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In order to quantify and classify ciliary changes of bronchial epithelium in chronic bronchitis, an ultrastructural study on biopsy specimens of 30 patients submitted to bronchoscopy was performed. Fourteen patients were affected by endoscopically and clinically confirmed chronic bronchitis, 8 showed an endoscopic picture of chronic bronchitis but without the clinical features of this pathology, 8 were free of chronic bronchitis. The mean percentage of abnormal cilia in control subjects (1.9 +/- 1.0%) was significantly lower than in patients either with only 'endoscopic' chronic bronchitis (5.7 +/- 2.6%; p less than 0.001) or with 'clinical' chronic bronchitis (14.8 +/- 15.9%; p less than 0.05). Like the ciliary body, the ciliary tip can also show some abnormalities. It is possible to subdivide them in two types: (1) ciliary membrane blebs; (2) ciliary membrane extrusions, variously shaped and sized, with homogeneous content. Bronchial cilia abnormalities become more serious as soon as the clinical picture worsens and this is likely to contribute to the impairment of the mucociliary clearance in chronic bronchitis patients.

It has been demonstrated that cyclosporin A (CyA) blocks the immune system, acts on cytoskeleton and stimulates the production of extracellular matrix (ECM) and transforming growth factor-beta1 (TGF-beta1). This cytokine, such as transforming growth factor-alpha (TGF-alpha), induces deposition of glycosaminoglycans (GAG), proteoglycans and collagen fibres in the ECM.In this work, we examined the effect induced by CyA, TGF-beta1 and TGF-alpha on cultures of healthy and overgrown human gingival fibroblasts in order to evaluate the glycosaminoglycan, cytoskeletal changes and the behaviour of fibroblasts after concanavalin A (Con A) treatment. Moreover, we examined gingival biopsies by Alcian blue histochemical staining and electron transmission microscopy.Total and extracellular sulphated GAG in overgrown gingiva specimens and in derived fibroblast cultures treated with CyA and cytokines were significantly higher than controls. The action of cytokines was increased (Por = 0.01) compared with CyA with a greater effect of TGF-alpha in comparison with TGF-beta1; the electron microscopy showed ECM accumulation. The agglutinations showed the heterogeneity of fibroblast populations.Stimulation with Con A showed that the fibroblast population had cell surface heterogeneity, and could respond in a different way to both CyA and cytokine stimulus. Moreover, increased synthesis of GAG in overgrown gingiva compared with synthesis in normal fibroblasts before CyA treatment suggests a possible genetic origin of damage. As not all CyA-treated patients develop gingival overgrowth, a genetic predisposition may explain the different responses of gingival fibroblast populations.

Serum and erythrocyte levels of the polyamines spermine, spermidine and putrescine, as well as ornithine decarboxylase in erythrocytes, were studied in patients with different neoplasms (breast, lung and colon cancer) and in those with a nonmalignant proliferative disease (familial polyposis). The blood levels of polyamines and the spermine/putrescine ratio were significantly higher in all tumors and in nonmalignant colon polyposis. In erythrocyte ornithine decarboxylase activity, spermine and spermidine levels, as well as spermidine/putrescine and spermine/putrescine ratios showed a significant decrease after surgery and chemotherapy. Our data suggest that high levels of blood polyamines and erythrocyte ornithine decarboxylase activity are related to cell proliferation and cancer treatment, but that levels of polyamines in serum and erythrocytes are still significantly high after cancer treatment and are similar to those in polyposis disease. Polyamines are related to nuclear activity during differentiation; therefore, the altered turnover of polyamines could be a sign of abnormal nuclear function. Since polyamines stimulate protooncogene expression, their high levels could be considered an important cofactor in malignant cell transformation.

The use of growth factors in oral tissue regeneration is currently under investigation. When growth factors are combined with commercial materials, the in vitro mechanisms of action still remain unclear. The present study first evaluated the capacity of barrier membranes, used in oral surgery, to sequester TGFbeta(1). Resorbable HYAFF, paroguide, poly DL-lactide and nonresorbable PTFE membranes were immersed in MEM containing 0.2 ng (125)I-TGFbeta(1) for different periods of time. It was found that HYAFF membrane and paroguide sequestered the most TGFbeta(1), which was then released in its active form (as shown by the CCL64 cell line bioassay). Untreated membranes and membranes enriched with TGFbeta(1) were then used as substrate for human bone cells to evaluate the synthesis of the osteoblast phenotype, as indicated by specific parameters. Results showed that membranes enriched with TGFbeta(1) increased alkaline phosphatase activity, collagen, and osteocalcin production more than untreated membranes. HYAFF and paroguide membranes, which sequestered the most of TGFbeta(1), were the most suitable for stimulating bone matrix proteins.

Cutaneous periarteritis nodosa (PAN) is a clinical feature characterized by chronic, benign course; its pathogenesis is unknown. In patients submitted to renal transplantation cutaneous PAN is a rare complication. We report a case of cutaneous PAN associated with the reappearance of hepatitis B antigen 16 years after kidney transplantation. A 44-year-old man underwent successful renal transplantation in June 1980. In December 1996 he presented multiple painful erythematous subcutaneous nodules on both legs. Skin lesion biopsy showed the presence of cutaneous PAN. Six months later laboratory data demonstrated the presence of HbsAg. HBeAg, HBcAb and detectable HBV-DNA serum by polymerase-chain-reaction (PCR) assay. Anti-HBs and anti-HBe proved negative. In July 1998 the laboratory tests showed an important increase of HBV-DNA (5.1 billion by Branched DNA), and so lamivudine (100 mg/day) was introduced. HBV-DNA became undetectable by PCR after 3 months of therapy. Seven months later a new skin biopsy was performed. The typical signs of PAN were no longer evident. As HBV infecion was demonstrated six months after the clinical appearance of the PAN, in a patient who was believed to be immune to the virus, it is possible that, in the early stages, the hepatitis B antigen title was methodologically indeterminable, but sufficient to form circulating immune complexes responsible for vasculitis primer. Lamivudine therapy resulted efficacious in favouring the regression of cutaneous PAN, but its long-term efficacy requires further evaluation as regards potential selection of drug resistant hepatitis B virus (HBV) mutants during treatment.

Lung branching morphogenesis is a result of epithelial-mesenchymal interactions, which are in turn dependent on extracellular matrix composition and cytokine regulation. Polyamines have recently been demonstrated as able to modify chick embryo skin differentiation. In this work we have examined the effects of putrescine and spermidine during chick embryo lung morphogenesis in organotypic cultures by morphological, histochemical and biochemical examination. To verify the role of polyamines, we used specific inhibitors, such as bis-cyclohexylammonium sulphate and alfa-difluoromethylornithine, and transforming growth factor beta1, an ornithine decarboxylase and polyamine stimulator. Our data show that lung morphogenesis is significantly altered following the induced mesenchymal glycosaminoglycan changes. The increase of mesenchymal glycosaminoglycans is correlated with a stimulation of lung development in the presence of polyamines, and with its inhibition when transforming growth factor beta1 is added to the culture medium. The morphometric data show a uniform increase of both the mesenchyme and epithelial branching with spermidine and putrescine stimulus, whereas the mesenchymal substance alone is significantly increased in apical-median lung sections with transforming growth factor beta1 and transforming growth factor beta1 + spermidine lung cultures. Transforming growth factor beta1 and transforming growth factor beta1 + spermidine confirm the blocking of epithelial branching formations and fibroblast activation, and show that polyamines are unable to prevent the blocking of epithelial cells due to the inhibitory effect of transforming growth factor beta1.

Implanting an expander in the subcutaneous layer causes gradual expansion and provides additional tissue for reconstruction of tissular defects. The force applied remodels the connective tissue and modifies dermis contractibility in additional tissue. Other authors confirm that parameters such as mitosis and hyaluronan influence the system in the tissue regeneration processes. We studied histochemical and morphological variations of tissue expanders before and 6 months after transplant. Our histochemical data do not show any changes in dermis glycosaminoglycans of the expanded and transplant-expanded skin when compared to controls. Morphological data demonstrate reorganization of connective fibers and disappearance of the papillar layer. The latter is not yet formed in the expanded skin 6 months after transplant. This suggests that a long time is required for biological reconstruction of epidermal-dermal interactions after transplant.

In order to have a model for studying the possible implications of 2-ethylhexyl-phthalate and dialysate on connective tissue, we evaluated their direct effects on the air pouch lining tissue and on fibroblast cultures. Air pouches were formed on the backs of 60 ten-week-old Wistar rats by subcutaneous injections of 10 ml sterile air. On the tenth day 2 ml sterile air, or 2 ml 5 microg/L or 2 ml 10 microg/L 2-ethylhexyl-phthalate in olive oil, or 2 ml olive oil alone, or 2 ml 5 mg/ml or 12 mg/ml lyophilized dialysate were injected into the air pouches. After sampling at seven or twenty-one days, the rats were killed. The biochemical data showed an increase in sulphated glycosaminoglycans with 2-ethylhexyl-phthalate and dialysate. Electron microscopy findings revealed cellular alterations such as vacuolation and cell remnants with 2-ethylhexyl-phthalate, while the cells of the air pouches treated with dialysate showed regular organelles with increased and dilated cisternae of rough endoplasmic reticulum. Moreover, an increase in collagen fibres surrounding the damaged zones was noticed in 2-ethylhexyl-phthalate and dialysate treated rats. The glycosaminoglycan modifications and collagen fibre increase seem to suggest that the morphological changes, with the features of fibrosis, could be the result of 2-ethylhexyl-phthalate and dialysate action on connective tissue. Moreover, the air pouch technique can be considered a good model for studying the direct effects of 2-ethylhexyl-phthalate and other substances, such as uremic toxins, on connective tissue.

Patients treated with immunosuppressive drugs can develop cancers. The authors present two cases of Kaposi's sarcoma in kidney transplant patients who had been treated with azathioprine, steroids and cyclosporin-A; during this treatment the Langerhans cells decreased and Kaposi's sarcoma appeared. Discontinuation or reduction of the dosage of cyclosporin-A led to complete regression of the illness, and the Langerhans cells reappeared. We suggest that cyclosporin-A damages the immunological function of the epidermal Langerhans cells, and that this is the primary factor in the development of Kaposi's sarcoma.

The direct effects of D(2-ethylhexyl)phthalate (DEHP), on subcutaneous tissue were studied in the rat, using the air pouch technique. Two ml of DEHP either undiluted or diluted in olive oil (10 microm/ml), were introduced into a previously created air pouch in the rats. The air pouch was removed and specimens of the lining tissue examined by optical and electron microscopy. The persistence of DEHP for 7 days is sufficient to cause alterations in the tissue. The major histological changes were the destruction of the tissue organization with cytoplasmic and nuclear alterations resulting in apoptotic bodies. The histochemical data showed an increase of sulphated glycosaminoglycans, showing proof of alterations in fibroblast functions. The authors suggest that the hyperplastic-type alteration induced by DEHP may be connected with the sclerosis of the peritoneal membrane described in uremic patients in CAPD.

In this study we have examined the relationship between growth factor-induced proliferation and ODC/polyamine levels. TGF beta promotes cell growth and enhances [3H]-thymidine incorporation in chick embryo fibroblasts maintained in a serum-depleted medium. The action on DNA synthesis declines in the second day of treatment. IL-1 does not affect proliferation or [3H]-thymidine incorporation either when it is added alone or in combination with TGF beta. The response of the cells to TGF beta is associated with a significant stimulation of ODC activity and Put, Spd levels together with an enhancement of the Spd/polyamines ratio. IL-1, which does not act on cell proliferation, fails to activate ODC and to increase polyamine levels, thus indicating that the ODC/polyamine system is most likely to be an important link in the chain of events that leads to growth factor-induced proliferation.

Placenta and amnion were analyzed to ascertain the presence of antigens in common with red blood cells (RBC) from cord or fetuses. The expression of distinct antigens displayed on a subpopulation of cord RBC and detected by anticord RBC membrane antibodies was particularly investigated, concomitantly with the presence of transferrin receptors (TR) by employing immunohistochemistry. The placenta showed both cord antigen and TR; on the contrary, amnion--which was labelled by anti-cord RBC membrane antibodies--was not stained by the anti-TR antibody. The results of inhibition and double labelling assays further excluded TR as the relevant antigen in the labelling of both placenta and amnion. The staining of fetal membranes disappeared after absorption of antibodies with cord RBC membranes. These results suggest that the antigens externally expressed on a subpopulation of cord RBC are shared by amnion and placenta.

A study was carried out to obtain a more detailed picture of the phenotypes of human otosclerotic and normal bone cells and to analyse the response of both populations to treatment with TGF beta 1. Total collagen synthesis was found to be decreased, but fibronectin secretion increased in otosclerotic with respect to normal cells. Although overall glycosaminoglycan (GAG) synthesis was lower in otosclerotic cells, the sulphated GAG to hyaluronic acid (HA) ratio was higher, in particular there was greater expression of chondroitin (CS) and dermatan sulphates (DS). TGF beta 1 induced a more marked increase in collagen and fibronectin release and greater production of sulphated GAGs as DS and heparan sulphate (HS) in the otosclerotic cells. The fact that the phenotype of the otosclerotic cells differed from that of the normal cells and could be modified by TGF beta 1 treatment, suggests that TGF beta 1 is implicated in the pathogenesis of otosclerosis.

Aliphatic polyamines, putrescine, spermine and spermidine present in bacteria and eukaryotic cells are essential for cell growth. Generally, polyamine levels are elevated in rapidly growing normal and pathological systems. Since polyamines belong to the category of molecules whose synthesis is strongly activated during the G1 period they have been implicated in the cell's preparation for DNA replication. In our experiments, we have differentiated by phorbol-ester, the human pre-B leukemic cell line KM-3. Biochemical and flow cytometric analysis show an increase, in treated cells, of polyamines pathway related to G1 and early S-phase of cell cycle during B cell differentiation.

The administration of bis-cyclohexylammonium sulphate (BCHS), an inhibitor of spermidine synthase, to cultured chick embryo fibroblasts provoked alterations in cell morphology, a marked disorganization of microfilaments and changes in microtubule network structure. In addition, the rate of microtubule reappearance, after disrupting them with colchicine, was impaired by BCHS. These responses to BCHS were prevented by spermidine addition, which thus suggests an involvement of spermidine in microtubule and microfilament organization.

Unsuspected abnormalities are occasionally detected during real-time examinations. To determine the frequency of such findings detected during the standard ultrasound examination of the entire abdomen, the records of 765 consecutive patients were reviewed. A significant number of occult abnormalities were detected: approximately one-fourth of all abnormalities were found in the abdominal survey. The influence of the abdominal survey on the clinical management and improved radiological consultation are discussed.

In order to define the lifespan of heterozygous beta thalassemic subjects as compared to non thalassemic subjects, the authors prospectively studied all patients deceased during a period of 76 months at St. Camillo Hospital of Comacchio (Ferrara), located in the Po delta, a region with high prevalence of thalassemia minor. No statistically significant differences were found in the average life expectancy between the thalassemic and non thalassemic groups.

The authors present a case of Henoch-Schönlein purpura in a young soldier (19 years old) which they consider important for its etiology and the length of its oligoanuric phase. The syndrome followed a Salmonella hirschfeldii infection, and a protracted oligoanuric phase was followed by nephrotic syndrome and selective glomerular proteinuria which lasted for 1 year. The young man recovered after the eradication of the Salmonella. It seems possible that there was an overall anomalous regulation of the 'lymphoid system of the mucosa', perhaps dependent on a genetic predisposition.

Information concerning the developmental mechanisms involved in skin differentiation have been mainly derived from in vitro experiments. We have previously observed that 6-day chick embryonic thigh skin keratinizes in vitro in chicken serum-containing medium, but does not do so in chick embryo extract-containing medium. This system seems adequate in many respects for investigating the mutual regulative relationship between epithelium and mesenchyme (so called epithelio-mesenchymal interactions). We have therefore attempted better to define the time course of differentiative events and their dependence upon the time of serum administration. Skin explants have been sequentially supplemented with differentiation-stimulating or non-stimulating nutrient, removed at different intervals and examined by histological and histochemical procedures. Different epithelial and mesenchymal behaviour results according to serum supplementation time. Serum administered in the first 48 h of in vitro maintenance is unable to stimulate either the subsequent epidermal keratinization or the correlated changes in dermal histochemical pattern. On the other hand, serum-containing medium induces keratinization and changes in dermal intercellular composition (glycoproteins accumulate to a greater extent than glycosoaminoglycans, as does hyaluronic acid relative to chondroitin sulphuric acids), if added for only the second two days. In the last 48 h, serum may promote epidermal keratinization provided that in vitro incubation has been prolonged. Administration of actinomycin D with serum in the second 48 h prevents epidermal keratinization and modifies dermal histochemical reactivity, but is ineffective if added in the last 48 h. The above findings demonstrate that serum factor(s) stimulating epidermal differentiation act(s) on skin explants in the central incubation period and that correlatively mesenchyme acquires a characteristic histochemical pattern, supporting the possibility that the effect of serum may be mediated by changes in the composition of dermal ground substance.

A rapid high-performance liquid chromatographic method for the direct assay of the taurine and glycine conjugated bile acids in human gastric juice is described. After extraction with Sep-Pak C(18) cartridges, compounds are baseline resolved on a reversed-phase column and detected by UV absorption. The procedure is linear from 10 micromol l(-1) to 1200 micromol l(-1), with recovery rates ranging from 87 to 100%. The present method is applicable to the quantification of bile acid conjugates in human gastric bile with satisfactory sensitivity, selectivity and precision. Intragastric bile acid compositions in 10 patients with bile reflux gastritis during Deursil or placebo treatment are presented.

Observation of a case of CREST syndrome associated with autoimmune chronic hepatopathy (so-called Reynolds syndrome) and Sjögren's syndrome brings to the fore the importance of symptomatological superimpositions within the autoimmune diseases. A number of diagnostic and pathogenetic aspects of this rare disease association are discussed.

Several reports have suggested that mesenchymal glycosaminoglycans (GAG) may be involved in the regulatory role of epithelial differentiation. Some researchers have pointed out that exogenous GAG affects extracellular GAG accumulation. We have therefore examined the effect of added GAG on two typical processes of avian skin differentiation: keratinization and feather formation. Glycosaminoglycans, either obtained from fibroblasts cultures (conditioned media) or purified commercially available GAG were administered to 5/6-day chick embryo back skin explants. Control cultures were supported with 199 synthetic medium, chick embryo extract or calf serum. Explants have been examined by histological and histochemical procedures. Skin explants maintained in vitro for 7 days exhibited an epithelial differentiation and a dermal histochemical reactivity which were related to the composition of the culture medium. In conditioned media from dermal fibroblasts, but not from heart or lung fibroblasts, explants always exhibited keratinization. In purified-GAG-containing media, keratinization was observed with condroitinsulphates and not with hyaluronic acid. Keratinization was always related to prevalent accumulation of hyaluronic acid in the underlying mesenchyme whereas feather formation was in relation to deposits of condroitinsulphates in dermis pulp. The above findings demonstrate that exogenous GAG is able to modulate avian skin differentiation and that this regulation is linked to an influence on the mesenchymal GAG pattern.

Cultured fibroblast derived from 7 and 14 days chick embryonic skin were tested with three fluorescent lectins (Con A, WGA and SBA). The result obtained shown that the percentage of cells that bind WGA decrease from 7 to 14 days, supporting the presence of an age-dependent heterogeneity at cell surface level. In both cellular population only the 50% of the cell bind to SBA, suggesting a more subtle heterogeneity into the same population.

This paper shows that cultured chick embryo fibroblasts derived from lung, heart and skin contain different amounts of sulphurated glycosaminoglycans (GAG). The sulphurated GAGs were detected by X-ray microanalysis at scanning electron microscopy. The results support the current hypothesis on the heterogeneity of the embryonic fibroblast populations.

The pH values in 203 patients with various gastroduodenal pathologies were measured by means of a glass electrode inserted into the bioptic channel of the gastroscope. Analysis of the results confirms what we know of pH variations in gastric or duodenal ulcer cases and shows where perendoscopic pH measurements may be used beneficially. In particular it was found that mucous pH is not affected by H2 antagonists if these are taken more that 12 hours before the examination. Perendoscopic pH measurement appears to be a valuable tool in screening stomach lesions at risk in cases of chronic gastritis, since it facilitates the identification of atrophic or metaplastic lesions.

The aim of our study is to evaluate the eventual activity of the total dialysate of two uremic nephrectomized patients in recirculating dialysis and the chromatographic peak of the dialysate fractionated by Sephadex column G 15 on PMN chemotaxis. Only the total dialysate and the chromatographic peak B showed inhibition of chemotaxis. On the contrary the commercial polyamines in the same concentration range, and the other chromatographic peaks, containing polyamines too, did not revealed inhibition. Our data show, therefore, that the chemotaxis inhibition could be due to the middle-molecules present in the peak B, rather than the polyamines itself. Polyamines were determined by dansylation method, separated by thin layer chromatography and quantified by spectrofluorimeter. Chemotaxis was evaluated using the modified Boyden chamber.

We studied the ornithine decarboxylase (ODC) activity and polyamines (PAs) content in the red cells lysate before and after haemodialysis of 15 uremic patients and 15 healthy subjects. ODC activity is significantly increased after haemodialysis but the PAs concentrations (particularly spermine and spermidine) do not increase. This could be because of a minimal loss of PAs surplus production. On the contrary the increase of ODC activity could be explained in several ways, e.g. by stimuli during haemodialysis (hypoxia of the first hour of treatment, decrease of plasmatic osmolarity and biological stimulation of the blood during its passage through the filter).

Total bile acid concentrations were estimated in fasting serum samples from 251 patients with chronic liver disease and 108 controls without liver disease, together with conventional liver tests. Serum bile acids level was significantly higher in patients with liver disease than in control group. Fasting serum bile acid concentration was raised in 172 of the 251 patients with impaired hepatic function (68.5%). Only gamma-glutamyltranspeptidase and transaminase tests exhibited a higher abnormality frequency. Two-hour postprandial serum bile acid concentrations were measured in 78 patients: the meal test shows a rise in sensitivity (+26.9%). Our results confirm that serum bile acids concentration represent a sensitive and reproducible test for hepatobiliary disorders.

We have carried out ultramicroscopical investigations on 30 patients with endoscopical aspects of congestive or erosive duodenitis associated or not with bulbar ulcer in relation to the presence of Campylobacter-like organisms (CLO), considered as a possible etiologic agent of peptic disease. Bacteria were found in all of the 14 patients with duodenal ulcer and in only five patients without ulcer. The ultrastructural studies provided elements for the evaluation of epithelial colonization of the duodenal bulb by CLO and which may lead to a new interpretation of gastric metaplasia.

We studied possible effects of uremic toxins and polyamines (PAs) on organ cultures of chick embryo. We added on culture media the lyophilized of total dialysate and its chromatographic peak II obtained with chromatography with Sephadex G 15. The total dialysate and peak II showed toxicity with degeneration of the culture, whereas the free PAs, we added, did not effects.