptional results suggested that berberine intervention mainly regulated forkhead box O (FOXO) signal pathway, prolactin signal pathway, neurotrophic factor signal pathway, and hypoxia-inducible factor 1 (HIF-1) signal pathway. The effect of berberine on RA was related to the regulation of RAS/mitogen-activated protein kinase/FOXO/HIF-1 signal pathway in RA-FLS cells.Cite this article: Bone Joint Res2023;12(2):91-102.

The Value of Interleukin-17A as a Prognostic Indicator in COVID-19 Patients

Background: SARS-CoV-2 infections (COVID-19) first occurred in Wuhan, China, in December 2019 and spread worldwide, causing significant mortality and morbidity. IL-17A may mediate numerous immunopathological effects secondary to cytokine release syndrome during SARS-CoV-2 infection. However, there has not been enough research on its effect on prognosis. Objectives: This study evaluated the predictive power of serum interleukin (IL)-17A level as a prognostic marker in COVID-19. Methods: The study included 152 patients diagnosed with COVID-19 by real-time polymerase chain reaction analysis of nasopharyngeal swab samples in the infectious diseases department and intensive care unit of our hospital between October 1 and December 31, 2020. The control group consisted of 40 asymptomatic healthcare workers who had negative RT-PCR results during routine COVID-19 screening in our hospital. Samples were collected in anticoagulant-free tubes and left at room temperature for 30 minutes. Afterward, it was centrifuged at 1000 × g for 15 minutes at 4°C per the instructions provided with the enzyme-linked immunoassay (ELISA) kit. Serum IL-17A levels were measured using the Human Interleukin 17A ELISA Kit. Results: Serum IL-17A levels were significantly higher in COVID-19 patients than in controls (P &lt; 0.001). IL-17A levels increased significantly in association with disease severity in patients with the moderate, severe, and critical disease, with a less pronounced difference between severe and critical patients (moderate vs. severe, P &lt; 0.001; severe vs. critical, P = 0.048). IL-17A levels at hospital admission and day 7 were significantly higher in non-surviving patients (P &lt; 0.001). At a cut-off value of 210.25 ng/L, IL-17A at admission had a predictive power of 0.792 (P &lt; 0.001). Compared to baseline, IL-17A values on day seven were significantly increased in non-survivors (P = 0.004) and decreased in survivors (P = 0.014). An increase of 26.17 ng/L or more on day 7 had a predictive mortality power of 0.634 (P = 0.005). Conclusions: The results of this study suggest that IL-17A, an important part of the immune system previously shown to be useful in the treatment and follow-up of COVID-19, may also help predict mortality in COVID-19 patients.

Serum s100b protein levels as a neuroinflammatory biomarker of acutely relapsed paranoid schizophrenia patients.

The s100b inflammatory protein is involved in schizophrenia pathophysiology. We aim at studying the evolution of the s100b serum levels in acutely relapsed paranoid schizophrenia patients at three different time points (admission, discharge and 3 months after hospital discharge 3MAHD). Twenty-three paranoid schizophrenia inpatients meeting DSM-IV criteria participated in the research. Twenty-three healthy subjects matched by age, gender and season acted as the control group. Psychopathology was measured with the Positive and Negative Syndrome Scale (PANSS). Serum s100b levels were determined at 12:00 and 24:00 h with an enzyme-linked immunoassay kit. Patients had significant higher serum s100b levels at admission and discharge (12:00 h) than the group of healthy subjects. At admission and discharge, s100b serum levels at 24 h had decreased compared to the 24:00 h s100b levels of the healthy subjects. At 3MAHD patients and healthy subjects had similar levels of serum s100b protein. Positive and negative PANSS scores decreased significantly between admission and discharge. Positive and negative PANSS scores decreased between discharge and 3MAHD, but these changes had no statistical significance. Our study confirms that the acute inflammatory response produced in acutely relapsed patients is reversed after 3 month of hospital discharge. The variations of serum s100b concentrations when the patients suffer from an acute relapse may be a useful predictor of disease evolution.

Low serum level of 25-OH vitamin D relates to Th17 and treg changes in colorectal cancer patients.

BackgroundSerum 25‐hydroxyvitamin D [25(OH)D] level alters in colorectal cancer (CRC) development. Regulatory T (Treg) cells and T‐ helper type 17 (Th17) cells are involved in immune response. Th17‐mediated proinflammatory responses contribute to tumorigenesis, and Treg plays different roles in different periods of CRC. Vitamin D deficiency is associated with significant variations in peripheral immune cells. This study investigated the relationship between Th17 and Treg cells and 25(OH)D level in CRC.MethodsNinety‐five CRC patients were included, as well as 80 healthy controls during the same period at the Affiliated Hospital of Jiangnan University. 25(OH)D level was analyzed through electrochemiluminescence (ECLIA). Th17 and Treg levels were evaluated through flow cytometry. Serum levels of interleukin (IL)‐10, IL‐17, IL‐23, and transforming growth factor‐β (TGF‐β), were analyzed through commercial enzyme‐linked immunoassay (ELISA) kits.Results25(OH)D levels were downregulated in the serum of CRC patients. Decreased 25(OH)D level contributed to CRC pathogenesis. Decreased 25(OH)D level in CRC correlated with increased Treg and Th17 cell ratios and TGF‐β1, IL‐10, IL‐17, and IL‐23 levels in peripheral blood.ConclusionDecreased 25(OH)D level in the serum of CRC patients had negative correlation with Treg and Th17 ratios and relative cytokines levels.

Heterologous vaccination of BNT162b2 in Ad26. COV2.S-vaccinated healthcare workers elicits long-term humoral immune response

Background. To date, there are no immunological data for the SARS-CoV-2 heterologous vaccination schedule in the South African (SA) population. Objectives. To assess and compare the immunogenicity and reactogenicity of the Jansen Ad26.COV2.S vaccine with the Pfizer/BioNTechBNT162b2 booster following prime Ad26.COV2.S in 65 SA healthcare workers. Methods. In a prospective, quantitative, cross-sectional trial on individuals &gt;18 years of age vaccinated with a single Ad26.COV2.S dose or single Ad26.COV2.S and a BNT162b2 single-dose/both doses booster, participants filled in a questionnaire on their demographics, type of vaccination, breakthrough infection/s (BTI/s), vaccine reactogenicity, prior SARS-CoV-2 infection and dates of vaccination. Qualitative analysis for presence/absence of anti-S (spike) immunoglobulin G (IgG) was performed using the Euroimmun anti-IgG enzyme-linked immunoassay kit, and anti-S IgG titres were quantitatively assessed using the Abbott IgG Quant II kit. Results. Between 28 October 2021 and 30 November 2021, 65 individuals were enrolled and assigned as either prime Ad26.COV2.S (n=18) or Ad26.COV2.S with a BNT162b2 supplement (n=47) at Charlotte Maxeke Johannesburg Academic Hospital, SA (mean age 45 years (95% confidence interval (CI) 29.5 - 58), 42 women (64.6%) and 23 men (35.4%)). The median IgG titre for the primed Ad26.COV2.S group was 4 272.55 (95% CI 68.40 - 10 351.40) and that for the BNT162b2 supplement group was 7 360.80 (95% CI 4 207.40 - 15 372.60). In the univariate model, the BNT162b2 supplement group showed a significant 1.99 times higher antibody titre factor (95% CI 0.045 - 5.553; p&lt;0.005) than the Ad26.COV2.S group. In both univariate and multivariate models, age, time since prime vaccination, BTI and prior infection failed to show any statistically significant association (p&gt;0.05) with antibody titres in both groups. However, sex (–55.381 (95% CI –76.984 - –13.498; p=0.018) in a multivariate model was found to have a statistically significant association with anti-S IgG titres observed in both groups. Participants who received their first dose of BNT162b2 9 - 10 months after their prime Ad26.COV2.S (n=44) had a higher degree of antibody response than those who received it earlier. Reactogenicity was observed to be manageable, with mild/moderate adverse effects in the study population. Conclusion. A BNT162b2 supplement given in single or two doses as booster in individuals primed with Ad26.COV2.S induced immunological response, with acceptable and manageable reactogenicity. This study provides novel evidence of the highest degree of antibody response in individuals who received a BNT162b2 first dose 9 - 10 months after prime Ad26.COV2.S, implying that a longer time gap between the two vaccines stimulates higher antibody response than a shorter gap, and that this antibody response can persist for as long as 6 months after the last BNT162b2 dose.

A novel method for detection of ochratoxin A in foods—Co-MOFs based dual signal ratiometric electrochemical aptamer sensor coupled with DNA walker

Ochratoxin A (OTA) is a naturally occurring mycotoxin that poses serious threats, such as kidney damage, to human health. Therefore, we developed a DNA walker-based dual-signal electrochemical ratiometric platform for OTA detection, which could overlook the variations in environmental and instrumental factors and DNA load densities. Cobalt metal–organic frameworks (Co-MOFs) and toluidine blue were used as the electrochemical signal tag and internal reference probe, respectively. In the presence of OTA, this developed machine resulted in the DNA labelled-Co-MOFs far away from the electrode. Thus, Co-MOFs signal at −1.18 V decreased, while toluidine blue at −0.28 V increased. This proposed strategy has displayed superior sensitivity (limit of detection = 0.31 fg/mL, linear range = 1–50 ng/mL) and high reproducibility. The sensor was also applied for determining OTA content in red wine samples and the results were comparable to those of commercial enzyme-linked immunoassay kits with satisfactory results.