Sulphurous thermal water increases the release of the anti-inflammatory cytokine IL-10 and modulates antioxidant enzyme activity.

The beneficial effects of hot springs have been known for centuries and treatments with sulphurous thermal waters are recommended in a number of chronic pathologies as well as acute recurrent infections. However, the positive effects of the therapy are often evaluated in terms of subjective sense of wellbeing and symptomatic clinical improvements. Here, the effects of an S-based compound (NaSH) and of a specific sulphurous thermal water characterized by additional ions such as sodium chloride, bromine and iodine (STW) were investigated in terms of cytokine release and anti-oxidant enzyme activity in primary human monocytes and in saliva from 50 airway disease patients subjected to thermal treatments. In vitro, NaSH efficiently blocked the induction of pro-inflammatory cytokines and counterbalanced the formation of ROS. Despite STW not recapitulating these results, possibly due to the low concentration of S-based compounds reached at the minimum non-toxic dilution, we found that it enhanced the release of IL-10, a potent anti-inflammatory cytokine. Notably, higher levels of IL-10 were also observed in patients' saliva following STW treatment and this increase correlated positively with salivary catalase activity (r2 = 0.19, *p less than 0.01). To our knowledge, these results represent the first evidence suggesting that S-based compounds and STW may prove useful in facing chronic inflammatory and age-related illness due to combined anti-inflammatory and anti-oxidant properties.

[SOSIA classification of the frail elderly in nursing homes of region of Lombardy]. / La Classificazione SOSIA degli anziani ospiti delle residenze sanitario-assistenziali lombarde.

Since 2001, the region of Lombardy has accreditated nursing homes named R.S.A. "Residenza Sanitario-Assistenziale", which make up Italy's principal RSA network with the annual turnover of approximately 60,000 people. The most noteworthy element of the reform introduced is a concentration on resident's frailty rather than disability. This is assessed by using SOSIA, a form for intermediate observation of assistance. Residents are classified into eight classes of frailty, called isofrailty classes of SOSIA, which are differentiated by how compromised their motor and cognitive skills are, and by the presence of comorbidity. The study presents the methodology used to identify and estimate cut-offs of three indicators employed in SOSIA classification. It also discusses their characteristics versus other evaluation systems, such as Resource Utilization Group RUG-III and Autonomie Gerontologique--Groupes Iso-Resources AGGIR.

Chondrocyte aggregation and reorganization into three-dimensional scaffolds.

Articular cartilage has a very limited self-repairing capacity; thus, chondral lesions normally result in chronic degeneration and, eventually, osteoarthritis development. Currently, tissue engineering offers a new tool for the clinical treatment of osteochondral defects. The present investigation aimed to develop an in vitro engineered cartilage using a new class of semisynthetic scaffolds. Two nonwoven meshes of hyaluronan esters (Hyaff(R) derivatives) were seeded with sternal chick embryo chondrocytes cultured for up to 21 days, after which time they were assessed for both the cellular growth profile and histological features. Avian chondrocytes easily adhered and proliferated onto hyaluronan-based scaffolds, demonstrating a significant preference for the fully esterified benzylic form. Histochemical staining revealed the presence of a neosynthesized glycosaminoglycan-rich extracellular matrix, and immunohistochemistry confirmed the deposition of collagen type II. Moreover, ultrastructural observations supported evidence that chondrocytes grown onto a hyaluronan-derived three-dimensional scaffold maintained their unique phenotype and organization in a cartilage-like extracellular matrix. These findings support the further pursuit of a transplantable engineered cartilage using human chondrocytes for the regeneration of chondral lesions.

[Hemostatic and structural effects of the lyophilized cellulose sponge]. / Efeitos hemostático e estrutural da esponja de celulose liofilizada.

Hemostatic effects of oxidized cellulose (Surgicel) are well known. Based on a possible similar effect of a sponge obtained after lyophilization of biosynthetic cellulose, two different experimental studies were planned. Phase I-Pieces of cellulose sponge were inserted into small provoked cortical wounds of twelve dogs. The time elapsed to obtain bloodstill after cortical damage and application of cellulose was observed in every dog, searching to detect any possible hemostatic effect of the material. The animals were sacrificed after 7, 30 and 90 days. An average time of 1 minute was elapsed until bleeding control was achieved. No clinical adverse effect was noticed. Microscopy showed histiocytic and mild foreign body reaction at 7 days, which diminished at 30 days. Almost no reaction surrounded the implant at 90 days. Lyophilized cellulose has a peculiar eosinophilic appearance, composed by thin irregular filaments which diminished their thickness with the time. At 90 days only sparse irregular cellulose filaments could be detected. Phase II-Small equal sponge fragments were inserted in the liver of twelve rats and observed 7, 30 and 90 days. At autopsy, small peritoneal adhesions were noticed at 30 and 90 days. Microscopy showed intense histioplasmocytic and foreign body reaction in all animals mainly at 7 days. In two animals, refringent intracellular cellulose particles were evident inside giant foreign body cells after 90 days. This fact evidences that cellulose can be reabsorbed by phagocytic phenomena when implanted in mammalians. A comparative group with other hemostatic material and the same method must be done to clarify the issue of hemostatic effects of this membrane.

In vitro engineering of human skin-like tissue.

Coverage of large, full-thickness burns presents a challenge for the surgeon due to the lack of availability of the patient's own skin. Currently, tissue engineering offers the possibility of performing a suitable therapeutic wound coverage after early burn excision by using cultured keratinocyte sheets supported by a dermal layer. The aim of this study was to develop and characterize a skin substitute composed of both epidermal and dermal elements. For this purpose we grew keratinocytes and fibroblasts separately for 15 days within two different types of biomaterials. Cells then were co-cultured for an additional period of 15 days, after which samples were taken and processed with either classic or immunohistochemical stainings. Results showed that (1) human fibroblasts and keratinocytes can be cultured on hyaluronic acid-derived biomaterials and that (2) the pattern of expression of particular dermal-epidermal molecules is similar to that found in normal skin. The data from this study suggest that our skin equivalent might be useful in the treatment of both burns and chronic wounds.

Cultured human Langerhans' cells are superior to fresh cells at presenting native HIV-1 protein antigens to specific CD4+ T-cell lines.

Cultured Langerhans' cells (CLC) exhibit enhanced antigen-presenting function compared to freshly isolated LC (FLC), but they are commonly believed to be inefficient at processing intact proteins. In this study, FLC and CLC from normal, human immunodeficiency virus (HIV) seronegative volunteers were compared for their ability to present the HIV-1 envelope glycoprotein gp120 or reverse transcriptase (p66) antigens to autologous, specific CD4+ T cell lines. Epidermal cell suspensions enriched for LC were prepared from suction blister roofs. FLC stimulated T cells at lower antigen concentrations compared to unfractionated peripheral blood mononuclear cells (PBMC). CLC were more potent on a per cell basis than FLC, PBMC or adherent monocytes at presenting native gp120, native p66 or immunogenic peptides. CLC were also more efficient than FLC or PBMC in terms of the amount of antigen required for T-cell activation. Chloroquine and leupeptin inhibited presentation of intact p66, but not of an immunodominant peptide, by FLC or CLC, thus indicating that both cells utilize antigen-processing mechanisms that are based on intracellular acidification and protease activity. Incubation of CLC with monoclonal antibodies against HLA-DR, CD11b, CD18, CD50, CD54, CD58 or CD80, but not anti-major histocompatibility complex class I (MHC-I), inhibited antigen-specific T-cell proliferation to varying degrees. We conclude that human CLC retain the ability to process and present protein antigens potently to CD4+ T cells. Thus, CLC have the capacity to participate actively in the generation and maintenance of T-helper cell immunity to viral antigens during HIV-1 infection.

Functional intercellular adhesion molecule-3 is expressed by freshly isolated epidermal Langerhans cells and is not regulated during culture.

Activation of T lymphocytes by antigen-presenting cells requires the interaction of major histocompatibility complex/antigen complexes with the T-cell receptor as well as the binding of co-stimulatory molecules to receptors on T cells. Freshly isolated epidermal Langerhans cells (LC) do not display a significant number of co-stimulatory molecules. After short-term culture, LC express and then upregulate intercellular adhesion molecule-1 (ICAM-1) (CD54), leukocyte function-associated antigen (LFA)-3 (CD58), and B7-1 (CD80) accessory molecules and exhibit an enhanced antigen-presenting function. The present study examined the presence on human LC of the LFA-1 ligands ICAM-2 (CD102) and ICAM-3 (CD50) and their functional role in the activation of allogeneic T cells. Immunohistochemistry of skin sections and flow-cytometry analysis of freshly procured epidermal cell suspensions showed that LC (CD1a+ or HLA-DR+) expressed ICAM-3 but not ICAM-2. After 48-72-h culture in the presence of granulocyte/macrophage colony-stimulating factor, LC did not stain for ICAM-2 but expressed ICAM-3 at the same level as fresh cells. Incubation of both freshly isolated and cultured LC with monoclonal antibodies directed against ICAM-3 reduced T-cell proliferation (25-75% inhibition) in the primary allogeneic mixed leukocyte reaction assay; incubation of cultured LC with anti-ICAM-1 and anti-ICAM-3 synergistically reduced T-cell response. The results indicate that ICAM-3 is constitutively expressed and represents an important costimulatory molecule on freshly isolated LC but, in contrast to other accessory molecules, is not subjected to regulation during LC culture.

Expression of B7 costimulatory molecule in cultured human epidermal Langerhans cells is regulated at the mRNA level.

Langerhans cells (LC) belong to the dendritic cell lineage and are the principal antigen-presenting cells of squamous epithelia. Short-term cultured LC (cLC) exhibit a marked augmented capacity to stimulate allogeneic T cells and acquire the ability to activate naive T cells, probably in relation to enhanced expression of accessory signals. In this study, we evaluated the expression of B7 costimulatory molecule (CD80) in human freshly isolated (fLC) and cLC at both the protein and mRNA level. Staining of frozen skin sections did not reveal any epidermal dendritic cell reactive with either of two different anti-B7 monoclonal antibodies. fLC in suspension did not exhibit any B7 staining as evaluated by two-color flow-cytometry analysis and immunoelectron microscopy. In contrast, LC that were cultured for 24-72 h displayed strong surface B7 reactivity with a characteristic patchy pattern. Treatment with dispase and trypsin did not reduce B7 staining of cLC. Following warming to 37 degrees C, cLC tagged with anti-B7 monoclonal antibody and gold-conjugated secondary antibody could internalize surface B7 by using the organelles of receptor-mediated endocytosis. B7 mRNA, detected by the reverse-transcriptase polymerase chain reaction technique, was expressed at a low level in purified (> 90% HLA-DR+) fLC but not in LC-depleted epidermal cells, and was markedly upregulated in purified cLC. The results indicate that 1) fLC do not express B7 protein on their surface, but acquire B7 during culture, 2) surface B7 is not sensitive to trypsin, 3) B7 expression is regulated primarily at the mRNA level, and 4) membrane B7 can be internalized within cLC. B7 molecule on CLC may be relevant to their increased antigen-presenting cell potency and ability to stimulate naive T lymphocytes.

[Transvaginal sonography as a screening method for the identification of patients at risk of postmenopausal endometrial pathology]. / La transvaginosonografia (TVS) come metodo di screening per l'identificazione delle pazienti a rischio per la patologia endometriale nella postmenopausa.

Starting from the anatomopathological assumption that endometrial thickness in postmenopausal women never exceeds 3 mm, and in view of the reliability of measurements made using an echographic probe, the authors evaluated the value of transvaginosonography (TVS) as a mass screening method for postmenopausal endometrial pathologies. A group of 74 patients were examined who were recruited from those attending the out-patient menopause clinic. All subjects conformed to the following admission criteria: amenorrhea for the past two years; absence of oestroprogestin therapy for at least six months; absence of vaginal blood loss. A 5 MHz probe was used to measure maximum endometrial thickness on the longitudinal plane; values were divided by two if the surfaces were adjacent. Patients were monitored according to the following protocol: endometrial thickness under 1 mm control every 12 months; thickness between 1-3 mm--control every 3 months; thickness equal to or over 4 mm--hysteroscopy, targeted biopsy, possible scraping and TVS control after 3 months. The group was subdivided as follows: 65 patients (87.8%) were without risk; 3 patients (4%) belonged to the intermediate risk group; 6 patients (8.2%) belonged to the high risk group. Of the latter, 4 revealed an endometrial polyp, one presented uterine polymyomatosis and one a proliferative-type endometrium. The authors' experience is still limited but the absence of false positives encourages them to continue their research using this simple and well tolerated method. It might represent a valid alternative to hysteroscopy as a screening method in the asymptomatic population at risk for endometrial carcinoma.

Phosphatidylserine enhances the ability of epidermal Langerhans cells to induce contact hypersensitivity.

Phosphatidylserine (PS) modulates several immune functions in vitro, including T cell activation, antibody and cytokine production, and macrophage growth. In the present work we studied the effects of PS on the induction of contact hypersensitivity (CH) in mice. BALB/c mice painted with PS (9.4-75 mg/kg) and with a sensitizing dose of DNFB or oxazolone on the same skin site exhibited a dose-dependent augmentation of CH reactions to either DNFB (> 60%) or oxazolone (> 35%), respectively. Bovine brain PS-enriched phospholipid mixture, lyso-PS, and dipalmitoyl-PS also induced similar enhanced CH responses, whereas phosphatidylglycerols had no effect. Increased CH was observed only when PS was applied from 2 days before to 12 h after DNFB. Immunization of naive syngeneic mice with skin grafts that were treated with PS and DNFB also led to enhanced (> 50%) CH responses. In addition, immunization by iv injection of epidermal cell suspensions enriched for Langerhans cells (LC) or of purified LC that were treated with PS (1-100 microM, 30 min, 37 degrees C), and then modified in vitro with DNBS (1 mg/ml, 30 min, 37 degrees C) led to increased (> 30-75%) CH responses in recipient syngeneic animals. Finally, adoptive transfer of DNFB-immune lymph node cells obtained from mice that were treated with PS induced augmented CH responses in recipient animals. The results suggest that PS is capable of up-regulating the induction of CH in mice by stimulating the APC function of epidermal LC.