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Journal Article 2014; 35(Suppl 2): 99-104 PubMed PMID: 25638373 Citation  Keywords:  Animals, Antibodies:immunology, Bacterial Infections:prevention & control, Cell Line, Chickens, Egg Yolk:immunology, Epithelial Cells:immunology, Humans, Immunoglobulin Fab Fragments:immunology, Immunoglobulins:immunology, Inflammation:immunology, Lung:im.   

OBJECTIVES: As an alternative therapeutic approach for the prevention and treatment of bacterial infections with P. aeruginosa of cystic fibrosis (CF) patients, chicken yolk antibodies (IgY) could be used. The most significant advantage of IgY, in contrast to mammalian IgG, consists in the fact, that when bound to the antigen, they usually do not induce inflammatory reaction. In addition, the simplicity of egg production and the ease of IgY preparation makes this kind of antibody an excellent tool for passive immunization. Thus, the aim of our project was to study the effect of IgY and its Fab fragment on the potential induction of pro-inflammatory reactions in lung epithelial cells.

METHODS: Chicken IgY were prepared from pooled egg yolks. Fab fragmens of IgY were purified from the papain digest of IgY using DEAE-Sephacel ion exchange chromatography. Their purity was verified by SDS electrophoresis in polyacrylamide gel. Immortalized human cell lines, CuFi (CF patient) and NuLi (healthy subject), and A549 (human adenocarcinoma cells) were exposed to IgY, Fab, OVA, LPS (positive control), PBS (negative control), and human and goat IgG for 24 hours. The concentration of pro-inflammatory cytokines TNF-α, IL-1β, IL-6 and GM-CSF were determined in cell media using the BioPlex method, which enables the quantification of multiple analytes simultaneously in one sample.

RESULTS: Our results show that i) the Fab fragment induced levels of some proinflammatory cytokines, when compared to the PBS control, whereas ii) chicken IgYs did not induce any notable production of pro-inflammatory cytokines in contrast to intense effect of LPS on TNF-α and GM-CSF. In summary, our data show that levels of all cytokines are comparable with physiological values in human serum except of IL-1β, which concentration in cell medium was markedly elevated by Fab fragment.

CONCLUSIONS: The present data indicate that IgY are not inflammatory for lung cells and thus they are possibly applicable for prevention of airway bacterial infections.

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Controlled Clinical Trial 2014; 35(7): 619-623 PubMed PMID: 25617886 Citation  Keywords:  Cell Adhesion Molecules:genetics, CpG Islands:genetics, DNA Methylation:genetics, Female, Humans, Immunoglobulins:genetics, Myelin and Lymphocyte-Associated Proteolipid Proteins:genetics, Papillomavirus Infections, Promoter Regions, Genetic, Sequence Anal.   

OBJECTIVE: Cervical cancer is the second most common cancer disease affecting the female population. A key factor in development of the disease is the human papillomavirus infection (HPV). The disease is also impacted by epigenetic changes such as DNA methylation, which causes activation or exclusion of certain genes, and simultaneously the hypermethylation of cytosines in the promoters and turn-off of previously active genes occur. In this study, we focused on the introduction of pyrosequencing for the detection of DNA methylation of the selected CADM1 and MAL genes.

METHODS: DNA was isolated from cytological cervical smear of patients with different types of dysplasia [L-SIL (n=14), ASC-US (n=15), H-SIL (n=1)] and four control samples from healthy women. Prepared samples were further analyzed by bisulfite conversion and subsequent pyrosequencing (Pyromark Q96 ID, Qiagen, Germany). We examined the extent of methylation of CpG islands and as control samples of this method we used a fully methylated and unmethylated DNA. Methylation level (Met level) from each sample was quantified as the mean value [sum of all methylated CpG islands in %/total number of CpG islands (MAL n=4; CADM1 n=3)].

RESULTS: In total, 30 clinical samples and 4 control samples from healthy women were analyzed. By means of the analysis of the CADM1promoter region, the values of the Met level were obtained [fully methylated DNA (94.83 and 88); completely unmethylated DNA (0 and 0); and control samples from healthy patients (6.825 and 0.825), L-SIL (2.107 and 2.778), ASC-US (7.313 and 3.626), H-SIL (0 and 0)]. By means of the analysis of the MAL promoter region, the values of Met level were obtained [fully methylated DNA (53.25); completely unmethylated DNA (0.875); and control samples from healthy patients (2.925), L-SIL (1.517), ASC-US (2.833), and H-SIL (4)].

CONCLUSION: We introduced a pyrosequencing method for quantification of methylation of CADM1, MAL promoter regions, and detected methylations in clinical samples and also some basal methylation in healthy women....

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Case Reports 2007; 28(6): 739-744 PubMed PMID: 18063928 Citation  Keywords:  Adolescent, Antibodies:blood, Antioxidants:therapeutic use, Biomarkers, Diet Therapy, Enterobacteriaceae:immunology, Enterobacteriaceae Infections:complications, Fatigue Syndrome, Chronic:etiology, Female, Humans, Immunoglobulins, Intravenous:therapeutic .   

: There is now evidence that chronic fatigue syndrome (CFS) is accompanied by an increased translocation of endotoxins from gram-negative enterobacteria through the gut wall, as demonstrated by increased prevalences and median values for serum IgM and IgA against the endotoxins of gram-negative enterobacteria. This condition can also be described as increased gut permeability or leaky gut and indicates intestinal mucosal dysfunction (IMD). Here we report a case of a 13 year old girl with CFS who showed very high values for serum IgM against the LPS of some enterobacteria and signs of oxidative and nitrosative stress, activation of the inflammatory response system, and IgG3 subclass deficiency. Upon treatment with specific antioxidants and a "leaky gut diet", which both aim to treat increased gut permeability, and immunoglobins intravenously, the increased translocation of the LPS of gram negative enterobacteria normalized and this normalization was accompanied by a complete remission of the CFS symptoms....

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Case Reports 2007; 28(5): 545-546 PubMed PMID: 17984955 Citation  Keywords:  Adolescent, Angelman Syndrome:complications, Child, Preschool, Chromosomes, Human, Pair 15:genetics, Humans, Hypothyroidism:blood, Immunoglobulins, Thyroid-Stimulating:blood, Sequence Deletion, Thyroxine:therapeutic use,.   

: Angelman Syndrome (AS, MIM 105830), classified among neurogenetic disorders, occurs with estimated frequency of 1:10 000 to 1:40 000. The characteristics features apart from neurodevelopmental impairment and seizures include peculiar face traits, absent speech, outburst of laughter, ataxia, stereotyped jerky (puppet-like) movements. The authors report three children with Angelman syndrome who were also diagnosed with hypothyroidism....

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Journal Article 2006; 27(1-2): 271-276 PubMed PMID: 16648800 Citation  Keywords:  Adult, Anaerobic Threshold:physiology, Complement C3:metabolism, Complement C4:metabolism, Complement System Proteins:metabolism, Exercise:physiology, Humans, Immunoglobulins:blood, Leukocyte Count, Leukocytes:physiology, Male, Nephelometry and Turbidimet.   

BACKGROUND: The objective of this study was to investigate the effect of various sports disciplines on basic elements of the acquired and natural defense systems and to compare their results with that of non-sporting controls.

MATERIALS/METHODS: The study included 15 sedentary (Group 1), 15 volleyball players (Group 2) and 15 long distance running athletes (Group 3). The VO2 Max for the groups was determined by subjecting the individuals to a 20 meter Shuttle run test. Immunoglobin and complement levels were measured by the turbidometric method. Leukocyte counts were made through peripheral blood smear examination.

RESULTS: The IgA levels in sedentary living individuals was found to be lower than that in volleyball and athlete groups while that in athletes was in turn lower than that in volleyball players (p<0.05). In volleyball players the IgG level was significantly higher than both the sedentary living individuals and the athletes (p<0.05). IgM however, showed a significantly higher level in the sedentary group than in both athletes and volleyball players. The C3 levels in athletes was significantly lower than that of volleyball players and sedentary individuals (p<0.05), while the C4 levels were higher in the volleyball group (p<0.05). The leukocyte, eosinophil, and monocyte counts in the athletes were also significantly lower than those in the volleyball and sedentary groups (p<0.05). The values for the sedentary group on the other hand however, was found to be lower than those of the volleyball players (p<0.05). The neutrophil counts was lower in the sedentary group than in the volleyball group (p<0.05). The levels in the athlete group was lower than that in the volleyball group, however (p<0.05). The lymphocyte counts in volleyball players was higher than that in the sedentary group (p<0.05).

CONCLUSIONS: In conclusion, changes in the immune parameters were found to be dependent on the type of physical exercise performed on regular basis. Considering the protective antibody response in those performing regular sports, moderate exercise carried out on regular basis can be said to affect positively mononuclear and polymorphonuclear phagocytic cells--the basic elements of the natural immune system....

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