Increased microRNA-323-3p in IL-22/IL-17-producing T cells and asthma: a role in the regulation of the TGF-ß pathway and IL-22 production.

BACKGROUND: IL-22- and IL-17-producing T cells have important roles in allergic diseases. MicroRNAs (miRNAs) are posttranscriptional regulators of gene expression and modulate numerous biological processes. Little is known about the functions of miRNAs in IL-22/IL-17-producing T cells. MATERIAL AND METHODS: IL-22- and IL-17-positive T cells were sorted from human peripheral blood mononuclear cells (PBMCs) by intracellular staining and dual-secretion assay. miRNA expression profiles were detected with TaqMan array microfluidic cards. T cells were transfected with miRNA mimics. Gene expression was analyzed using RT-qPCR and/or enzyme-linked immunosorbent assay in T-cell subsets and PBMCs from patients with asthma and atopic dermatitis. RESULTS: The increased expression of miR-323-3p and noncoding RNA nc886 and reduced expression of miR-93, miR-181a, miR-26a, and miR-874 were detected in IL-22-producing T cells. The pathway analysis of the putative targets suggested that these differentially expressed miRNAs could impact the proliferation, differentiation, and effector functions of T cells. Further analyses showed the highest expression for miR-323-3p in IL-22- and IL-17-double-positive T cells and its capacity to suppress multiple genes from the transforming growth factor-ß pathway and the production of IL-22 in T cells. An increased expression of miR-323-3p in PBMCs from patients with asthma and reverse correlation between miR-323-3p levels and IL-22 production in PBMCs cultured in T-cell growth conditions was observed. CONCLUSIONS: Our data suggest that miR-323-3p acts in a negative feedback loop to control the production of IL-22 in IL-22/IL-17-producing T cells and might thus impact the T-cell responses in asthma.

The XRF mapping of archaeological artefacts as the key to understanding of the past.

The article describes the X-ray fluorescence (XRF) studies on the chemical composition of archaeological artefacts. The mapping of the concentration of selected elements has been used to recognise the way of object production and the use. The obtained data allowed to obtain the new information, which is impossible to gain by use of different methods. 'The data obtained from the chemical composition of the particular parts of the objects may be used for the interpretation of the manufacturing technology or the primal form of the objects. Additionally, the knowledge obtained from the chemical composition of the different parts of the artefacts may be essential for the selection of the protection and conservation methods. The present studies can be useful to improve knowledge about the level of former craftsmanship. These knowledge allow us to exam archaeological artefacts in a new light, and these findings can also broaden the archaeological knowledge horizons and provide good bases for further detailed studies.

A novel, dual cytokine-secretion assay for the purification of human Th22 cells that do not co-produce IL-17A.

BACKGROUND: Interleukin-22 is produced by certain T helper cells subsets (Th17, Th22) and at lower levels by γ-δ T cells, NKT and innate lymphoid cells. Th22 cells are unique immune cells that regulate tissue responses by IL-22 production. The exact discrimination between Th17 cells that co-produce IL-22 and single IL-22-producing Th22 cells has not been possible until the present study. Isolation of pure Th22 cells without co-expression of cytokines of other T-cell subsets is essential to better understand their function in humans. The aim of this study is the isolation and characterization of viable, human IL-22-producing CD4+ T cells that do not produce IL-17A. METHODS: Isolation of viable Th22 cells was performed with the combination of two cytokine secretion assays detecting IL-17A- and IL-22-producing cells in a single purification step. RESULTS: The newly developed cytokine secretion assay consists of anti-IL-22 and anti-IL-17A catch antibodies, which via biotin-streptavidin interaction are bound to the biotinylated surface of the target cell, and anti-IL-22 and IL-17A detection antibody labelled with a fluorescent dye, which detects cytokines bound to these catch antibodies. A unique population of human Th22 cells, which do not produce IL-17A, was sorted, and cytokine expression pattern was confirmed by quantitative PCR analysis and ELISA. The presented technique allows the detection and isolation of pure human Th22 cells. CONCLUSIONS: This technique may allow the purification of any single cytokine-producing cell subset, and the combination of several different cytokine secretion assays can be used to purify and characterize novel and unique cell subsets.

Regular atomic narrowing of Ni, Fe, and V nanowires resolved by two-dimensional correlation analysis.

We present a novel statistical method for the study of stable atomic configurations in breaking nanowires based on the 2D cross-correlation analysis of conductance versus electrode separation traces. Applying this method, we can clearly resolve the typical evolutions of the conductance staircase in some transition metal nanojunctions (Ni, Fe, V) up to high conductance values. In these metals our analysis demonstrates a very well ordered atomic narrowing of the nanowire, indicating a very regular, stepwise decrease of the number of atoms in the minimal cross section of the junction, in contrast to the majority of the metals. All these features are hidden in traditional conductance histograms.

[Relation between specific IgE antibodies to grass pollens and to wheat, rye, soya in children with allergic disease]. / Wzpólzaleznosc mieczy obecnoscia swoistych przeciwcial klasy ige dla pylków traw, pszenicy, zyta i soi u dzieci z choroba alergiczna.

The aim of this study was to investigate the relationship between the hypersensitivity to grass pollens (gxl) and to wheat (f4), rye ff5) and soya (fl 4) in children with allergic disease. Specific IgE was determined by the FEIA CAP- System. Hypersensitivity was expressed in classes from 1 to 6. Specific IgE antibodies against grass (gxl) in 944 children with allergic disease, aged from 6 months to 18 years were detected. At the same time in 532 of these children - specific IgE to wheat and rye and in 122 children specific IgE to soya were also estimated. We detected very high correlation between the hypersensitivity to grass and the presence of specific IgE to wheat, rye (r=0.96, p<0.01) and soya (r=0.90, p<0.05).

[Changes in spontaneous blast transformation of children's lymphocytes which have been observed in vitro in the last years]. / Zmiany w spontanicznej transformacji blastycznej limfocytów dzieci obserwowane in vitro w ciagu ostatnich lat.

In the Immunology Department of the Institute of Mother and Child in Warsaw we have used lymphocyte blastic transformation test (LETT) over 30 years to study lymphocyte function. During the last 10 years we have noticed progressive elevation of lymphocyte percentage undergoing spontaneous blast transformation (autotransformation) in culture of these cells. The percentage rose from mean value of 2.5% to 14.7% in year 2000 with statistically significant difference already between 1985 and 1990 (2.5% versus 9.3%). The reason for this phenomenon is unknown and needs more investigations.

[Nitric oxide (NO)--Nobel prize in medicine and physiology for 1998]. / Tlenek azotu (no)--nagroda nobla z dziedziny medycyny i fizjologii za rok 1998.

On October 12, 1998. The Nobel Assembly announced the award of the Nobel Prize in Medicine and Physiology to pharmacologists Robert Furchgott, Louis Ignarro, and Ferid Murad. The Nobel Committee decided to award the prize for their discoveries concerning--nitric oxide as a signalling molecule in the cardiovascular system. Nitric oxide (NO) has a key importance for vascular tonus, acts as a signal molecule in the nervous system and plays an important function in the immunological system. Nitric oxide is a multifunction molecule which controls the blood pressure, modulates gastrointestinal motility. It is produced in abnormal level intensifies septic shock and destruction of nervous tissue. NO is important in different branches of medicine. For instance NO gas has been used to reduced high blood pressure in the lung of infants. Several unknown NO applications in medicine are waiting for discovery.

[Efficacy of specific immunotherapy in children with bronchial asthma, sensitised to dermatophagoides pteronyssinus]. / Skutecznosc immunoterapii swoistej u dzieci chorych na asthme oskrzelowa, uczulonych na roztocza kurzu domowego.

The aim of study was the evaluation of efficacy of specific immunotherapy using Alavac S HDM for 2-4 years in children with atopic bronchial asthma induced by Deramtophagoides pteronyssinus. The study was carried out in a group of 33 children aged 5-18 years, from the Outpatient Department of Immunology at the National Research Institute of Mother and Child. The children demonstrated clinical symptoms of asthma and had a medical history typical for this disease. Diagnostic procedures including skin prick tests and estimation of specific IgE to Dermatophagoides pteronyssinus by Pharmacia-CAP system gave positive results. The efficacy of therapy was monitored by a clinical score of symptoms with 0-10-20-30 points, recording symptoms relating to intensity of dyspneaa, wheezing, cough, value of PEF and concomitant medications for example corticoids and beta-antagonists. In the study group, 31 children (94%) with asthma treated by specific immunotherapy (AlavacS HDM) improved. This was confirmed by a statistically significant difference. The results indicate that specific immunotherapy is effective in the treatment of asthma in children sesitized to Dermatophagoides pteronyssinus.

[Sensitivity to cow's milk proteins and gluten in children's allergic disease]. / Uczulenia na bialka mleka krowiego i gluten w chorobie alergicznej u dzieci.

Aim of the study was the evaluation of sensitivity to cow's milk and gluten participation in children's allergic disease. Our studies were carried out in a group of 191 children, aged between 7 months and 18 years, with sensitivity to cow's milk proteins and gluten in type I and/or IV according to Gell and Coombs. Sensitivity to these allergens was confirmed in 88.4% of the 216 examined children with allergic disease. We showed a prevalent frequency of allergen specific T-cell responses in mechanism IV (in type IV - 79.58%, in type I - 10.99%, in I+IV - 9.42%). Evidence is presented that sensitivity to cow's milk proteins and gluten in I and IV Gell's and Coombs' mechanisms is more likely in small children and that it decreases with age.

[Evaluation of thyroid structure and function in patients with Turner syndromes]. / Ocena struktury i czynnosci tarczycy u chorych z zespolem Turnera.

The aim of our study was an estimation of thyroid structure and function in 37 patients with Turner syndrome aged from 19 to 60 years and in control group of healthy women. In each case the following studies were performed: cytogenetic examination, thyroid ultrasonography, serum total and free thyroid hormones, TSH, thyroglobulin (Tg), thyroid hormones binding globulin (TBG), antithyroglobulin and antithyroperoxidase antibodies (anti-Tg and anti-TPO) levels. In Turner syndrome ultrasonographic volume of the thyroid was significantly lower than in control group (11.03 vs 16.98 cm3). Abnormalities of thyroid function were found in 8 (22%) studied cases (subclinical primary hypothyroidism in 16%, full-clinical primary hypothyroidism in 3% and hyperthyroidism in course of Graves disease in 3%). Serum elevated antithyroid antibodies were present in 62% cases of Turner syndrome and were significantly higher than in control group (16%). In Turner syndrome thyroid diseases are more frequent than in healthy population. Every patient with Turner syndrome needs routine diagnostics of the thyroid structure and function.

[The effect of tobacco smoking on the human immune system]. / Wplyw palenia tytoniu na organizm czlowieka ze szczególnym uwzglednieniem ukladu odpornosciowego.

The influence of tobacco smoking in development of respiratory, cardiovascular diseases and impairment of foetal development is well documented. Recent studies have shown adverse effects of tobacco smoke on the immune system. Particular place of impact of tobacco smoke is BALT (bronchus-associated lymphoid tissue). Tobacco smoking increases the inflammatory processes and also has an immunosuppressive effect. It is very important in relation to more frequent youth and children diseases linked with the immune system. In school education programmes to prevent tobacco smoking carried out by the Institute of Mother and Child it is also very important to include this aspect.

[Bacterial and clinical evaluation of axetil cefuroxime in suspension]. / Bakteriologiczna i kliniczna ocena aksetylu cefuroksymu w zawiesinie.

For determination of cefuroxime activity, MIC was determined for 320 strains and by a diffusion-disc method susceptibility to this antibiotic of 3321 microorganisms isolated from children treated in Institute of Mother and Child was tested. Therapeutic value of axetil cefuroxime in suspension (Zinnat-Glaxo) was determined basing on specific and bacteriologically monitored treatment of 30 children (22 with respiratory tract infections and 8 with urinary tract infections). Etiological agents of these infections were: M. catarrhalis, H. parainfluenzae, S. aureus, P. rettgeri, E. coli and K. pneumoniae. MIC 50 of cefuroxime in mg/l was following: K. pneumoniae and S. pyogenes--0.12; M. catarrhalis--1; S. aureus and P. mirabilis--2; E. coli--4; K. pneumoniae--8, C. freundii--16; P. vulgaris--32, S. marcescens--128 and E. cloacae--256. Correlation was found between MIC values and percentages of susceptible strains. In 25 children full therapeutic effect was obtained. In 3 children partial recovery was noted, but they required application of an additional antibiotic. These studies demonstrated that majority of microorganisms responsible for infection of respiratory and urinary tract is susceptible to cefuroxime and that axetil cefuroxime in suspension presents as a very useful antibiotic, especially in pediatric ambulatory treatment.

Experimental ocular siderosis after extrabulbar administration of iron.

The authors studied the penetration of iron administered extrabulbarly into the ocular tissues of rabbits. It was found that iron passes from the orbit into the eyeball and accumulates in considerable quantities in the sclera, choroid, retina, ciliary body, and even in the vitreous and corneal epithelium. However, light microscopy failed to show any damage to the ocular tissues. The mechanism by which iron penetrates into the eyeball is discussed, and comparison is made between changes in the tissues, which characterise siderosis produced by an intrabulbar iron foreign body, and those in which an extrabulbar foreign body is involved.

The locomotor and stereotype response to dopaminergic drugs and caffeine after intracerebroventricular kainic acid in rats.

The effects of single kainic acid (KA) (0.1 microgram) intracerebroventricular injection on locomotor and stereotypy responses to amphetamine and apomorphine, and on locomotor response to caffeine were measured at different intervals (24, 120, 240, 480 h). The responses to amphetamine and caffeine were markedly elevated during the all period. In contrast, the responses to apomorphine remained unchanged. Histological examination showed severe destructions of neurons and fibres within the nucleus caudatus and the permanent loss of hippocampal pyramidal cells.

The haloperidol effect on the histofluorescence in the caudate nucleus of the rat brain treated with agonistic and antagonistic acetylcholine compounds.

The aim of the paper was to trace the haloperidol effect in the acute and chronic experiments on the histofluorescence in the caudate nucleus of the rat and in conditions of pretreatment of those animals with agonistic and antagonistic compounds of acetylcholine. The experiments were performed on 35 Wistar rats of 180 g mean body weight. According to the experimental model the following were administered: alpha-MT; alpha-MT + haloperidol; Haloperidol in single dose and in repeated multiple doses; Atropine: Atropine + haloperidol; carbachol and carbachol + haloperidol. The Falck et al. [5] histofluorescence method was applied to demonstrate catecholaminergic structures. Changes in the fluorescence of nucleus caudatus of the rat after single or repeated administration of haloperidol and after combinations with cholinergic compounds were always accompanied by the decrease of histofluorescence. We assume that it can be accounted for the existence of functional bonds between the dopaminergic and the cholinergic systems. Those bonds can be explained on the structural grounds, that is by a dense network of nerve endings and in bundles of preterminal nerve fibres. Neurocytes bodies were not sensitive to the administered drugs. The dense network of nerve endings proved to be most sensitive to the action of haloperidol given by itself and in combination with other drugs. A single large dose of haloperidol produced more pronounced effects than several repeated doses of that neuroleptic.

The locomotor and exploratory activities in rats after lesion of hippocampal pyramidal cells with kainic acid.

The effect of single kainic acid (KA) intracerebroventricular(icv) injection on spontaneous locomotor, exploratory, basal and total activities in rats measured a different times was tested. Whereas at 3 hr after KA icv administration an increase of the total, basal and spontaneous activities of rats were observed, a great decrease of the exploratory activity was noted. KA at 24 hr after its administration depressed all types of rats activities. From 72nd hr after KA icv injection a significant increase in the total, exploratory and spontaneous locomotor activities without changes in the basal activity was observed. The above mentioned changes persisted up to 10-20 days after KA administration. The histological examinations showed evident neuropathologic changes in hippocampal stratum pyramidale. The most evident changes concerned the hippocampal areas CA3/CA4 and CA1. There were observed shrinking, narrowing and fragmentation of pyramidal cells. An intense progressive gliotic reaction was present among the fragmented cells, the layer of pyramidal cells being considerably diminished.

Activity of acetylcholine hydrolase (EC 3.1.1.7) in the pig brain; spectrophotometric and histochemical studies in the light and electron microscope.

AChE activity (EC 3.1.1.7) in the pig brain was examined by spectrophotometric method and histochemically in the light and electron microscope. The results obtained correlate to a high degree with the existing data concerning the activity of the studied enzyme in other mammals. In the pig, the highest activity was encountered in the striatum nucleus accumbens and nucleus amygdale lateralis: the lowest activity was found in encephalon and cerebellum. In comparison with other mammals AChE activity in the pig brain is considerably lower. No comparable proportions have been encountered between the activities of AChE and ChAT in particular brain regions in the mammals examined so far and also in the pig. The results of histochemical examination both in the light and in the electron microscope correspond to the results obtained by other authors. The discussion deals with the problem of correlation between the level of AChE activity in particular regions of the pig brain with respect to their phylogenetic growth.

Interaction between cholinotropic and adrenotropic drugs on the histofluorescence of the central catecholamine structures in the rat.

Male Wistar rats were given phentolamine or phenoxybenzamine in the lateral ventricles in doses of 1 mg/kg. After 30 minutes they received in the same way atropine in doses of 1 mg/kg or carbachol in doses of 0.05 mg/kg. The control group was given physiological saline. The animals were decapitated 30 minutes after drug administration. The Falck and Hillarp histofluorescence method was applied. The areas of DA (nigrostriatal and meso-limbic) and NA systems were examined. It was found, that atropine increased the intensity of fluorescence in comparison with the control group, in all areas of DA structures. The action of carbachol was more differentiated. In the substantia nigra (A8 and A9) respectively in the globus pallidus and the nucleus arcuatus (A12) its effect was the same as that of atropine. In other areas it caused weakening of fluorescence or showed no effect. In the NA system atropine weakened the fluorescence considerably while carbachol increased it in five out of eleven areas. The interaction of cholinotropic and adrenotropic drugs is disscused.