The methylation profile of IL4, IL5, IL10, IFNG and FOXP3 associated with environmental exposures differed between Polish infants with the food allergy and/or atopic dermatitis and without the disease.

Objectives: Epigenetic dynamics has been indicated to play a role in allergy development. The environmental stimuli have been shown to influence the methylation processes. This study investigated the differences in CpGs methylation rate of immune-attached genes between healthy and allergic infants. The research was aimed at finding evidence for the impact of environmental factors on methylation-based regulation of immunological processes in early childhood. Methods: The analysis of methylation level of CpGs in the IL4, IL5, IL10, IFNG and FOXP3 genes was performed using high resolution melt real time PCR technology. DNA was isolated from whole blood of Polish healthy and allergic infants, with food allergy and/or atopic dermatitis, aged under six months. Results: The significantly lower methylation level of FOXP3 among allergic infants compared to healthy ones was reported. Additional differences in methylation rates were found, when combining with environmental factors. In different studied groups, negative correlations between age and the IL10 and FOXP3 methylation were detected, and positive - in the case of IL4. Among infants with different allergy symptoms, the decrease in methylation level of IFNG, IL10, IL4 and FOXP3 associated with passive smoke exposure was observed. Complications during pregnancy were linked to different pattern of the IFNG, IL5, IL4 and IL10 methylation depending on allergy status. The IFNG and IL5 methylation rates were higher among exclusively breastfed infants with atopic dermatitis compared to the non-breastfed. A decrease in the IFNG methylation was noted among allergic patients fed exclusively with milk formula. In different study groups, a negative correlation between IFNG, IL5 methylation and maternal BMI or IL5 methylation and weight was noted. Some positive correlations between methylation rate of IL10 and child's weight were found. A higher methylation of IL4 was positively correlated with the number of family members with allergy. Conclusion: The FOXP3 methylation in allergic infants was lower than in the healthy ones. The methylation profile of IL4, IL5, IL10, IFNG and FOXP3 associated with environmental exposures differed between the studied groups. The results offer insights into epigenetic regulation of immunological response in early childhood.

The Lack of Influence of Homozygous Long Allele of the 5-HTTLPR Gene on the Severity of Alcohol Craving During 6 Weeks of Rehab Hospitalisation in Comparison to Not Homozygous and Homozygous Short Alleles - Preliminary Report.

Purpose: The aim of this study was to assess changes in the severity of alcohol craving according to allelic variants of the 5-HTTLPR gene polymorphism during hospitalisation and their association with selected clinical variables in alcohol-dependent patients. Patients and Methods: The study is exploratory. Participants were investigated at the 2nd and 6th week of alcohol-dependence therapy in the addiction treatment unit. Recruitment was conducted among alcohol-dependent patients from several Polish drug treatment centres. The total sample size was 130 persons (12 females and 118 males). Study subjects' mean age was 43.0 years. Patients were investigated twice by using the Penn Alcohol Craving Scale (PACS) and Beck Depression Inventory (BDI), and once by using Short Alcohol Dependence Data Questionnaire (SADD) and taking a swab for genetic testing. The polymorphism of the gene encoding the serotonin transporter 5-HTTLPR (SLC6A4) was determined from isolated DNA and its homozygous variants of short/short or long/long alleles and heterozygous short/long alleles were analysed. Results: At 6th week of the follow-up, there was a decrease in the severity of alcohol craving in half of subjects with the short/short allele (p = 0.033) and in one-fifth of subjects with the long/short allele (p = 0.002) of the 5-HTTLPR gene. In subjects with long/long allele of the 5-HTTLPR gene, there was no change in the severity of alcohol craving between 2nd and 6th weeks of the study (p = 0.242). Conclusion: There was no statistical influence of the homozygous long allele of the 5-HTTLPR gene on severity of alcohol craving during 6 weeks of rehab hospitalisation in comparison to not homozygous and homozygous short alleles. The s-allele was associated with decrease of alcohol craving. It may point on the potential need for differentiated rehabilitation methods depending on the genetic diversity of addicted patients and its role in the severity of alcohol craving.

Initial Study on COMT and DRD2 Gene Polymorphisms as Well as the Influence of Temperament and Character Trait on the Severity of Alcohol Craving in Alcohol-Dependent Patients.

The main aim of this work was to determine the impact of COMT and DRD2 gene polymorphisms together with temperament and character traits on alcohol craving severity alcohol-dependent persons. The sample comprised of 89 men and 16 women (aged 38±7). For the sake of psychological assessment various analytic methods have been applied like the Short Alcohol Dependence Data Questionnaire (SADD), Penn Alcohol Craving Scale (PACS) or Temperament and Character Inventory (TCI) test. The SNP polymorphism of the analyzed genes was determined by Real Time PCR test. The results showed, that the COMT polymorphismmay have an indirected relationship with the intensity and changes in alcohol craving during abstinence. The DRD2 receptor gene polymorphisms are related with the intensity of alcohol craving. It seems that the character traits like "self-targeting", including "self-acceptance", are more closely related to the severity of alcohol craving and polymorphic changes in the DRD2 receptor than temperamental traits. Although this is a pilot study the obtained results appeared to be promising and clearly indicate the link betweengene polymorphisms alcohol craving and its severity.

Mother's Milk Microbiome Shaping Fecal and Skin Microbiota in Infants with Food Allergy and Atopic Dermatitis: A Pilot Analysis.

The child microbiome, including gut and skin communities, is shaped by a multitude of factors, and breastfeeding is one of the most essential. Food allergy (FA) and atopic dermatitis (AD) are among the most common diseases in pediatrics, with the prevalence of each up to 6% and 20%, respectively. Therefore, we aimed at finding differences between the fecal and skin microbiomes of FA and AD patients in the context of breastfeeding, by means of the Illumina sequencing of 16S rRNA gene fragment libraries amplified from the total DNA isolated from samples collected from allergic and healthy infants. We also analyzed milk samples from the mothers of the examined children and searched for patterns of incidence suggesting milk influence on an infant's allergy status. Here we show that a mother's milk influences her child's fecal and skin microbiomes and identify Acinetobacter as the taxon whose abundance is correlated with milk and child-derived samples. We demonstrate that breastfeeding makes allergic children's fecal and skin communities more similar to those of healthy infants than in the case of formula-feeding. We also identify signature taxa that might be important in maintaining health or allergy development.

Variability of the rs333 in Polish patients with lupus erythematosus.

INTRODUCTION: Lupus erythematosus (LE) is an autoimmune disease with a strong influence of genetic and environmental factors. C-C motif chemokine receptor 5 (CCR5) gene expression may affect the development and intensity of LE. AIM: To evaluate the possible association between the 32bp deletion in rs333 locus located within the CCR5 gene and the development of LE or the occurrence of various clinical symptoms in the course of the disease. MATERIAL AND METHODS: One hundred and twenty patients with LE (77 with systemic lupus erythematosus (SLE) and 43 with discoid lupus erythematosus (DLE)) and 100 healthy controls from the Polish population were genotyped for deletion in rs333. RESULTS: 32 bp deletion in the rs333 was significantly more frequent among healthy individuals than DLE patients. Moreover, heterozygotes and homozygotes with deletion in rs333 were significantly more frequent within the control group than the group of patients with discoid lupus erythematosus. In contrast, any statistically significant differences in allele or genotype frequencies between healthy persons and SLE patients were observed. Furthermore, nucleotide sequence variability of rs333 was not associated with certain clinical symptoms of LE patients. CONCLUSIONS: Deletion in the rs333 might be a protective factor for DLE, but not SLE in the Polish population. Nevertheless further studies performed on larger populations are needed to confirm these observations.

A Combined Analysis of Gut and Skin Microbiota in Infants with Food Allergy and Atopic Dermatitis: A Pilot Study.

The gut microbiota in patients with food allergy, and the skin microbiota in atopic dermatitis patients differ from those of healthy people. We hypothesize that relationships may exist between gut and skin microbiota in patients with allergies. The aim of this study was to determine the possible relationship between gut and skin microbiota in patients with allergies, hence simultaneous analysis of the two compartments of microbiota was performed in infants with and without allergic symptoms. Fifty-nine infants with food allergy and/or atopic dermatitis and 28 healthy children were enrolled in the study. The skin and gut microbiota were evaluated using 16S rRNA gene amplicon sequencing. No significant differences in the α-diversity of dermal or fecal microbiota were observed between allergic and non-allergic infants; however, a significant relationship was found between bacterial community structure and allergy phenotypes, especially in the fecal samples. Certain clinical conditions were associated with characteristic bacterial taxa in the skin and gut microbiota. Positive correlations were found between skin and fecal samples in the abundance of Gemella among allergic infants, and Lactobacillus and Bacteroides among healthy infants. Although infants with allergies and healthy infants demonstrate microbiota with similar α-diversity, some differences in ß-diversity and bacterial species abundance can be seen, which may depend on the phenotype of the allergy. For some organisms, their abundance in skin and feces samples may be correlated, and these correlations might serve as indicators of the host's allergic state.

[Polymorphism of helicobacter pylori and the presence of genes babA2 and sabA and endoscopic and histopathological changes in patients infected with Heicobacter pylori]. / Polimorfizm Helicobacter pylori i obecnoss genów babA2 i sabA a obraz endoskopowy i histopatologiczny u chorych zakazonych Helicobacter pylori.

UNLABELLED: The infection of Helicobacter pylori is the main reason of a duodenal and gastric ulcer disease. Among other virulence factors of Helicobacter pylori, there are outer membrane proteins (OMPs), such as babA2 and sabA. THE AIM OF THE STUDY: An assessment of a relationship between the presence of genes babA2 and sabA and endoscopic and histopathologic changes during gastritis, duodenitis and an ulcer disease. MATERIAL AND METHODS: We included 119 patients aged from 3 to 17 (average 13.6) with gastritis and duodenitis and the infection of Helicobacter pylori. The endoscopy was conducted with taking samples of the mucosa for the histopathologic and genetic examination. The degree of endoscopic and histopatologic changes were determined according to Sydney's classification. The patients were devided in the extra groups with a small level (without erosion) and with a large level (with erosion) of endoscopic changes. To identify the infection of Helicobacter pylori, the PCR technique was used and then the presence of the babA2 and sabA genes of Helicobacter pylori was verified. The genetic confirmation of Helicobacter pylori infection was obtained in 88 patients and material was directed to the further examination. RESULTS: Not statistically significant differences were determined between endoscopic and histopathologic pictures and either the presence or absence of the genes babA2 and sabA. CONCLUSION: The presence of the genes babA2 and sabA is not related with level of endoscopic and histopathologic changes in pediatrics patients.

[The prevalence of dupA (duodenal ulcer-promoting gene) of Helicobacter pylori in children and adolescents--own observation]. / Czestosc wystepowania genu dupA (duodenal ulcer-promoting gene) Helicobacter pylori u dzieci i mlodziezy--obserwacje wlasne.

UNLABELLED: The strains of Helicobacter pylori are described by many common features which determine their virulence. The genes which are connected with much higher virulence of some strains are vacA, cagA, oipA, dupA. Duodenal Ulcer Promoting Gene--dupA is the new virulence factor coexisting with a duodenum ulcer. There is a rationale that shows a protective character of dupA with reference to a stomach cancer. The dupA gene probably causes increasingly higher releasing of pro-infectious IL-8 via stomach cells and it influences the production of IL-12 and other cytokines. The aim of the study was to determine the frequency of dupA gene's appearance in the Polish children's group and in the Polish teenagers' group infected with H. pylori. The research was also aimed to determine the coexistence of dupA gene and duodenum ulcer disease or erosion infection of duodenum's mucous membrane. MATERIAL AND METHODS: The endoscopic examination of the upper part of digestive duct was performed in 119 qualified patients with dyspeptic symptoms and with suspicion of stomach and duodenum's mucous membrane infection. The segments were taken for histopathological identification of H. pylori and for genetic indicating via PCR method. To confirm the presence of H. pylori in the extract the amplification of DNA fragment sized 860 pz was used. The presence of dupA gene was detected by PCR reaction with using the starters which include the fragment of jhp0917-jhp0918 sequence in the plastic H. pylori's genome area. To confirm the infection the urea breathing test was taken. RESULTS: 88 patients confirm the infection of H. pylori. The presence of dupA gene was found in 20 patients--a group A (22.7%), whereas in 68 patients dupA gene was not found--a group B (77.2%). Pathological changes in duodenum was found in 20 patients infected with H. pylori (22.7%), included 4 patients in the group A (20%) and 16 in the group B (23.5%). There was an infection (swelling, redness, congestion) in duodenum was found in the group A in all cases and there was an erosion presented in 3 patients. In the group B in 2 patients the duodenum ulcer disease was diagnosed. The infectious changes in duodenum were found in 7 patients but they were not infected with H. pylori; 1 patient was diagnosed with the duodenum ulcer disease. CONCLUSION: The presence of dupA gene in the Polish children population infected with H. pylori is quite frequent but there is no clinical correlation with the duodenum ulcer disease.

[The application of mitochondrial genomics to forensic investigations based on human mitochondrial DNA testing]. / Wykorzystanie osiagniec genomiki mitochondrialnej w badaniach genetyczno-sadowych opartych na analizie sekwencji ludzkiego mitochondrialnego DNA.

In this study we present two forensic cases where mitochondrial DNA HVS I and HVS II haplotypes of evidentiary hairs match reference samples. Based on the information retrieved from mtDNA coding region of reference material, we selected single nucleotide polymorphisms (SNPs) located outside the HVS I and HVS II regions that could increase the informativeness of mtDNA analysis. The SNPs were typed via SNaPshot or dideoxy sequencing technology. In both cases the SNP results allowed for unambiguous exlusion of the evidence and for determining that reference samples originated from the same person.

[Genetic type of Helicobacter pylori and the efficacy of eradication therapy]. / Typ genetyczny Helicobacter pylori a skutecznosc leczenia eradykacyjnego.

UNLABELLED: Helicobacter pylori is one of the most popular bacteria in the world. The H. pylori infection is an etiological factor of permanent changes in inflammatory of stomach mucous membrane, peptic ulcer of the stomach and duodenum disease and stomach cancer or mucosa associated lymphoid tissue from lymphoid tissue of mucous membrane. The strain bacteria which produce the protein CagA and cytotoxin VacA belong to the more pathogenic strains. The most successful method of treatment for H. pylori infection is an eradication of the bacteria. THE AIM OF THE STUDY: Was an evaluation of the influence which H. pylori genetic type (type I: CagA-positive, CagA-negative, VacA-positive, VacA-negative) has on efficacy of eradication therapy. MATERIAL AND METHODS: 214 of patients over the third year of life with symptoms of dyspepsia, of the upper part of the gastrointestinal tract was performed and H. pylori infection was proved in histopathological or (and) urease test and urea breath test. H. pylori identification was performed using PCR method for biopsy specimens of the gastric mucosa, estimating genetic type of the bacteria (CagA-positive, CagA-negative, VacA-positive, VacA-negative). Triple-drug eradication therapy was introduced. The efficiency of this treatment was checked after 6 weeks with the breath test. RESULTS: The H. pylori infection was found in 101 patients (47.2%), 33 patients were infected with the strain type I (32.7%) and 68 patients (67.3%) with the strain type II. After the treatment the eradication of the infection was found at 71 patients (70.3%), lack of efficacy in H. pylori infection treatment was found at 30 patients (29.7%). Considerably higher percentage of eradicative infection was shown in the group of patients infected with the type II of H. pylori (76.5% vs. 58.8%, p < 0.04). CONCLUSIONS: The effectiveness of eradication can be influenced by the genetic type of H. pylori. The better effects of eradicative treatment can be expected if one is infected with the strains of smaller virulence.

[Identification of semen in bloodstains with the use of alternative light source and biochemical screening tests]. / Identyfikacja nasienia w zaplamieniach krwawych z uzyciem alternatywnego zródla swiatta i przesiewowych testów biochemicznych.

The objective of the investigation was the verification of the presence of semen in stains constituting mixtures of semen and blood employing alternative light source (ALS) and commercially available biochemical screening tests based on the activity of acid phosphatase (AP) and prostate-specific antigen (PSA). The tests demonstrated that discrimination between particular components of a blood-semen mixture was impossible either with the naked eye, as well as with the use of ALS. White fluorescence was observed only in stains consisting of pure semen and semen-blood mixtures at a ratio of 100:1. The assay for PSA was positive in the case of all the examined semen dilutions and semen-blood mixtures, whereas the sensitivity of the AP-based test assay was lower by one order of magnitude.