The relationship between aflatoxin M1 and immunoglobulin levels in cows' colostrum.

The purpose of the present research was to assess the amounts of aflatoxin M1 (AFM1) and immunoglobulin (IgA, IgG, and IgM) in cow colostrum samples, as well as their relationship. The sampling involved 90 cows (54 Montofon and 36 Simmental) from 15 independent farms. An appropriate number of samples from the total mixed ration (TMR) used in feeding the cows were collected simultaneously with the colostrum samples. AFB1 in feed, AFM1, and immunoglobulin (IgA, IgG, and IgM) levels in colostrum were evaluated using the ELISA method. The AFM1 level in colostrum samples exceeded the maximum allowed values for raw milk. Ig levels were higher in Montofon-breed cows' colostrum than in Simmentals. However, it was determined that neither the breed nor the lactation number significantly affected the colostrum Ig level. It was determined that there was no linear relationship between the number of lactations and immunoglobulin levels or between AFM1 and immunoglobulin levels in colostrum. Similarly, it was determined that there was no linear relationship between the AFB1 level in the feed and the AFM1 level in the colostrum of animals consuming these feeds.

Frequency and genotyping of group A rotavirus among Egyptian children with acute gastroenteritis: a hospital-based cross-sectional study.

BACKGROUND: This hospital-based cross-sectional study aims to investigate the epidemiologic and clinical characteristics of rotavirus group A (RVA) infection among children with acute gastroenteritis and to detect the most common G and P genotypes in Egypt. METHODS: A total of 92 stool samples were collected from children under five who were diagnosed with acute gastroenteritis. RVA in stool samples was identified using ELISA and nested RT-PCR. Common G and P genotypes were identified utilizing multiplex nested RT-PCR assays. RESULTS: RVA was detected at a rate of 24% (22 /92) using ELISA and 26.1% (24 /92) using VP6 nested RT-PCR. The ELISA test demonstrated diagnostic sensitivity, specificity, and accuracy of 91.7%, 100%, and 97.8%, respectively. G3 was the most prevalent G type (37.5%), followed by G1 (12.5%), whereas the most commonly detected P type were P[8] (41.7%) and P[6] (8.2%). RVA-positive samples were significantly associated with younger aged children (p = 0.026), and bottle-fed (p = 0.033) children. In addition, RVA-positive samples were more common during cooler seasons (p = 0.0001). Children with rotaviral gastroenteritis had significantly more frequent episodes of diarrhea (10.87 ± 3.63 times/day) and vomiting (8.79 ± 3.57 times/day) per day (p = 0.013 and p = 0.011, respectively). Moreover, they had a more severe Vesikari clinical score (p = 0.049). CONCLUSION: RVA is a prevalent cause of acute gastroenteritis among Egyptian children in our locality. The discovery of various RVA genotypes in the local population, as well as the identification of common G and P untypeable strains, highlights the significance of implementing the rotavirus vaccine in Egyptian national immunization programs accompanied by continuous monitoring of strains.

Severe fever with thrombocytopenia syndrome (SFTS) in Thailand: using a one health approach to respond to novel zoonosis and its implications in clinical practice.

Severe fever with thrombocytopenia syndrome (SFTS), a tick-borne disease caused by Dabie bandavirus (SFTSV) is an emerging infectious disease of substantial concern in East Asia. In 2019, Ongkittikul S et al. reported the first case of SFTS in Thailand. Our report describes a One Health investigation of SFTS zoonosis examining the index case and suspected animal reservoirs using real-time RT-PCR and immunoassays. We add to the report on the first confirmed case of SFTSV infection in a human in Thailand by conducting a limited but informative One Health surveillance study. Dogs and cats tested positive for SFTSV antibody using IgG ELISA. We conclude that domestic dogs and cats might serve as potential reservoirs for SFTSV spread due to their closer proximity to the index case than other non-domestic animals. Notably, we did not detect SFTSV in synanthropic cats or dogs-nor did we detect SFTSV in Rhipicephalus sanguineus ticks-using RT-PCR. We propose that One Health investigations coupling genomic and serologic assays in response to new SFTS cases could play a pivotal role in preventing and managing SFTS among humans and animals in East Asia. As such, we are establishing a collaborative response to SFTS in Thailand through human outbreak investigations that align with principles of One Health, through environmental surveys and animal RT-PCR and immunoassays. Our investigation highlights the importance of coupling RT-PCR with seroprevalence assays as principal elements of One Health surveillance for SFTS in order to shed light on potential animal reservoirs and track emerging zoonosis.

Exploring the Association of VEGF-A and ANGPTL2 with the Prognosis of Non-proliferative and Proliferative Diabetic Retinopathy.

Introduction Diabetic retinopathy (DR) is a microvascular ailment that can arise from the long-term effects of diabetes mellitus. It can potentially cause retinal damage that could endanger vision and cause blindness. The worsening of DR is mainly linked to poor glycemic control, uncontrolled hypertension, and dyslipidemia. There is a need for alternative and clinically significant novel molecules involved in the pathogenesis of DR because the diagnostic and prognostic markers have reached a limit. Materials and method This study included sex and age-matched diabetic patients with proliferative stage (N = 70), non-proliferative stage (N = 80), and control (N = 80, without the sign of DR). These patients were recruited from outpatients in the Department of Ophthalmology, Sri Ramachandra Institute of Higher Education and Research, Chennai, India. A random blood sample was collected from each study participant, and the serum was separated after centrifugation and stored at -80 °C for batch analysis. The biomarkers vascular endothelial growth factor (VEGF-A) and angiopoietin-like protein-2 (ANGPTL2) were measured using a sandwich enzyme-linked immunosorbent assay (ELISA) technique, and the laboratory parameters such as fasting blood sugar (FBS), lipid profile, blood urea nitrogen (BUN), creatine, and glycated hemoglobin (HbA1C) were also assessed. Results We observed statistically significant differences in the duration of diabetes, FBS, total cholesterol (TC), triglyceride level (TGL), BUN, and creatine (p<0.05), and the mean age of study participants was 52.95±8.20 years in the control group, 53.85±10.20 years in the proliferative diabetic retinopathy (PDR) group, and 55.02±7.65 in the non-proliferative diabetic retinopathy (NPDR) group. Furthermore, ANGPTL2 levels were statistically significant according to the severity of the disease (p<0.001*), and they were also linked (p<0.05) with established markers such as VEGF-A. Conclusion Thus, our research implies that the up-regulated markers might be linked to the disease's advancement and could serve as a prognostic indicator or therapeutic target for DR.

Early antibody responses to lipid antigens in red deer infected with Mycobacterium bovis.

THE PROBLEM: Early and rapid diagnosis of bovine tuberculosis remains an issue of great interest. AIM: The aim of this study was to evaluate the use of synthetic lipid antigens for diagnosis of tuberculosis in red deer (Cervus elaphus). The proposition: Synthetic mycolic acid derivatives, identical to components of mycobacterial cells, bind to antibodies to lipids produced in active human tuberculosis. Experimental infection studies in red deer (Cervus elaphus) allow the evaluation of such antigens for the serodiagnosis of bovine tuberculosis. RESULTS: Antibody levels in plasma from deer experimentally infected with Mycobacterium bovis were evaluated in ELISA using synthetic antigens based on several classes of mycolic acid, using protein G as conjugate. All antigens gave significantly increased responses 60 days post-infection, when all animals had active disease. A significantly increased response was also observed with four antigens 15 days after infection. CONCLUSION: ELISA using synthetic lipid antigens not only detects antibodies in the plasma of deer experimentally infected with M. bovis, but a strong response occurs early in the infection. With a full analysis of responses with naturally infected animals, this may offer a useful supplement to current diagnostic methods.

Characterization of the immunodominant regions of Senecavirus A-VP1 structural protein via ELISA epitope mapping.

Senecavirus A (SVA) is an RNA virus in the family Picornaviridae that has been detected in swine-production systems and is associated with vesicular disease and neonate mortality. The viral capsid is composed of four structural proteins: VP1-VP4. Although the VP1 protein has been reported to be the most immunogenic protein in vivo, no information on the immunodominant regions of the SVA polyprotein is available. The objective of this study was to identify the immunodominant regions of SVA polyprotein using an enzyme-linked immunosorbent assay (ELISA) epitope-mapping approach. The binding effect of SVA polyclonal antibody (SVA-pAb), SVA-VP1 monoclonal antibodies (SVA-mAb), and SVA-positive sera from clinically affected animals were characterized using a set of 18 overlapping SVA VP1-derived peptides by indirect and blocking ELISAs. All VP1 peptides yielded significant signal against SVA-pAb and SVA-VP1-mAb upon indirect ELISA. One peptide (aa 1-20) showed significantly high optical density on SVA recombinant VP1 protein (rVP1) and whole-virus-based indirect ELISAs. The blocking ELISA results demonstrated that peptides spanning aa 165-185 and 225-245 had a 50 % or greater inhibitory effect on SVA-pAb, while six groups of overlapping peptides spanning aa 1-35, 45-80, 90-140, 150-170, 195-230, and 240-264 and two groups of overlapping peptides spanning aa 1-50 and 60-264 showed a 50 % inhibitory effect or greater on swine VP1-mAb and SVA-seropositive swine serum, respectively, against SVA rVP1. Three-dimensional protein homology modeling showed that the peptides binding SVA-pAb are located on the outer surface of the viral capsid, while SVA mAbs and swine-positive sere can bind to epitopes located in both the inner and outer surfaces of the capsid. These linear epitopes showed differential binding and inhibitory activity on mAb and pAb; however, further studies will be necessary to evaluate whether they can act as decoy or neutralizing epitopes. Because mAb antibodies demonstrated a high binding affinity for this set of peptides, this information could lay the foundation for generating and screening specific antibodies for therapeutic potential.

Matrix metalloproteinases in dentin: Assessing their presence, activity, and inhibitors - a review of current trends.

INTRODUCTION: Dentin integrity is a critical aspect of tooth structure, with matrix metalloproteinases (MMPs) playing a crucial role in dentinogenesis, caries formation, and dental bonding. It is crucial to accurately assess MMP activity to understand dentin pathophysiology and develop effective clinical strategies. OBJECTIVES: The study aimed to conduct a thorough review and comprehensive summary of diverse techniques employed in assessing MMPs in dentin. DATA AND SOURCES: To conduct the research, electronic databases were systematically searched and manual citation searches were performed. A total of 621 articles were identified. After eliminating duplicates and irrelevant studies, 70 articles were included in the review. 25 articles with overlapping methodologies were also excluded. STUDY SELECTION: The selection criteria were based on the relevance of the studies to MMPs and MMP inhibitors in dentin without regard to the study design. Only peer-reviewed articles published in English were included. The search was restricted to studies published until November 2022. CONCLUSION: The comprehensive analysis of various studies has yielded 37 techniques for evaluating MMPs and MMP inhibitors, which hold significant promise in creating diagnostic markers and devising targeted therapeutic interventions.

Serosurvey and associated risk factors for bovine viral diarrhea virus infection in cattle in Egypt.

Bovine viral diarrhea virus (BVDV), is widely spread, poses a considerable risk of infection in the majority of dairy farms, causing respiratory, gastrointestinal, and reproductive problems. The aim of this study was to determine the seroprevalence and the risk variables associated with the seroprevalence of BVDV infection in cattle in four Egyptian governorates. A total of 680 blood samples were collected from cattle and examined for the presence of antibodies against BVDV using indirect ELISA (iELISA). Reproductive and management factors were considered, and epidemiological surveys were conducted. The total seroprevalence of BVDV in cattle was 18.24% (124/680) and it was significantly higher in females 19.66% (116/590), cattle older than 8 years 22.14% (62/280), dairy animals 22.65% (94/514), introduction of new animals to herd 21.39% (89/416), breeding with artificial insemination 28.46% (74/260), animals with history of abortion 28.76% (49/357), or during lactation stage 23% (89/387). The present findings suggest that BVD is prevalent in Egyptian dairy cattle and has an impact on farm productivity and production. Therefore, older, lactating, and aborted animals should also be identified for the disease, pose a risk of infection, and be handled appropriately.

Insulin-like growth factor (IGF) levels in pre-treatment plasma identifying breast cancer: A case control study.

Background: Diabetes (primarily type 2) is linked to a higher risk of breast cancer. Insulin-like growth factor (IGF) is one of the most important factors that affects mitosis and thus inhibits apoptosis. The purpose of this study was to compare the pre-treatment insulin-like growth factor (IGF) levels in breast cancer against normal population. Methods: In this case-control study, 60 patients with breast cancer and 60 healthy controls were enrolled in 2017 and 2018 at Tehran's Shahid-Modarres Hospital. In this study, the blood sugar of the patients was examined before entering the study, and the age of the patients was also within the age limit of 18 to 70 years. They were studied to determine the relationship between insulin-like growth factor (ELISA method) and breast cancer. Results: Both groups have similar IGF-1 levels (Ctrl and Case) (P= 0.188). But, IGF-2 levels were significantly higher in breast cancer patients (373.4 vs. 317.3 ng/ml), (P=0.0001). Conclusion: According to our study, IGF-2 may serve as a prognostic biomarker and potential therapeutic target for breast cancer. However, further investigation is needed to validate this claim.

Two metabolic enzymes, LDH and FASN, serum levels in Bladder cancer patients.

Background: Bladder cancer is one of the most common cancers in the world and is associated with high treatment costs and mortality. The role of different enzymes and molecules in this cancer has been the subject of extensive research in recent years. Among these, the role of metabolic enzymes such as FASN and LDH has been studied less than others. Therefore, the present study was designed to investigate the role of FASN and LDH in bladder cancer patients. Methods: One hundred cases diagnosed with bladder cancer and 50 sex-age- matched healthy individuals as control were examined. FASN and LDH serum levels in both patients and controls were determined by human-specific sandwich ELISA kits. Results: Serum levels of FASN and LDH elevated in bladder cancer patients in comparison to healthy individuals (P= 0.03, P= 0.01, respectively). We also found that than higher stages of bladder cancer (III-IV) had higher serum levels of LDH and FASN compared to early stages (I-II) (P= 0.007 and P= 0.006, respectively). Moreover, there was a statistically significant association between smoking history and serum FASN levels in bladder cancer patients (P=0.015). However, there were no remarkable associations between the serum levels of LDH and FASN with other clinicopathological features including sex, age, tumor grade, and tumor size. Conclusion: The data indicate that LDH and FASN may be good and useful biomarkers in the diagnosis and clinical management of bladder cancer. However, further studies are needed.

Molecular characterization of Peste des petits ruminants virus (PPRV) in sheep and goats and risk factors associated with it in selected districts of Khyber Pakhtunkhwa-Pakistan.

BACKGROUND: Peste des Petits Ruminants (PPR) is an economically significant transboundary viral disease of sheep and goats caused by the PPRV virus, affecting annual losses of 1.45-2.10 billion US dollars globally. We designed the current study to evaluate the positive cases, molecular characterization, phylogenetic analysis, and risk factors correlated with the disease in various districts of Khyber Pakhtunkhwa, Pakistan, with the aim of contributing to these strategies. METHODS AND RESULTS: A total of 384 samples from three selected districts, i.e., Peshawar, Charsadda and Chitral (n = 128 each), were collected, and the virus was investigated by using the sandwich ELISA, while the N gene of the virus was used as a target for molecular detection via RT-PCR. The confirmed samples were then sequenced, and phylogenetic analysis was performed. According to our findings, the highest positive cases was found in district Peshawar (50.87%), followed by Charsadda and Chitral (24.56%), respectively, while risk factor analysis showed that certain categories, such as species, sex, and age less than two years, have higher risk (P < 0.05) in contrast to their respective categories. Furthermore, sequencing and phylogenetic analysis of representative samples showed that the PPRV strains in the current study clustered in lineage IV, which is circulating in the small ruminant population of Asia, the Middle East, and African countries. Comparative residue analysis highlighted the mutation by representing 242 variable sites out of 371 locations. CONCLUSIONS: PPRV has foremost importance in Pakistan because the virus was detected in a considerable number of samples, and most of which were sourced from subsidiary areas where veterinary services are not prioritized.

Development of an indirect ELISA for the serologic detection of bovine viral diarrhea virus based on E2 antigen sub-genotypes 1b, 1e, and 1d.

Bovine viral diarrhea virus (BVDV) causes ongoing economic losses to cattle industries, directly through reduced herd performance or indirectly through control program costs. ELISA assays, one of the most widely used techniques due to their ease of implementation, have been a valuable tool for mass surveillance and detection of BVDV. In this study, we developed a new indirect ELISA (rE2-ELISA) for serologic detection of BVDV. The assay considers three recombinant E2 protein subtypes as antigens, allowing serologic diagnosis of BVDV-1b (high prevalence worldwide), BVDV-1d and 1e (high prevalence in southern Chile) sub-genotypes. Recombinant E2 (rE2) proteins were successfully expressed in stably transfected CHO cells. Conditions for rE2 ELISAs were established after determining appropriate concentrations of antigen, blocking agent, secondary antibody, and serum dilutions to achieve maximum discrimination between positive and negative serum samples. The developed rE2-ELISA showed a sensitivity of 92.86% and a specificity of 98.33%. Clinical testing of 180 serum samples from herds in southern Chile showed high accuracy (kappa > 0.8) compared to the commercial BVDV Total Ab kit (IDEXX), with 95.37% positive and 87.5% negative predictive value. In addition, the rE2 ELISA has shown the capability to detect anti-BVDV antibodies from naturally infected animals with sub-genotypes 1b, 1e, or undetermined. These results indicate that the developed indirect ELISA could serve as a valid, and efficient alternative for identifying BVDV-infected animals, thus contributing to the success of disease control and eradication programs.

Seroprevalence and assessment the associated risk factors for Fasciola hepatica infection in water buffaloes in Egypt.

Fasciolosis is a zoonotic neglected parasitic disease that affects a variety of hosts, resulting in substantial economic losses. The epidemiological information about fasciolosis in water buffaloes in Egypt is very scarce. Hence, the present study aimed to determine the seroprevalence of F. hepatica in water buffaloes using commercial ELISA kits in three governorates at north of Egypt and to estimate the associated risk factors for F. hepatica infection. The total seroprevalence of F. hepatica in buffaloes was 15.4% (63/410), with a higher seroprevalence in Kafr Elsheikh governorates 17.9% (25/140) than in other areas. Fasciolosis was more likely in older buffaloes (OR = 3.4, 95%CI:1.5-7.8), throughout the winter season (OR = 5.3, 95%CI:1.9-14.7). Moreover, the absence of prophylactic treatment (OR = 2.3, 95%CI:1.2-4.2) increased the risk of F. hepatica infection in buffaloes, particularly in animals suffered from diarrhea (OR = 3.8, 95%CI:1.4-10.6). The present study confirmed the prevalence of F. hepatica in water buffaloes in north of Egypt. Consequently, the implementation of preventive and control for the parasite and its intermediate host are very necessary to decrease the economic losses and public health hazard.

Carbohydrate antigens Lewis a and Lewis b act as tumor markers cooperating with CA19.9 in the management of PDAC patients.

BACKGROUND: CA19.9 is the unique marker recommended for the preoperative staging and the follow-up of patients suffering from pancreatic ductal adenocarcinoma (PDAC) but up to 30% of PDAC patients maintain normal CA19.9 values and cannot be monitored in this way. Lewis a (Lea Galß1,3[Fucα1,4]GlcNAc) and b (Leb, Fucα1,2Galß1,3[Fucα1,4]GlcNAc) are antigens which are structurally similar to sialyl-Lewis a (Siaα2,3Galß1,3[Fucα1,4]GlcNAc), the epitope of CA19.9. METHODS: We set an ELISA procedure determining the levels of Lea, Leb, and CA19.9 in the blood of healthy individuals or PDAC patients. Moreover, such antigens were also detected in cancer resections by immunofluorescence microscopy, and the levels of glycosyltransferase transcripts involved in Lewis antigen biosynthesis were determined by RT-qPCR. RESULTS: In our cohort of 116 healthy individuals, the distribution of circulating Lea and Leb was similar to that of CA19.9, allowing us to set putative cutoff values for both antigens. In a cohort of 115 PDAC patients, the differential distribution with respect to the controls was statistically significant for both antigens (p < 0.001). Out of 37 patients presenting normal CA19.9 values, 15 patients presented Lea or Leb above the cutoffs. By immunofluorescence, Lea, Leb and CA19.9 were all detected in cancer resections and expression levels were heterogeneous among patients in terms of intensity, localization and diffusion. The levels of relevant glycosyltransferase transcripts were found to be heterogeneous between cancers of different patients and no association was detectable with the levels of any circulating antigen. CONCLUSIONS: The concurrent quantification of Lea and Leb together with CA19.9 improves the management of PDAC patients.

Role of bone marrow mesenchymal stem cell-derived exosomes in reducing neurotoxicity and depression-like behaviors induced by doxorubicin in rats.

Background: Doxorubicin (DOX) is a broad-spectrum antitumor drug while its use is limited nowadays due to its neurobiological side effects associated with depression. Bone marrow mesenchymal stem cells (BM-MSCs) derived exosomes are a promising regenerative therapy. In this study, we investigated the therapeutic potentiality of BM-MSCs derived exosomes against the neurotoxicity induced by DOX. Methods: Twenty-four male albino rats were divided equally in to three groups as follow: group 1 (control), group 2 (rats injected intraperitoneally (i.p|) with DOX at a dose 2.5mg/Kg), and group 3 (rats injected with DOX and BM-MSCs derived exosomes i.p at a dose 1.5ml/Kg). During the experiment the behavior tests were noted, after three weeks rats were sacrificed, serum and brain samples were collected for biochemical, molecular and histopathological examinations. Results: The results revealed that DOX causing impairment of the locomotor and increasing the anxiety like behavior of rats, marked neuropathological changes, significant elevation of MDA content and TNF-α concentration, reduction of phospholipase (PLD) and acetylcholinesterase (AChE) protein concentration in addition, there were up regulation of JNK, NF-κB and p38 genes and down regulation of Erk1. Conclusion: Exosomal therapy improved the substantial neurotoxicity of DOX through modulating the markers involved in the neurotoxic signalling pathway of DOX that resulting in improving the pathological lesions and the animal behaviours.

Assessment of salivary and serum leptin in tobacco smokers and oral squamous cell carcinoma-A case-control study.

OBJECTIVES: Oral squamous cell carcinoma (OSCC) is the most common head and neck malignancy and tobacco-related cancers account for a significant portion of all oral cancers. Cancer patients often suffer from cachexia, which contributes significantly to mortality. Leptin is a protein released by adipocytes identified to play an important role in obesity and inflammation. The present study aimed to quantify and compare salivary and serum leptin in tobacco smokers and OSCC. MATERIALS AND METHODS: The present prospective case-control study enrolled 42 subjects divided equally among OSCC and tobacco smokers without oral lesions (TS). Both saliva and blood were collected from each subject and leptin levels were determined using enzyme-linked immunosorbent assay. The data obtained were analysed using Mann Whitney U, Kruskal Wallis, Friedman, and Spearman correlation tests (p < 0.05). RESULTS: A significant reduction in both salivary and serum leptin levels in OSCC was observed (p < 0.001, 0.002 respectively). In addition, significant reductions in weight and body mass index were also observed during follow-ups at 3, 6, and 9 months (p < 0.001 for both). CONCLUSIONS: Reduced salivary and serum leptin levels in OSCC proved that it is an important diagnostic marker, with non-invasive saliva measurement being more patient-friendly. Future multicentric studies with higher samples in OSCC subgroups are warranted. CLINICAL RELEVANCE: Leptin reduction in oral squamous cell carcinoma proved to be an important diagnostic marker. Non-invasive salivary techniques could be employed in mass screening programmes. The significant correlation between leptin and BMI also shed insight into the overall well-being of the patient.

Evaluation of the focus reduction neutralization and ELISA tests compared to the plaque reduction neutralization test for the detection of antibodies against measles virus.

Measles is an infectious febrile sickness caused by the measles virus (MeV). Despite an effective vaccine, regional elimination of measles remains a global priority and still faces challenges. To estimate community protection against measles, sensitive tests are needed to identify measles-specific antibodies. The enzyme-linked immunosorbent assay (ELISA) is the standard test for assessing immunity but may fail to detect weak antibody responses in vaccinated populations. The plaque reduction neutralization test (PRNT), is the gold standard test for the assessment of protective antibody levels, however, it is not suitable for routine use. This study validated the focus reduction neutralization test (FRNT) as an alternative. In eight assay runs, fifty serum samples were analyzed in triplicate using PRNT, FRNT, and ELISA. Data analysis revealed that 38 samples were positive by PRNT, 37 by FRNT, and 19 by ELISA. The results showed that ELISA was not sensitive enough to identify low levels of anti-measles antibodies and showed weak agreement with neutralization assays. In contrast, the two neutralization assays had a perfect correlation and similar sensitivity. FRNT appears to be a suitable alternative to PRNT for characterizing immunological responses and vaccination efficacy. Our results highlight the necessity of validating negative and equivocal ELISA results through neutralization methods, during the elimination phases.

A longitudinal study of the dynamics of Mycoplasma bovis antibody status in primiparous cows and bulk tank milk in Swedish dairy herds.

Mycoplasma (M.) bovis is an important pathogen causing pneumonia, mastitis, and arthritis in cattle all over the world entailing reduced animal welfare and economic losses. In this longitudinal study, we investigated the presence of M. bovis antibodies in bulk tank milk (BTM) and in milk from primiparous (PP) cows at 4 sampling occasions over 2 years. Herd characteristics associated with a positive antibody test result in PP cows were investigated. The participating dairy herds (n = 149) were situated in southern Sweden, samples were collected and analyzed with ID Screen antibody ELISA. Information on herd characteristics was retrieved from the national Dairy Herd Improvement database. To identify herd characteristics associated with the presence of antibodies in PP cows, mixed linear regression with herd and sample as random factors were used. The apparent herd-level prevalence of M. bovis infection based on antibodies in BTM was 17% but with the addition of PP cows the prevalence increased to 28%. The results showed that larger herds and introduction of cattle was associated with higher antibody levels in PP cows. In conclusion, this study showed a clear difference in the apparent prevalence of M. bovis infection based on antibodies in BTM or in PP cows, the no. of positive herds were almost doubled when including PP cows. This motivates repeated sampling of a few PP cows to find newly infected herds in an early stage. Finally, the results showed that introduction of cattle influences the level of M. bovis antibodies. This is important in the control and prevention of further spread of the infection. It is essential for free herds to know their M. bovis status and antibody testing is highly recommended if introducing cattle.

Lateral flow test versus enzyme-linked immunosorbent assay to measure infliximab trough concentrations: A head-to-head comparison.

OBJECTIVES: Infliximab is an antitumour necrosis factor agent used to treat inflammatory bowel disease (IBD). Measurement of infliximab trough concentrations (C-troughs) are used to optimize drug exposure and improve outcomes. Currently, enzyme-linked immunosorbent assays (ELISAs) are used predominantly for this purpose. Novel lateral flow immunoassays provide a rapid result. METHODS: We collected 100 paired serum samples of adolescents and young adults with IBD, who were treated with infliximab maintenance infusions. C-troughs were measured with the Quantum Blue® lateral flow test (QB) with ELISA. Results were categorized as low-range (mean C-trough ≤5 µg/mL) or high-range (>5 µg/mL). A Bland-Altman plot was created with limits of clinical acceptability set at ≤2 µg/mL for low-range and ≤40% for high-range C-troughs. A concordance matrix was created to evaluate the C-trough-based clinical scenario (whether or not to escalate infliximab) using a cutoff value of 5 µg/mL. RESULTS: Agreement between QB and ELISA was good (intraclass correlation coefficient: 0.85). In the low-range, 90% (95% confidence interval [CI]: 79-96) of measurements were within the limits of clinical acceptability. In the high-range this was 67% (95% CI: 53-79). QB provided higher results than ELISA. The concordance matrix showed 81% agreement (95% CI: 72-88, κ: 0.62). CONCLUSIONS: Lateral flow- and ELISA-based infliximab C-trough measurements were in agreement. The swift establishment of infliximab C-troughs matters for patients experiencing increased disease activity. In the event of a low C-trough, prompt dose escalation can be initiated.

Serum Concentration of MMP-9 as a Predictive Biomarker for the Progression of Oral Cancer.

Background: Oral cancer is the most prominent cancer subtype among all head and neck cancers, the incidence and prevalence of which has been consistently increasing in past years around the globe. At advanced stages, oral cancer imparts significant mortality, morbidity, and mutilation among the patients, and therefore, diagnosis and treatment of the disease at early stages are considered the optimum strategy for the management of the disease. Since molecular changes appear earlier than physical symptoms, several molecular biomarkers are currently being investigated for their role in diagnosing and treating disease. MMP-9 belongs to the family of proteinases that are involved in cytoskeletal degradation, which is a crucial phase of cancer progression. Objective: In the present study, we analyzed the serum concentration of MMP-9 in oral cancer patients and tried to establish MMP-9 as a predictive biomarker for the progression of oral cancer. We also correlated the clinical, sociodemographic and biochemical parameters with the serum concentration of MMP-9 in oral cancer patients. Methods: Serum was extracted from the blood sample of 38 oral cancer patients and was analyzed for the concentration of MMP-9 using sandwiched ELISA. Predesigned proforma was used to capture the clinical, sociodemographic and biochemical parameters. Unpaired t-test was used to compare two means, one way ANOVA was used to compare more than two means and Pearson's correlation was used to correlate the variables. Results: The mean concentration of MMP-9 in patients of oral cancer was 816.9 ± 236.1 ng/mL. The MMP-9 expression level was higher at advanced oral cancer stages than in the early stages. No significant difference in the MMP expression was found in terms of sociodemographic risk factor and tumor site. MMP-9 exhibit significant negative correlation with the HDL and significantly positive correlation with the PTI. Rest of the biochemical parameters does not exhibit significant correlation. Conclusion: The present study suggests that serum concentration of MMP-9 can be a predictive biomarker for the progression of oral cancer, which needs to be validated by performing further longitudinal cross-sectional studies by taking ample sample size.