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.

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.

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.

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.

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.

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.

Role of FOXP3 expression and serum vitamin D and C concentrations when predicting acquisition of tolerance in infants with cow's milk allergy

BACKGROUND: 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

ANTECEDENTES: Las células Treg y los factores dietéticos pueden desempeñar un papel importante en la adquisición natural de tolerancia en niños con alergia a la leche de vaca (CMA). El mejor marcador de linfocitos Treg es el factor de transcripción Forkhead box P3 (FOXP3). OBJETIVOS: El artículo examina la relación entre la expresión de mRNA específico para FOXP3 y la concentración sérica de vitaminas D y C, así como el desarrollo de diferentes fenotipos de tolerancia en niños con CMA. Material y métodos: El grupo de estudio estaba compuesto por 138 bebés con CMA y 101 sanos. Todos los niños tomaron primero un producto lácteo hervido vía oral, y posteriormente productos lácteos sin calentar. Se evaluó la expresión de ARNm para FOXP3 y la concentración sérica de vitamina C y D. RESULTADOS: A los dos años de vida, 54 (39,1%) de los niños aún mostraban CMA, 43 (31,2%) eran reactivos a la leche sin calentar y tolerantes a la leche caliente, mientras que 41 (29,7%) habían superado la alergia. El nivel medio de expresión de FOXP3 en el grupo CMA fue de 2,07 ± 1,23; inferior al obtenido en el grupo control de 2,98 ± 1,52 (p < 0,001). Un valor por debajo de 1,45 indica alergia. El nivel sérico medio de vitamina D en el grupo de estudio (29,67 ± 7,09 ng/ml) fue más bajo que en el grupo control, 33,35 ± 4,13 ng/ml (p < 0,001). No se encontraron diferencias significativas en el contenido medio de vitamina C en suero. CONCLUSIONES: El aumento de la expresión de FOXP3 mRNA puede predecir la adquisición de tolerancia más rápida en los bebés con CMA. Estos niños tienen niveles séricos más bajos de vitamina D que los niños sanos. No se encontró relación entre la historia natural de CMA y la concentración de vitamina C en suero

Interactions of ferrous ions with peroxidized lecithin-liposomes.

Oxidation of Fe2+ in the presence of peroxidized lecithin-liposomes small unilamellar vesicles suspended in various media (pH = 5.5) is described. Peroxidation of lecithin was induced by UV light and assayed by three methods: estimation of conjugated dienes, accumulation of hydroperoxides and by the thiobarbituric acid test. The oxidation of Fe2+ was accelerated by phosphate and citrate buffers, but in acetate buffer it was dependent on lipid peroxidases concentration and exhibited saturation kinetics.

Changes in the distribution of blood vessels in the rat cerebral hemisphere injured during early postnatal period.

A response of the vascular network of rat cerebral hemisphere to injury inflicted neonatally and on the postnatal day 6 was examined. After the neonatal injury, no change could be recorded, however, following similar lesion made in 6-day-old rats zones of significantly increased vascular density were observed. The data indicate a significant change in the reactivity of cerebral blood vessels to injury occurring during the earliest period of postnatal development. This phenomenon is considered to be parallel to a similar developmental increase in the ability of astroglia to form a reactive gliosis.

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.

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.

Influence of kinetin (6-furfurylo-amino-purine) on human fibroblasts in the cell culture.

Influence of kinetin on human fibroblasts cultured in vitro was studied. Investigations were carried out on I passage fibroblasts, taken from the human skin. Feulgen's method and radioautography were used. Quantitative estimations of DNA in nuclei of fibroblasts were done on cytophotometer. An increase in amount of DNA in nuclei of fibroblasts cytophotometer. An increase in amount of DNA in nuclei of fibroblasts cultured in the presence of kinetin was found. Stronger 3H uridine incorporation was recorded in cultures after administration of the lowest dosis of kinetin. Kinetin, derivative of adenine (6-furfurylo-amino-purine) belongs to cytokinins and it regulates the growth of plants.

Influence of 3-indoleacetic acid on cytochemical changes in nuclei and cytoplasm of human fibroblasts in the cell culture.

The performed studies covered the action of 3-indoleacetic acid (3IAA) on human fibroblasts cultured in vitro. The 3IAA implemented in the studies was provided by Chemapol Firm (Czechoslovakia). All the investigations were carried out on I passage fibroblasts, taken from the human skin in form of monolayer culture. Feulgen's method and autographic one were resorted to in the studies. The optimal time of acid hydrolysis was 10 min. Quantitative measurements of DNA in nuclei of fibroblasts were done on integrating cytophotometer. Incubation with labelled 3H uridine as RNA precursor was employed for demonstrating the activity of cell nucleus transcription. As concerns the quantitative changes in nucleic acids, it was disclosed that DNA and RNA synthesis was intensified. Nuclei were seen to appear with polydiploidal DNA amount in cultures being influenced by 3IAA action. The smallest number of mitoses was revealed in the cultures exposed to the action of the highest dose of the said compound. Stronger 3H uridine incorporation was recorded in cultures 24 and 72 hours after the administration of 10 mg/1000 ml of 3IAA. The results were presented in the form of diagrams, nucleinogram and tables.

[Diagnostic yield of electrophysiological and immunological studies in inflammatory myopathies]. / Zastosowanie badan elektrofizjologicznych i immunologicznych do diagnostyki miopatii w chorobach tkanki lacznej.

The aim of the study was to estimate the value of the immune markers in defining distinct subsets of inflammatory myopathies. The series of 76 patients was divided, on the basis of the clinical data, into 5 groups (polymyositis (PM), dermatomyositis (DM), scleromyositis (Scm), mixed connective tissue disease (MCTD), unclassified). In all cases detailed clinical, electrophysiological (concentric needle electromyography (CNEMG) and skin sympathetic response (SSR)) were performed as well as immunologic studies: the anti Mi-2, anti RNP, PM-Scl, Jo-1. The findings indicate that immune markers have a diagnostic value in differentiating the defined subsets of patients different in respect of course, prognosis and therapeutic indications. The authors stressed the value of the Jo-1 antibody in detecting the subset of polymyositis with coexisting interstitial lung disease. Electrophysiological data do not differentiate the groups of patients with different clinical syndrome and different immune markers. EMG results seem however to be useful in monitoring the course of the disease and response to the therapy.

[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.

[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.