Involvement of cell surface TG2 in the aggregation of K562 cells triggered by gluten.

Gluten-induced aggregation of K562 cells represents an in vitro model reproducing the early steps occurring in the small bowel of celiac patients exposed to gliadin. Despite the clear involvement of TG2 in the activation of the antigen-presenting cells, it is not yet clear in which compartment it occurs. Herein we study the calcium-dependent aggregation of these cells, using either cell-permeable or cell-impermeable TG2 inhibitors. Gluten induces efficient aggregation when calcium is absent in the extracellular environment, while TG2 inhibitors do not restore the full aggregating potential of gluten in the presence of calcium. These findings suggest that TG2 activity is not essential in the cellular aggregation mechanism. We demonstrate that gluten contacts the cells and provokes their aggregation through a mechanism involving the A-gliadin peptide 31-43. This peptide also activates the cell surface associated extracellular TG2 in the absence of calcium. Using a bioinformatics approach, we identify the possible docking sites of this peptide on the open and closed TG2 structures. Peptide docks with the closed TG2 structure near to the GTP/GDP site, by establishing molecular interactions with the same amino acids involved in stabilization of GTP binding. We suggest that it may occur through the displacement of GTP, switching the TG2 structure from the closed to the active open conformation. Furthermore, docking analysis shows peptide binding with the ß-sandwich domain of the closed TG2 structure, suggesting that this region could be responsible for the different aggregating effects of gluten shown in the presence or absence of calcium. We deduce from these data a possible mechanism of action by which gluten makes contact with the cell surface, which could have possible implications in the celiac disease onset.

Colloidal mercury (Hg) distribution in soil samples by sedimentation field-flow fractionation coupled to mercury cold vapour generation atomic absorption spectroscopy.

Diverse analytical techniques are available to determine the particle size distribution of potentially toxic elements in matrices of environmental interest such as soil, sediments, freshwater and groundwater. However, a single technique is often not exhaustive enough to determine both particle size distribution and element concentration. In the present work, the investigation of mercury in soil samples collected from a polluted industrial site was performed by using a new analytical approach which makes use of sedimentation field-flow fractionation (SdFFF) coupled to cold vapour generation electrothermal atomic absorption spectroscopy (CV-ETAAS). The Hg concentration in the SdFFF fractions revealed a broad distribution from about 0.1 to 1 µm, roughly following the particle size distributions, presenting a maximum at about 400-700 nm in diameter. A correlation between the concentration of Hg in the colloidal fraction and organic matter (O.M.) content in the soil samples was also found. However, this correlation is less likely to be related to Hg sorption to soil O.M. but rather to the presence of colloidal mercuric sulfide particles whose size is probably controlled by the occurrence of dissolved O.M. The presence of O.M. could have prevented the aggregation of smaller particles, leading to an accumulation of mercuric sulfides in the colloidal fraction. In this respect, particle size distribution of soil samples can help to understand the role played by colloidal particles in mobilising mercury (also as insoluble compounds) and provide a significant contribution in determining the environmental impact of this toxic element.

New online management software and DICOM viewer for dentistry.

The way we collect, store, and share dental records of our patients is rapidly becoming digital. Many programs have been designed to run on a single computer or local network to handle various tasks, so selecting a system can be complex; it can require high acquisition costs, update fees, and installation difficulties. The purpose of this article is to explain the architecture, characteristics, and advantages related to Web-based Management Software. In particular, this article describes the first Web-based Electronic Medical Record (DocSapiens.com), which is able to view and edit DICOM files directly online.

Responses of Trapa natans L. floating laminae to high concentrations of manganese.

The present study focuses on the responses of floating laminae of the Mn-tolerant hydrophyte Trapa natans L. to 1 mM Mn and their ability to accumulate the metal. Studies were carried out first on young floating laminae belonging to the second verticil of 30-day-old plants which originated from fruits that had been maintained in a 1 mM Mn-treated environment and again on the young floating laminae after 10 days of further treatment with 1 mM Mn. Mn storing was observed from the first days after germination, but only 10-day-treated laminae showed the capability to hyperaccumulate the element inside specialised cells (>20000 microg/g [dry weight]). Electron microscopy and the Folin-Ciocalteu reaction for phenolics revealed deposits of chelated material inside vacuoles of the first palisade layer and of idioblasts in the spongy tissue. X-ray microanalysis indicated that the deposits were Mn chelated with phenolic compounds. Numerous trichomes were observed at the lower epidermis of 10-day-treated laminae. They were rich in phenolics and characterised by Mn concretions at their base. As they are associated with a high concentration of the metal in culture water and sediments, trichomes may constitute a morphological differentiation for the secretion of Mn-chelating molecules into the culture water, as a probable "avoidance" mechanism. Finally, monitoring of the photosynthetic apparatus showed that photosynthetic function was not impaired, though differences in development occurred.

Development of an SdFFF-ETAAS hyphenated technique for dimensional and elemental characterization of colloids.

Direct hyphenation of electrothermal atomic-absorption spectroscopy (ETAAS) to sedimentation field-flow fractionation (SdFFF) has been developed to enable elemental characterization of submicron particles as a function of size. This hyphenation is particularly suitable for characterizing colloidal particles of environmental interest, for example water-borne particles. The interface is an automatic capillary injection device (CID) which enables direct introduction of large and variable volumes of colloidal particle suspensions into a hot graphite furnace, thus preconcentrating the colloidal particles on the furnace walls. The method was validated by determination of Fe in certified submicron Fe2O3. The procedure was set up by first optimizing the SdFFF fractionation under programmed field conditions, thus enabling optimum fractionation of particle size. The ETAAS procedure was then tested to determine whether it could be used for direct analysis of Fe2O3 slurries without the need for a mineralization step. CID coupled to ETAAS was subsequently exploited for its ability to enhance the sensitivity, because of the increased injection volume. Statistical tests and data handling were conducted to prove the suitability of the ETAAS-CID module. Finally, off-line and on-line ETAAS-CID-SdFFF hyphenation were investigated. These experiments emphasized the advantages of the on-line coupling, because it enables synchronized sampling, enrichment, and elemental analysis of the flowing eluate. The benefits of the proposed hyphenation are the high specificity of analytical detection, increased sensitivity, reduction of analysis time, and minimum sample handling and contamination.

Adaptive modifications of the photosynthetic apparatus in Euglena gracilis Klebs exposed to manganese excess.

Asynchronous cultures of wild-type Euglena gracilis were tested for their morphophysiological response to 10 mM MnSO4. Growth was only moderately slowed (15%), while oxygen evolution was never compromised. Inductively coupled plasma analyses indicated that the Mn cell content doubled with respect to controls, but no signs of localised accumulation were detected with X-ray microanalysis. Evident morphological alterations were found at the plastid level with transmission electron microscopy and confocal laser scanning microscopy. An increase in the plastid mass, accompanied by frequent aberrations of chloroplast shape and of the organisation of the thylakoid system, was observed. These aspects paralleled a decrease in the molar ratio of chlorophyll a to b and an increase in the fluorescence emission ratio of light-harvesting complex II to photosystem II, the latter evaluated by in vivo single-cell microspectrofluorimetry. These changes were observed between 24 and 72 h of treatment. However, the alterations in the pigment pattern and photosystem II fluorescence were no longer observed after 96 h of Mn exposure, notwithstanding the maintenance of the large plastid mass. The response of the photosynthetic apparatus probably allows the alga to limit the photooxidative damage linked to the inappropriately large peripheral antennae of photosystem II. On the whole, the resistance of Euglena gracilis to Mn may be due to an exclusion-tolerance mechanism since most Mn is excluded from the cell, and the small amount entering the organism is tolerated by means of morphophysiological adaptation strategies, mainly acting at the plastid level.

Specific intra-tissue responses to manganese in the floating lamina of Trapa natans L.

Plant tolerance to heavy metals requires morpho-physiological mechanisms that are still poorly understood, especially in hydrophytes. This study focuses on the young floating lamina of the rhyzophyte Trapa natans exposed for 10 d to 130 microM Mn. The lamina has the ability to bioaccumulate Mn (> 3000 microg g(-1)). X-ray microanalysis of Mn cellular distribution revealed accumulation in the upper epidermis, in the first palisade layer, and in the idioblasts of the spongy tissue, which were shown with electron microscopy to contain osmiophilic vacuolar deposits, also observed to a minor extent in the control leaves. On the basis of biochemical and histochemical tests, these deposits were attributed to phenolic compounds that were probably able to chelate Mn. Net photosynthesis, photosynthetic pigments, room temperature microspectrofluorimetric analyses, and ultrastructural studies of plastids were performed to evaluate the status of the photosynthetic apparatus. A greater development of thylakoid membranes was observed in plastids of the second palisade and spongy tissue, which, however, did not accumulate Mn. Only the spongy tissue experienced inadequate assembly of PS II, but this did not significantly influence the photosynthetic yield of the whole lamina. It was concluded that T. natans can optimise productivity in the presence of Mn by means of specific intra-tissue responses within the framework of the floating lamina.

Lack of burning and stinging from a novel first-aid formulation applied to experimental wounds.

Antiseptic-anesthetic first aid formulations typically produce a burning or stinging sensation when used on wounds. The aim of this study was to assess the lack of sting/burn potential of a prototype first-aid wipe when applied to superficial wounds. This was a one-day, double-blind, randomized study, evaluating the stinging/burning sensation from a prototype first-aid wipe with a cellulose base containing 1.0% pramoxine HCL and 0.13% benzalkonium chloride. The study followed a direct comparison test design of four test formulations. The remaining three articles were: (a) sterile 0.9% sodium chloride (no-sting/no-burn control), (b) 3% hydrogen peroxide (first-aid sting/burn control), and (c) 70% isopropyl alcohol (second sting/burn control). The test was performed on 24 subjects. The tape stripping method was used to create four standardized wounds on the volar forearms, reaching the glistening layer. Each test article was applied to the wound for 15 seconds. The subjects were asked to report the intensity of the stinging/burning sensation during the application. The prototype pramoxine-benzalkonium chloride wipe produced significantly less stinging/burning than both 70% isopropyl alcohol and 3% hydrogen peroxide. Also, the prototype wipe did not produce more stinging and burning in superficial wounds compared to the saline control.

A multi-sample design for assessing the comedolytic activity of topical products.

BACKGROUND: Facial comedolytic studies allow testing of a maximum of two products. The upper back provides a larger area with a more uniform distribution of microcomedones for comedolytic studies. OBJECTIVE: To design a multi-sample method for assessing comedolytic activity of topical products on the back. The effect of season on product discrimination was also explored. METHODS: Three cleansing formulations (products B, C and D), a negative water control and 0.025% Retin-A cream (positive control) were tested. Seven subjects were recruited in summer and nine in fall. Products were applied for 8 weeks, comedolysis was assessed by visually evaluating cyanoacrylate follicular biopsies taken at baseline and post-treatment. RESULTS: In all data sets (summer, fall and combined), sites treated with Retin-A had a significantly lower number of microcomedones as compared to the negative water control. In addition, cleansers B and D showed a significant reduction from baseline in the fall and combined (summer and fall) data, but not in summer data alone indicating different responses to treatment during the year. CONCLUSIONS: The design was sensitive enough to detect differences between cleansing formulations under normal washing conditions. Reduced sensitivity observed during summer suggests hot humid conditions may decrease the comedolytic performance of topical products.

Analysis of patient flows via data mining.

The paper presents DoMiner, a data mining tool for the analysis of patient flows among public hospitals and care units. DoMiner is based on the theory of rough sets, and allows for the extraction of association rules from data base tables. The paper describes both the clustering and rule extraction algorithm of DoMiner, and illustrates an introductory analysis example.

Digital fluorescence photography can assess the suppressive effect of benzoyl peroxide on Propionibacterium acnes.

BACKGROUND: Porphyrins produced by Propionibacterium acnes exhibit an orange-red fluorescence under UVA light. The amount of fluorescence can be estimated by digital fluorescence photography. OBJECTIVE: We thought that digital fluorescence photography would be a quicker and simpler method than bacteriologic culture to demonstrate depopulation of P acnes in sebaceous follicles. We used benzoyl peroxide to bring about rapid suppression of P acnes. METHODS: Benzoyl peroxide 10% was applied twice daily for 7 days to the faces of 9 subjects. Five subjects were untreated controls. Digital fluorescence photographs of cheek and nose, and scrub samples for quantitative recovery of P acnes from the cheek were taken at baseline, day 3, day 7 (end of treatment), and day 16 (regression phase). RESULTS: The effect of benzoyl peroxide against P acnes was clearly demonstrated both by culture and by fluorescence photography after only 3 days. Image analysis of porphyrin fluorescence correlated well with the decrease in P acnes density from scrub cultures. No further decrease was observed at day 7 (end of therapy). Ten days later there was a return to baseline values, although in some subjects these remained lower. CONCLUSION: Digital fluorescence photography is a reliable, fast, and easy screening technique to demonstrate the suppressive effect of topical antibacterial agents on P acnes.

Hypopigmented macules of photodamaged skin and their treatment with topical tretinoin.

Hypopigmented macules are frequently observed in the photodamaged skin of elderly people. We undertook to study and treat 2 types of hypomelanosis of photoaged skin. These lesions were: 1) idiopathic guttate hypomelanosis; and 2) macular hypomelanosis. Comparative studies included: 1) high-resolution photography using parallel polarized light, ultra-violet (UVA) and epiluminescence; 2) Silflo replicas for microtopography; and 3) suction device (Cutometer) for elasticity. Macular hypomelanosis was distinguishable from idiopathic guttate hypomelanosis because the macules were less white and less well demarcated. Glyphic markings were essentially absent in macular hypomelanosis, but variably effaced in idiopathic guttate hypomelanosis. Distensibility of the macules was characteristically low in proportion to the loss of glyphic markings. The chief histologic finding was the absence of melanin in basal keratinocytes. Macular hypomelanosis and idiopathic guttate hypomelanosis are probably related disorders along a spectrum of depigmentation. Treatment with tretinoin for 4 months restored the elasticity, the glyphic markings, with a partial restoration of pigmentation.

Lack of correlation of skin thickness with bone density in patients receiving chronic glucocorticoid.

The objective was to test the hypothesis that there is a correlation between thinning of the skin and bone in patients on chronic oral glucocorticoids (GCs). This was a one-time cross-sectional analysis performed in an academic referral center. The study group consisted of 14 patients on GCs for a variety of disorders, including dermatomyositis, pemphigus vulgaris, pyoderma gangrenosum, and urticarial vasculitis. Skin thickness was compared with that of 24 sex- and age-matched controls. The main outcome measures were the bone density of the lumbar spine (L2-L4) and the skin thickness. The skin thickness (mm, mean +/- SEM) in GC-treated (n = 7) vs unmedicated age-matched Caucasian women (n = 20) was 0.84 +/- 0.04 vs 1.02 +/- 0.04 (t = 3.07, P < 0.01) in the upper arm, 1.13 +/- 0.09 vs 1.49 +/- 0.05 (t = 3.65, P < 0.002) in the dorsal forearm, and 0.96 +/- 0.07 vs 1.17 +/- 0.02 (t = 2.92, P < 0.01) in the ventral forearm. L2-L4 bone densities averaged 106 +/- 2% in the GC-treated female patients relative to the age and sex-matched controls. There was no correlation between skin thickness and bone density. In GC-treated (n = 4) vs unmedicated Caucasian men matched for age (n = 4), skin thickness was 1.09 +/- 0.4 vs 1.33 +/- 0.05 (t = 3.51, P < 0.02) in the upper arm, but was not significantly different at the two forearm sites. No correlation between skin thickness and bone density was observed. The level of type I procollagen mRNA in skin from three GC-treated patients was 45% of the value in three age-matched controls. In conclusion, GCs cause statistically significant thinning of skin independently of the effects on bone.

A model to assess the efficacy of moisturizers--the quantification of soap-induced xerosis by image analysis of adhesive-coated discs (D-Squames).

Clinical grading of dry skin syndromes (xerosis) is beset by many variables which limit its reliability. Removing scales onto adhesive coated discs (D-Squames) has provided a means for objective measurement of xerotic states. Using image analysis we have modified the basic procedure to assure greater accuracy and reproducibility. The distribution of grey values on the discs was determined, from which a single value of the degree of scaling was calculated. We term this the scaling index. To induce xerosis, the lower legs of 22 women, aged 26-64, were washed daily for 10 days with soap. After washing, one side received a moisturizing lotion, the other served as an untreated control. At specified intervals, scaling was graded clinically, electrical conductance was measured and D-Squames were obtained. On the moisturizer-treated legs there was a significant decrease in dryness grades and scaling indices at all time points. Conductance was significantly increased on days 8 and 11. The untreated washed legs showed no changes in clinical grades, although conductance significantly decreased and the scaling indices increased in the last stage of the study. D-Squame analysis unequivocally revealed the beneficial effect of a moisturizer on soap-induced xerosis and the expected increase in scaliness without treatment.

Determination of density of follicles on various regions of the face by cyanoacrylate biopsy: correlation with sebum output.

The density of follicles on various regions of the human face has received scant attention. We used cyanoacrylate follicular biopsies to determine the number of follicles on the forehead, cheek, chin and nose of 12 healthy adult white women. Sebum output was assessed on the same regions by means of Sebutape. The density of follicles and sebum output followed a centrolateral decreasing gradient. There was no correlation between these two parameters. These regional patterns are important for studies of the pathogenesis of common disorders of the face and their response to treatment.

Echographic evaluation with image analysis of normal skin: variations according to age and sex.

In order to identify and characterize the sonographic variations between different age groups, 48 subjects, 24 aged 27-30, and 24 over 60, were studied with a 20-MHz B scanner on six skin sites. Images were evaluated by the instrument's standard programme and by a new image analysis software package enabling the characterization of steady structures or transitory functional aspects of skin reactions, by highlighting areas in which the echo amplitudes are included within selected values, and by calculating their extension. Three bands were selected as intervals of interest, respectively, highlighting hyporeflecting parts of the dermis, tissue reflecting with intermediate amplitude values and hyperreflecting epidermis and dermis. This method was employed to assess skin thickness, demonstrating its decrease in elderly skin, to characterize and quantify the hypoechogenic subepidermal band appearing in the elderly at volar and dorsal forearm skin, and to evaluate echogenicity of the upper and lower dermis. Our data show that there is a great regional variation in the behaviour of ultrasound reflection of elderly skin in respect to the skin of young subjects. However, a general trend can be identified, consisting in a shift from low-intensity ultrasound echoes, characteristic in the dermis of young subjects, to intermediate or high reflection amplitudes, which are more frequent in elderly skin. Thus, the echographic method provides two parameters for the evaluation of skin aging.(ABSTRACT TRUNCATED AT 250 WORDS)

Long-term results of combined chemotherapy-radiotherapy approach in Hodgkin's disease: superiority of ABVD plus radiotherapy versus MOPP plus radiotherapy.

In an attempt to reduce some of the delayed sequelae associated with combined modality therapy in Hodgkin's disease, we randomly tested stages IIB, IIIA, and IIIB MOPP (mechlorethamine, vincristine, procarbazine, and prednisone) v ABVD (Adriamycin, bleomycin, vinblastine, and dacarbazine). In 232 previously untreated patients, three cycles of either combination preceded and followed extensive irradiation. The complete remission rate was 80.7% following MOPP and 92.4% following ABVD (P less than .02). The 7-year results indicated that ABVD was superior to MOPP in terms of freedom from progression (80.8% v 62.8%; P less than .002), relapse-free survival (87.7% v 77.2%; P = .06), and overall survival (77.4% v 67.9%; P = .03). Moreover, the comparative iatrogenic morbidity showed that irreversible gonadal dysfunction as well as acute leukemia occurred only in patients subjected to MOPP, while cardiopulmonary studies failed to document significant laboratory differences between the two treatment groups. Present findings indicate that ABVD followed by extensive irradiation represents a valid therapeutic alternative to the widely used alkylating agent-containing regimens plus radiotherapy.

Ambroxol and pulmonary toxicity induced by antineoplastic drugs.

The authors describe the potential effects of ambroxol on the pulmonary disorders induced by antineoplastic agents (in particular, bleomycin and the nitrosureas). An experimental stage focussed attention on the early modifications occurring in the alveolar surfactant and in the afflux of inflammatory and immune-effector cells following bleomycin-induced lung fibrosis in the rat (by intratracheal instillation). The ambroxol-protected rats showed a slower drop of alveolar lecithins in the first few hours after bleomycin administration and a lower afflux of neutrophils, macrophages and lymphocytes. In the clinical stage, respiratory function was studied in two groups of cancer patients treated with nitrosureas or bleomycin. Preliminary findings indicate a rapid worsening of some functional parameters--maximal expiratory flow at 25% vital capacity, diffusing capacity for carbon monoxide and diffusing capacity/ventilation--in controls, while no such changes occurred in the ambroxol-protected subjects. The possible pathogenetic implications of these results and perspective for future investigations are discussed.