Enzyme-linked Immunosorbent Assay Test Research Articles

The Association between Vitamin D, Interleukin-4, and Interleukin-10 Levels and CD23+ Expression with Bronchial Asthma in Stunted Children.

Children with stunted growth have an increased risk of wheezing, and studies have shown that low levels of vitamin D and interleukin (IL)-10, along with increased IL-4 levels and CD23+ expression, are present in stunted and asthmatic children. To date, it is not known whether these factors are related to the incidence of asthma in stunted children. This case-control study investigated the association between vitamin D, IL-4, and IL-10 levels and CD23+ expression with bronchial asthma in stunted children. The study included 99 children aged 24-59 months, i.e., 37 stunted-sthmatic children (cases), 38 stunted children without asthma, and 24 non-stunted children with asthma. All children were tested for their 25(OH)D levels using chemiluminescent immunoassay (CLIA), IL-4 and IL-10 levels were measured through enzyme-linked immunosorbent assay (ELISA) testing, and CD23+ expression was measured through flow cytometry bead testing. The data were analyzed using chi-squared, Kruskal-Wallis, and Mann-Whitney tests. The results showed that stunted asthmatic children had a higher incidence of atopic family members than those without asthma. Additionally, stunted asthmatic children had a higher prevalence of vitamin D deficiency (48.6%) than the control group (44.7% and 20.8%). Furthermore, stunted asthmatic children had significantly lower levels of 25(OH)D [20.55 (16.18-25.55), p = 0.042] and higher levels of IL-4 [1.41 (0.95-2.40), p = 0.038], although there were no significant differences in IL-10 levels and CD23+ expression. The study concluded that low vitamin D and high IL-4 levels are associated with bronchial asthma in stunted children, while IL-10 and CD23+ do not show a significant association.

Exposure to farm environment and its correlations with total IgE, IL-13, and IL-33 serum levels in patients with atopy and asthma.

The aim of the study was to evaluate total immunoglobulin E (IgE), IL-13, and IL-33 serum level in people with bronchial asthma and atopy, and in healthy control group depending on their exposure to farm animals currently and in the first year of life. The study included 174 individuals living in rural areas and in a small town. Standardized questions from the International Study of Asthma and Allergy in Childhood and The European Community Respiratory Health Survey (ECRHS) questionnaires were used to define asthma. Atopic status was verified by skin prick tests. Rural exposure including contact with livestock was verified by adequate questionnaire. Total serum IgE, IL-13, and IL-33 levels were assessed by ELISA (enzyme-linked immunosorbent assay) tests. Participants with atopy and bronchial asthma were characterized by high level of immunoglobulin E. Tendency to lower serum IgE level was observed among people reporting present contact with farm animals. Also, among those having contact with livestock in their first year of life, the analogous tendency was noticed. No difference in serum IL-13 levels in participants with asthma and atopy, and controls was observed, and there was no effect of exposure on farm animals on the concentration of IL-13. The highest IL-33 level was found in the atopic group, and the lowest in the control group. Participants currently exposed to farm animals were predisposed to have lower IL-33 serum level. Exposure of farm animals currently and in first year of life may result in a lower level of total IgE. Correlation between IL-13 and IL-33 serum levels and contact with livestock was not confirmed.

Possibility of using mathematical analysis methods to evaluate the potency of antibodies to hepatitis B virus surface antigen in human immunoglobulin preparations

Scientific relevance. Anti-hepatitis B virus surface-antigen (HBsAg) immunoglobulins are used to prevent hepatitis B in adults and children after exposure and to treat mild to moderate acute viral hepatitis B. The clinical effectiveness of human immunoglobulin preparations is determined by their potency, which is assessed by the content of antibodies to hepatitis B virus surface antigen (anti-HBs antibodies). Currently, this assessment involves using immunoassay techniques, such as enzyme-linked immunosorbent assay (ELISA).Aim. This study examined several mathematical methods for analysing the experimental data obtained in ELISA-based anti-HBs antibody potency assays of human immunoglobulin preparations.Materials and methods. This study used the international standard for human anti-HBs immunoglobulin, two immunoglobulin preparations, and an ELISA test kit for the detection and quantification of anti-HBs antibodies in serum or plasma samples.Results. Using sandwich ELISA, the authors ascertained that the measured anti-HBs antibody concentration depended on the choice of calibration curve calculation method (i.e. manual analysis, parallel-line analysis using PARALINE software, linear regression, and 4-parameter logistic regression). The measured anti-HBs antibody concentrations varied by ± 19 IU/mL. According to the study results, an incorrectly selected method of data analysis can lead to an erroneous calculation of the analyte potency (concentration) in the test sample.Conclusions. The study demonstrated the need for improved mathematical methods for the evaluation of experimental data used to determine the anti-HBs antibody concentration in human immunoglobulin preparations. It is essential to switch from manual to automated calculation (for example, using a 4-parameter logistic model), taking into account the requirements for bioanalytical methods and the capabilities of the available equipment.

Prevalence of asymptomatic strongyloidiasis co-infection in COVID-19 patients residing in endemic areas

BackgroundFatal forms of strongyloidiasis, hyperinfection syndrome (HS) and disseminated strongyloidiasis (DS), are caused by exaggerated autoinfection of the intestinal nematode, Strongyloides stercoralis (S. stercoralis). Corticosteroids, frequently administered to patients with severe COVID-19, can transform chronic asymptomatic strongyloidiasis into the above-mentioned fatal diseases. This study aimed to investigate the prevalence of strongyloidiasis in COVID-19 patients receiving corticosteroids in a hypoendemic region.MethodsThe present cross-sectional study enrolled 308 COVID-19 patients admitted to two hospitals in Ahvaz and Abadan in the southwest of Iran between 2020 and 2022. A real-time reverse transcription polymerase chain reaction (RT-PCR) test and chest computed tomography (CT) scan were employed to detect and monitor the disease’s severity in the patients, respectively. All patients were evaluated for IgG/IgM against S. stercoralis using Enzyme-linked immunosorbent assay (ELISA) test. Subsequently, individuals with a positive ELISA test were confirmed using parasitological methods, including direct smear and agar plate culture (APC).ResultsThe patients were between 15 and 94 years old, with a mean age of 57.99 ± 17.4 years. Of the 308 patients, 12 (3.9%) had a positive ELISA test, while 296 (96.1%) had negative results. Three of the 12 patients with a positive ELISA result died, and three failed to provide a stool sample. To this end, only six cases were examined parasitologically, in which S. stercoralis larvae were observed in five patients. Significant differences were found between S. stercoralis infection with sex (p = 0.037) and age (p = 0.027). Binary regression analysis revealed that strongyloidiasis was positively associated with sex (odds ratio [OR]: 5.137; 95% confidence interval [CI]: 1.107–23.847), age (OR: 5.647; 95% CI 1.216–26.218), and location (OR: 3.254; 95% CI: 0.864–12.257).ConclusionsOur findings suggest that screening for latent strongyloidiasis in COVID-19 patients in endemic areas using high-sensitivity diagnostic methods, particularly ELISA, before receiving suppressive drugs should be given more consideration.

Measurements and Accuracy of IgM and IgG Anti Phenolic Glycolipid-1 Levels in Blood Serum for Early Detection Mycobacterium leprae by using Enzyme-Linked Immunosorbent Assay (ELISA): A Reality of a Laboratory

Indonesia was the third most recent case of leprosy globally in 2020 with 11,173 people, after India and Brazil. Most of the leprosy manifestations are asymptomatic. This is possibly as subclinical leprosy which individuals without leprosy symptoms but have leprosy specific antibodies high levels, so it has the potential to become a transmission and disability. Therefore, an ELISA test need for early detection in preventing leprosy transmission. This study aims to measure IgM and IgG antibody levels in leprosy patients and assess the accuracy of the measurement results. This research is a cross-sectional study. Five patients' blood samples have analyzed for IgM and IgG anti-PGL-1 antibody levels by ELISA. Accuracy interpretation of this measurement based on the %CV. Antibody levels were classified based on the cut-off <605 u/ml as IgM seronegative or <630 u/ml as IgG seronegative, 605–1000 u/ml as low seropositive IgM or 630-1000 u/ml as low seropositive IgG, and >1000 u/ml as high seropositive IgM and IgG. Among five patients examined, 40% had high seropositive leprosy with anti-PGL-1 IgM and IgG antibody titers>1000 u/ml, and 60% of patients had seronegative leprosy. Accuracy in this ELISA test shows high accuracy with %CV <10% in the conversion of OD to antibody titer levels. IgM and IgG Anti PGL-1 antibody titers by ELISA as one of the parameters in identifying patients at higher risk of leprosy. A significant portion of patients with high seropositive leprosy with high accuracy.

Seroprevalence and incidence of primary dengue infection and its correlation with fetomaternal prognosis in Western Rajasthan.

Viral hemorrhagic fevers are becoming increasingly common in the tropics and subtropics. Dengue fever is currently the most important arthropod-borne viral disease because of its widespread distribution in more than 100 countries and its potential for extensive outbreaks of life-threatening disease. This study was a hospital-based cross-sectional study conducted in the Microbiology Laboratory of Maternal and Child Tertiary Care Hospital in Western Rajasthan, India, between January 2021 and December 2021. Institutional Ethical Committee permission was obtained. All patients with clinical suspicion of dengue-like illness (DLI), attending outpatient department (OPD) or inpatient department (IPD), were included in the study after obtaining their written consent. A blood sample was collected, and the Dengue Duo rapid card test was conducted for the detection of nonstructural protein 1 (NS1) antigen and immunoglobulin (Ig) M or IgG antibody estimation. All positive samples were tested for IgM enzyme-linked immunosorbent assay (ELISA) test using MAC-ELISA. Of 250 positive sample, the distribution of cases as per clinical features was as follows: all cases presented with fever (100%) followed by myalgia (24.5%), headache (16.06%), hemorrhagic manifestation (13.25%), rash (8.84%), and bleeding gums (2.01%). Thrombocytopenia was seen in 30.40% (76/250) of dengue fever cases. NS1 antigen was detected in 157 cases (62.80%) followed by IgG in 84 cases (33.60%), IgM in 77 cases (30.80%), NS1+IgG in 27 cases (10.80%), NS1 + IgM in 16 cases (6.40%), and NS1 + IgM + IgG in five cases (2%). Of 250 samples, 77 cases were IgM positive and 173 were IgM negative by the Dengue Duo card test. Among the 173 Dengue Duo IgM card negative, 131 cases (79.39%) were also detected negative by IgM ELISA and 42 cases (49.41%) were detected positive by IgM ELISA. The sensitivity was 50.59%, the specificity was 79.39%, the positive predictive value (PPV) was 55.84%, the negative predictive value (NPV) was 75.72%, and the diagnostic accuracy was 69.90%. The case fatality of the cases was 2.35%. Early diagnosis and treatment can prevent mortality in pediatric and pregnant females suffering from dengue and dengue-like illness. Facility and availability of ELISA kits should be adequate for early confirmation of suspected dengue patients by ELISA test.

Examination of the cross-reactivity between vaccinia virus Tiantan strain and monkeypox virus

AimTo investigate the cross-reactivity between the sera collected from Vaccinia Virus Tiantan Strain vaccinated rabbits and viral antigens of monkeypox virus. MethodsVaccinia viruses were prepared on chicken embryo fibroblasts (CEF) and Vero cells respectively named as CEF-VTT NVSI-1 and Vero-VTT NVSI-1. Rabbits were inoculated with a total of three doses of adjuvanted 1.3 × 108 PFU CEF-VTT NVSI-1 each dose or adjuvanted 3.9 × 107 PFU Vero-VTT NVSI-1 (Freunds complete adjuvant) via the subcutaneous route. We then performed the enzyme-linked immunosorbent assay (ELISA) and bio-layer interferometry (BLI) for determination of the binding activity and affinity of immune sera to five crucial surface antigens on monkeypox virus including A35, B6R, H3 and to corresponding homologous antigens A33R, B5 and L1R of vaccinia virus. For comparison, plaque reduction neutralizing tests were used to evaluate the neutralization of immune sera against vaccinia virus. ResultsBoth CEF-VTT NVSI-1 and Vero-VTT NVSI-1 vaccinations following planned schedule could induce neutralizing antibody titers greater than 1:2048 in rabbit sera. Binding antibodies targeting monkeypox viral antigens were confirmed by both indirect ELISA and BLI methods. Indirect ELISA for rabbit sera revealed 1:51200 binding antibody titers to A35/B6R/H3 monkeypox virus antigens while BLI tests yielded affinities at 2 × 10−6 to 8 × 10−7 between the sera and the three antigens. Similarly, such sera showed binding strength to vaccinia virus antigens A33R/B5/L1R consistent with that to three preceding monkeypox virus antigens. These results demonstrated the cross-reactivity between the sera of vaccinia virus vaccinated animals and monkeypox virus antigens. Traditional ELISA test and BLI method displayed a high consistency in antigen screening and they were further proved to correlate to the results of plaque reduction neutralizing test, which indicates that BLI could be utilized as an indirect alternative for assessment of neutralizing activity of samples in response to live virus. ConclusionsSera of vaccinia virus-vaccinated rabbits exhibited cross-reactivity with viral antigens of monkeypox virus. Potential in improving the accuracy of antigen discovery while reducing the lengthy work needed for the screening as BLI method possesses, it contributes greatly to the rapid preliminary evaluation of immune response generated by vaccines.

Advancements in Lily Viruses Management: Challenges and Solutions in Elimination and Detection

Lilies are important crops that are commonly used as cut flowers (Lilium spp.) and edible bulb crops (Lilium davidii var. unicolor). However, virus infections can significantly impact the quantity and quality of lily production. Various methods have been developed to eliminate viruses in lilies, including in vitro culture and virus detection techniques. Meristem culture is the most effective method, which can be combined with other techniques such as thermotherapy and chemotherapy. Nonetheless, virus elimination is affected by several factors, including cultivar, explants used, virus type, and duration of treatments. Efficient diagnostic methods, such as serological and molecular techniques, have been developed to detect viral infections in lilies, including enzyme-linked immunosorbent assay (ELISA) and real-time reverse transcriptase polymerase chain reaction (real-time RT-PCR). However, cross-contamination and multiple-virus contamination can lead to unreliable results, and more sophisticated protocols and systems have been developed to address these issues. The objective of this review is to provide a comprehensive overview of the development of lily virus eradication, detection strategies, challenges, and solutions associated with these procedures, and how more sophisticated approaches such as multiplex RT-PCR, indirect ELISA (ID-ELISA), immunocapture RT-PCR (IC-RT-PCR), and immunochromatographic test strips (ICSs) can alleviate some of these setbacks.

The neuroprotective effects of Chalcones from Ashitaba on cuprizone-induced demyelination via modulation of brain-derived neurotrophic factor and tumor necrosis factor α.

Multiple sclerosis (MS) is the most common demyelinating disease of the central nervous system. However, the limitations of available therapeutic strategies are frustrating, both in terms of their low efficacy and multiple side effects. Previous studies showed that natural compounds such as Chalcones possess neuroprotective effects on neurodegenerative disorders. However, few studies have so far been published on the potential effects of Chalcones on treating demyelinating disease. The present study was designed to investigate the effects of Chalcones from Ashitaba (ChA) on cuprizone-induced noxious changes in the C57BL6 mice model of MS. The mice received normal diets (Control group: CNT), or Cuprizone-supplemented diets either without ChA (Cuprizone group: CPZ) or with low or high (300, 600mg/kg/day) doses of ChA (ChA-treated groups: CPZ+ChA300/600). Brain-derived neurotrophic factor (BDNF) and tumor necrosis factor alpha (TNFα) levels, demyelination scores in the corpus callosum (CC), and cognitive impairment were evaluated using the enzyme-linked immunosorbent assay, histological, and Y-maze tests, respectively. The findings showed that ChA Co-treatment significantly reduced the extent of demyelination in the CC and the serum and brain levels of TNFα in the ChA-treated groups compared to the CPZ group. Besides, treatment with a higher dose of ChA significantly improved the behavioral responses and BDNF levels in the serum and brain of the CPZ+ChA600 group when compared with the CPZ group. The present study provided evidence for the neuroprotective effects of ChA on cuprizone-induced demyelination and behavioral dysfunction in C57BL/6 mice, possibly by modulating TNFα secretion and BDNF expression.

Hypericum perforatum ekstresinin siyatik sinir hasarı ile indüklenen periferik nöropati üzerindeki düzenleyici etkisi: fareler üzerinde deneysel bir çalışma

Purpose: The effect of Hypericum perforatum (HP), which is a medicinal plant, on sciatic nerve injury-induced peripheral neuropathy has been less studied so far. The current experimental study aimed to investigate the neuroprotective and antinociceptive effects of Hypericum perforatum (HP) extract on sciatic nerve injury-induced peripheral neuropathy in mice.
 Materials and Methods: In the present study, 18 Balb/C albino mice were allocated equally into three groups. The first group was determined as controls, and no procedure was performed on these mice. Neuropathy was generated by the partial sciatic nerve ligation method on mice allocated to the second and third groups. Mice in the third group received HP extract at a dose of 70 mg/kg per day for fourteen days. Nociception (cold allodynia) was evaluated using the cold plate test at the end of the experimental period. Tumor necrosis factor –αlpha (TNF-α) and interleukin-6 (IL-6) in plasma; inducible nitric oxide synthase (iNOS), phospholipase A2, cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-κB), caspase-3, Bcl-2, and Bax levels in sciatic nerve were measured by enzyme-linked immunosorbent assay test. 
 Results: Cold plate latencies (sec) of the neuropathy + HP, neuropathy, and control groups were 8.33 ± 0.67, 5.17 ± 0.60, and 13 ± 0.73, respectively. Plasma TNF-α, IL-6 levels, and sciatic nerve iNOS, COX-2, NF-κB, caspase-3, and Bax levels were significantly decreased after HP supplementation. Bcl-2 levels of the neuropathy + HP, neuropathy, and control groups were 9.92 ± 0.71, 5.37 ± 0.53, and 13.65 ± 0.68, respectively.
 Conclusion: HP has improved oxidative, inflammatory, and apoptotic responses, as well as cytokine levels in plasma and sciatic nerves of mice. It has been concluded that HP provided neuroprotective, anti-inflammatory, and antinociceptive effects in experimental mice with sciatic nerve injury models, which is suggested to guide future studies on neuropathic pain management.

Molds and Mycotoxins in Ration Wheat Flour in Sulaimani Provence

Wheat flour is one of the main food sources of mycotoxin exposure, hence its safety warrants investigation. Aflatoxin B1 (AFB1) and Deoxynivalenol (DON), specifically, and their levels of contamination in wheat flour samples from ration wheat flour samples in Sulaimani City, Kurdistan, Iraq, were examined in the current study. According to the findings of the fungal study, Penicillium (35%) infected the bulk of the samples, with Aspergillus (27%) and Rhizopus (17%) coming in second and third, respectively, and Alternaria (9%), Fusarium (6%), Helminthosporium (4%), Mucor (1%), and Cladosporium (1%).In November and December, the number of molds reached intolerable heights. The findings of the enzyme-linked immunosorbent assay (ELISA) test revealed that AFB1 was present in varying amounts in 100% of the wheat flour samples. AFB1 surpassed the 2 μgkg-1 maximum permissible quantity in 37.5% of samples, particularly in November and December. In terms of DON, 100% of samples were contaminated with DON without going over the 750 μgkg-1 EU limit.

DP21 Thymoma and the skin: manifestations of multiorgan autoimmunity

Abstract We present three patients who developed distinct cutaneous manifestations of thymoma: a rare mediastinal tumour. A 25-year-old man was treated for myasthenic crisis. Past medical history included thymectomy in 2016. Admission imaging confirmed recurrence of thymoma. He developed a red–brown macular rash affecting the trunk, back and thighs. Biopsy showed a florid lichenoid dermatitis consistent with graft-versus-host disease (GVHD). After resolution with steroids and mycophenolate mofetil, he subsequently developed a Stevens–Johnson syndrome/toxic epidermal necrolysis-like eruption with erythroderma and epidermal loss. Unfortunately, he developed multiorgan failure and died shortly afterward. A 43-year-old man presented to the emergency dermatology clinic with a 4-month history of a blistering rash affecting the chest and limbs. His previous medical history was significant for thymoma in 2008 and a recurrence in 2013. Skin biopsy demonstrated changes suggestive of bullous pemphigoid, and repeat computed tomography confirmed the recurrence of thymoma. Enzyme-linked immunosorbent assay testing was positive for bullous pemphigoid 180. He has responded well to prednisolone and doxycycline. A 51-year-old woman was reviewed in 2020 and 2021 with pruritic hyperpigmented patches affecting the face, trunk and limbs. Her past medical history included several relapses of thymoma with occurrence in 2002 and relapses in 2007, 2008, 2011, 2015 and 2021. Skin biopsy in June 2021 showed features of cutaneous GVHD. She received ruxolitinib and tapering steroids for thymoma-associated multiorgan autoimmunity (TAMA)-like GVHD and has had a remarkable improvement in her skin disease. Thymoma is described as a rare neoplasm, and up to 34% of thymomas are thought to be accompanied by symptoms of autoimmunity. The most frequently described syndromes include myasthenia gravis, pure red blood cell aplasia and hypogammaglobulinaemia. Other reported presentations include panencephalitis, enterocolitis and autoimmune hepatitis. Thymoma and cutaneous autoimmune presentations have been previously described, often in association with two rare syndromes: TAMA and paraneoplastic autoimmune multiorgan syndrome (PAMS). Both conditions demonstrate variability in clinical phenotype, driven by the activation of humoral and cellular immune responses. Clinically, patients may present with a spectrum of skin disease, including GVHD-like exanthems, lichen planus-like disease and pemphigus-like and pemphigoid-like lesions. Our case series demonstrates the heterogeneous cutaneous manifestations of thymoma-induced autoimmunity. We also describe two extremely rare phenomena accompanying thymomas: TAMA and PAMS. We also suggest that there is much crossover between these syndromes and previous definitions may require further review. This case series also highlights the importance of the wider multidisciplinary team in managing these patients and the complexities of their disease.

Comparison of Three Serologic Tests for the Detection of Anti-Coxiella burnetii Antibodies in Patients with Q Fever.

The performance of a commercial immunofluorescence assay (IFA commercial), an in-house immunofluorescence assay (IFA in-house) and an indirect enzyme-linked immunosorbent assay (ELISA) were evaluated in the detection of antibodies anti-C. burnetii in the serum of Q fever patients and persons without the disease. For the study, seropositive and seronegative samples for Q fever (n = 200) from a serum bank of the Instituto Adolfo Lutz in Brazil were used. Commercial IFA was considered in this study as the gold standard for diagnosing Q fever. The in-house IFA demonstrated good agreement with the commercial test, showing high sensitivity (91%) and specificity (97%) compared to the gold standard, with a Kappa coefficient of 0.8954. The indirect ELISA test showed lower agreement with the gold standard, showing low sensitivity (67%), although the specificity of the technique was high (97%) and the Kappa coefficient was moderate (0.6631). In-house IFA is an excellent alternative for diagnosing Q fever.

Recurrent cardiac hydatid cysts with pericardial protrusion; a rare case presentation of Echinococcus granulosus

Hydatid cyst (HC) is a rare pathology mostly found in less developed, livestock-raising countries. HC can be found anywhere in the body, however it is a rare finding in the heart, and its recurrence in the heart and pericardium is very rare. A 54-year-old woman who had undergone surgery for the removal of HC in another country 11 years before presented with shortness of breath and heaviness in the chest. The examinations and computerized tomography (CT) scan revealed a large mass measuring 60×50 mm in the pericardium cavity originating from the right atrial wall. Infection with Echinococcus granulosus was confirmed with serological enzyme-linked immunosorbent assay (ELISA) test and the patient underwent cardiac surgery. The patient underwent cardiopulmonary bypass, nevertheless did not suffer cardiac arrest, and aortic cross-clamping was not performed for her. The removed mass was filled with daughter cysts. The endocysts were removed and intramural cysts were drained. Following the surgery, the patient received treatment with albendazole for six months. Cardiac hydatid cyst recurrence is a rare but possible incident, and annual follow-ups and echocardiography are recommended.

Gut barrier protein levels in serial blood samples from critically ill trauma patients during and after intensive care unit stay.

In an effort to better manage critically ill patients hospitalised in the intensive care unit (ICU) after experiencing multiple traumas, the present study aimed to assess whether plasma levels of intestinal epithelial cell barrier proteins, including occludin, claudin-1, junctional adhesion molecule (JAM-1), tricellulin and zonulin, could be used as novel biomarkers. Additional potential markers such as intestinal fatty acid-binding protein (I-FABP), D-lactate, lipopolysaccharide (LPS) and citrulline were also evaluated. We also aimed to determine the possible relationships between the clinical, laboratory, and nutritional status of patients and the measured marker levels. Plasma samples from 29 patients (first, second, fifth and tenth days in the ICU and on days 7, 30 and 60 after hospital discharge) and 23 controls were subjected to commercial enzyme-linked immunosorbent assay (ELISA) testing. On first day (admission) and on the second day, plasma I-FABP, D-lactate, citrulline, occludin, claudin-1, tricellulin and zonulin levels were high in trauma patients and positively correlated with lactate, C-reactive protein (CRP), number of days of ICU hospitalisation, Acute Physiology and Chronic Health Evaluation II (APACHE II) score and daily Sequential Organ Failure Assessment (SOFA) scores (P < 0.05-P < 0.01). The results of the present study showed that occludin, claudin-1, tricellulin and zonulin proteins, as well as I-FABP, D-lactate and citrulline, may be used as promising biomarkers for the evaluation of disease severity in critically ill trauma patients, despite the complexity of the analysis of various barrier markers. However, our results should be supported by future studies.

An ELISA Test Able to Predict the Development of Oral Cancer: The Significance of the Interplay between Steroid Receptors and the EGF Receptor for Early Diagnosis.

Oral disorders including non-homogeneous leukoplakia, erythroplakia, erosive lichen planus, and many others can potentially progress to oral squamous cell carcinoma (OSCC). Currently, the late diagnosis of OSCC contributes to high mortality rates, emphasizing the need for specific markers and early intervention. In this study, we present a novel, quick, sensitive, and non-invasive method for the early detection and screening of oral cancer, enabling the qualitative assessment of neoplastic forms even before the onset of symptoms. Our method directly examines the expression of oral cancer biomarkers, such as the epithelial growth factor receptor (EGFR), and steroid receptors, including the androgen receptor (AR) and the estrogen receptor (ER). The crosstalk between sexual hormones and the EGF receptor plays a crucial role in the progression of different types of cancers, including head and neck squamous cell carcinoma. To implement our method, we developed a kit box comprising nine wells or stations, each containing buffers, lysis systems, and dried/lyophilized antibodies stored at room temperature. The kit includes instruments for sample collection and a PVDF strip (Immobilon) with specific primary antibodies immobilized on it. These antibodies capture the target proteins from cytological samples. Additionally, complementary tools are provided to ensure efficient utilization and optimal test performance. The technique can be performed outside the laboratory, either "patient side" with an instant chemocolorimetric response or with a digital reader utilizing the enzyme-linked immunosorbent assay (ELISA) method.

Clinical and Epidemiological Characteristics of 30 Fatal Cases of Crimean-Congo Hemorrhagic Fever in Kabul, Afghanistan: A Retrospective Observational Study.

Crimean-Congo hemorrhagic fever (CCHF) is a zoonotic disease associated with a high fatality rate. CCHF is endemic in Afghanistan, and its morbidity and mortality have increased recently but there is limited data about the characteristics of fatal cases. We aimed to report the clinical and epidemiological features of fatal CCHF cases who were admitted to Kabul Referral Infectious Diseases (Antani) Hospital. This is a retrospective cross-sectional study. The demographic and presenting clinical and laboratory features of 30 fatal CCHF cases diagnosed by reverse transcription polymerase chain reaction (RT-PCR) or enzyme-linked immunosorbent assay (ELISA) tests were collected from the patients' records between March 2021 and March 2023. During the study period, a total of 118 laboratory-confirmed CCHF patients were admitted to Kabul Antani Hospital of whom 30 patients (25 males, 5 females) consequently died, indicating a 25.4% case fatality rate (CFR). The age of the fatal cases ranged from 15 to 62 years and their mean age was 36.6 ± 11.7 years. Concerning occupation, the patients were butchers (23.3%), animal dealers (20%), shepherds (16.6%), housewives (16.6%), farmers (10%), student (3.3%), and others (10%). The clinical symptoms of the patients on admission were fever (100%), generalized body pain (100%), fatigue (90%), bleeding (any type) (86.6%), headache (80%), nausea/vomiting (73.3%), and diarrhea (70%). The initial abnormal laboratory findings were leukopenia (80%), leukocytosis (6.6%), anemia (73.3%), and thrombocytopenia (100%), raised hepatic enzymes (ALT & AST) (96.6%) and prolonged prothrombin time/international normalized ratio (PT/INR) (100%). The hemorrhagic manifestations associated with low platelet and raised PT/INR levels are linked with fatal outcomes. A high index of clinical suspicion is required to recognize the disease at an early stage and to begin the treatment promptly for reducing mortality.

Mechanisms of Wuyao (Linderae Radix) in treating chronic pelvic inflammatory disease based on network pharmacology and animal experiments

ObjectiveTo predict the active components and action targets of Wuyao (Linderae Radix) in the treatment of chronic pelvic inflammatory disease (CPID) based on network pharmacology, explore possible mechanisms of the treatment through animal experiments, and provide a scientific basis for clinical applications of Wuyao (Linderae Radix). MethodsPossible active components and targets of Wuyao (Linderae Radix) in the treatment of CPID were obtained applying network pharmacology and molecular docking technology. CPID rat models were established using the mixed Escherichia coli, Staphylococcus aureus, and Ureaplasma urealyticum plus the performance of mechanical injury. Hematoxylin-eosin (HE) staining was applied to observe the pathological changes in the uterus, fallopian tube, and spleens of rat models. The contents of nitric oxide (NO), superoxide dismutase (SOD), and malondialdehyde (MDA) in the serum of rats were determined with the use of corresponding detection kits. Enzyme-linked immunosorbent assay (ELISA) test was used to measure the expression of interleukin (IL)-6 and IL-10 in the serum of rat models. Flow cytometry was used to determine the percentage of CD4+ and CD8a+ T cells as well as CD4+ CD25+ regulatory T cells (Tregs) in the spleen of rat models. ResultsA total of nine potential active components and four core therapeutic targets related to inflammatory response in Wuyao (Linderae Radix) were obtained. The animal experiments showed that Wuyao (Linderae Radix) markedly inhibited uterus swelling, regulated morphological changes in the fallopian tube and spleen, effectively reduced inflammatory infiltration and injuries in the uterus and fallopian tube, and improved spleen functions in CPID rats. Moreover, Wuyao (Linderae Radix) markedly reduced the levels of NO, IL-6, and MDA, and increased the levels of IL-10 and SOD in the serum of rats. Wuyao (Linderae Radix) also elevated the percentage of CD4+ T cells and the CD4+ T/CD8a+ T cell ratio, reduced the percentage of CD8a+ T cells, and raised the percentage of CD4+ CD25+ Tregs that had been abnormally decreased in rat models (P < 0.05). ConclusionWuyao (Linderae Radix) could have therapeutic effects on CPID rats by relieving oxidative stress, mitigating inflammatory levels, and regulating the immuno-function of T cell subgroups to improve the pathological changes in CPID rats. It is a medicinal herb worth being further explored for its clinical values.

Evaluation of Immunogenicity of Clostridium perfringens Type B Toxoid and Inactivated FMD (O) Virus with Adjuvant (ISA70-MF59).

Foot and mouth disease (FMD) and enterotoxemia are important diseases of hoofed animals. Vaccination against livestock pathogens, especially these two diseases, plays a key role in the prevention and control of these diseases. The use of combined vaccines with the aim of creating a better immune response and producing cheaper vaccines is a great contribution to Vaccine industry. This research aimed to compare the immunogenicity of FMD (O) and Clostridium perfringens type B toxoid along with adjuvant (MF59) and Montanide (ISA70) to create the best immunogenicity. To investigate the immune responses of vaccines, it was injected into an animal model, and the antibody titer was measured by enzyme-linked immunosorbent assay (ELISA) test and VN antibody titer. The results showed that the formulation with MF59 adjuvant brought more stable immunogenicity against FMD and Clostridium perfringens type B, and the length of the immunogenicity period also increased significantly. Therefore, the combined vaccine (Clostridium perfringens + FMD) could play a major role invaccine industry as an alternative vaccine against Clostridium perfringens and FMD in livestock.

Comparative Evaluation of Specific Antibody against Mycobacterium tuberculosis ESAT-6 Recombinant Antigen in Healthy Subject with Positive and Negative Skin Test.

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (M. tuberculosis). The laboratory diagnosis of the disease includes various bacteriologic and immunologic methods. Despite the effectiveness of many of these methods in diagnosing active TB, their high cost and time-consuming nature have led researchers to adopt more accurate and rapid screening methods based on specific antigens for M. tuberculosis. The present study aimed to measure specific antibody serum levels against the early secretory antigenic target 6 kDa (ESAT-6) recombinant protein in healthy people and compare it to TB patients. The target population included 27 TB patients and 87 healthy individuals with no clinical TB symptoms. The healthy population was divided into two groups, including positive purified protein derivative (PPD) and negative PPD (35 and 52 people, respectively), using the Tuberculin skin test. The specific antibody level against the ESAT-6 recombinant antigen and the PPD protein was measured using an indirect Enzyme-Linked Immunosorbent Assay (ELISA) test. The results of the study showed that the majority of the healthy population with no symptoms of clinical TB and having negative skin test results did not have antibodies against the recombinant ESAT-6 (98%) and PPD (96%) antigens. On the other hand, there was a high level of the specific antibody of the ESAT-6 recombinant and PPD antigens in TB patients (77%). It is notable that in people with positive skin test results, the level of the antibody against the ESAT-6 recombinant antigen and PPD antigen was 94%. The results demonstrated that the ELISA method based on the measurement of antibodies against the ESAT-6 recombinant antigen can be a proper diagnostic method for rapid and accurate screening of healthy from infected people.