Search Results (6,034)

Hepatitis E virus (HEV) is a zoonotic pathogen with multiple hosts, posing significant public health risks, especially in regions like Japan where game meat consumption is prevalent. This study investigated HEV infection and viral shedding in wild boars, sika deer, and Japanese serows across Japan. A total of 1896 serum samples were tested for anti-HEV antibodies, 1034 for HEV RNA, and 473 fecal samples for viral shedding. Anti-HEV antibodies were detected in wild boars from all seven prefectures studied, while HEV RNA was detected in wild boars from Fukuoka, Oita, and Miyazaki in southern Japan, as well as Yamaguchi prefecture. Genetic analysis revealed subtypes 3b, 4a, and 4g, with 3b being the most prevalent. Subtype 3b exhibited distinct geographical clustering, whereas 4g persisted exclusively in Yamaguchi for over 12 years. Infectious HEV particles were confirmed in wild boar feces, highlighting the risk of environmental contamination and zoonotic transmission. Sika deer showed no evidence of HEV infection, and only one Japanese serow tested positive for antibodies without detectable RNA. These findings underscore the importance of ongoing surveillance to assess the zoonotic risks from game meat consumption and prevention of HEV transmission to humans. Full article

Insulin-like growth factor-1 (IGF-1) is a regulatory factor closely associated with diabetes, obesity, and breast cancer, and it also acts as one of the most abundant growth factors in bovine colostrum. Current methods generally have the problem of low sensitivity, a time-consuming nature, and low stability, which makes it difficult to crack down on the false advertising of IGF-1 content in dairy products. In this work, an ultrasensitive fluorescent enzyme-linked immunosorbent assay (ELISA) is proposed, where the antibody and the target are combined in the form of a “sandwich” to ensure the accuracy and specificity of the assay. IGF-1 is quantified based on an effective hydrogen peroxide (H₂O₂) probe with 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) as the fluorogenic substrate. The proposed fluorescent sandwich ELISA has a low limit of detection (LOD) of 77.29 pg/mL, fast experimental process within 1 h, and stable signal of 1 h. Furthermore, multi-step pretreatment methods for bovine colostrum powders are established to remove interfering substances, including fat, casein, and binding proteins, achieving the accurate and specific detection of IGF-1. IGF-1 recovery studies on treated bovine colostrum powders exhibit good recovery rates ranging from 91.71% to 102.32%, which proves the feasibility of detecting IGF-1 in real bovine colostrum. Full article

Surface-imprinted polymers (SIPs) represent an exciting and cost-effective alternative to antibodies for electrochemical impedance spectroscopy (EIS)-based biosensing. They can be produced using simple printing techniques and have shown high efficacy in detecting large biomolecules and microorganisms. Stamp imprinting, a novel SIP method, creates the target analyte’s imprint using a soft lithography mask of the analyte matrix, thereby reducing material complexities and eliminating the need for cross-linking, which makes the process more scalable than the conventional SIPs. In this work, we demonstrate a stamp-imprinted EIS biosensor using a biocompatible polymer, polycaprolactone (PCL), for quantifying amyloid beta-42 (Aβ-42), a small peptide involved in the pathophysiology of Alzheimer’s disease. The evaluated SIP-EIS biosensors showed a detection limit close to 10 fg/mL, and a detection range covering the physiologically relevant concentration range of the analyte in blood serum (from 10 fg/mL to 10 μg/mL). The device sensitivity, which is found to be comparable to antibody-based EIS devices, demonstrates the potential of SIP-EIS biosensors as an exciting alternative to conventional antibody-based diagnostic approaches. We also evaluate the viability of analyzing these proteins in complex media, notably in the presence of serum albumin proteins, which cause biofouling and non-specific interactions. The combination of high sensitivity, selectivity, and ease of fabrication makes SIP-EIS biosensors particularly suited for portable and point-of-care applications. Full article

Background: Myasthenia gravis (MG) and idiopathic inflammatory myopathy (IIM) are autoimmune diseases that affect the musculoskeletal system. The association of the two diseases is rare. Their management is different, so it is important to recognize the concomitant presentation. Methods: In this cross-sectional study, we study the presence of CK elevation, myositis-specific and myositis-associated antibodies (MSA/MAA), and vitamin D levels in a cohort of 101 MG patients. Electromyography, limb magnetic resonance imaging (MRI), and, in some cases, muscle biopsy were performed when IIM was suspected. We reviewed the patients’ medical records to access the results of these tests if they had been performed previously. Results: CK elevation was detected in 10 patients (9.9%). We identified one case of anti-Jo-1 antibody-positive polymyositis and two cases of possible myositis. MSA/MAA antibodies were not found in the patients with high CK levels, except for the one with anti-Jo-1-positive IIM. One patient with elevated CK levels had an overlapping muscular dystrophy. MSA/MAA antibodies were detected in 19 patients (18.8%). A total of 37% had high-titer antibodies and concomitant systemic autoimmune diseases, while 63% had low-titer antibodies, most of whom had no systemic autoimmune disease. Low serum vitamin D levels were found in 67.3% of patients. Comparison of myasthenia gravis composite (MGC) scores between patients with low and normal vitamin D levels did not show a statistically significant difference. Conclusions: Our results may raise awareness among neuromuscular specialists caring for MG patients of the possibility of associated myositis or other neuromuscular diseases and the need to assess vitamin D levels. Although deficiency was frequent, its impact on MG severity remains unclear, necessitating further investigation into its immunological relevance. Full article

CDD plays a pivotal role within the pyrimidine salvage pathway. In this study, a novel, rapid method for the identification of cell lines lacking functional cytidine deaminase was developed. This innovative method utilizes immunocytochemical detection of the product of 5-fluorocytidine deamination, 5-fluorouridine in cellular RNA, enabling the identification of these cells within two hours. The approach employs an anti-bromodeoxyuridine antibody that also specifically binds to 5-fluorouridine and its subsequent detection by a fluorescently labeled antibody. Our results also revealed a strong correlation between the 5-fluorouridine/5-fluorocytidine cytotoxicity ratio and cytidine deaminase content. On the other hand, no correlation was observed between the 5-fluorouridine/5-fluorocytidine cytotoxicity ratio and deoxycytidine monophosphate deaminase content. Similarly, no correlation was observed between this ratio and equilibrative nucleoside transporters 1 or 2. Finally, concentrative nucleoside transporters 1, 2, or 3 also do not correlate with the 5-fluorouridine/5-fluorocytidine cytotoxicity ratio. Full article

Leptospirosis is one of the world’s major neglected tropical zoonotic diseases (NTZDs), implicated in animal health and welfare with economic consequences for livestock production. This study aims to estimate the seroprevalence of Leptospira spp. and identify potential risk factors in small ruminant herds. This epidemiological cross-sectional study was conducted in Nuevo León, a semi-arid region of Mexico. A total of 389 blood samples from goats and 385 from sheep older than eight months were randomly collected from 128 herds. Anti-Leptospira antibodies were detected using the microscopic agglutination test (MAT), and univariate and multivariate logistic regression analyses were performed to determine their association with leptospirosis infection. The overall prevalence was 13.5% (105/774), with 14.4% (56/389) in goats and 12.7% (49/385) in sheep. Sejroe was the most predominant serogroup. The main risk factors in sheep were contact with domestic cattle, ≥100 animals per herd, congenital abnormalities, contact with feral pigs, meat production system, absence of veterinary care, and abortions with odds ratios (OR) between 1.7 and 4.1. In goats, the main risk factors included lack of quarantine measures, contact with feral pigs, absence of veterinary care, and abortions where the OR ranged from 1.7 to 3.3. These findings indicate that Leptospira spp. is present in small ruminant herds. This is the first study aimed at understanding leptospirosis epidemiology in the northeastern region of Mexico, as goats and sheep may act as potential reservoirs. Continuous monitoring of Leptospira infections is imperative, as well as developing educational initiatives for farmers to implement biosecurity and prevention measures to prevent infections within herds and protect public health. Full article

Ricin (RT) and abrin (AT) are plant toxins extracted from Ricinus communis and Abrus precatorius, respectively, and both have N-glycosidase activity. The detection of these toxins is vital because of their accessibility and bioterrorism potential. While ricin can be effectively detected based on its depurination activity, only a few tests are available for detecting the depurination activity of abrin. Therefore, it is unclear whether they share the same optimal reaction substrate and conditions. Here, we established optimum depurination conditions for ricin and abrin, facilitating the in vitro detection of their depurination activity using high-performance liquid chromatography–tandem mass spectrometry. The parameters optimized were the reaction substrate, bovine serum albumin (BSA), buffer, pH, temperature, time, antibodies, and magnetic beads. Both toxins showed better depurination with single-stranded DNA. However, substrate length, adenine content, BSA concentration, buffer concentration, reaction temperature, and reaction time differed between the two toxins. The optimal conditions for ricin depurination involved a reaction in 1 mM ammonium acetate solution (0.5 μM DNA15A, 20 μg/mL BSA, and 1 mM Zn²⁺, with pH 4.0) at 55 °C for 1 h. The optimal conditions for abrin depurination involved a reaction in 1 mM ammonium citrate solution (0.2 μM DNA20A, 10 μg/mL BSA, 1 mM Mg²⁺, and 0.5 mM EDTA, with pH 4.0) at 45 °C for 2 h. After optimization, the limits of detection (LOD) for ricin and abrin were 0.506 ng/mL and 0.168 ng/mL, respectively. The detection time was also significantly reduced. Full article

China’s livestock industry grapples with challenges posed by infectious diseases and the misuse of antibiotics, resulting in a heightened risk of drug-resistant pathogens. This study explored the immunomodulatory effects of co-expressing porcine interleukin 3, 7, and 15 in Yarrowia lipolytica, denoted as Po1h-IL-3/7/15. A 42-day experiment involving mouse immunization and pathogen challenge was conducted, during which in vivo assessments of antibodies, immune-related cells, and gene expression were detected following oral administration of Po1h-IL-3/7/15. Immunological alterations in mice were analyzed using flow cytometry, qRT-PCR, ELISA, and HE staining. Notably, the serum IgG and intestinal sIgA levels in the Po1h-IL-3/7/15 group were substantially elevated compared to the control groups (p < 0.01), so were the contents of IL-7, IL-15, IFN-γ, IL-22, IL-23, and TNF-α. Furthermore, there was a marked increase in naïve T cells and central memory T cells, accompanied by a significant decrease in regulatory T cells in peripheral blood. Post-challenge with Staphylococcus aureus or Salmonella typhimurium, the expression levels of BD2, IL-1β, IL-8, Jak1, RegⅢ, S100A8, STAT1, and TNF-α genes in the intestines of the Po1h-IL-3/7/15 group were markedly higher than those in the control groups (p < 0.01). Following the challenges, the survival rate of the Po1h-IL-3/7/15 group was 100%, a significant increase compared to the 20% and 40% survival rates observed in the control groups (p < 0.05). These results confirm that IL-3/7/15 significantly boosts innate immunity, humoral and cell-mediated immune responses, and intestinal mucosal immunity in mice, enhancing resistance to bacterial infections and exhibiting potent protective effects. Full article

Antibody-mediated rejection is a critical factor in acute and chronic allograft rejection, with Human Leukocyte Antigen as the primary target of the humoral immune response in kidney transplants. In addition to HLA antibodies, non-HLA Abs also play a significant role in AMR. These non-HLA Abs, which can target either autoantigens or alloantigens, may be present pre-transplantation or develop post-transplant. They are associated with various types of allograft injury. The major non-HLA Abs include those directed against the angiotensin II type 1 receptor, endothelin type A receptor, and MICA, as well as other antigens such as vimentin, collagens, and anti-endothelial cell antibodies. Factors such as ischemia, reperfusion injury, and calcineurin inhibitor toxicity can trigger the pathogenic activity of these Abs. The mechanisms underlying non-HLA Ab production are not yet fully understood but are thought to involve endothelial injury and the exposure of neoantigens. Research indicates that these non-HLA Abs can cause graft injury through both complement-dependent and complement-independent pathways. However, detecting non-HLA Abs remains a challenge due to the lack of reliable diagnostic tools. Current treatment strategies for managing the effects of pathogenic non-HLA Abs include intravenous immunoglobulin, plasmapheresis, rituximab, and bortezomib. Early identification of high-risk patients and timely intervention are crucial to preventing graft failure. This review examines the development, mechanisms, and clinical significance of non-HLA Abs in kidney transplantation, highlighting the need for improved diagnostic methods and tailored therapeutic approaches. Full article

Aethina tumida, commonly known as the small hive beetle, poses a threat to honeybee populations, particularly Apis mellifera, across several European regions. Originating in sub-Saharan Africa, there is a risk of the infestation spreading from Calabria, a region in the south of Italy. The essential role played by Apis spp. in pollination biodiversity preservation, agricultural productivity, and the overall economy is related to the dangers posed by the invasion of Aethina tumida. Current detection methods often fail to identify infestations early, leading to significant colony losses. We focused on creating a biosensor to improve the detection of Kodamaea ohmeri, a recently identified yeast that coexists symbiotically with Aethina tumida. The biosensor was designed to exploit the highly sensitive quartz crystal microbalance to identify a specific peptide linked to Kodamaea ohmeri in honey specimens. Its gold-plated surface over quartz was functionalized with an antibody effective in recognizing the peptide associated with Kodamaea ohmeri, a potential warning for detecting Aethina tumida. Preliminary results support the possibility of using such biosensor technology to detect infestation and enhance colony management techniques for honeybees, enabling beekeepers to implement prompt and focused treatments. In addition, reducing the size and cost of these biosensors and offering user training would be very helpful in having them used in beekeeping. Full article

Background: Monoclonal antibodies against the sodium-dependent phosphate transporter NaPi2b (SLC34A2) represent a promising approach in the treatment of ovarian and lung cancer. Of particular interest is the potential cancer-specific MX35 epitope of NaPi2b, as it serves as a target for monoclonal antibodies studied at various stages of preclinical and clinical trials. However, variations in the NaPi2b protein structure may limit the efficacy of therapeutic antibodies by affecting the accessibility of the MX35 epitope. Methods: An in silico analysis was performed using data from 101,562 tumor samples. Genomic DNA sequencing was conducted on blood samples from patients with ovarian carcinoma, breast cancer, and renal carcinoma to access the frequency of germline mutations in the SLC34A2 gene region encoding the MX35 epitope. To assess the impact of the selected mutation, we generated a model cell line through site-directed mutagenesis carrying the mutant NaPi2b variant. Results: Using in silico analysis, we identified 17 unique variants in the SLC34A2 gene leading to amino acid substitutions within the MX35 epitope of the NaPi2b. Among these, the most prevalent mutation, c.989C>T, resulting in p.T330M substitution, was detected in 5 out of 64 patients through genomic DNA sequencing. Using site-directed mutagenesis, we created the OVCAR-8/NaPi2b^p.T330M model cell line. L3 (28/1) monoclonal antibodies specific to the MX35 epitope failed to recognize the mutant NaPi2b^p.T330M variant compared to the wild-type of the NaPi2b in both Western blot and confocal microscopy experiments. Conclusions: The obtained data may serve as a basis for predicting the efficacy of monoclonal antibody-based targeted therapy binding to the MX35 epitope of NaPi2b in the treatment of oncological diseases. Full article

Background/Objectives: Primary Sjogren’s syndrome (pSS) is an autoimmune disease caused by chronic inflammation of the exocrine glands. We aimed to determine the immunohistochemical parameters that may help in the diagnosis of pSS and to determine histopathological markers for the early diagnosis of patients. Methods: Hematoxylin–eosin-stained preparations of salivary gland tissue samples of a control group consisting of 9 healthy patients and 12 patients diagnosed with non-specific chronic sialadenitis (NSCS) and a study group consisting of 39 patients diagnosed with pSS were evaluated. SSA/Ro (TRIM21), SSB/la, BAFF, enolase (ENO1), and MMP-9 antibodies were applied to the samples to assess the presence of staining in the ductal, acini, and inflammation regions. Results: In our study, mild staining with the SSA/Ro (TRIM21) antibody was observed in the ductal areas of all pSS cases, while no staining was detected in the healthy and NSCS cases (p < 0.01). Additionally, stronger staining was identified in the ductal and inflammatory areas of the pSS cases with BAFF compared to the control group. The staining for the ENO1 antibody was higher and more intense in the NSCS and pSS cases compared to those with normal salivary glands, and this difference was found to be statistically significant (p < 0.01). While mild staining was detected in the ductal areas with MMP-9 antibody in most of the NSCS and pSS cases, it was detected in 11% of the healthy cases. Conclusions: Our study suggests that SSA/Ro (TRIM21), ENO1, MMP9, and BAFF can be used to confirm the diagnosis in cases of suspicion. Full article

Background: We performed a phase I experiment in a healthy teenage population in Sui County, Henan Province, China. The trial was randomized, double-blind, and placebo-controlled. Methods: Ninety-six adolescents were randomly assigned in three groups (high-dose, medium-dose, and low-dose) to receive a dose of the vaccine or the placebo. The patients were monitored for adverse events (AEs) for up to 30 days after each dose of the vaccine and for up to 6 months after all doses of serious AEs (SAEs). All observed AEs and SAEs were reported. Microneutralization assays were used to measure geometric mean titers (GMTs) and seroconversion rates for neutralizing antibodies. IgA and IgG antibodies specific to the rotavirus were detected. Results: The rates of total AEs in these groups were 8.33%, 37.50%, 12.50%, and 4.17%, respectively. The neutralizing antibody test revealed that the teenage groups with low, medium, and high doses of the vaccine had geometric mean titers of 424.32, 504.63, and 925.45, respectively, at 28 days following complete vaccination. The GMT of serum IgG at final immunization was 6501.86, 6501.82, and 10,173.3, in the low-dose, medium-dose, and high-dose groups, respectively. The GMT of serum IgA at final immunization was 2733.64, 2233.29, and 3596.66 in the low-dose, medium-dose group, and high-dose groups, respectively. Conclusions: The majority of adverse events (AEs) were deemed Grade 1 or 2, suggesting that the vaccine’s safety profile is suitable for healthy adolescents. For the primary immunogenicity endpoints, a preliminary examination of the GMTs and the positive transfer rate of neutralizing antibodies in the different experimental groups revealed that, in adolescents aged 6–17, the high-dose group displayed significantly higher levels of neutralizing antibodies compared to the medium- and low-dose groups. Adolescents had few side effects from the new inactivated rotavirus vaccination, and it elicited an immune response. Full article

Background/Objectives: This is the first systematic review and meta-analysis investigating early myocardial dysfunction in children/adolescents with celiac disease and the effect of a gluten-free diet by comparing early echocardiographic markers between patients and healthy individuals and between compliant and non-compliant celiac disease patients (based on serum antibody titers). Methods: A systematic literature search was conducted across major electronic databases, with data collection extending up to 3 March 2024. Results: In total, 15 studies with 916 children/adolescent patients with celiac disease and 569 healthy individuals were included. Our results showed a trend toward reduced myocardial function in all echocardiographic parameters (conventional and advanced), with statistical significance in fractional shortening and the myocardial performance index. However, these parameters did not differ significantly after adherence to a gluten-free diet. Conclusions: Therefore, we recommend that an examination of the cardiovascular system should be incorporated into the routine investigations of children with celiac disease in order to detect early subclinical myocardial dysfunction based on echocardiography. Although the results of our meta-analysis indicate that the myocardial performance index may serve as a useful, non-invasive marker for assessing myocardial function in children and adolescents with celiac disease, further research is needed in order to confirm its reliability and clinical applicability in this population. The improvement of echocardiographic parameters after long-term compliance to a gluten-free diet is yet to be evaluated. Full article

The introduction of bispecific antibodies (BsAbs) has led to significant improvements in survival for patients with relapsed and refractory B-cell lymphomas. Despite these advances, there remains a significant number of patients who experience disease progression after these novel therapies. Predicting which patients may respond to certain treatments and the durability of their responses remains challenging. Measurable residual disease (MRD) has become easier to detect and quantify through the use of genomic next-generation sequencing tools and has been studied as a possible biomarker to predict long-term outcomes and risk-stratify patients after BsAb therapy in several lymphoma subtypes. Here, we review recent data demonstrating that MRD negativity is associated with radiographic response and improved progression-free survival. Because of heterogeneity in assay choice, assessment timing, and technical parameters, further work is needed before MRD testing is ready to be incorporated into clinical practice in the context of BsAb treatment for B-cell lymphomas. Full article