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.

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.

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.

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.

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.

[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.

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.

[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.

Attempts at differentiation of the structure of neurons in the corpus geniculatum laterale of the pig by the immunocytochemical technique.

The investigations were carried out on the corpus geniculatum laterale of the pig with the aim to differentiate the structure of its neurons on the basis of the antigenic properties of the nervous tissue. The antigenic material consisted of the homogenate of the corpus geniculatum laterale (H) and of its nuclear (P1) and mitochondrial (P2) fractions. The antigens were used to immunize 3 groups of rabbits, the 4th one serving as control. The antigens were prepared according to requirements of the technique. By immunization of the rabbits with the antigens immune sera were obtained. These sera were processed in accordance with the requirements of the technique. The authors conducted the studies employing the immunofluorescent technique after Coons with the use of the anti-rabbit IgG goat serum conjugated with fluorescein, as well as the immunohistochemical method according to Nakane with the use of the antibodies labelled with horseradish peroxidase. The investigations led the authors to conclude that H as well as P1 and P2 possess distinct antigenic traits. Moreover, the findings of the immunofluorescent and immunohistochemical tests in which the anti-H serum was used point to the localization of the antigen in the following nerve structures: neuron perikaryons, glia cells, and nerve fibres. On the other hand, the application of anti-P1 and anti-P2 sera permitted the antigens to be localized in the peripheral areas of the neuron nuclei and in the mitochondria. The results obtained with the immunofluorescent technique were in agreement with those produced immunohistochemically.

Effect of dehydration of the cornea with experimentally induced ulcer on collagenase activity.

In the course of healing of experimental ulcer one could observe a distinct difference between a cornea, subjected to the action of glycerol and a control one. The damage to the chemical structure of collagenous fibers was much smaller than in the control. On the basis of the investigations performed one may conclude that glycerol, through dehydration of the corneal tissue and stimulation of mucopolysaccharide synthesis, suppresses the activity of collagenase. In polarized light this process was manifested by the double refraction phenomenon and in clinical observation--by an accelerated healing of ulcer.

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.

Influence of atropine and carbachol on the fluorescence of catecholaminergic structures in selected areas of the rat brain.

Using the formaldehyde-fluorescence technique, the authors studied the influence of atropine and carbachol, administered intraventricularly to Wistar rats, on the fluorescence of catecholaminergic structures in 20 areas of the CNS, situated within the range of the 10th-46th frontal plane according to KONIG and KLIPPEL. 1. A confirmation of the antagonistic action of atropine and carbachol was obtained. It was expressed by mutually opposed occurrence of the specific fluorescence of the catecholaminergic structures. 2. In 16 out of 20 studied areas of the CNS, carbachol abolished or considerably weakened the specific fluorescence. In 3 areas it was increased by this drug, and one area proved insensitive. 3. Atropine increased the specific fluorescence in the DA (dopaminergic system) areas, while it had varying effects in the NA (noradrenergic system areas. In some areas of the CNS it increased and in others reduced the specific fluorescence of the catecholaminergic structures. 4. An interference between atropine and carbachol is observed, but it seems that the results of the present experiment speak in favour of an interaction between the catecholamine transmitters and ACh in the particular areas of the CNS under the influence of atropine and carbachol. 5. The authors discuss in detail the reactions of the catecholaminergic structures in the particular areas of the CNS, in which, as compared with the control, an increase or a decrease of the specific fluorescence under the influence of the administered drugs was observed.

The influence of phentolamine on the fluorescence of catecholaminergic structures in selected areas of rat brain.

Wistar rats were given phentolamine into the ventriculus lateralis. The D1 group of rats received a larger dose than the D2 group. The animals were decapitated within 2 hours after phentolamine injection. The FALCK fluorescence technique was applied to demonstrate the fluorescence of catecholaminergic structures. The results in rat brain areas selected according to KONIG and KLIPPEL are shown in figs 10-46. Whereas earlier biochemical experiments did not show any phentolamine-induced changes in the NA level of the whole brain, the present histochemical experiments carried out with the fluorescence technique revealed the influence of phentolamine on individual structures and areas of the NA system. In comparison with the control material in 5 out of 11 areas the fluorescence was much weaker, in 3 it was similar to the control group, and in 3 (1 with a larger dose and 2 with a smaller dose of phentolamine) it was slightly stronger. The simultaneous increase of fluorescence in 6 out of 7 areas of the DA system may indicate a compensatory interaction of these areas as a response to the neuromediator decrease in the NA system. The paper discusses the increase in the intensity of fluorescence induced by a small dose of phentolamine in some brain areas, or by a large dose in others, both these alternatives depending on the neuromediator turnover.