N-glycomic analysis of Z(IgA1) partitioned serum and salivary immunoglobulin A by capillary electrophoresis..

Author(s):  
Brigitta Meszaros ◽  
Zsuzsanna Kovacs ◽  
Eniko Gebri ◽  
Hajnalka Jankovics ◽  
Ferenc Vonderviszt ◽  
...  

Aims: To apply capillary electrophoresis with laser induced fluorescence detection (CE-LIF) to identify the N-glycosylation structures of serum and saliva IgA from healthy controls and patients with malignant hematological diseases having cytostatic treatment induced mild oral mucosal lesions. Background: Altered N-glycosylation of body fluid glycoproteins can be effective indicators of most inflammatory processes. Immunoglobulin A (IgA) is the second highest abundant of the immunoglobulins and has a major role in the immune-defense against potential pathogen attacks. While IgA is abundant in serum, secretory immunoglobulin A (sIgA) is one of the most prevalent proteins in mucosal surfaces such as in saliva. Objective: Our aim was to investigate the changes of IgA glycosylation in serum and saliva as a response to an administered cytostatic treatment in patients with malignant hematological disorders. Materials: Capillary electrophoresis with laser induced fluorescent detection (CE-LIF) was used to analyze the N-glycosylation profiles of Z(IgA1) partitioned immunoglobulin A in pooled serum and saliva of 10 control subjects and 8 patients with malignant hematological diseases having cytostatic treatment induced mild oral mucosal lesions. Results: Eight of 31 and four of 38 N-glycans in serum and saliva, respectively, showed significant (p<0.05) differences upon comparison to the control group. Thirteen glycans were present in the saliva but not in the serum, on the other hand six structures were found in the serum samples not present in the saliva. Conclusion: The developed Z(IgA1) partitioning and the high resolution CE-LIF based glyocoanalytical methods provided an efficient and sensitive workflow to detect and monitor IgA glycosylation alterations in serum and saliva with the scope for widespread molecular medicinal use.

2010 ◽  
Vol 67 (9) ◽  
pp. 766-770 ◽  
Author(s):  
Milos Cankovic ◽  
Marija Bokor-Bratic

Bacground/Aim. Systemic candidiasis in intensive care units remains an improtant problem due to antifungal resistance. Patients undergoing radiotherapy for head and neck cancer are at increased risk of developing oral candidiasis and they more frequent have prior fungi colonization. Due to identification of specific risk factors predisposing to fungal infection in order to threat such patients the aim of this study was to determine the presence of Candida species in patients with oral squamous cell carcinoma and compare it to the control subjects (patients with benign oral mucosal lesions). Methods. A total number of 30 consecutive oral cancer examined patients were included in this prospective study (24 men and 6 women with a mean age of 61.47 years, range 41-81 years). The control group consisted of 30 consecutive patients with histologically proven benign oral mucosal lesions (16 men and 14 women with a mean age of 54.53 years, range 16- 83 years). The samples for mycological examination were obtained by using sterile cotton swabs from the cancer lesion surface and in the patients of the control group from the benign mucosal lesion surface. Samples were inoculated in Sabouraud' dextrose agar. For identification purposes, Mackenzie germ tube test was performend on all isolates. Results. The prevalence of Candida was significantly higher in oral cancer patients than in control subjects (?2 = 5.455, p = 0.020). Candida was found on nine of the 30 cancer surfaces; 5 (16.7%) were identified as non-albicans Candida and 4 (13.3%) as Candida albicans. In the control group, only Candida albicans was isolated from 2 (6.7%) patients. In this study, no statistically significant differences in the presence of Candida species was found with respect to gender, age, smoking, alcohol consumption, wearing of dental protheses and the site of cancer lesion. Conclusion. The increased prevalence of yeasts on the surfaces of oral carcinoma indicates a need for their suppression before any cancer treatment. There was no evidence for an association between gender, age, smoking, alcohol consumption, wearing of dental protheses, the site of cancer lesion and the yeast presence.


2012 ◽  
Vol 1 (2) ◽  
pp. 28
Author(s):  
H. Kaur ◽  
Swati . ◽  
N. Puri ◽  
A. Vashist ◽  
H. P. Singh ◽  
...  

As oral cavity is an integral part of general health, persistent and unrelenting stress often becomes a danger affecting the oral health and overall health causing various psychosomatic disorders. Present study aimed to determine the prevalence of oral mucosal lesions and bruxism in psychiatric patients dependent on psychotropic drugs. Dental examination was done and questionnaire administered to 150 psychiatric patients and 150 control participants. In dental examination recurrent apthous stomatitis, burning mouth syndrome, oral lichen planus, bruxism, temporomandibular disorders were evaluated. Results showed that psychiatric patients had higher prevalence of bruxism, oral mucosal lesions in general, than the control group.


2017 ◽  
Vol 100 (2) ◽  
pp. 485-491 ◽  
Author(s):  
Laura K Allred ◽  
Katherine Lesko ◽  
Diane McKiernan ◽  
Cynthia Kupper ◽  
Stefano Guandalini

Abstract Enzymatic digestion, or hydrolysis, has been proposed for treating gluten-containing foods and beverages tomake them safe for persons with celiac disease (CD). There are no validated testing methods that allow the quantitation of all the hydrolyzed or fermented gluten peptides in foods and beverages that might be harmful to CD patients, making it difficult to assess the safety of hydrolyzed products. This study examines an ELISA-based method to determine whether serum antibody binding of residual peptides in a fermented barley-based productis greater among active-CD patients than a normal control group, using commercial beers as a test case. Sera from 31 active-CD patients and 29 nonceliac control subjects were used to assess the binding of proteins from barley, rice, traditional beer, gluten-free beer, and enzymatically treated (gluten-removed) traditional beer. In the ELISA, none of the subjects' sera bound to proteins in the gluten-free beer. Eleven active-CD patient serum samples demonstrated immunoglobulin A (IgA) or immunoglobulin G (IgG) binding to a barley extract, compared to only one nonceliac control subject. Of the seven active-CD patients who had an IgA binding response to barley, four also responded to traditional beer, and two of these responded to the gluten-removed beer. None ofthe nonceliac control subjects' sera bound to all three beer samples. Binding of protein fragments in hydrolyzed or fermented foods and beverages by serum from active-CD patients, but not nonceliac control subjects, may indicate the presence of residual peptides that are celiac-specific.


2000 ◽  
Vol 7 (2) ◽  
pp. 192-196 ◽  
Author(s):  
Harry E. Prince ◽  
Gary L. Norman ◽  
Walter L. Binder

ABSTRACT Immunoglobulin A (IgA) deficiency occurs more frequently in patients with celiac disease (CD) than in the general population and can lead to false-negative results in the best serologic test for CD, endomysial IgA (EMA). To evaluate the impact of IgA deficiency on serologic detection of CD in a reference laboratory setting, IgA levels were measured in 510 consecutive serum specimens submitted for testing for EMA; 510 consecutive serum specimens submitted forHelicobacter pylori IgG testing served as a gastrointestinal symptom control group. The frequency of IgA deficiency was significantly higher among the specimens submitted for testing for EMA (5.1%) than among the specimens from the symptom control group (1.4%). Three subsets of sera from the group of specimens submitted for testing for EMA were then tested by additional serologic assays for CD; these subsets were EMA-positive sera (n = 25), EMA-negative, IgA-deficient sera (n = 26), and control sera (from EMA-negative, IgA-nondeficient patients age matched to IgA-deficient patients; n = 26). The proportions of EMA-positive sera positive by other assays for CD were 92% for transglutaminase IgA (TG-IgA), 80% for gliadin IgA, 84% for gliadin IgG, 60% for endomysial IgG (EMG), and 32% for transglutaminase IgG (TG-IgG). Very low proportions (0 to 8%) of IgA-deficient sera and control sera were positive for TG-IgA, gliadin IgA, EMG, and TG-IgG. Eight of 26 (31%) IgA-deficient serum samples were positive for gliadin IgG, whereas 3 of 26 (12%) control serum samples were positive for gliadin IgG, but this difference was not statistically significant. Physicians supplied clinical data for 18 of 26 patients with IgA deficiency; only 4 patients had undergone small-bowel biopsy, and 0 of 4 patients showed villous atrophy. These findings show that IgA deficiency is found more frequently among sera submitted for testing for EMA in a reference laboratory setting, but there was no clear-cut serologic or clinical evidence of CD in EMA-negative, IgA-deficient patients.


2009 ◽  
Vol 13 (4) ◽  
pp. 119-126
Author(s):  
Yoshimitsu Bamba ◽  
Tetsunari Nishikawa ◽  
Akio Tanaka

2017 ◽  
pp. 85-88
Author(s):  
O.I. Ostapenko ◽  
◽  
V.P. Kvashenko ◽  
I.K. Akimova ◽  
I.N. Nosova ◽  
...  

The objective: the study of immunomodulatory effects of a probiotic, which contains lyophilized Lactobacillus (Lactobacillus rhamnosus) – 13 mg (2,0ґ109 CFU) and lyophilized bifidobacteria (Bifidobacterium lactis) – 4 mg (2,0ґ109 CFU) the level of serum immunoglobulin IgA as a marker of local immunity in the plasma of women of reproductive age with the violation of the biocenosis of the vagina. Patients and methods. The study involved 86 patients of reproductive age with the violation of the vaginal biocenosis, which were divided into two groups according to received treatment. A survey was conducted for all patients in both groups: determine the level of serum IgA, measuring pH of vaginal environment and the quantification of lactobacilli and pathogenic flora with the help of test-system «Florotsenoz» before treatment and in 6 weeks after treatment. The state of vaginal microbiocenosis in both groups before treatment was homogeneous. Patients in both groups as therapy at the first stage of treatment received, if necessary antimicrobial therapy depending on the selected flora. In the second stage (restoration of microflora) patient of the main group received systemic probiotic combined with a complex prebiotic local action, patients in the control group, the probiotic localy in the form of the vaginal candles or tablets. Results. The research stated the increasing level of serum IgA in blood plasma of patients of the main group compared to control group at 20%, normalizing the pH of the vaginal environment in the main group in 94% of cases, which indicates an increase of immunity in mucosal. Conclusion. The inclusion of the systemic probiotic in the scheme of treatment of disorders of biocenosis of the vagina system enhances the increasing of immunity of the mucous membranes, and the vaginal tablets prebiotic of local action restores the own normal microflora of the vagina. Key words: serum immunoglobulin A, local immunity, vaginal dysbiosis, probiotics, prebiotics, vaginal microbiocenosis, the pH of the vaginal environment.


2001 ◽  
Vol 47 (1) ◽  
pp. 110-117 ◽  
Author(s):  
Magnus Jonsson ◽  
Joyce Carlson ◽  
Jan-Olof Jeppsson ◽  
Per Simonsson

Abstract Background: Electrophoresis of serum samples allows detection of monoclonal gammopathies indicative of multiple myeloma, Waldenström macroglobulinemia, monoclonal gammopathy of undetermined significance, and amyloidosis. Present methods of high-resolution agarose gel electrophoresis (HRAGE) and immunofixation electrophoresis (IFE) are manual and labor-intensive. Capillary zone electrophoresis (CZE) allows rapid automated protein separation and produces digital absorbance data, appropriate as input for a computerized decision support system. Methods: Using the Beckman Paragon CZE 2000 instrument, we analyzed 711 routine clinical samples, including 95 monoclonal components (MCs) and 9 cases of Bence Jones myeloma, in both the CZE and HRAGE systems. Mathematical algorithms developed for the detection of monoclonal immunoglobulins (MCs) in the γ- and β-regions of the electropherogram were tested on the entire material. Additional algorithms evaluating oligoclonality and polyclonal concentrations of immunoglobulins were also tested. Results: CZE electropherograms corresponded well with HRAGE. Only one IgG MC of 1 g/L, visible on HRAGE, was not visible after CZE. Algorithms detected 94 of 95 MCs (98.9%) and 100% of those visible after CZE. Of 607 samples lacking an MC on HRAGE, only 3 were identified by the algorithms (specificity, 99%). Algorithms evaluating total gammaglobulinemia and oligoclonality also identified several cases of Bence Jones myeloma. Conclusions: The use of capillary electrophoresis provides a modern, rapid, and cost-effective method of analyzing serum proteins. The additional option of computerized decision support, which provides rapid and standardized interpretations, should increase the clinical availability and usefulness of protein analyses in the future.


2021 ◽  
Vol 10 (8) ◽  
pp. 1771
Author(s):  
Violetta Opoka-Winiarska ◽  
Ewelina Grywalska ◽  
Izabela Korona-Glowniak ◽  
Katarzyna Matuska ◽  
Anna Malm ◽  
...  

There is limited data on the effect of the novel coronavirus disease (COVID-19) caused by severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) on pediatric rheumatology. We examined the prevalence of antibodies against SARS-CoV-2 in children with juvenile idiopathic arthritis (JIA) and a negative history of COVID-19 and the correlation of the presence of these antibodies with disease activity measured by juvenile arthritis disease activity score (JADAS). In total, 62 patients diagnosed with JIA, under treatment with various antirheumatic drugs, and 32 healthy children (control group) were included. Serum samples were analyzed for inflammatory markers and antibodies and their state evaluated with the juvenile arthritis disease activity score (JADAS). JIA patients do not have a higher seroprevalence of anti-SARS-CoV-2 antibodies than healthy subjects. We found anti-SARS-CoV-2 antibodies in JIA patients who did not have a history of COVID-19. The study showed no unequivocal correlation between the presence of SARS-CoV-2 antibodies and JIA activity; therefore, this relationship requires further observation. We also identified a possible link between patients’ humoral immune response and disease-modifying antirheumatic treatment, which will be confirmed in follow-up studies.


Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1394
Author(s):  
Jianyong Zhou ◽  
Jingyi Yao ◽  
Luhong Bai ◽  
Chuansong Sun ◽  
Jianjun Lu

EGF has been shown to stimulate the growth of animals. In this study, the content of EGF in chicken embryos (gallus EGF, gEGF) aged from 1 to 20 days of incubation were determined by ELISA kit, and the 5-day-old chicken embryos with the highest content of 5593 pg/g were selected to make gEGF crude extracts. A total of 1500 1-day-old Xianju chickens were randomly divided into five groups with six replicates of 50 chickens each. The control group was fed a basal diet, and other treatment diets were supplemented with 4, 8, 16 and 32 ng/kg gEGF crude extract, respectively. The experiment lasted for 30 days. Chicks were harvested at the end of the experiment, and liver, spleen, thymus, bursa and serum samples were collected. Results showed that average daily gain (ADG) and average daily feed intake (ADFI) of 16 ng/kg group were higher than those in the control group (p < 0.05). The serum uric acid (UA) of the 16 ng/kg group was reduced (p < 0.01), and the serum alkaline phosphatase (AKP) of the 16 ng/kg group increased (p < 0.01). The gEGF extract also increased chick’s antioxidant capacity, decreased malondialdehyde (MDA) and increased catalase (CAT) in the liver and serum of 16 ng/kg groups in compared to the control group (p < 0.01). Furthermore, immunity was improved by the addition of gEGF to broiler diets. The serum immunoglobin A (IgA) content of 8 and 16 ng/kg groups and the serum immunoglobin M (IgM) content of 4 and 8 ng/kg groups were increased (p < 0.05) compared to the control group. The bursa index of each experimental group was higher than the control group (p < 0.01). These findings demonstrate that the crude extract of gEGF prepared in this experiment could improve the growth performance, antioxidant capacity and immunity of broilers.


2004 ◽  
Vol 50 (9) ◽  
pp. 1607-1617 ◽  
Author(s):  
Ville Väisänen ◽  
Susann Eriksson ◽  
Kaisa K Ivaska ◽  
Hans Lilja ◽  
Martti Nurmi ◽  
...  

Abstract Background: Free and total human kallikrein 2 (hK2) might improve the discrimination between prostate cancer and benign prostatic hyperplasia. Concentrations of hK2 are 100-fold lower than concentrations of prostate-specific antigen (PSA); therefore, an hK2 assay must have a low detection limit and good specificity. Methods: PSA- and hK2-specific monoclonal antibodies were used in solid-phase, two-site immunofluorometric assays to detect free and total hK2. The total hK2 assay used PSA-specific antibodies to block nonspecific signal. The capture antibody of the free hK2 assay did not cross-react with PSA. To determine the hK2 concentrations in the male bloodstream, total hK2 was measured in a control group consisting of 426 noncharacterized serum samples. Free and total hK2 were measured in plasma from 103 patients with confirmed prostate cancer. Results: All 426 males in the control group had a total hK2 concentration above the detection limit of 0.0008 μg/L. The median total hK2 concentration was 0.022 μg/L (range, 0.0015–0.37 μg/L). hK2 concentrations were 0.1–58% of total PSA (median, 3.6%). hK2 concentrations were similar in men 41–50 and 51–60 years of age. The ratio of hK2 to PSA steadily decreased from 5–30% at PSA &lt;1 μg/L to 1–2% at higher PSA concentrations. In 103 patients with prostate cancer, the median hK2 concentration in plasma was 0.079 μg/L (range, 0.0015–16.2 μg/L). The median free hK2 concentration was 0.070 (range, 0.005–12.2) μg/L. The proportion of free to total hK2 varied from 17% to 131% (mean, 85%). Conclusions: The wide variation in the free-to-total hK2 ratio suggests that hK2 in blood plasma is not consistently in the free, noncomplexed form in patients with prostate cancer. The new assay is sufficiently sensitive to be used to study the diagnostic accuracies of free and total hK2 for prostate cancer.


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