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Molecular characterization of IgE reactivity of specific individual components of allergenic extracts is now possible due to the technology of recombinant allergens derived from studies of molecular biology of allergic pathology. The identification of the immunoreactivity to single allergenic components in allergic subjects allows to specifically define her/his allergic profile and obtain the so-termed Component Resolved Diagnosis (CRD). Molecular allergens can be classified into those that induce the respiratory allergic reactivity and those that identify the food-related allergic pathology. It is also essential to identify those molecular allergens whose immunoreactivity is able to connect the two clinical conditions: respiratory symptoms and food allergy symptoms. The present study was conducted on 50 patients with a clinical history of hypersensitivity to pollen and/or allergy and positivity to Skin Prick Test. The sera were analyzed in our laboratories and the panel of recombinant allergens was applied in the case of positivity of the specific IgE. Of the 50 patients enrolled, 31 were selected as positive to 4 main pan-allergen Bet v1, Par j2, Art v1 and Phl p1; among these, 14 subjects showed one allergen-specific IgE towards natural extracts of tested foods even in absence of clinical history. CRD allows for an increased accuracy in allergy diagnosis and prognosis and plays an important role in: a) resolving genuine vs cross-reactive sensitization in poly-sensitized patients, b) assessing, in selected cases, the risk of severe, systemic vs mild, local reactions in food allergy, and c) identifying patients and triggering allergens for specific immunotherapy (ITS). In light of our results, we believe that the transition from a diagnostic based on the use of allergenic extracts to another one based on the use of single allergenic molecules that is able to define the specific allergenic profile of each patient, seems to be able to revolutionize the allergy diagnosis.

Vitiligo is a multifactorial polygenic disorder with a complex pathogenesis. It is related to both genetic and no genetic factors. The role of genetics is currently studied with several analytical approaches, such as genetic linkage, candidate gene association studies, genome-wide association studies (GWAS), deep DNA re-sequencing and gene expression studies. To date, there are no genetic traits directly related to vitiligo pathogenesis.43 cases of vitiligo patients and 30 healthy donors recruited as control, were screened by assaying the biochemical molecules involved in the self-cells cytotoxicity (haptoglobin and homocysteine) and candidate genes involved in the regulatory process of the re-methylation cycles and transsulfuration. Candidate genes and their polymorphisms screened are methylene-tetrahydrofolate-reductase (MTHFR) C677T and A1298C; cystathionine-beta-synthase enzyme (CBS) I278T and Ins68bp; and methionine-synthase-reductase (MTRR) A66G.A peculiar genetic profile in vitiligo patients are defined: 11.6% of vitiligo patients shown polymorphic variant MTHFR 677TT vs. 3.3% of healthy donor MTHFR 677CC profile (p=0.0017); 14.0% of vitiligo patients shown CBS polymorphic variant 278TT vs. 3.3% of healthy donor 278II profile (p=0.0012); and 11.6% of vitiligo patients shown MTRR 66GG vs. 3.3% of healthy donor MTRR 677AA profile (p0.0001).This is the first study reporting the correlation between the polymorphic status of MTHFR C677T, CBS I278T, and MTRR A66G and vitiligo. The genetic screening of these polymorphisms could be useful for early detection of the inheritance risk factor in a subject carrying relatives with vitiligo. Although these data could suggest a kind of dysregulation, genetically based, of thiols production mechanisms. Based on these results, we have not been able to get hypothesis about the putative pathogenesis of vitiligo, and the precise cause remains unclear.

Rheumatoid Arthritis (RA) is an autoimmune inflammatory disease that leads to local and systemic arthritis and bone loss. Exploring genetic markers of candidate genes in osteoporosis and inflammatory cytokine genes could be a useful tool for the early identification of bone loss and fracture risk in RA patients. The target of this study is the evaluation and correlation between of Single Nucleotide Polymorphisms (SNPs) of Vitamin D Receptor (VDR) and possible effects on bone loss in RA.40 Caucasian patients with RA (26 of them with a severe bone loss) and 40 healthy donors as control samples were genotyped for the VDR SNPs (called BsmI, ApaI, TaqI and FokI). The detection method is based on Restriction Fragment Length Polymorphism (RFLP).Genotyping profile shown no difference between RA patients and controls. Only VDR-TaqI genotype (TT vs. tt) seem to influence the bone density in females, but not in males. The mean differences of Bone Mass Density (BMD) at the lumbar spine in RA women with the tt allele were 4.7% compared to 0.1% in women with the TT allele (p0.05).The results of these studies support an association between specific VDR alleles and bone loss in RA. The TaqI t and BsmI B alleles were associated with an accelerated bone loss in RA, but not with a focal bone loss. These effects of VDR genotypes and vitamin D supplementation are not unexpected, given that the central pathological feature in RA is bone and joint destruction. The VDR SNPs genotyping should be a useful tool to screen early women RA patients with the bone loss.

Burley tobacco (Nicotiana tabacum L.) plants were grown in the field with or without irrigation and fertilized with 0, 120, 240 or 360 kg N ha−1 over two growing seasons to assess nitrogen use under Mediterranean climate conditions. Kjeldahl-N and NO3-N in leaves and stems and NO3-N and NH4-N in the soil at two depths (0–0.3 and 0.3–0.6 m) were determined. The effect of N fertilization on total N accumulated in the canopy biomass was markedly different between irrigated and non-irrigated plants. Under non-irrigated conditions N accumulated in the plant did not depend on the amount of N applied. In both years, the amount of N in irrigated plants increased in response to the amount of N applied, starting from 49 to 56 days after transplanting (DAT). The average amount of total N in the canopy of irrigated plants, measured across all sampling dates of both years, ranged from 30 kg ha−1 of the unfertilized control to 88 kg ha−1 of the 360 kg ha−1 of N applied. The average amount of plant NO3-N was 2.6 and 4.4 kg ha−1 for non-irrigated and irrigated plots across all N treatments (means of 1996 and 1997). Nitrogen uptake rate (NUR) of non-irrigated plants was high between seedling establishment and the period of rapid stem elongation in 1996 (from 36 to 50 DAT) and until flowering in 1997 (from 42 to 71 DAT), but much less or negligible at later stages of plant development. Irrigation increased NUR dramatically in the second part of the growing season. Maximum NUR was estimated for plants receiving 240 or 360 kg N ha−1 in both years. The year of study did not affect the recovery fraction (RF), physiological efficiency (PE) or agronomic efficiency (AE). Irrigation and N fertilization had significant effects on both RF and AE, but not on PE. Maximum values of RF were 45 and 22% for irrigated and non-irrigated treatments, respectively. In irrigated plots there was a negative relationship between RF and increasing N levels at all sampling dates.

Despite the economic importance of tobacco, there is limited field study on the quantitative response of growth and yield to increasing soil salinity. The effects of irrigation with saline water on yield components of field-grown tobacco ( Nicotiana tabacum L.) “Burley” type plants were studied over two growing seasons. Growth, dry matter partitioning and gas exchange were measured either in rainfed or fully irrigated plants growing in a clayey–sandy–loam soil. The four fully irrigated treatments received amounts of saline waters at 0.54, 2.5, 5.0 or 10 dS m −1 electrical conductivity (EC w ) equal to crop evapotranspiration. In both years, the electrical conductivity of the saturation phase (EC e ) across the 0.6 m topsoil profile increased with increasing salinity of the irrigation water. Soil moisture was markedly lower in the rainfed treatment than in fully irrigated treatments. Different saline concentrations of irrigation water had virtually no effect on soil moisture. Carbon assimilation rate, stomatal conductance and water use efficiency of the saline treatments were lower than the fully irrigated plants at 0.54 dS m −1 (NW treatment) in 1996, but not in 1997. Transpiration rates were unaffected by salinity in both years. The highest yield was produced by plants irrigated with good quality water. The number of leaves per unit land area was greater for the NW plants, whereas there were no differences between the other four treatments. Salinity decreased plant dry matter and height at harvest, increased dry matter partitioning into leaves and decreased that into stems in both years. Dry matter partitioning to leaves was also greater for the rainfed plants than for the NW plants. Tobacco plants grown under field conditions showed a maximum reduction of relative yield at the highest salinity level of only 31%. The threshold values (0.56 and 0.96 dS m −1 ) and the EC e at which a 10% yield reduction was obtained (3.12 and 2.55 dS m −1 ) calculated from the linear model of response of relative yield to increasing EC e were typical of moderately sensitive crops. The EC e values at which 50% yield was reduced (13.34 and 8.91 dS m −1 ) were indicative of moderate tolerance to salinity.

Two clones of white clover (Trifolium repens L.) differing in ozone tolerance were grown in southern Italy during 1997 and 1998 to study the effects of ambient ozone exposure on yield, leaf morphology and water use. Ambient ozone levels were high in both years with values exceeding the threshold for leaf injury reported in the literature. In both years ozone injury was observed on the sensitive clone (NC-S) but not on the resistant one (NC-R), and leaf and stolon dry matter production was significantly lower in NC-S than in NC-R. However, it cannot be excluded that other factors, such as high temperature, interacted with the effect of ozone on biomass production. The clones differed in morphological characteristics. Lower total leaf area in NC-S plants was due to a smaller number of leaves per plant, but the average area per leaf was higher in NC-S. Specific leaf weight and net assimilation rate were higher in the more productive clone (NC-R). Cumulative plant water use was higher in NC-R in each growth period because of the larger leaf area; by contrast, water use per unit leaf area was higher in NC-S, indicating higher leaf conductance to water vapour. The results suggest that ozone significantly reduces the yield of sensitive white clover plants under well-watered conditions, and that the difference in ozone tolerance between clover clones is related to differences in leaf morphology and water use.

The aim of the present study was to evaluate in vitro the biological behavior of human gingival fibroblasts cultured on two different titanium surfaces.Titanium test disks were prepared with a machined, relatively smooth (S) surface or a rough surface (O) obtained by a double acid etching procedure. Primary cultures of human gingival fibroblasts were plated on the experimental titanium disks and cultured up to 14 days. Titanium disk surfaces were analysed by scanning electron microscopy (SEM). Cell proliferation and a quantitative analysis by ELISA in situ of ECM components as CoI, FN and TN were performed.Results have shown different effects of titanium surface microtopography on cell expression and differentiation. At 96 hours of culture on experimental surfaces human gingival fibroblasts displayed a favourable cell attachment and proliferation on both surfaces although showing some differences.Both the relatively smooth and the etched surfaces interacted actively with in vitro cultures of human gingival fibroblasts, promoting cell proliferation and differentiation. Results suggested that the microtopography of a double acid-etched rough surface may induce a greater Co I and FN production, thus conditioning in vivo the biological behaviour of human gingival fibroblasts during the process of peri-implant soft tissue healing.

Hashimoto's thyroiditis (HT) is an autoimmune disorder characterized by the presence of specific antibodies and by a lymphocytic infiltration of the thyroid secreting inflammatory cytokines. Macrophages, lymphocytes, and cytokines play a pivotal role in both development and progression of Th1-mediated autoimmune diseases, and a direct role in the destruction of thyroid follicles and follicular cell function in autoimmune thyroiditis. Integrins are integral membrane receptors involved in cell-extra-cellular matrix (ECM) interaction with both structural and signaling functions. The integrin- ECM interaction is necessary for the correct function and survival of thyroid follicular cells. The purpose of this study was to determine the effect of cytokine stimulation on integrin expression and signaling in the thyroid cell. Primary cultures from normal thyroids were treated with interferon-γ (IFN-γ), INF-α, tumor necrosis factor-α, interleukin 1a or these cytokines all together. Integrin expression, cell adhesion to fibronectin (FN) and FN-stimulated extracellular signal-regulated kinase (ERK) phosphorylation were determined after cytokine treatment. IFN-γ and IFN-α were the most effective, reducing the expression of the integrin αvβ3 and slightly increasing the α3β1. Cell treatment with IFN-γ strongly impaired cell adhesion to FN. At the same time, the treatment with IFN-γ dramatically inhibited the stimulation of ERK phosphorylation induced by cell adhesion to FN. In conclusion, IFN-γ inhibits the expression of the integrin αvβ3, reducing the cell adhesion to FN and the following intracellular signaling in thyroid cells in culture. These results suggest that integrins may be a target of the infiltrating lymphocytes and have a role in the pathogenesis of autoimmune thyroiditis.

The calcium/calmodulin-dependent kinase II (CaMKII) participates with Ras to Raf-1 activation, and it is necessary for activation of the extracellular signal-regulated kinase (ERK) by different factors in epithelial and mesenchimal cells. Raf-1 activation is a complex multistep process, and its maximal activation is achieved by phosphorylation at Y341 by Src and at S338 by other kinase/s. Although early data proposed the involvement of p21-activated kinase 3 (Pak3), the kinase phosphorylating S338 remains to be definitively identified. In this study, we verified the hypothesis that CaMKII phosphorylates Raf-1 at Ser338. To do so, we determined the role of CaMKII in Raf-1 and ERK activation by oncogenic Ras and other factors. Serum, fibronectin, Src (Y527) and Ras (V12) activated CaMKII and ERK, at different extents. The inhibition of CaMKII attenuated Raf-1 and ERK activation by all these factors. CaMKII was also necessary for the phosphorylation of Raf-1 at S338 by serum, fibronectin and Ras. Conversely, inhibition of Pak3 activation by blocking phosphatidylinositol 3-kinase was ineffective. The direct phosphorylation of S338 Raf-1 by CaMKII was demonstrated in vitro by interaction of purified kinases. These results demonstrate that Ras activates CaMKII, which, in turn, phosphorylates Raf-1 at S338 and participates in ERK activation upon different stimuli.

Hashimoto's thyroiditis is the most frequent autoimmune disorder, characterized by the presence of a large lymphocytic infiltration and secretion of inflammatory cytokines in the thyroid. Infiltrating lymphocytes and cytokines play a pivotal role in the progression of HT, characterized by the progressive destruction of the normal follicular architecture of the gland and death of follicular cells, ending with loss of thyroid function. Integrins are plasma membrane receptors for the cell–extra-cellular matrix components, with both structural and signaling functions. Integrin-mediated fibronectin (FN) binding is necessary for the correct function and survival of thyroid follicular cells. The purpose of this study was to determine the effect of interferon-γ (IFN-γ) stimulation on integrin expression and signaling in the thyroid cell. Cytotoxicity, integrin expression, cell adhesion to FN, and FN-stimulated ERK and AKT phosphorylation were determined in a normal human thyroid cell line treated with IFN-γ. IFN-γ induced apoptosis and reduced the expression of the integrin αvβ3. Integrin-mediated cell adhesion to FN was strongly impaired. Similarly, FN-stimulated ERK and AKT phosphorylation were inhibited. In conclusion, our study in a thyroid cell model demonstrates that IFN-γ induces apoptosis and inhibits the expression of the integrin αvβ3, reducing cell adhesion to FN and the succeeding outside-in signaling. These results suggest that integrins mediate the cytotoxic effect of IFN-γ and are involved in the destructive mechanism of autoimmune thyroiditis.

Differentiation toward the osteoblastic phenotype is a complex phenomenon regulated by means of several factors. Numerous studies in vitro et in vivo showed that surface properties of titanium dental implants modulate cell proliferation and osteoblastic differentiation, affecting bone healing processes. Optimal superficial morphology is still controversial. The aim of the present study was to evaluate in vitro the effects of two different titanium surfaces on biological behaviour of human osteoblast-like cells SaOS-2 with regard to production of extra-cellular matrix (ECM) proteins.Human osteoblast-like cells SaOS-2, cultured on commercially pure titanium disks with two different surface topographies, smooth and microstructured with sand-blasting and acid-etching treatment, were evaluated by investigating adhesion, proliferation and deposition of extra-cellular matrix (ECM) proteins Fibronectin, Tenascin and Collagen I.The different values of cellular adhesion at three hours and the increase of SaOS-2 proliferation values at the different experimental times on both evaluated surfaces didn't result statistically significant. ECM deposition analysis showed that Fibronectin, Tenascin and Collagen I were gradually produced, with not statistically significant differences for Fibrone-ctin and Tenascin and statistically significant differences for Collagen I.Implant surface properties modulate in vitro the biological behavior of osteoblasts-like cells SaOS-2. SaOS-2 cells proliferate on both evaluated surfaces, showing a more organized differentiation towards an osteoblastic phenotype in ECM proteins production when cultured on microstructured surface.

We investigated the effects of human granulocyte macrophage- colony stimulating factor (GM-CSF) on the relation between differentiation and apoptosis in SaOS-2 cells, an osteoblast-like cell line. To determine the relationship between these cellular processes, SaOS-2 cells were treated in vitro for 1, 7 and 14 days with 200 ng/mL GM-CSF and compared with untreated cells. Five nM insulin-like growth factor (IGF-I) and 30 nM okadaic acid were used as negative and positive controls of apoptosis, respectively. Effects on cell differentiation were determined by ECM (extracellular matrix) mineralization, morphology of some typical mature osteoblast differentiation markers, such as osteopontin and sialoprotein II (BSP-II), and production of bone ECM components such as collagen I. The results showed that treatment with GM-CSF caused cell differentiation accompanied by increased production of osteopontin and BSP-II, together with increased ECM deposition and mineralization. Flow cytometric analysis of annexin V and propidium iodide incorporation showed that GM-CSF up-regulated apoptotic cell death of SaOS-2 cells after 14 days of culture in contrast to okadaic acid, which stimulated SaOS-2 apoptosis only during the early period of culture. Endonucleolytic cleavage of genomic DNA, detected by “laddering analysis”, confirmed these data. The results suggest that GM-CSF induces osteoblastic differentiation and long-term apoptotic cell death of the SaOS-2 human osteosarcoma cell line, which in turn suggests a possible in vivo physiological role for GM-CSF on human osteoblast cells.

Commercially pure titanium implants presenting a relatively smooth, machined surface or a roughened endosseous surface show a large percentage of clinical success. Surface properties of dental implants seem to affect bone cells response. Implant topography appears to modulate cell growth and differentiation of osteoblasts affecting the bone healing around the titanium implant. The aim of the present study was to examine the effects of 1cm diameter and 1mm thick titanium disks on cellular morphology, adhesion and bone phenotypic expression of human osteoblast-like cells, SaOS-2. SaOS-2 cells were cultured on commercially 1 cm pure titanium disks with three different surface roughness: smooth (S), sandblasted (SB) and titanium plasma sprayed (TPS). Differences in the cellular morphology were found when they were grown on the three different surfaces. An uniform monolayer of cells recovered the S surface, while clusters of multilayered irregularly shaped cells were distributed on the rough SB and TPS surfaces. The adhesion of SaOS-2 cells, as measured after 3h of culture, was not affected by surface roughness. ECM components such as Collagen I (CoI), Fibronectin (FN), Vitronectin (VN) and Tenascin (TN) were secreted and organized only on the SB and TPS surfaces while they remained into the cytoplasm on the S surfaces. Osteopontin and BSP-II were largely detected on the SB and TPS surfaces, while only minimal production was observed on the S ones. These data show that titanium surface roughness affects bone differentiation of osteoblast like-cells, SaOS-2, indicating that surface properties may be able to modulate the osteoblast phenotype. These observations also suggest that the bone healing response around dental implants can be affected by surface topography.

We investigated the effects of human granulocyte macrophage-colony stimulating factor (GM-CSF) on the relation between differentiation and apoptosis in SaOS-2 cells, an osteoblast-like cell line. To determine the relationship between these cellular processes, SaOS-2 cells were treated in vitro for 1, 7 and 14 days with 200 ng/mL GM-CSF and compared with untreated cells. Five nM insulin-like growth factor (IGF-I) and 30 nM okadaic acid were used as negative and positive controls of apoptosis, respectively. Effects on cell differentiation were determined by ECM (extracellular matrix) mineralization, morphology of some typical mature osteoblast differentiation markers, such as osteopontin and sialoprotein II (BSP-II), and production of bone ECM components such as collagen I. The results showed that treatment with GM-CSF caused cell differentiation accompanied by increased production of osteopontin and BSP-II, together with increased ECM deposition and mineralization. Flow cytometric analysis of annexin V and propidium iodide incorporation showed that GM-CSF up-regulated apoptotic cell death of SaOS-2 cells after 14 days of culture in contrast to okadaic acid, which stimulated SaOS-2 apoptosis only during the early period of culture. Endonucleolytic cleavage of genomic DNA, detected by "Aúladdering analysis"Aù, confirmed these data. The results suggest that GM-CSF induces osteoblastic differentiation and long-term apoptotic cell death of the SaOS-2 human osteosarcoma cell line, which in turn suggests a possible in vivo physiological role for GM-CSF on human osteoblast cells.

Yield losses from ozone pollution can be estimated by two methods: one involves the use of sensitive (S) and resistant (R) biotypes of white clover (Trifolium repens L., cv. Regal) exposed in ambient air, the other is based on the use of open-top chambers (OTC) supplied either with charcoal-filtered (CF) or non-filtered (NF) air. In southern Italy the two methods have been compared using the clover biotypes. The aim was (1) to compare the extent of ozone-induced yield reductions estimated by the two methods, (2) to evaluate the effect of the chamber enclosure on the growth of both biotypes, and (3) to compare plant water consumption in the different environments. On the average, the yield reduction was 23% when derived from the S/R yield ratio in ambient air, and 18% obtained by the CF/NF yield ratio of the S-type, without a significant difference between the two values. The slightly lower value for the OTC-based system may be due to the lower ozone levels in NF chambers due to losses in the ventilation system. Thus, both methods yielded equivalent yield reductions of about 20% due to ozone at this Mediterranean site. However, the higher air temperature inside OTCs influenced the plant growth, and this effect was stronger in the case of the R type. Therefore, R/S yield ratios in NF chambers differed from ambient air. Also, plant water consumption was higher in OTCs than in ambient air. The results suggest that the OTC-based method enables yield loss estimates at this Mediterranean site, in spite of chamber effects on plant growth and water use.

The Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) is a hematopoietic growth factor that regulates the in vitro and in vivo proliferation and differentiation of hematopoietic cells through the interaction with a specific heterodimeric receptor complex (GM-CSFR), consisting of an alpha and a beta chain with molecular weights of 80 and 120 KDa, respectively. We have studied the expression of the GM-CSFR (alpha chain) on the surface of the human osteosarcoma cell line SaOS-2 and the in vitro effects of different concentrations (10, 100, and 200 ng/ml) of GM-CSF on GM-CSFR expression and the biological activity of SaOS-2 cells. Our data show that SaOS-2 cells express GM-CSFR and that GM-CSF can down-regulate the expression of its own receptor on these cells. Furthermore, to evaluate the biological effects of GM-CSF on SaOS-2 cells, we have investigated cell proliferation and differentiation of these cells treated with different doses of the growth factor through: (1) a morphological analysis of typical osteoblast differentiation markers such as osteopontin and BSP-II; (2) measurement of alkaline phosphatase (ALP) activity; (3) production of bone ECM components (collagen I, fibronectin, tenascin, and laminin); (4) production of interleukin-6 (IL-6) and osteocalcin in the culture medium. The results show that the in vitro treatment of SaOS-2 cells with recombinant human GM-CSF causes a decreased cell proliferation and an increased production of osteopontin, BSP-II, ALP, IL-6, and most but not all ECM components. These findings suggest that GM-CSF can regulate proliferation and differentiation of osteoblast-like SaOS-2 cells and could also play an unexpected role in the maturation of bone tissue.

Surface properties may affect the clinical outcome of titanium dental implants. The aim of the present study was to investigate the effects of 3 different titanium surfaces—smooth (S), sandblasted (SB), and titanium plasma-sprayed (TPS)—on proliferation, differentiation, and apoptosis of human osteoblast-like cells, SaOS-2. Cell proliferation was significantly (p < 0.05) higher on the S surface, and synthesis of extracellular matrix proteins was more abundant on TPS and SB than on S surfaces. Analysis of integrin receptors showed a higher expression of α2, α5, αVβ3, and ß1 on TPS as compared with SB and S surfaces. An increase in alkaline phosphatase activity was detected only on SB and TPS surfaces. Analysis of cell apoptosis did not demonstrate any significant difference among the 3 different surfaces. The results indicate that titanium surface topography affects proliferation and differentiation of osteoblast-like SaOS-2 cells, suggesting that surface properties might be important for bone response around dental implants in vivo.

Interleukin-6 (IL-6) and its soluble receptor (sIL-6R) stimulate osteoclast formation and activity. The primary cell abnormality in Paget's disease of bone (PDB) involves osteoclasts. Pagetic osteoclasts overproduce IL-6 and IL-6 receptor in vitro. In vivo, IL-6 serum levels are very high in the acute phase of PDB. The aim of this study was to evaluate the modification in the serum levels of IL-6, sIL-6R and osteotropic hormones (parathormone, 25OHD3 and 1,25(OH)2D3) as a in long-term response to clodronate treatment in patients with PDB.16 patients (8 females) with polyostotic PDB were studied. IL-6, sIL-6R and osteotropic hormones serum levels were evaluated in active PDB and after clodronate treatment (300 mg injected intravenously for 5 consecutive days). The sequential changes in total alkaline phosphatase (tALP) serum levels were used to assess the maximal pharmacological response to treatment.In untreated pagetic patients, mean serum levels of IL-6 (3.20+/-1.18 pg/ml) and sIL-6R (35.02+/-8.33 ng/ml) were significantly increased. Serum osteotropic hormone levels fell within the normal range. Eight weeks after treatment, the maximal pharmacological response to clodronate was associated with a significant reduction of sIL-6R serum levels in all patients, without a significant variation in serum IL-6 and osteotropic hormone levels. Moreover, we observed a correlation between lower sIL-6R serum levels before clodronate therapy and complete remission of PBD, defined as a decrease of tALP serum levels within the normal range.The decrease in serum sIL-6R levels could be one of the molecular mechanisms that play a role in the clinical response to clodronate treatment in PDB.

We report on a survey of apolipoprotein A-I and B measurements among 11 laboratories using a lyophilized serum to be evaluated for its possible use as “reference serum” in a large prospective study. Using a single method, radial immunodiffusion (RID), and a common antibody the partecipating laboratories evaluated the apo A-I and apo B concentrations in the proposed reference serum. Analyses were performed in quadruplicate in four consecutive days to estimate the day by day variation in a given laboratory. This value resulted to be 2% for apo A-I and 1.9% for apo B.

A new enzymatic colorimetric method for phospholipids determination, has been employed to find out "reference ranges" of phospholipid concentration in 300 healthy subjects of Neapolitan area (200 males, 100 females). Linearity of response, precisior and accuracy of this method has been previously demonstrated. Mean, standard deviation and frequency distribution of two sex groups are reported. Moreover phospholipids fraction (VLDL,LDL,HDL) on 30 healthy subjects have been evaluated.

A growing body of experience has demonstrated that physical activity may be protective against C.H.D.1.

Linearity of response "day-to-day" and "within-day" variation of a simple method for serum monoaminoxidase (SMAO) activity have been evaluated. SMAO determination in 209 clinically healthy subjects was performed to find out the "reference values" for the method.

Enzymatic methods have progressively substituted extractive procedures in plasma lipids determination. A new enzymatic method for serum phospholipids evaluation has been recently introduced in Italy as a kit of reagents. In this method phospolipase D hydrolyzes phospholipids with free choline production which, in presence of choline-oxidase and peroxidase, is estimated by colorimetric reaction. Linearity of response and analytical variations "within-run" and "between-run" during six month observations on serum pools have been preliminarily verified.

Significant changes of alpha 2-ceruloplasmin serum concentration and decreased number of T-lymphocites have been found only in 15 R F positive out of 25 patients with definite R.A. Possible correlation between these findings in discussed.

Serum cholesterol and triglyceride levels have been evaluated in samples from fasting males aged 20--59 in Northern (Brisighella), Central (Rome) and Southern (Pozzuoli) Italy. Regularly performed quality controls between laboratories assured comparability of data. A statisitically significant difference of mean serum cholesterol and triglyceride levels was observed for most age-groups in the 3 different areas, lower values being found in the southern population as compared to the central and northern ones. These results support previous findings and the thesis that large differences in blood lipid levels may still exist even within the same country and that they at least in part may be culturally determined in connection with different dietary habits.

Increased serum copper (sCu) and alpha 2 caeruloplasmin (alpha 2 sCP) concentrations have been reported in patients with rheumatoid arthritis and other inflammatory conditions. Close parallel changes of sCu and alpha 2 sCP in rheumatoid arthritis have recently been observed by us in 30 subjects. The 'IgM rheumatoid factor positive (IgM-RF)' patients showed higher sCu and alpha 2 sCP concentrations when compared with 'IgM-RF negative'. Moreover a significant correlation was found between sCu alpha 2 sCP and other humoral indexes altered in rheumatoid arthritis. Therefore it seems useful, as further index of inflammation, to test both sCu and alpha 2 sCP.

In a group of 25 patients with rheumatoid arthritis, the percent of erythrocyte rosettes forming cells (ERFC) was found significantly decreased in patients with rheumatoid factor positive (RF+), whereas ERFC were normal in the RF negative patients (RF--). Likely, T-lymphocytic depression in the RF+ patients could be related to a T-suppressor lymphocytes deficit and therefore, B-lymphocytes would be abnormally stimulated. Moreover, in the RF-- patients, B-lymphocytes would not be enhanced for the normal T-suppressor lymphocytes activity.