Role of FOXP3 Expression and Serum Vitamin D and C Concentrations When Predicting Acquisition of Tolerance in Infants With Cow's Milk Allergy.

BACKGROUND AND OBJECTIVE: Treg cells and dietetic factors may play a significant role in the natural acquisition of tolerance in children with cow's milk allergy (CMA). The best marker for Treg lymphocytes is the transcription factor forkhead boxP3 (FOXP3). Objective: We examine the relationship between FOXP3 mRNA expression and serum concentrations of vitamins D and C and the development of different phenotypes of tolerance in children with CMA. MATERIAL AND METHODS: The study group comprised 138 infants with CMA and 101 healthy infants. All children underwent oral food challenge, first with an extensively heated milk product and then with unheated products. FOXP3 mRNA expression and serum vitamin C and D concentrations were evaluated. RESULTS: At 2 years of life, 54 children (39.1%) still had CMA, 43 (31.2%) were unheated milk-reactive and heated milk-tolerant, while 41 (29.7%) had outgrown their allergy. The mean (SD) level of FOXP3 expression in the study group was 2.07 (1.23), which was lower than the control group value of 2.98 (1.52) (P<.001). A value below 1.45 indicated allergy. The mean serum level of vitamin D in the study group was lower than in the control group (29.67 [7.09] vs 33.35 [4.13] ng/mL; P<.001). No significant differences were found in mean serum vitamin C content. CONCLUSIONS: Increased FOXP3 mRNA expression can predict faster acquisition of tolerance in infants with CMA. These children have lower serum vitamin D levels than healthy children. No relationship was found between the natural history of CMA and serum vitamin C concentration.

Silver-modified octahedral anatase particles as plasmonic photocatalyst.

Octahedral anatase particles (OAPs) were modified with silver nanoparticles (NPs) by photodeposition method. The properties of OAPs influenced the properties of silver deposits, and thus the photocatalytic activity of the obtained silver-modified OAPs. Photocatalytic activities were tested under UV and vis irradiation for oxidative decomposition of acetic acid and oxidation of 2-propanol, respectively. The properties of silver-modified OAPs were investigated by XRD, STEM, DRS, XPS and time-resolved microwave conductivity (TRMC) method. It was found that electron traps (ETs) worked as nucleation sites for silver, resulting in formation of smaller silver NPs on smaller OAPs with larger content of ETs. The modification with silver resulted in enhanced photocatalytic activity under both UV and vis irradiation. It was found that larger crystallite size of silver NPs, and thus larger polydispersity of silver deposits resulted in broad and intense plasmon resonance peak causing enhanced visible activity. The correlation between photocatalytic activity and TRMC data, e.g., slower decay of TRMC signal for more active samples, allowed discussion on property-governed photocatalytic activities of silver-modified titania.

Noble metal-modified octahedral anatase titania particles with enhanced activity for decomposition of chemical and microbiological pollutants.

Octahedral anatase particles (OAPs) were prepared by hydrothermal (HT) reaction of titanate nanowires (TNWs). OAPs were modified with noble metals (Au, Ag, Cu and Pt) by two photodeposition methods: in the absence and in the initial presence of oxygen in the system. Photocatalytic activities for oxidative decomposition of acetic acid and anaerobic dehydrogenation of methanol under UV/vis irradiation and for oxidation of 2-propanol under visible light irradiation were investigated. Antibacterial activities for bacteria (Escherichia coli) and fungi (Candida albicans) were investigated in the dark and under UV irradiation and/or visible light irradiation. It was found that the kind of metal deposition significantly influenced the properties of photocatalysts obtained and thus their photocatalytic and antimicrobial activities. Modification of OAPs with metallic deposits resulted in enhanced photocatalytic activities for all tested systems. Pt-modified OAPs showed the highest activity for dehydrogenation of methanol due to their highest work function and lowest activation overpotential of hydrogen evolution. Cu-modified OAPs exhibited the highest activity for oxidative decomposition of acetic acid under UV/vis irradiation, probably due to the heterojunction between Cu oxides and TiO2. On the other hand, Au-modified OAPs showed the highest photocatalytic activity under visible light irradiation due to their plasmonic properties. Bare OAPs, prepared with various durations of the HT reaction, did not have any antibacterial properties in the dark, while their activity under UV/vis irradiation was correlated with their photocatalytic activities for dehydrogenation of methanol and decomposition of acetic acid. Antimicrobial activity of modified OAPs in the dark and under visible light irradiation was the highest for Ag-modified OAPs. Under UV irradiation, Cu-modified OAPs showed the highest activity for inactivation of both bacteria and fungi.

Switching Between Infliximab Originator and Biosimilar in Paediatric Patients with Inflammatory Bowel Disease. Preliminary Observations.

BACKGROUND AND AIMS: The growing incidence of inflammatory bowel disease (IBD) in children necessitates the use of biological treatments. Recently, an infliximab biosimilar was authorized in the European Union, which may result in switching patients. We present our preliminary experiences with such switches. METHODS: The prospective study included 32 paediatric patients diagnosed with Crohn's disease (CD) and 7 children with ulcerative colitis (UC) at 3 academic hospitals, who were switched from infliximab originator to its biosimilar (Remsima). Patient characteristics, disease severity, laboratory parameters and adverse events were recorded. Means, medians and ranges were calculated. RESULTS: Mean age at diagnosis of CD and UC was 11.1 (2.7-15.3) and 12.3 years (8.5-14.8), respectively. Mean number of infliximab originator infusions before switching to the biosimilar was 9.9 (median 8, range 4-29) and 5.1 (5, 1-12) for the CD and UC group, respectively. Evaluation efficacy of last biosimilar doses of all patients revealed rates of clinical remission of 88 and 57% for CD and UC patients, respectively. Last follow-up assessment of patients who continued with biosimilar therapy showed that 16/20 (80%) CD patients and all 4 UC individuals were in remission. One infusion reaction to infliximab biosimilar was observed in a CD patient, which led to treatment discontinuation. The incidence of sporadic mild adverse events prior to and after switching did not differ significantly and was consistent with the safety profile of the infliximab molecule. CONCLUSION: Switching from infliximab originator to its biosimilar seems to be a safe option in children with CD. After the switch the biosimilar was just as effective as the originator.

The impact of immobilized metal affinity chromatography (IMAC) resins on DNA aptamer selection.

DNA aptamers are single-stranded oligonucleotides which can form various secondary and tertiary structures. They can recognize a broad range of targets ranging from small molecules, such as ions, vitamins, antibiotics, to high molecular weight structures, including enzymes and antibodies. DNA aptamers are extensively studied as a potential source of new pharmaceutical drugs due to their inexpensive synthesis, low immunogenicity, and high specificity. The commonly used aptamer selection procedure is systematic evolution of ligands by exponential enrichment (SELEX) where the target molecule is immobilized on an appropriate chromatography resin. For peptide/protein targets, immobilized metal affinity chromatography (IMAC) resins are frequently used. There is a broad range of commercially available resins which can be used for IMAC. They are characterized by different metal ions, linker types, and bead materials. In this study, we tested the impact of different IMAC resins on the DNA aptamer selection process during eight SELEX cycles. A histidine-tagged 29 amino acid peptide corresponding to the interdomain connecting loop of human proliferating cell nuclear antigen was used as a selection target. Different resin materials containing the same metal ion (Co(2+)) were tested. Simultaneously, agarose resins containing identical linkers, but different metal ions (Co(2+), Cu(2+), Ni(2+), and Zn(2+)) were analyzed. The results of this study clearly demonstrated the impact of the metal ion and resin material on the DNA aptamer selection progress. The presented data indicate that for successful IMAC resin-based SELEX, the determination of the optimal resin might be crucial.

Diagnosis of gastroesophageal reflux and anti-reflux procedures among Polish children with gastrostomies: a 10-year nationwide analysis.

BACKGROUND/OBJECTIVES: To analyse the approach to diagnose gastroesophageal reflux (GER) and the qualification criteria for anti-reflux (AR) procedures in Polish children fed via gastrostomy between 2000 and 2010. SUBJECTS/METHODS: An electronic questionnaire containing questions on the demographic and clinical data of patients with gastrostomies was distributed to six Polish centres of nutritional therapy. The portion pertaining to GER included data on clinical exponents, diagnostic procedures (pH-metry, pH-impedance, scintigraphy and upper gastrointestinal (GI) series) and AR. RESULTS: In total, 348 children (M199/F149; age at gastrostomy 5.78±5.49 years) were included. Data on the diagnosis of GER and the AR criteria were available for 343 and 336 subjects, respectively. Percutaneous endoscopic gastrostomy was performed in 258/348 patients (74.1%), while surgery was performed in 80/348 patients (23%). The data from 10/348 (2.9%) cases were unavailable. At least one of the tests for GER was conducted in 177/343 (51.6%) of patients: pH-metry in 74/343 (21.6%), pH-impedance in 17/343 (5.0%), scintigraphy in 60/343 (17.5%) and upper GI series in 102/343 (29.7%). GER was reported in 114/343 cases (33.2%), and fundoplication was performed in 87 children (76.3% of patients with GER). The highest congruence between a positive test result and the decision to perform fundoplication was documented in cases of scintigraphy and upper GI series (P=0.00000 and P=0.00191, respectively). A significant increase in the prevalence of simultaneous gastrostomy and AR was observed over the decade analysed (r=0.8, P=0.009). This study revealed a centre-specific attitude towards the diagnosis of GER and the assessment of qualifications for fundoplication in Polish gastrostomy-fed children. CONCLUSIONS: The unified diagnostic algorithm of GER and the universal qualification criteria for AR procedures need to be defined for gastrostomy-fed children.

Insulin sensitivity in Type 1 diabetic children and adolescents.

AIM: To estimate insulin sensitivity in Type 1 diabetic children and adolescents, and assess the relationship between insulin sensitivity and clinical markers of adiposity and parameters of the metabolic syndrome. METHODS: A total of 202 patients aged 8-18 years with Type 1 diabetes and disease duration 1.5-15 years participated. Insulin sensitivity was estimated by glucose uptake during an euglycaemic-hyperinsulinaemic clamp and was calculated as the average amount of glucose (M(lbm) = mg/kg(lbm)/min) required to maintain euglycaemia. Blood pressure, glycated haemoglobin (HbA(1c)) and lipid concentrations were measured. RESULTS: The M(lbm )value ranged from 4.14 to 25.25 mg/kg(lbm)/min (mean 9.81 +/- 3.34 mg/kg(lbm)/min). There was a significant relationship between M value and patients' age (r = -0.38, P < 0.0001). Insulin sensitivity decreased significantly with the onset of puberty; hence, it was significantly lower in pubertal and post-pubertal adolescents. Girls were significantly more insulin resistant than boys (9.01 +/- 0.32 vs. 10.43 +/- 0.29 mg/kg(lbm)/min, P = 0.005). Insulin sensitivity correlated with body mass index (r = -0.29, P < 0.001), waist circumference (r = -0.35, P < 0.001), triceps skin fold (r = -0.17, P = 0.018), subscapular skin fold (r = -0.23, P = 0.001) and body fat (r = -0.19, P = 0.006). There was a relationship between M(lbm) value, cholesterol (r = -0.18, P = 0.012), high-density lipoprotein cholesterol (r = 0.15, P = 0.035), low-density lipoprotein cholesterol (r = -0.22, P = 0.002), triglycerides (r = -0.32, P < 0.001) and systolic blood pressure (r = -0.15, P = 0.029). Insulin resistance was related to HbA(1c) (r = -0.18, P = 0.012). Additionally, there was a correlation between M(lbm) value and insulin dose. CONCLUSIONS: Children and adolescents with Type 1 diabetes mellitus have a very wide range of insulin sensitivity, which is determined by sex, age, amount of adipose tissue and glycaemic control.

Common variants of DNA repair genes and malignant melanoma.

In the current study, we evaluated the possible associations of seven common variants of the DNA repair and cell cycle control genes BRCA2 and CHEK2 with malignant melanoma (MM). We genotyped 630 unselected MM patients and over 3700 controls (newborns, age- and sex-matched healthy adults with negative cancer family histories, and the adults selected at random by family doctors) for the prevalence of three common variants of the BRCA2 (T1915M, N991D and N372H) and four common variants of the CHEK2 (1100delC, VS2+1G --> A, I157T and del5395). Our study strongly suggests that the common variant of the BRCA2 gene -- the N991D variant is associated with malignant melanoma risk (OR=1.8, p=0.002 after Bonferroni correction). Patients homozygote for the N991D variant were present in 0.32% of cases and only 0.13% of controls. The other variants studied were not over-represented among MM patients when compared to the general population. In conclusion, we report an increased melanoma risk among carriers of the N991D change of the BRCA2 and no association of the CHEK2 changes with malignant melanoma.

Peripheral clocks: keeping up with the master clock.

Circadian clocks influence most aspects of physiology and behavior, so perhaps it is not surprising that circadian oscillators exist in nearly all mammalian cells. These cells remain synchronized to the outside world in hierarchical fashion, with a "master clock" tissue in the suprachiasmatic nucleus of the hypothalamus receiving light input from the retina and then conveying timing information to "slave" clocks in peripheral tissues. Recent research has highlighted both the similarities and differences between central and peripheral clocks and provided new insight into their communication. Above all, however, this parallelism of clockwork has provided a unique opportunity to study at the cellular level a regulatory mechanism that affects complex behaviors.

Germline MSH2 and MLH1 mutational spectrum including large rearrangements in HNPCC families from Poland (update study).

Germline mutations in the DNA mismatch repair genes MSH2 and MLH1 account for a significant proportion of hereditary non-polyposis colorectal cancer (HNPCC) families. One approach by which development of an efficient DNA-testing procedure can be implemented is to describe the nature and frequency of common mutations in particular ethnic groups. Two hundred and twenty-six patients from families matching the Amsterdam II diagnostic criteria or suspected HNPCC criteria were screened for MSH2 and MLH1 germline mutations. Fifty different pathogenic mutations were found, 25 in MSH2 and 25 in MLH1. Twenty-four of these had not previously been described in other populations. Among our 78 families with MSH2 or MLH1 mutations, 54 (69.2%) were affected by recurrent mutations including 38 found at least twice in our own series. Two of the most frequent alterations were a substitution of A to T at the splice donor site of intron 5 of MSH2 and a missense change (A681T) of MLH1 found in 10 and eight families, respectively. Among large deletions detected by the multiplex ligation-dependent probe amplification assay, exon 9 deletions in the MSH2 gene were found in two families. Our results indicate that a screening protocol specific for the Polish population that is limited to the detection of all reported mutations will result in the identification of the majority of changes present in MLH1 and MSH2 genes in Polish HNPCC kindreds.

Breast cancer predisposing alleles in Poland.

Mutant alleles of several genes in the DNA repair pathway have been found to predispose women to breast cancer. From a public health perspective, the importance of a given allele in a population is determined by the frequency of the allele and by the relative risk of breast cancer that it confers. In Poland founder alleles of the BRCA1, CHEK2 and NBS1 genes have been associated with an increased risk of breast cancer, but the relative contribution of each of these alleles to the overall breast cancer burden has not yet been determined. We screened 2012 unselected cases of breast cancer and 4000 population controls for 7 different mutations in these genes. Overall, a mutation was found in 12% of the cases and in 6% of the controls. Mutations in BRCA1 and CHEK2 contributed in approximately equal measure to the burden of breast cancer in Poland. A BRCA1 mutation was present in 3% of the cases. The missense BRCA1 mutation C61G was associated with a higher odds ratio for breast cancer (OR=15) than were either of the truncating BRCA1 mutations 4153delA (OR=2.0) and 5382insC (OR=6.2). In contrast, a higher odds ratio was seen for truncating CHEK2 mutations (OR=2.1) than for the missense mutation I157T (OR=1.4). This study suggests that cancer risks may be specific for particular alleles of a susceptibility gene and that these different risks should be taken into account by genetic counselors.

A common variant of CDKN2A (p16) predisposes to breast cancer.

BACKGROUND: A common missense variant of the CDKN2A gene (A148T) predisposes to malignant melanoma in Poland. An association between malignant melanoma and breast cancer has been reported in several families with CDKN2A mutations, OBJECTIVE: To determine whether this variant also predisposes to breast cancer. METHODS: Genotyping was undertaken in 4209 cases of breast cancer, unselected for family history, from 18 hospitals throughout Poland and in 3000 controls. RESULTS: The odds ratio (OR) associated with the CDKN2A allele for women diagnosed with breast cancer before the age of 50 was 1.5 (p = 0.002) and after age 50 it was 1.3 (p = 0.2). The effect was particularly strong for patients diagnosed at or before the age of 30 (OR = 3.8; p = 0.0002). CONCLUSIONS: CDKN2A appears to be a low penetrance breast cancer susceptibility gene in Poland. The association should be confirmed in other populations.

H2O2/UV enhanced degradation of pesticides in wastewater.

Photodegradation of organic pesticides in industrial wastewater was examined in a UV/H2O2/air system. An experimentally determined optimal amount of hydrogen peroxide (0.008% v/v) indicates that hydrogen peroxide concentration controlled the efficiency of photodegradation. Pre-treatment operations such as sedimentation, filtration and coagulation were used to obtain better efficiency of pesticide removal and to cut down on irradiation time. Finally, scale-up experiments in the air-sparged hydrocyclone (ASH) reactor were carried out. After 5 min irradiation of 100 dm3 industrial wastewater almost all pesticides were destroyed. Thus the ASH reactor proved to be an effective contactor for carrying out photochemical reactions.

[Biochemical characteristics of fungi isolated from ontocenoses of digestive tract in children]. / Wlasciwosci biochemiczne grzybów izolowanych z ontocenoz przewodu pokarmowego dzieci.

Fermentation of carbohydrates and assimilation of carbon compounds were evaluated. The utilization of 18 specific carbon compounds was estimated in the investigation of each strains for differentiation of the species. From among 125 strains of fungi the following were found: Candida albicans (112 strain), C. famata (2), C. glabrata (1), C. guilliermondii (1), C. kefir (4), C. tropicalis (2). The activity of 19 hydrolases was investigated using API ZYM. Biotyping of Candida strains was done according to the Williamson classification (1986), modified by Kurnatowska (1998). All strains were isolated from the oral cavity, rectum, as well as from the materials collected during endoscopy in children.

[Fungal invasions in the digestive tract in children]. / Inwazje grzybami przewodu pokarmowego dzieci.

The aim of presented study were fungal invasions of the oral cavity and gastrointestinal (GI) tract in children suspected of the inflammation of gastric and duodenal mucosa and of absorption disturbance. The fungal strains (125) were identified using API 20C and API 20C AUX (bio Me'rieux); they were as follows: Candida albicans, C. famata, C. glabrata, C. guillermondii, C. kefyr and C. tropicalis.

[Evaluation of selected X syndrome factors during hormonal replacement therapy]. / Zachowanie sie wybranych czynników zespolu X w czasie stosowania hormonalnej terapii zastepczej.

UNLABELLED: Estrogen deficiency after menopause leads to characteristics changes in the hormonal profile, which may influence lipid carbohydrate and calcium-phosphate metabolism and some elements of homeostasis. AIM OF STUDY: To evaluate the influence of hormone replacement therapy with 2 mg estradiol valerate and 0,15 mg levonorgestrel on carbohydrate and lipid metabolism in women after menopause during 12 months of follow-up. We examined 101 women, mean age 52.9 +/- 4.6 years (range from 44 to 65). HRT was applied in 67 women whereas 34 women were without treatment. All of them had no carbohydrate disturbances. All women underwent clinical examination, and body mass index (BMI), fasting blood glucose, insulin, total cholesterol and triglycerides levels were obtained. At 1 and 2 hours after 75 g glucose challenge (75OGTT) glucose and insulin levels were obtained. During hormone replacement therapy all women noted release or significant decrease of climacteric symptoms. Total cholesterol level was decreased, whereas triglycerides did not change. After 12 months of treatment there was also a significant decrease of all factors relating to carbohydrate metabolism--fasting glucose and insulin, insulin/glucose ratio and area under glucose and insulin curves. In our study--after 6 months of follow-up fasting insulin level, area under glucose and insulin curves were increased whereas fasting blood glucose level remained unchanged. Among women without HRT there were no significant changes in selected lipid parameters and BMI. Triglyceride levels decreased (albeit insignificantly) but, total cholesterol levels did not change. After 12 months glucose level did not change, although other carbohydrate parameters were increased. 1) In the study group there was a statistically significant decrease in total cholesterol levels whereas triglycerides remained unchanged. 2) HRT we significantly decreased of insulin resistance and fasting blood glucose levels as compared with non-group HRT. 3) The present results indicate HRT-induced improvement of lipid and carbohydrate metabolism. 4) Long-term HRT is necessary to improve carbohydrate metabolism.

[Levels of tumor necrosis factor-alpha and interleukin-2 in sera of twins delivered prematurely and at term]. / Stezenie czynnika martwicy guza-alpha i interleukiny-2 w surowicy krwi blizniat urodzonych w terminie i przedwczesnie.

The concentration of tumor necrosis factor-alpha (TNF-alpha) and interleukin-2 (IL-2) was estimated in first day of live in sera of premature (n = 8 pair) and in term delivered twins (n = 12 pair). One of premature delivered newborns was born vaginally 48 hours after first twin. Concentrations of TNF-alpha were approximately 4-times higher in sera of premature delivered in comparison to term delivered twins and a highest, more than 5-times higher level, was detected in serum of premature newborn born a 48 hours after first twin. Concentrations of IL-2 in sera of premature delivered were approximately 2.5-times higher than in term delivered twins and not significantly variations were observed in delayed delivered twin.

Estimation of antithyroid antibodies occurrence in children with coeliac disease.

INTRODUCTION: The coexistence of coeliac disease (CD) and the diseases of autoimmune origin is often discussed in literature. In this study the evaluation of antithyroid antibodies (antimicrosomal-TMA, anti-thyreoglobulin-ATG, thyroid peroxidase antibodies anti-TPO) occurrence against clinical and laboratory determinants of thyroid function is demonstrated in children with coeliac disease. MATERIAL AND METHODS: The study was conducted on 34 IgA-EmA positive children; control group consisted of 28 children with negative screening tests for coeliac disease. RESULTS: In both groups, the level of antithyroid antibodies (TMA, ATG, anti-TPO) and determinants of thyroid gland function (TSH, fT3, fT4) were evaluated; USG examination of thyroid gland was also performed. Elevated titres of antithyroid antibodies observed in children with coeliac disease (41.1%) in comparison to control group (3.56%) indicate the need for performing the screening tests for antithyroid antibodies in children with CD.

The amount of DNA in cells of the granular layer of the cerebellum and their susceptibility to hypoxia.

Purkinje cells of the cerebellum are particularly susceptible to hypoxia. In these cells tetraploidy has been demonstrated. Therefore, a link between the susceptibility of cells of the cerebellum to hypoxia and the amount of DNA seems probable.