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This document is a systematic review that discusses the challenges and inconsistencies in diagnosing cold agglutinins (CA), which are IgM autoantibodies that can cause hemolysis. The review aims to inform future research on the diagnostic investigation and interpretation of CA by describing serological techniques and cut-offs for diagnosing cold agglutinin disease (CAD) or cold agglutinin syndrome (CAS). The authors conducted a literature search and included 56 articles in their analysis, which revealed a lack of standardized definitions and thresholds for diagnosing clinically significant CA. The document provides a summary of various studies and case series related to cold agglutinins (CA), examining different aspects such as the number of participants with CA, the underlying clinical diseases studied, and the definition of CA. It presents a range of perspectives and findings from different studies. The article discusses the lack of consistency in defining the characteristics of cold agglutinin disease (CAD) in autoimmune hemolytic anemia (AIHA). The most commonly used laboratory test for diagnosing CAD is the direct antiglobulin test (DAT), which detects complement deposition on red blood cells. However, the clinical utility of DAT testing and the interpretation of results are variable. CA titers, which reflect the concentration and avidity of cold agglutinins, are also widely used but do not always correlate with disease severity. The determination of thermal amplitude [Extracted from the article]

Background: Cold agglutinins (CAs) in blood samples can cause a reversible agglutination of red blood cell (RBC) which result in an incorrect complete blood count (CBC). So, it is important to explore new simple and feasible treatment conditions for clinical work. Methods: The CAs group included 32 samples with CAs. The parameters of CBC at room temperature or after prewarming at 37°C or 41°C for different time periods were compared. The consistency and correlation of those parameters were analyzed. The morphology of erythrocytes in the CAs group was observed manually. The control group included 45 samples without CAs and prewarmed at 37°C or 41°C for different time periods. The differences were also analyzed. Results: CAs have a significant effect on CBC. After prewarming at 37°C or 41°C the interferences are all corrected. Consider prewarming at 37°C for 120 minutes as the standard procedure. The consistency and correlation analysis showed there was no statistical difference between the results of each subgroup and standard group, except the MCHC of group 41°C 10 minutes. The correlation of parameters between all subgroups and the standard group is satisfied. Microscopic examination showed no RBC aggregation or fragmentation after prewarming at 41°C or 37°C. According to the maximum bias requirements for expert performance in Validation, Verification, and Quality Assurance of Automated Hematology Analyzers, 2nd Edition (CLSI H26-A2), the differences in over-all results in control group are negligible. Conclusions: The 41°C 2 minutes prewarming method is a rapid and effective way for treating samples with CAs. It is an efficient way to obtain more reliable CBC results, without specific instruments. [ABSTRACT FROM AUTHOR]

The kidney is a highly complex organ equipped with a multitude of miniscule filter-tubule units called nephrons. Each nephron can be subdivided into multiple segments, each with its own morphology and physiological function. To date, conventional manual approaches to isolate specific nephron segments are very laborious, time-consuming, often limited to only a specific segment, and typically have low yield. Here, we describe a novel, unconventional method that is superior in many aspects to previous protocols by combining low-cost fluorophore-conjugated lectins or agglutinins (Flaggs) with flow sorting. This allows the simultaneous separation of different nephron segments with preserved 3D morphology from mouse or human samples in under 3 h. Using a 200-µm nozzle and 5 psi, glomeruli, proximal, or distal convoluted tubules are sorted with Cy3-labeled Sambucus Nigra agglutinin (SNA-Cy3), Fluorescein-labeled Lotus Tetragonolobus lectin (LTL-FITC), or Pacific Blue-labeled soybean agglutinin (SBA-PB), respectively. Connecting tubules and collecting ducts are sorted by double-positive SBA-PB and SNA-Cy3 signals, while thick ascending limb segments are characterized by the absence of any Flaggs labeling. From two mouse kidneys, this yields 37–521 ng protein/s or 0.71–16.71 ng RNA/s, depending on the specific nephron segment. The purity of sorted segments, as assessed by mRNA expression level profiling of 15 genes, is very high with a 96.1-fold median enrichment across all genes and sorted segments. In summary, our method represents a simple, straightforward, cost-effective, and widely applicable tool yielding high amounts of pure and morphologically largely intact renal tubule materials with the potential to propel nephron segment-specific research. [ABSTRACT FROM AUTHOR]

The self-assembly of carbohydrate-based amphiphiles can lead to colloidal soft materials such as supramolecular gels featuring highly desirable characteristics like biodegradability and biocompatibility. The report herein presents the synthesis, characterization and supramolecular self-assembly, physical gelation and wheat lectin binding of two structurally related amphiphilic compounds having β-S-N-acetylglucosamine residues linked to a 2,3-diacyl-N,N′-dipropargylated-l-tartaric diamide. A 1-thio-β-N-acetyl-d-glucosamine precursor attached to a conveniently functionalized linker with an azido group was synthesized by means of a one-pot procedure followed by deprotection. A click reaction successfully led to the two amphiphiles, which differed in length of the fatty acid attached to the tartaric acid scaffold. Although both compounds are poorly soluble in water and organic solvents, the difference in terms of hydrophilic moieties provided them with distinct supramolecular gelation properties. While the presence of an octadecyl chain produced a hydrogelator, the dodecadecyl homologue would only form weak gels in DMSO. SEM and rheology experiments confirmed the characteristic fibrillar morphology and viscoelastic properties, in agreement with the presence of physical gels. Both amphiphiles were able to interact reversibly with wheat germ agglutinin (WGA), a lectin that specifically recognizes GlcNAc residues, indicating a potential use in the food industry, as a gluten sensitivity manager, as well as in health-related industries, for example, for drug delivery systems. [ABSTRACT FROM AUTHOR]

Autoimmune hemolytic anemias (AIHAs) are hemolytic anemias characterized by the production of autoantibodies against surface antigens on red blood cells. Cold agglutinin disease (CAD) is a relatively rare subcategory of AIHA, affecting mainly the middle-aged with anemia often being the presenting symptom. We present a case of a middle-aged female who was admitted for severe anemia under evaluation. A discrepancy in blood grouping was noted when request for blood transfusion was received in a blood bank, owing to autoantibodies interfering in testing. Routine complete blood counts showed temperature-based variations. A detailed workup confirmed a diagnosis of CA type of AIHA. The case reiterates the importance of good interdepartmental coordination between clinicians, laboratory, and blood bank, which helped clinch the diagnosis of CAD through the correlation of clinical findings, blood counts, and immunological and biochemical parameters.

Autoimmune hemolytic anemias (AIHAs) are hemolytic anemias characterized by the production of autoantibodies against surface antigens on red blood cells. Cold agglutinin disease (CAD) is a relatively rare subcategory of AIHA, affecting mainly the middle-aged with anemia often being the presenting symptom. We present a case of a middle-aged female who was admitted for severe anemia under evaluation. A discrepancy in blood grouping was noted when request for blood transfusion was received in a blood bank, owing to autoantibodies interfering in testing. Routine complete blood counts showed temperature-based variations. A detailed workup confirmed a diagnosis of CA type of AIHA. The case reiterates the importance of good interdepartmental coordination between clinicians, laboratory, and blood bank, which helped clinch the diagnosis of CAD through the correlation of clinical findings, blood counts, and immunological and biochemical parameters. [ABSTRACT FROM AUTHOR]

Malaria and typhoid fevers are twin diseases that plague many Sub-Saharan African and Asian countries at epidemic proportions with almost a third of all deaths occurring in Nigeria in the last decade. Diagnosis of malaria in Nigeria relies heavily on the simple and inexpensive microscopy and the Widal agglutination test is commonly used in resource-poor settings where cultures fail or unavailable for typhoid fever diagnosis. The study was conceived to determine the relevance of the Widal agglutination test in a malaria endemic region. Patients who had previously received artemisinin combination therapy (ACT) with unremitting febrile illness who gave consent were enlisted into the study. Blood samples were collected for blood films, examined microscopically for the presence of malaria parasites. Widal agglutination test was carried out on each separated serum with the rapid slide agglutination test and titre was determined for sera showing agglutination with tube dilution assay. Malaria parasites were present in 236/429(55.0%) of the blood samples of which 41.7% had parasitemia density of =5000/mL of blood. Parasitemia density higher than 5000/μL of blood accounted for 13.4%. Widal agglutination titres as high as 320 were recorded in patients with malaria parasites as wells as those without malaria parasitemia. Malaria is a major cause of febrile illness in the study population which can be carefully excluded with the inexpensive blood film microcopy to enable proper evaluation for typhoid fever to be undertaken. [ABSTRACT FROM AUTHOR]

Background. Mixed warm/cold autoimmune hemolytic anemia (AIHA) is a rare diagnostic entity with limited therapeutic options. Previous literature has described the diagnostic difficulty in this pathology and the limited response rates to corticosteroids. Furthermore, there is limited evidence regarding the use of rituximab in this condition. Methods. Alongside our case report, we conducted a scoping review of case reports/case series describing mixed AIHA, their treatment, and clinical outcomes since 2000. Inclusion criteria included a confirmed diagnosis of mixed AIHA (confirmed warm antibodies and cold agglutinins based on DAT). Case Summary/Results. We present a case of mixed AIHA in an 83-year-old female presenting with extensive, bilateral pulmonary embolisms and left renal vein thrombosis. The patient underwent extensive workup with no identifiable provoking etiology. Initial treatment involved prednisone therapy was transitioned to rituximab upon diagnosis of mixed AIHA. The patient demonstrated a mixed response with stable hemoglobin and transfusion independence; however, with persistently elevated hemolytic indices following completion of rituximab treatment. Our literature review identified 16 articles; two were excluded for unavailable clinical details. The most commonly associated conditions included autoimmune conditions (n = 5, 26%) and lymphoproliferative disorders (n = 3, 12%). The most common treatment involved corticosteroids; seven studies involved the use of rituximab. Conclusion. Mixed AIHA represents a complex diagnosis and optimal management is not well established. Consistent with our case, recent literature suggests a promising response to rituximab and a limited response to steroid treatment. Given the limited literature, additional studies are required to elucidate optimal management of this unique pathology. [ABSTRACT FROM AUTHOR]

The term "cold agglutinin (CA)" refers to a group of disorders caused by anti-erythrocyte autoantibodies that preferentially bind RBCs at cold temperatures (4°C-18°C). CAs contribute to 10 to 15% of autoimmune hemolytic anemia. We report a case of CAs diagnosed intraoperatively during emergency mitral valve replacement. [ABSTRACT FROM AUTHOR]

The influence of the composition of the nutrient medium on the yield of biomass and the biosynthesis of agglutinins of the mold fungus strain Rhizoctonia solani RS was studied. At the first stage of research, the agglutinin-producing micromycete strain was grown on liquid nutrient media (potato-glucose (PG), Chapek, Sabouraud) at a temperature of 28°C for 8 days. At the next stage of research, the composition of the culture medium was modified in order to select sources of nitrogen (NH4NO3, (NH4)2SO4, NaNO3, arginine, tryptophan, asparagine, threonine, peptone) and carbon (sucrose, glucose, starch) nutrition. Cultivation of the Rh. solani RS on a liquid PG medium, containing glucose in an amount of 20.0 g/l led to a maximum yield of fungal biomass of 23.58 ± 1.30 g/l and the production of its agglutinins (titer 16384). The studied micromycete strain was characterized by the ability to actively use the amino acids threonine or asparagine when added to the above nutrient medium in an amount of 0.1 mg/ml. At the same time, the hemagglutination activity of the micromycete agglutinins increased twofold (titer 32768) compared to the activity of agglutinins on the PG medium without the addition of amino acids (titer 16384).

Background: Diagnosis of prolonged febrile illness of unknown origin (FUO) is challenging even with the advances in the diagnostic techniques. As common as the infection with cytomegalovirus (CMV) is, most health care providers would not suspect CMV infection as a cause of FUO unless mononucleosis syndrome is evident. The aim of this study is to investigate the rate of CMV infection among patients with FUO and shed light on IgG avidity as a diagnostic tool. Patients and methods: Two hundred and twenty three (223) immune competent patients with FUO were included in our study. They were subjected to all routine laboratory investigations, fever agglutinins, tuberculin and abdominal ultrasound along with IgG and IgM for CMV and IgG avidity test. Results: This study shows that the 92.8% of the overall studied population were positive for CMV IgG. However, only 74(33.2%) of the studied population was found positive for IgM. Only one patient had positive IgM with negative IgG. IgG avidity was high in almost all of them. Only 3 patients showed low IgG avidity denoting that they have primary infection. Conclusion: CMV infection was found to be the cause of 33.2% of prolonged febrile illness experienced by immuncompetent adults. Only 1.8% of patients had primary CMV infection and the majority of them had detectable IgG level and were diagnosed with primary infection depending mainly on IgG avidity test.

Mixed autoimmune hemolytic anemia (AIHA) represents <5% of pediatric AIHA cases. Spontaneous agglutination is known in patients with cold agglutinins reacting at room temperature. They often interfere with serological tests; generally, they can be dispersed by simple warming techniques. Rarely, in severe cases, as seen in the index case, dithiothreitol treatment of the red cells is needed to eliminate autoagglutination. [ABSTRACT FROM AUTHOR]

Background Hand arm vibration syndrome (HAVS) is a condition caused by hand transmitted vibration from the use of hand-held vibrating tools or workpieces. The disease affects the vascular, neurological and musculoskeletal systems. The vascular component of HAVS is a form of secondary Raynaud's phenomenon. Other causes of disease must be excluded before attributing the cause to hand transmitted vibration. Aims To evaluate the prevalence, and utility of testing for, cryoglobulins and cold agglutinins in patients with HAVS symptoms. Methods A retrospective cohort study of 1183 patients referred for HAVS clinical assessment at St. Michael's Hospital, Toronto, Canada, between 2014 and 2020. The standard operating procedure at the clinic includes a detailed clinical and exposure history, physical examination, objective investigations and blood tests. Data were retrieved from patient chart review and laboratory investigation results for all cases with cryoglobulin and cold agglutinin testing. Results A total of 1183 patients had a serum cryoglobulin measurement. Eleven patients (1%) were positive. Seven positive results were 'low titre' (1% positive) and the other four results were 2%, 6%, 9% and 18%. The patient with a 9% positive cryoglobulin titre had previously diagnosed Sjögren's syndrome. There were no positive cold agglutinin tests in the 795 patients tested. Conclusions Routine testing for cryoglobulins and cold agglutinins in patients with HAVS symptoms is not recommended because test positivity rates are negligible. Testing may be considered if the clinical history or routine blood investigations suggest evidence of underlying cryoglobulinaemia or cold agglutinin disease. [ABSTRACT FROM AUTHOR]

Background: Diagnosis of prolonged febrile illness of unknown origin (FUO) is challenging even with the advances in the diagnostic techniques. As common as the infection with cytomegalovirus (CMV) is, most health care providers would not suspect CMV infection as a cause of FUO unless mononucleosis syndrome is evident. The aim of this study is to investigate the rate of CMV infection among patients with FUO and shed light on IgG avidity as a diagnostic tool. Patients and methods: Two hundred and twenty three (223) immune competent patients with FUO were included in our study. They were subjected to all routine laboratory investigations, fever agglutinins, tuberculin and abdominal ultrasound along with IgG and IgM for CMV and IgG avidity test. Results: This study shows that the 92.8% of the overall studied population were positive for CMV IgG. However, only 74(33.2%) of the studied population was found positive for IgM. Only one patient had positive IgM with negative IgG. IgG avidity was high in almost all of them. Only 3 patients showed low IgG avidity denoting that they have primary infection. Conclusion: CMV infection was found to be the cause of 33.2% of prolonged febrile illness experienced by immuncompetent adults. Only 1.8% of patients had primary CMV infection and the majority of them had detectable IgG level and were diagnosed with primary infection depending mainly on IgG avidity test. [ABSTRACT FROM AUTHOR]

Agglutinins/lectins are conventionally defined as proteins/ glycoproteins of non-immune origin with a remarkable ability to specifically and reversibly interact with carbohydrate ligands. Lectins from different sources may essentially exhibit common biological activities. This study was therefore undertaken to survey naturally occurring agglutinins in two species of marine crabs by hemagglutination assay using mammalian erythrocytes. Hemagglutination assay results showed that the hemolymph of the marine crab Grapsus albolineatus showed the highest HA titer with rat erythrocytes. HA titer of the crab Leptodius sanguineus varied from 0 to 32 with all the tested erythrocytes. Among the various tissues of Grapsus albolineatus analyzed for the presence of agglutinins, hemagglutination activity was observed in the hemolymph and hepatopancreas with rat erythrocytes. HA was determined by both male and female crabs of Grapsus albolineatus. HA activity increased with an increase in animal size. Biochemical factors like water, protein and calcium content of the hemolymph did not have any influence on the HA titer. [ABSTRACT FROM AUTHOR]

Background: Cold agglutinin disease (CAD) is a rare disorder, affecting 15% of patients with autoimmune hemolytic anemia. Few studies have assessed CAD symptoms and their impact on daily life, but these studies did not address the patients' perspectives. Objective: The aims of this study were to increase the knowledge about CAD through a patient-centric survey and to gain a better understanding of the burden of this disease. Methods: We conducted an internet-based survey in September 2020 among American patients registered on the CAD Unraveled website and members of the Cold Agglutinin Disease Foundation. Results: A total of 50 respondents were included in this study. Totally, 90% (45/50) of the patients reported having experienced fatigue. Fatigue was mainly reported on a daily basis, and approximately one-third of these patients (13/45, 29%) said that their fatigue was constant throughout the day. It has also been shown that CAD has a great impact on patients' physical well-being, emotional well-being, social life, and household finances. The disease varies over time, with or without symptoms. A total of 88% (44/50) of the patients reported previous episodes of the increased intensity or sensitivity of their CAD symptoms, with a mean of 4.5 (SD 5.4) episodes reported during the past year. More than half of the patients (27/50, 54%) considered their disease to be moderate or severe, and 42% (21/50) of the study group reported that their symptoms had worsened since the time of diagnosis. Conclusions: Our study has provided new data on CAD symptoms, particularly data on the importance and type of fatigue and the fluctuation of CAD symptoms. [ABSTRACT FROM AUTHOR]

Patents of lectins with antiviral, antibacterial and antifungal applications were searched and reviewed. Lectins are proteins that reversibly bind to specific carbohydrates and have the potential for therapy of infectious diseases as biopharmaceuticals, biomedical tools or in drug design. Given the rising concerns over drug resistance and epidemics, our patent review aims to add information, open horizons and indicate our view of the future perspectives about the antimicrobial applications of lectins. Patents with publications until December 2020 were retrieved from Espacenet using defined search terms and Boolean operators. The documents were used to identify the geographical and temporal distribution of the patents, characterize their lectins, and classify and summarize their antiviral, antibiotic and antifungal applications. Lectins are promising antiviral agents against viruses with epidemics and drug resistance concerns. Mannose‐binding lectins were the most suggested antiviral agents since glycans with mannose residues are commonly involved in viral entry mechanisms. They were also immobilized onto surfaces to trap viral particles and inhibit their spread and replication. Many patents described the extraction, isolation, amino acid and nucleotide sequences, and expression vectors of lectins with antibiotic and/or antifungal activities in terms of MIC and IC50 for in vitro assays. The inventions also included lectins as biological tools in nanosensors for antibiotics susceptibility tests, drug‐delivery systems for the treatment of resistant bacteria, diagnostics of viral diseases and as a vaccine adjuvant. Although research and development of new medicines is highly expensive, antimicrobial lectins may be worth investments given the emergence of epidemics and drug resistance. For this purpose, less invasive routes should be developed as alternatives to the parenteral administration of biologics. While anti‐glycan neutralizing antibodies are difficult to develop due to the low immunogenicity of carbohydrates, lectins can be produced more easily and have a broad‐spectrum activity. Protein engineering technologies may make the antimicrobial applications of lectins more successful. [ABSTRACT FROM AUTHOR]

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Carrageenan is a substance widely used as an additive in the food industry. Among other things, it is often added to processed cheese, where it has a positive effect on texture. Processing of such cheese involves grinding, melting and emulsifying the cheese. There is currently no official method by which carrageenan can be detected in foodstuffs, but there are several studies describing its negative health impact on consumers. Lectin histochemistry is a method that is used mainly in medical fields, but it has great potential to be used in food analysis as well. It has been demonstrated that lectin histochemistry can be used to detect carrageenan in processed cheese by Human Inspection and Computer-Assisted Analysis (CIE L*a*b*). The limit of detection (LoD) was established at 100 mg kg−1 for Human Inspection and 43.64 for CIE L*a*b*. The CIE L*a*b* results indicate that Computer-Assisted Analysis may be an appropriate alternative to Human Inspection. The most suitable parameter for Computer-Assisted Analysis was the b* parameter in the CIE L*a*b* color space. [ABSTRACT FROM AUTHOR]

The paper demonstrates examination results of antigenic properties of Avibacterium paragallinarum reference strain and ten isolates thereof recovered in the Russian Federation and Republic of Belarus. Nine isolates and the reference strain demonstrated type L hemagglutinin (thermoliable, trypsin-sensitive, active against fresh and glutaraldehyde-treated RBC). One isolate was marked by type Hl hemagglutinin (thermoliable, trypsin-resistant, active only against glutaraldehydetreated RBC). The reference strain antigen proved to be inagglutinable by homologous antiserum, and unable to agglutinate RBCs due to hyaluronic acid in the capsular substance. Serological and hemagglutination non-reactivity was removed through the cell treatment with hyaluronidase. The agglutination test demonstrated that seven out of ten tested isolates belonged to the same serological group; herewith, the proportion of the bilateral antigenic relatedness amounted to ≥78.4%. HI results demonstrated ≥92.6% antigenic relatedness of the tested isolates being indicative of the fact that they belonged not only to the same serological group but also to the same serotype. No serological relatedness was identifed between the tested isolates and reference strain of serogroup A both using agglutination test (≤23.6%) and HI (≤12.2%). Polymerase chain reaction demonstrated that all the isolates recovered in the Russian Federation and Republic of Belarus in 2014-2016 belonged to serogroup B.

VicRK (WalRK or YycFG) is a conserved 2-component regulatory system (TCS) that regulates cell division, cell wall biosynthesis, and homeostasis in low-GC Gram-positive bacteria. VicRK is also associated with biofilm formation of Streptococcus mutans on the tooth surface as it directly regulates the extracellular polysaccharide (EPS) synthesis. Of the 2 components, VicK possesses both autokinase and phosphatase activities, which regulate the phosphorylation and dephosphorylation of the regulator VicR in response to environmental cues. However, the dual mechanism of VicK as the autokinase/phosphatase in regulating S. mutans ' responses is not well elucidated. Previously, it has been shown that the phosphatase activity depends on the PAS domain and residues in the DHp domain of VicK in S. mutans. Specifically, mutating proline at 222 in the PAS domain inhibits VicK phosphatase activity. We generated a VicKP222A mutant to determine the level of VicR-P in the cytoplasm by Phos-tag sodium dodecyl sulfate polyacrylamide gel electrophoresis. We show that in VicKP222A phosphatase, attenuation increased phosphorylated VicR (VicR-P) that downregulated glucosyltransferases, gtfBC, thereby reducing the synthesis of water-insoluble polysaccharides (WIS-EPS) in the biofilm. In addition, VicKP222A presented as long-rod cells, reduced growth, and displayed asymmetrical division. A major adhesin of S. mutans, SpaP was downregulated in VicKP222A, making it unable to agglutinate in saliva. In summary, we have confirmed that VicK phosphatase activity is critical to maintain optimal phosphorylation status of VicR in S. mutans, which is important for cell growth, cell division, EPS synthesis, and bacterial agglutination in saliva. Hence, VicK phosphatase activity may represent a promising target to modulate S. mutans ' pathogenicity. [ABSTRACT FROM AUTHOR]

Introduction: Cryoproteins, such as cryoglobulins, cryofibrinogens and cold agglutinins, precipitate at low temperatures or agglutinate erythrocytes and dissolve again when warmed. Their pathogenetic and diagnostic importance in cold urticaria (ColdU) is unclear. In this study, we aimed to characterize the prevalence of cryoproteins in patients with ColdU. Methods: We conducted 3 analyses: i) a systematic review and meta-analysis of published data using an adapted version of the Joanna Briggs Institute's critical appraisal tool for case series, ii) a retrospective analysis of 293 ColdU patients treated at our Urticaria Center of Reference and Excellence (UCARE) from 2014 to 2019, and iii) a prospective observational study, from July 2019 to July 2020, with 49 ColdU patients as defined by the EAACI/GA2LEN/EDF/UNEV consensus recommendations. Results: Our systematic review identified 14 relevant studies with a total of 1151 ColdU patients. The meta-analyses showed that 3.0% (19/628), 1.1% (4/357) and 0.7% (2/283) of patients had elevated levels of cryoglobulins, cryofibrinogens and cold agglutinins, respectively. Our retrospective analyses showed that cryoproteins were assessed in 4.1% (12/293) of ColdU patients. None of 9 ColdU patients had cryoglobulins, and one of 5 had cold agglutinins. In our prospective study, none of our patients had detectable cryoglobulins (0/48) or cryofibrinogens (0/48), but 4.3% (2/46) of patients had cold agglutinins (without any known underlying autoimmune or hematological disorder). Conclusion: Our investigation suggests that only very few ColdU patients exhibit cryoproteins and that the pathogenesis of ColdU is driven by other mechanisms, which remain to be identified and characterized in detail. [ABSTRACT FROM AUTHOR]

Mast cell-activating signals in cold urticaria are not yet well defined and are likely to be heterogeneous. Cold agglutinins and cryoglobulins have been described as factors possibly associated with cold urticaria, but their relevance has not been explained. We performed a single-center prospective cohort study of 35 cold urticaria patients. Cold agglutinin and cryoglobulin test results, demographics, detailed history data, cold stimulation test results, complete blood count values, C-reactive protein, total immunoglobulin E levels, and basal serum tryptase levels were analyzed. Forty six percent (n = 16) of 35 tested patients had a positive cold agglutinin test and 27% (n = 9) of 33 tested patients had a positive cryoglobulin test. Cold agglutinin positive patients, when compared to cold agglutinin negative ones, were mainly female (P = 0.030). No gender-association was found for cryoglobulins. A positive cold agglutinin test, but not a positive cryoglobulin test, was associated with a higher rate of reactions triggered by cold ambient air (P = 0.009) or immersion in cold water (P = 0.041), and aggravated by increased summer humidity (P = 0.007). Additionally, patients with a positive cold agglutinin test had a higher frequency of angioedema triggered by ingestion of cold foods or drinks (P = 0.043), and lower disease control based on Urticaria Control Test (P = 0.023). Cold agglutinin levels correlated with erythrocyte counts (r = −0.372, P = 0.028) and monocyte counts (r = −0.425, P = 0.011). Cryoglobulin concentrations correlated with basal serum tryptase levels (r = 0.733, P = 0.025) and cold urticaria duration (r = 0.683, P = 0.042). Results of our study suggest that cold agglutinins and cryoglobulins, in a subpopulation of cold urticaria patients, are linked to the course and possibly the pathogenesis of their disease. [ABSTRACT FROM AUTHOR]

Sexual agglutinins of the budding yeast Saccharomyces cerevisiae are proteins mediating cell aggregation during mating. Complementary agglutinins expressed by cells of opposite mating types "a" and "α" bind together to promote agglutination and facilitate fusion of haploid cells. By means of an innovative single-cell manipulation assay combining fluidic force microscopy with force spectroscopy, we unravel the strength of single specific bonds between a- and α-agglutinins (~100 pN) which require pheromone induction. Prolonged cell–cell contact strongly increases adhesion between mating cells, likely resulting from an increased expression of agglutinins. In addition, we highlight the critical role of disulfide bonds of the a-agglutinin and of histidine residue H273 of α-agglutinin. Most interestingly, we find that mechanical tension enhances the interaction strength, pointing to a model where physical stress induces conformational changes in the agglutinins, from a weak-binding folded state, to a strong-binding extended state. Our single-cell technology shows promises for understanding and controlling the complex mechanism of yeast sexuality. Mathelié-Guinlet et al. investigate the molecular binding mechanisms of sexual agglutinins in budding yeast Saccharomyces cerevisiae. They report that mechanical tension enhances the strength of agglutinin interactions, supporting a new model in which physical stress induces conformational changes in the binding sites of agglutinins. [ABSTRACT FROM AUTHOR]

Cold agglutinin are erythrocyte antibodies which possess the property of agglutinating red blood cells at temperatures of below 37°C, this phenomenon is reversible after heating. This is usually immunoglobulin M (IgM) class. Their pathogenicity is much more related to their temperature range of activity than their title. As we report in this observation, cold hemagglutination makes it difficult to interpret certain immunological tests such as ABO Rh blood grouping or searching for irregular antibodies (SAI). The discovery of cold agglutinins can be fortuitous revealing itself by disturbances and aberrations in the results of blood count or as part of a suggestive clinical or laboratory table cold hemagglutinin disease. The search for a lymphoid hematological at their diagnosis should be systematic. [ABSTRACT FROM AUTHOR]

Soybean agglutinin (SBA) was purified using ammonium sulfate precipitation and liquid chromatography. Purified SBA was used to produce monoclonal antibodies through hybridoma technology. SBA secondary structure was studied using circular dichroism. pH-stressed (pHs 3.0, 7.2, 8.5, and 9.6) SBA physical properties (particle size, ζ-potential, and aggregation temperature) were investigated. Gel electrophoresis (non-native and native) was used to study heat-induced structural configuration changes in SBA. The effect of pH and temperature on the immunoreactivity of SBA was analyzed using enzyme-linked immunosorbent assay and immunoblots probed with two anti-SBA monoclonal antibodies with either linear or conformational epitopes. The hemagglutinating activity of heated SBA was measured by hemagglutination assay. Our results indicated that SBA had the least thermostability at pH 3.0 and the highest at pH 8.5. Temperature-induced structural configuration change on pH-stressed SBA led to immunoreactivity change. Heat-induced (70 and 80 °C) soluble SBA aggregation was proportionally related to hemagglutinating activity reduction., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The present review summarizes the current knowledge concerning provenance, properties, physiological and therapeutic actions of elderberry and the bioactive molecules present in the plant, with emphasis on their action on female reproduction. Elderberry or black elder (Sambucus nigra L.) attracts attention due to its easy cultivation and high availability of bioactive compounds. Most of the available data concerning black elder’s therapeutic action are focused on its effects such as activation of immune processes and anti-inflammatory processes (cytokine production, etc.) and regulation of hormones and their receptors in cancer cells. The effects of elderberry on reproduction have been poorly investigated so far. Nevertheless, conducted studies so far demonstrate the stimulatory influence of black elder extract and its constituents, such as rutin, anthocyanins and agglutinins, on the viability and steroidogenesis of healthy ovarian cells as well as their ability to promote apoptosis and reduce the viability and proliferation of ovarian cancer cells. Furthermore, the action of black elder extract and its constituent biomolecules, such as anthocyanins and lectins, on embryogenesis and the embryonal estradiol-estradiol receptor system have also been reported. The available information, despite limitations, suggest the applicability of black elder constituents for improvement of reproductive processes in animal biotechnology, animal production and assisted reproduction, as well as for prevention and treatment of reproductive disorders (including cancer) in veterinary and human medicine.

Leptospiral agglutinins have been measured in several species of chelonians in studies dating back to the1960s. However, despite the fact that sea turtles are one of the most charismatic and endangered groups of chelonians, only a single study has measured leptospiral antibodies in this group. Serum samples collected from hawksbill sea turtles (Eretmochelys imbricata) (n = 10) and green sea turtles (Chelonia mydas) (n = 10) from two locations from the Gulf of Mexico were analyzed to detect Leptospira interrogans using the microagglutination test. Hawksbill and green sea turtles tested positive to five and seven serovars, respectively, with the serovars Grippotyphosa, Bratislava, and Canicola presenting a prevalence ≥50% in both species. Titers ranged from 1:50 to 1:400. These and previous results demonstrate that terrestrial, freshwater, and sea turtles are exposed to Leptospira at some stage of their life. The role of reptiles in the epidemiology, persistence, and transmission of leptospirosis is still controversial; however, there is evidence that some species of turtles can disseminate them in the environment. The detection of Leptospira is of sanitary importance as it is a cosmopolitan zoonotic disease. Preventive measures should be taken by people working in sea turtle camps and aquariums to control and reduce their risk of exposure. [ABSTRACT FROM AUTHOR]

Fluoroalcohols like 1,1,1,3,3,3-hexafluoroisopropanol (HFIP) are widely used as cosolvent along with the biological solvent water to perturb the native protein molecules. The non native states obtained are of immense importance in the field of protein structure and folding, since these states may be present in the protein folding pathway or in the off pathway which leads to amyloid formation. In this study, HFIP-induced structure perturbation at secondary level of the tetrameric legume lectin, soybean agglutinin (SBA) is examined by far-UV circular dichroism (CD) spectroscopy. Like other member of the legume lectin family, native SBA is also an all β-sheet protein. Analysis of the far-UV CD spectra shows formation of α-helix rich conformations at the expense of native β-sheet in presence of higher concentration (50% or more) of HFIP. Visible aggregation is noticed at lower HFIP concentration (~10%) which disappears at higher concentration of HFIP with concomitant induction of the α-helical secondary structure. The results confirm about the helix propensity of amino acid sequence of SBA and helical intermediates may be involved in the early stage of its folding process. [ABSTRACT FROM AUTHOR]

Cold agglutinin disease (CAD) is a rare form of autoimmune haemolytic anaemia, and because of its rareness, there is no standard treatment for CAD patients. We retrospectively analysed the response to rituximab-containing therapy in CAD patients at our hospital. All patients received rituximab-containing therapy for at least 1 month. A total of 16 patients (11 males and 5 females) were included. The median age at the onset of the disease was 63.5 years (range 41–79). Most patients had manifestations including anaemia (81.3%) or cold-induced circulatory symptoms (75.0%). The median haemoglobin level was 72 g/L (range 29–101), and the median cold agglutinin titre was 1,024 (range 64–2,048). Thirteen of 16 patients (81%) responded to the therapy. Responders achieved a median increase in haemoglobin levels of 45 g/L. Grade 3–4 neutropenia occurred in 3 patients (19%), but only 1 (6%) of them experienced infection. Anaphylaxis related to rituximab occurred in 1 patient. During follow-up, five patients experienced relapse, and two patients died. The estimated median progression-free survival was 36 months, and median overall survival was not yet reached. In conclusion, A rituximab-based therapy in accordance with individual patient characteristics may be a reasonable choice for CAD patients. [ABSTRACT FROM AUTHOR]

In this paper we will present a novel inflection learning method using lattice based algorithms. The inflection grammar system is represented with a lattice of elementary inflection rules, enabling the description of prefix, infix and suffix transformations as well. The proposed grammar system is ideal for highly agglutinative languages like Hungarian. We created a learning algorithm to generate the rules in an automated way from training word pair sets. Our evaluation shows that this novel method can learn both suffix and infix transformations unlike our baseline algorithm called TASR, and has a better correctness ratio than finite state transducers, too. [ABSTRACT FROM AUTHOR]

Biofilm formation and hyphal growth are considered to be the most serious virulence factors of Candida species in blood causing candidemia infections, which are difficult to treat due to the spread of resistant Candida isolates to most antifungal drugs. Therefore, in this study, we investigated the effect of different types and concentrations of selected macroalgal extracts from Cladostephus spongiosus (Phaeophyta), Laurencia papillosa (Rhodophyta), and Codium arabicum (Chlorophyta) in inhibiting those virulence factors of the isolated Candida. Acetone extract of C. spongiosus (AECS) showed a stronger anticandidal activity against the selected strains than ethanol extract. Candida krusei was the highest biofilm producer among the selected isolates. AECS showed an inhibition of C. krusei biofilm formation as well as a reduction in the viability of preformed biofilms. Also, AECS reduced various sugars in the candidal exo-polysaccaride layer (EPS). Scanning electron microscopy (SEM) and light microscopic images revealed an absence of hyphae and an alteration in the morphology of biofilm cells when treated with AECS. Moreover, AECS downregulated the expression of hyphal specific genes, hyphal wall protein 1 (HWP1), Agglutinin-like protein 1 (ALS1) and fourth secreted aspartyl proteinase (SAP4), which confirmed the inhibitory effect of AECS on hyphal growth and biofilm formation. Gas chromatography-mass spectrophotometer (GC-MS) analysis of AECS showed three major compounds, which were non-existent in the ethanol extract, and might be responsible for the anticandidal activity; these revealed compounds were 4-hydroxy-4-methyl-2-pentanone, n-hexadecenoic acid, and phenol, 2-methoxy-4-(2-propenyl). These active compounds of AECS may be promising for future pharmaceutical applications in the treatment of candidemia. [ABSTRACT FROM AUTHOR]

The purpose of this study is to evaluate the prognostic value of preoperative Wisteria floribunda agglutinin-positive Mac-2 binding protein (WFA+-M2BP) in predicting overall survival for patients with hepatitis B- and hepatitis C-related early-stage hepatocellular carcinoma (ESHCC) after liver resection. Post-operative survival rates were compared according to WFA+-M2BP level and tumor stage. Six hundred and ten patients were identified and 198 were removed after application of the exclusion criteria; the median follow-up time was 4.33 years, and cancer-related death occurred in 117 (28.4%) patients. Age (p = 0.03), fibrosis grade (p = 0.042), cancer stage (p = 0.01), and WFA+-M2BP level (p = 0.001) were identified as independent risk factors for poor overall survival. The overall survival rates at 3 and 5 years for patients with WFA+-M2BP ≤ 1.12 were 0.92 and 0.90, respectively, and 0.76 and 0.61 for patients with WFA+-M2BP > 1.12 (p < 0.001). During the analysis of survival prediction, serum WFA+-M2BP level exhibited a higher log-likelihood and a lower AIC value compared to TNM stage (log likelihood: −638; AIC: 1279). Pre-operative serum WFA+-M2BP level provided important prognostic information after curative hepatic resection in our study. [ABSTRACT FROM AUTHOR]

When consumed as whole grain, wheat has a high nutrient density that contributes to a healthy diet. Yet, products labeled as whole wheat can still contain a substantial amount of refined grain leading to the confusion for consumers, so a method was designed to determine the whole grain status within wheat‐based foods. Wheat germ agglutinin (WGA), a lectin found in the germ tissue of wheat kernels, was evaluated as a biomarker of whole grain wheat. WGA content strongly correlated with the percentage of whole wheat within premade mixtures of whole and refined (white) flours. Then, commercial flours labeled as whole wheat were tested for WGA content and found to contain up to 40% less WGA compared to a whole grain standard. Commercial pasta products labeled as whole wheat were also tested for WGA content and found to contain up to 90% less WGA compared to a whole grain standard. The differences in WGA content were not likely due to varietal differences alone, as the WGA content in common varieties used in domestic wheat flour production varied less than 25%. The levels of other constituents in wheat kernels, including starch, mineral, phytate, and total protein, were not different among the commercial whole wheat flours and pasta products. WGA is a unique biomarker that can identify wheat products with the highest whole grain content. Whole grain wheat has a high nutrient density that can be part of a healthy diet, but products labeled as whole wheat can still contain some refined grain. Wheat germ agglutinin (WGA) was tested as a biomarker to measure whole grain status in wheat‐based foods and revealed that some commercial whole wheat flour and pasta contained unexpectedly lower levels of the WGA biomarker compared to a whole grain standard. WGA may therefore be a useful way to test for whole grain wheat content. [ABSTRACT FROM AUTHOR]

A functional simulation approach has been tempted between clinically proven lapin and human septic arthritis, in which twenty experimentally induced lapin S aureus septic arthritis and 20 natural human S.aureus septic arthritis were the human simulation materials. Lapin synovial fluids were aspirated by the veterinarian of the team and human synovial fluids were aspirated by the rheumatologist of the team. Direct synovial fluid Giemsa stained smears were showing neutrophilic infiltration responses both for lapin and human. Cell free synovial supernatants for lapin and human were found positive for S.aureus agglutinins up to the titre of 32. Sandwich eliza for TNF alpha, INF gamma and IL4 for lapin synovial fluid and same for human synovial fluid TNF alpha, INF gamma and IL4. Lapin knee joint synovial mucosal compartment was found valid model for human synovial mucosal compartment during the course of S, aureus septic arthritis. Likewise, the lapins knee joints synovial mucosal cytokine responses simulate that of man. Herd immunity concept was found operable in both lapin and human populations. Herd tripartite nature of the immune responses as low moderate and high responders both for rabbit and man. The herd immune plots were of skewd type for TNF alpha, normal distribution curves to INF gamma and IL4 both for lapin and human. This functional simulation study appear to be novel contribution in knee joint synovial mucosal cytokine responses. [ABSTRACT FROM AUTHOR]

Neuronal circuits of the spinal dorsal horn integrate sensory information from the periphery with inhibitory and facilitating input from higher central nervous system areas. Most previous work focused on projections descending from the hindbrain. Less is known about inputs descending from the cerebral cortex. Here, we identified cholecystokinin (CCK) positive layer 5 pyramidal neurons of the primary somatosensory cortex (CCK+ S1-corticospinal tract [CST] neurons) as a major source of input to the spinal dorsal horn. We combined intersectional genetics and virus-mediated gene transfer to characterize CCK+ S1-CST neurons and to define their presynaptic input and postsynaptic target neurons. We found that S1-CST neurons constitute a heterogeneous population that can be subdivided into distinct molecular subgroups. Rabies-based retrograde tracing revealed monosynaptic input from layer 2/3 pyramidal neurons, from parvalbumin positive cortical interneurons, and from thalamic relay neurons in the ventral posterolateral nucleus. Wheat germ agglutinin-based anterograde tracing identified postsynaptic target neurons in dorsal horn laminae III and IV. About 60% of these neurons were inhibitory and about 60% of all spinal target neurons expressed the transcription factor c-Maf. The heterogeneous nature of both S1-CST neurons and their spinal targets suggest complex roles in the fine-tuning of sensory processing. [ABSTRACT FROM AUTHOR]

Cold agglutinins (CA) are auto-antibodies that adhere to erythrocytes in cold temperatures, and can result in agglutination of red blood cells. This process can cause complementmediated intravascular hemolysis, which can be catastrophic. We describe a patient who developed CA during initiation of deep hypothermic circulatory arrest for emergent repair of Type A aortic dissection. The patient was found to have anti-I and anti-C antibodies and a positive direct Coombs test. CA resolved with re-warming, and resulted in no adverse events. [ABSTRACT FROM AUTHOR]

Carrageenan is a substance widely used as an additive in the food industry. Among other things, it is often added to processed cheese, where it has a positive effect on texture. Processing of such cheese involves grinding, melting and emulsifying the cheese. There is currently no official method by which carrageenan can be detected in foodstuffs, but there are several studies describing its negative health impact on consumers. Lectin histochemistry is a method that is used mainly in medical fields, but it has great potential to be used in food analysis as well. It has been demonstrated that lectin histochemistry can be used to detect carrageenan in processed cheese by Human Inspection and Computer-Assisted Analysis (CIE L*a*b*). The limit of detection (LoD) was established at 100 mg kg−1 for Human Inspection and 43.64 for CIE L*a*b*. The CIE L*a*b* results indicate that Computer-Assisted Analysis may be an appropriate alternative to Human Inspection. The most suitable parameter for Computer-Assisted Analysis was the b* parameter in the CIE L*a*b* color space.

Objectives: Crohn's disease patients, who are prone to develop periodontal diseases, may carry genetic defects in their Th17 cytokine, human beta-defensin (hBD) 1-3, and salivary and scavenger agglutinin (SALSA) expressions. Biochemical composition of saliva reflects the oral consequences of systemic immune response modifications. Our aim was to evaluate the salivary Th17 cytokine, epithelial hBD 1-3, and SALSA levels in relation to Crohn's disease., Materials and Methods: This cross-sectional study included 42 Crohn's disease patients and 34 systemically healthy controls. Periodontal and dental indexes were measured, and stimulated saliva samples were collected. Salivary Th17 cytokine levels were analyzed by multiplex technique, and hBD 1-3 and SALSA levels by enzyme-linked immunosorbent assay., Results: There were 19 gingivitis and 11 initial periodontitis patients in the Crohn's disease group, and 15 gingivitis and 4 initial periodontitis in the control group. In comparison to controls, higher salivary Th17 cytokine levels were observed in Crohn's disease patients. No statistical difference was observed between Crohn's disease and control groups in terms of their salivary hBD 1-3 and SALSA levels. Based on the regression analysis, there is no independent association between Crohn's disease and salivary Th17 cytokine levels., Conclusions: Crohn's disease does not relate to salivary antimicrobial hBD 1-3 or SALSA levels. While Crohn's disease patients have higher salivary Th17 cytokine levels in comparison to systemically healthy controls, an independent association between Crohn's disease and Th17 cytokine profile is still missing., Clinical Relevance: Diminished Th17 cytokine response in Crohn's disease, which might be related to genetic susceptibility, can be also visualized in saliva., (© 2024. The Author(s).)

Carrageenan is a polysaccharide that is widely used in the food industry. Due to its water holding capacity, there is a higher risk of adulteration for economic reasons related to it. A verifiable method for detecting carrageenan is still missing in the food inspection sector. The detection of carrageenan in meat products is not well described. Our study describes lectin histochemistry as a novel approach for carrageenan detection. Within this study, the detection of carrageenan in meat products by lectin histochemistry is validated. Lectins of Arachis hypogaea (PNA) and Bandeiraea simlicifolia (BSA), specific for galactose units of carrageenan, were used. The samples included model meat products (ground chicken-meat products) and meat products from retail markets (chicken and pork hams, sausages, salami, and dried sausages). The limit of determination (LoD) of this method was set at 0.01 g kg−1. The method sensitivity for lectin PNA reached 1, and, for lectin BSA, it reached 0.96. Method specificity for lectin PNA was 1, and, for lectin BSA, it was 1.33. Cross-reactivity with other hydrocolloids tested was not confirmed. The results confirm that lectin histochemistry is suitable for detecting carrageenan in meat products.

Streptococcus mutans SpaP mediates the binding of this cariogenic bacteria to tooth surfaces. It was reported that the SpaP of S. mutans clinical isolates could be classified to 2 genotypes, type A and B. Our aims are to examine spaP genotypes in often-used S. mutans laboratory strains as well as clinical isolates and to explore the relationship between the genotypes of S. mutans strains and their adherence to salivary-agglutinin (SAG). The sequences of SpaP of 11 S. mutans strains were analyzed with alignment tools. Out of these strains, 9 strains were examined for their adherence to SAG-coated surfaces. The SpaP expression on the cell surfaces and in the spent media of 9 strains were examined by a dot-blot assay. Based on the alignment of the variable V region of SpaP, 9 strains were classified as previously-defined type-A and 3 strains type-B. Among type-B strains, the SpaPs of GS5 and HG723 contain a premature stop codon which resulted in loss of adherence and absence of SpaP expression on the cell surfaces. However, clear SpaP expression was observed in the spent media of both strains. The type-B strain UA159 demonstrated low SpaP expression on the cell surface, but it showed similar adherence ability as the type-A strains. In conclusion, the presence of SpaP on the cell surface determines the adherence of S. mutans to SAG. No difference in SAG-mediated adherence could be seen between type A and B strains, probably due to the limited number of type B strain tested. [ABSTRACT FROM AUTHOR]

Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening disease eventually caused or reactivated by a viral infection, which can also lead to the production of cold agglutinins (CA). The nature of these autoantibodies is usually an IgM, less frequently an IgA or IgG, they agglutinate red blood cells at low temperatures. They can interfere with hematological parameters causing interpretation difficulties. We report a case of a 4-year-old boy who developed an IgG CA during recurrent HLH reactivated by EBV infection. The purpose of this observation is to underline HLH criteria and to analyze CA interference as well as its biological and clinical characteristics. [ABSTRACT FROM AUTHOR]

This study aimed to isolate and identify Candida spp. that cause mycotic mastitis in cows by using molecular techniques for genotype and virulence genes detection. Two hundred and fifty mastitis milk samples from cows in province of Basrah were collected and examined using rapid diagnostic tests and confirmed by polymerase chain reaction (PCR) assay, Candida species was identified in 116/250 (46.4%) of the mastitis milk samples. Based on conventional method and ID - Yst card system Vitek 2, Candida albicans was predominant 60/116 (51.7%), followed Candida parapasilosis 15/116 (12.9%). Concerning the results PCR amplification of 18S rRNA gene for identification of C. albicans and C. parapasilosis, this gene were present 60 samples in C. albicans, while C. parapasilosis was appeared in 15 samples. Secreted aspartyl proteinase 1 like (SAP1) gene was detected in 57/60 (95%) and agglutinin-like sequence (ALS1) gene was detected in 55/60 (91.6%) in C. albicans. While secreted aspartyl proteinase 1 like (SAP1) and agglutininlike sequence (ALS1) genes were detected in 15/15 (100%) and 14/15 (93.3%) in C. parapasilosis isolates, respectively. [ABSTRACT FROM AUTHOR]