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Biomedical subjects

C Palermo

Publications and source records attributed to C Palermo.

At least 19 recordsLinked to original sources
PubMed2005

Effects of alpha-interferon on insulin-like growth factor-I, insulin-like growth factor-II and insulin-like growth factor binding protein-3 secretion by a human lung cancer cell line in vitro.

The aim of our study was to evaluate the effect of alpha-interferon (alpha-IFN) on cell growth and on the different IGF system components in a human non-small cell lung cancer line (Calu-6) in vitro. Our results confirm the release of IGF-I and IGF-II by these cells. The amount of IGF-II in conditioned media (10.25 +/- 3.95 nM/10(6) cells, mean +/- SE) was more than 10-fold higher than that of IGF-I. alpha-IFN treatment reduced IGF-II levels in the media, with a maximal effect between 1 and 10 U/ml (delta% of control: -31 and -55%, respectively, p < 0.05). IGF-I was significantly reduced at 0.5 U/ml (p < 0.01). No difference, however, was observed in IGF mRNA expression between untreated and alpha-IFN treated cells. An increase in IGF-I and IGF-II intracellular levels in alpha-IFN treated cultures was observed, suggesting that alpha-IFN can regulate the transfer of these peptides into the cells. Furthermore, IGF type-I and particularly type-lI receptor expression was increased after alpha-IFN treatment. IGFBP-3 was detected only in trace amounts in the conditioned media; however, it showed an increase after alpha-IFN treatment (+110% at 1 U/ml). IGFBP-3 mRNA expression showed a slight increase after treatment with 1 and 10 U/ml. alpha-IFN (1-10 U/ml) reduced the stimulatory effect of IGF-I on cell replication (p < 0.01), inhibited (p < 0.01) cell replication in untreated and in fetal calf serum (FCS)-stimulated cells, and increased apoptosis in Calu-6 cells. Our data suggest that alpha-IFN may exert its effects at the cellular level in part through modification of the local IGF system.

Antineoplastic Agents↗
PubMed2005

FMS*Calciumfluor specifically increases mRNA levels and induces signaling via MAPK 42,44 and not FAK in differentiating rat osteoblasts.

The homeopathic compound of resonance FMS*Calciumfluor (FMS*) reportedly promotes osteogenic differentiation of rat pre-osteoblasts in vitro. Here, we show that the continuous exposure of differentiating rat osteogenic cells (ROB) to FMS* modulates the level of expression of mRNAs for 7 of the 8 osteogenic markers tested. Alkaline phosphatase (AP), osteocalcin (OC), metalloproteinases (MMP-2 and -14), procollagenase C (BMP-1), biglycan (BG) and integrin 1 are expressed at higher levels in FMS*-treated osteoblasts than in control cultures. MMP-2 and -14 mRNA are not down-modulated at mineralization. Also, the pattern of expression induced by FMS* for some of these genes (BMP-1, BG and integrin 1) is changed, but collagen type I (Coll I) mRNA levels are not affected by treatment with FMS*. This suggests that FMS* modulates mRNA levels and that this is not generalized, but gene(s) specific. We also report that exposure to FMS* rapidly and transiently induces activation of mitogen-activated protein kinases (MAPKs) 42,44 in populations of early osteoblasts, but not in pre-osteoblasts, with a cell differentiation stage-dependent and pertussis toxin (PTX)-sensitive response. Subsequent to FMS* MAPK signaling activation, an increase in AP and MMP-14 mRNA is detected, which is also inhibited by PTX, suggesting that FMS* activation of MAPK signaling could be an early event required for the induction of these genes. Exposure to FMS* does not cause changes in the activity of p125 (FAK)-mediated signaling.

Alkaline Phosphatase↗
PubMed2005

The IGF system and bone.

All components of the IGF systems and all regulatory mechanisms known are present and active in bone tissue. Osteoblasts synthesize IGF-I and IGF-II which have mitogenic effects on bone cells. Although the relative productions of IGF-I and IGF-II by osteoblasts are different depending on different bone districts and experimental conditions, it seems now that IGF-II is, in general, more expressed than IGF-I. The synthesis of IGFs is down-regulated by many locally produced growth factors, particularly trasforming growth factor beta (TGF-beta) and cortisol, and this probably accounts for the osteoporotic effects of this steroid, whereas PTH is stimulatory. Similarly to other tissues, IGFs action on bone is not only limited to cell replication but also to differentiated functions, such as production of collagen and matrix apposition. Binding proteins 2-5 have been demonstrated to be present in bone, the most expressed being IGFBP-4 and -5. Unlike IGFBP-4 which has an inhibiting effect on IGF actions, IGFBP-5 has a potentiating effect, both in vivo and in vitro, probably by binding directly to sites which are independent of the IGF receptor. The involvement of IGFBP proteases has also been demonstrated in human osteoblasts which are stimulated by IGF-II and TGF-beta. In a rat osteoblast cell culture that we studied, IGF-II and IGFBP-2 were the most abundant peptides of the IGF system released in cell medium, IGFBP-5 was abundantly expressed but bound to cell surface and cell matrix and IGF-I and IGFBP-3 were found at very low concentrations. All these peptides reach the maximum concentration in the first stage of maturation and gradually decrease in the following stages. Also IGFBP proteases, namely MMP-2, are important actors of the systems, being involved in the inactivation of IGFBPs in the late stages of maturation.

Animals↗
PubMed2004

Rapid multiresidue extraction method of organochlorinated pesticides from fish feed.

A rapid multiresidue extraction method for organochlorinated pesticides from fish feed was developed, which is based on the extracted fat treatment by n-hexane, concentrated sulphuric acid and ENVI-carb, a graphitized non-porous carbon material. The final residue, obtained in about 50 min, was dissolved in isooctane and analysed by gas chromatography with an electron capture detector (GC/ECD). The presence of the extracted pesticides was confirmed by gas chromatography-mass spectrometry (GC/MS). Concentration of sulphuric acid and amount of ENVI-carb were optimized in order to improve analytes recovery, accuracy and detection limits. This simple and relatively fast method allowed a high recovery of the HCB, Lindane, HEPO, p,p'-DDE, p,p'-DDD, p,p'-DDT residues, with mean values in the range 68-124% at four fortification levels (12.5, 25.0, 50.0, 100.0 ng/g), and coefficients of variation between 1.9 and 20.2%. Detection limit were equal to 3.0 ng/g, related to fat, for all pesticides, and calibration curves were linear (r > 0.999) in the range of explored concentrations from the detection limit to 100 ng/g. For all pesticides a good repeatability was obtained (CV% values in the range 0.23-4.16%) when a sequence of six injections of the isooctane extraction solution was performed. The usefulness of the proposed method has been tested by the analysis of fish feed samples.

Animal Feed↗
PubMed2003

Survey of total mercury and methylmercury levels in edible fish from the Adriatic Sea.

Total mercury and methylmercury concentrations were measured in the muscle tissue of different fish species from the Adriatic Sea to ascertain whether the concentrations exceeded the maximum level fixed by the European Commission. Large species-dependent variability was observed. The highest total mercury mean concentrations were in benthic (0.20-0.76 microg g(-1) wet wt) and demersal fish (0.22-0.73 microg g(-1) wet wt), while pelagic species showed the lowest levels (0.09-0.23 microg g(-1) wet wt). In 15% of frost fish, in 42% of skate and in 30% of angler fish samples total mercury concentrations exceeded the maximum level fixed by the European Commission (Hg = 1 microg g(-1) wet wt); for the species for which the maximum level was set to 0.5 microg g(-1) wet wt, concentrations exceeding the prescribed legal limit were observed in 6.4% of bokkem, in 6.6% of pandora, in 20% of megrin, in 12.5% of four-spotted megrim, in 16% of striped mullet, in 5.0% of forkbeard and in 5.3% of picarel samples. In all the different species, mercury was present almost completely in the methylated form, with mean percentages between 70 and 100%. Weekly intake was estimated and compared with the provisional tolerable weekly intake recommended by the FAO/WHO Expert Committee on Food Additives. A high exposure was associated with the consumption of only skates, frost fish and angler fish, thought the consumption of the other species, such as, megrim, four spotted megrim, red fish striped mullet and forkbeard, resulted in a weekly intake slightly below the established provisional tolerable weekly intake.

Animals↗
PubMed2000

Nitric oxide and salt sensitivity.

Studies in laboratory animals suggest that altered nitric oxide (NO) production may be associated with salt sensitivity. In this investigation we determined whether the endogenous NO production was altered in salt-sensitive human subjects when salt intake was changed. Salt sensitivity was assessed from the magnitude of the blood pressure (BP) lowering obtained when the salt intake was reduced from high to a low intake. The combined urinary excretion of nitrites and nitrates, the major metabolites of NO, was employed as an index of endogenous NO production. Salt-sensitive subjects (n = 23) were older, heavier, and had greater waist-to-hip ratios and higher baseline BP than salt-resistant individuals (n = 25). In salt-sensitive subjects, mean blood pressure (MBP) decreased 11.8+/-0.7 mm Hg, and NO metabolite excretion increased from 823+/-102 to 1530+/-148 mmol/24 h, when salt intake was reduced from 316 to 28 micromol/day. NO metabolite excretion was 45% lower during high salt (0.66+/-0.1 micromol/mg creatinine) than during low salt intake (1.12+/-0.1 micromol/mg creatinine) (P < .001). In contrast, when salt intake was reduced, salt-resistant subjects exhibited no significant mean changes in BP or NO metabolite excretion. During low salt intake, NO metabolite excretion (micromol/ day) was significantly higher in salt-sensitive individuals. The magnitude of decrease of systolic blood pressure, diastolic blood pressure, or MBP induced by reducing salt intake was not related to the increase in urinary excretion of NO metabolite levels (r2 = 0.009; P = .66). In summary, to the extent that urinary NO metabolite levels reflect the activity of the endogenous NO system, our results support the view that salt sensitivity may in part be determined by an inability to increase or to sustain NO production in response to high salt. Insufficient NO production during high salt may in turn lead to altered pressure-natriuresis relationships and to an increase in BP. The possibility that the increase in BP induced by high salt intake in salt-sensitive individuals could be the key factor in reducing NO metabolite levels can not be ruled out.

Adult↗
PubMed2000

Insulin and blood pressure responses to changes in salt intake.

In this study we evaluated the role of insulin in hypertension and on salt sensitivity. The study was conducted in 47 consecutive patients attending the Center for the Detection and Treatment of Cardiovascular and Metabolic Risk factors. The relationships between fasting and post-glucose load insulin levels and the blood pressure (BP) responses to changes in salt intake, were investigated. No correlation was observed between fasting or 2-h post-load insulin levels and mean BP (MBP), systolic BP (SBP) or diastolic BP (DBP). The plasma concentrations of insulin were not significantly related to body mass index (BMI) (r2 = 0.05; P = 0.135). Neither fasting nor 2-h post-load insulin predicted the BP response to changes in salt intake. A reduction in salt intake from 316 +/- 13 to 26 PM 3 mmoles/day, produced similar BP lowering in subjects with fasting insulin >15 microU/ml and in subjects with normal fasting insulin levels (<15 microU/ml). In addition, no relationship was observed between the magnitude of the BP responses to salt and the levels of insulin, either fasting (r2 = 0.007; P = 0.86) or 2-h after a glucose load (r2 = 0.01; P = 0.213). A very strong association was found between body weight or BMI and MBP (r2 = 0.443; P< 0.0001). In conclusion, our results are against the view of a cause-effect relationship between insulin and BP levels. In addition, the insulin status of a patient does not predict (nor determines) his (her) vascular reactivity to changes in salt intake. Finally, our findings further support the existence of a strong and direct association between body weight and hypertension, and speak against a major role of insulin in the pathogenesis of hypertension associated with obesity.

Adult↗
PubMed2000

The expression of metalloproteinase-2, -9, and -14 and of tissue inhibitors-1 and -2 is developmentally modulated during osteogenesis in vitro, the mature osteoblastic phenotype expressing metalloproteinase-14.

During osteogenesis, in vitro, of tibial-derived rat osteoblasts (ROB) and derived clones, changes occur in the interactions of mature osteoblasts with the endogenous extracellular matrix (ECM) and these culminate in the formation of tridimensional nodules, which become sites of mineral deposition. We investigated if these changes might be mediated by remodeling of ECM, and we focused our study on the neutral metalloproteinases (MMPs), known agents of matrix remodeling, and on their tissue inhibitors (TIMPs). We report that during in vitro differentiation, osteoblasts express the secreted MMP-2 and -9 and the membrane gelatinase MMP-14. These, along with the tissue inhibitors TIMP-1 and -2, are developmentally regulated according to the maturation stage of osteoblasts. Their levels change in a similar association with osteoblast phenotypic maturation in different populations of ROB, which take different times to complete osteogenesis in vitro. MMP-14 expression coincides in both cell populations with the mature osteoblastic phenotype and is localized in the cells forming nodules. MMP-2 and -9 are expressed diffusely in the osteoblast population. Developmentally associated changes in the activation of MMP-2 are detected, associated in their timing with the expression of MMP-14 in both populations of ROB, and MMP-14 activates pro-MMP-2 in vitro. Expression of messenger RNAs (mRNAs) for the three MMPs increases up to the time of nodule formation. At this stage, TIMP-1 mRNA levels are lowest. TIMP-2 mRNA decreases throughout osteogenesis. In situ hybridization in 7-day-old rat tibias shows the strongest expression of MMP-14 among osteogenic cells, in lining osteoblasts on the newly formed trabeculae under the growth plate, and on the endosteal surface of cortical bone. Our data support the concept that the developmentally regulated expression of MMP-14 triggers localized proteolysis within the osteogenic population, concomitant in vitro to nodule formation.

Animals↗
PubMed1999

Osteogenesis in vitro in rat tibia-derived osteoblasts is promoted by the homeopathic preparation, FMS*Calciumfluor.

We studied the effects of in vitro treatment of differentiating osteogenic cells with FMS*Calciumfluor, to determine whether it caused changes in proliferative or differentiation potential of osteoblasts. FMS*Calciumfluor was developed for the therapy of post-menopausal and age-related osteoporosis on the basis of the principles of resonance homeopathy and VTR Vega test. Its daily prescribed therapeutical usage is about 30,000-fold less in fluoride concentration than that recommended for NaF associated with calcium monophosphate. Rat tibial osteoblast (ROB) primary cultures represent populations of early osteoblasts and their derivative cultures of more than 60 cumulative population doubling (CPD) represent more mature osteogenic cells. Both these populations were shown to undergo in vitro differentiation, as monitored by the sequential expression of markers that define the stages of the osteogenic progression. Here we report that continual treatment of ROB during osteogenesis with FMS*Calciumfluor modulated the expression of critical osteogenic markers: alkaline phosphatase (AP), an indicator of osteoblast maturation, and(45)Ca incorporation into the matrix and nodule formation, events of the last phase of osteogenesis and a measure of osteoid mineralization. Treatment did not affect proliferation, or expression and activation of metalloproteinases (MMP). AP activity and levels of AP mRNA were increased by treatment with FMS*Calciumfluor; the incorporation of radiolabelled Ca into the matrix was also increased and the formation of nodules occurred in a shorter time and with higher frequency than in untreated control cultures. The effects of FMS*Calciumfluor were concentration dependent and specific for its modalities of preparation, and were observed at a concentration about three orders of magnitude lower than similar effects reported in the literature by treatment of osteoblast cultures in vitro with NaF.

Animals↗
PubMed1997

Rat tibial osteoblasts III: propagation in vitro is accompanied by enhancement of osteoblast phenotype.

Postproliferative confluent cultures of primary rat tibial osteoblasts (ROB), cultured in medium supplemented with ascorbic acid and beta-glycerophosphate (AS-bGP, differentiation medium) express, in sequence, specific bone markers which identify a succession of maturation stages, and eventually form mineralized noduli. We report an investigation on the effect of extensive proliferation in vitro in unsupplemented medium on the osteogenic potential of mass cultures of ROB. The growth rates of the populations, derived from two independent primary cultures, was constant throughout 110 cumulative population doublings (CPD) in culture. Propagated cells maintained features similar to osteoblasts in primary cultures with respect to serum and anchorage dependence for growth and to the chemokinetic effect on endothelial cells exerted by their conditioned media (CM). Propagated populations, set at confluence in differentiation medium, were tested for the expression of early [alkaline phosphatase (AP)] and late [osteocalcin (OC); bone sialoprotein (BSP); 45Ca incorporation and mineralization] osteogenic markers. We observed an increase, parallel to the increase in CPD, in both the level of maximal expression of AP (enzyme/microgram cellular DNA) and in the frequency of nodules, reaching five- to sixfold (at 78 CPD) and eightfold (at 60 CPD), respectively, the levels of primary cultures. AP expression (enzyme and mRNA) persisted during mineralization and 45Ca incorporation. The time required by propagated cultures for the formation of nodules decreased with increase of CPD, and was reduced to less than one third at 87 CDP. Nodules became mineralized over a similar lapse of time as in primary cultures and were positive by histochemistry for BSP and OC. We also obtained osteogenic clones from two independent cultures after 72 CPD. 90% of these showed an osteoblast phenotype, expressing AP and forming nodules positive for OC and BSP, which mineralized. Timing of formation and frequency of nodules/plated cells in clones was similar to that found in propagated cultures of equivalent CPD. In summary, propagated ROB populations and derived clones showed enhanced osteoblast phenotype, possibly due to an increase in osteogenic cells and enrichment of proliferating mature osteoblasts, consequent to extended propagation in culture.

Alkaline Phosphatase↗
PubMed1995

The diagnosis value of beta 2-microglobulin and immunoglobulins in primary Sjögren's syndrome.

Salivary and serum concentrations of beta 2-microglobulin, salivary levels of the immunoglobulins, and salivary flow (as measured by a gravimetric method) were assessed in 34 patients and 11 controls. Of the 34 study patients, 16 had a primary form of Sjögren's syndrome (pSS) and 18 had sicca syndrome. The salivary and serum concentrations of beta 2-microglobulin and salivary levels of IgA and IgG were much higher in the patients with pSS than in controls or those with sicca syndrome (p < 0.001). Similarly, the salivary IgM levels of patients with pSS were higher (although at a lower degree of statistical significance) as compared to those of patients with sicca syndrome (p < 0.01) or controls (p < 0.05). No correlation was observed among the salivary flow and the salivary IgG and IgM levels, while there was a negative relationship (p < 0.01 with salivary IgA levels in all 45 patients considered as a whole. In 12 patients with pSS, the coefficient of correlation between the salivary gland biopsy focus score and the salivary (but not serum) concentration of beta 2-microglobulin was highly significant (p < 0.001). A similar relationship was observed with the IgG (p < 0.001) and IgM (p < 0.05) levels, but not IgA. In the diagnosis of pSS, the salivary IgA level demonstrates high sensitivity and low specificity. In contrast, serum and salivary concentrations of beta 2-microglobulin, salivary IgG, and to a lesser degree, salivary IgM have high specificity and positive predictive value.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1993

[Salivary immunoglobulins in the diagnosis of primary Sjögren's syndrome].

The salivary concentrations of immunoglobulins and total proteins and the salivary flow rate (measured by the method of Deniker et al.) were determined in 29 patients with keratoconjunctivitis sicca and xerostomia and in 11 healthy subjects. According to Fox criteria the group pf 29 patients was divided into two groups: 14 patients with primary Sjögren's syndrome (pSS) and 15 patients with sicca syndrome (SS). The IgA salivary levels were significantly higher in the patients with pSS than in the patients with SS (p < 0.05) and in the healthy subjects (p < 0.001). Moreover the IgG and IgM salivary levels in the first group of patients were higher than in the group of patients suffering from sicca syndrome (p < 0.05) and in the control group (p < 0.02). The salivary flow rate of the patients with pSS and sicca syndrome was lower when compared with the control group (p < 0.005). No significant correlation was found between the salivary flow rate and the immunoglobulins in the three groups of patients. The salivary flow was inversely correlated (p < 0.01) only with the IgA levels in all 40 subjects. The salivary total proteins were significantly elevated in pSS patients when compared with the SS patients (p < 0.04) and with the control group (p < 0.01). The immunoglobulins/total proteins ratios in saliva were significantly higher in the first group than in the other groups (p < 0.005) for the IgG and in the first group than in the SS patients (p < 0.05) for the IgM; no significant differences were noted for the IgA. The bioptic focus scores, graded according Greenspan's criteria, of minor salivary glands of 20 patients with pSS or sicca syndrome were directly correlated with a high significance (p < 0.001) to IgG salivary levels and with a lower significance (p < 0.05) with IgM salivary levels. No significant correlation was found between the bioptic focus scores and the IgA levels. The present results demonstrated a high specificity (100%) and a good sensitivity (86%) of the assessment of salivary IgG for the diagnosis of pSS. The sensitivity of the salivary levels of IgA was higher than that of IgG (93%) but the specificity was low (50%); on the contrary the specificity of the assessment of salivary IgM was good (92%) but the sensitivity was low (60%). This study therefore indicated that the assessment of salivary IgG has a great value for the diagnosis of pSS because IgG levels are related to the amount of lymphoplasmocytic infiltration.(ABSTRACT TRUNCATED AT 400 WORDS)

Female↗
PubMed1992

[Salivary and serum beta 2-microglobulin in the diagnosis of primary Sjögren's syndrome].

beta 2-microglobulin (beta 2m) serum and salivary levels and total protein salivary levels were determined in 15 patients with primary Sjögren's syndrome (pSS), 14 patients with sicca syndrome and 11 healthy subjects. beta 2m serum and salivary levels were higher (p < 0.005) in the patients with pSS than in the healthy subjects and in the patients with sicca syndrome. The bioptic focus scores of minor salivary glands were correlated to beta 2m salivary levels with a high significance (p < 0.001) but were not correlated to beta 2m serum levels. The assessment of beta 2m serum and salivary levels is a non-invasive clinical investigation, which may be easily repeated. It is a very sensitive and highly specific test for the diagnosis of pSS. It may be therefore a useful mean to follow the natural course of pSS and to evaluate the effects of therapy.

Diagnosis, Differential↗
PubMed1990

Superoxide anion production by circulating polymorphonuclear leucocytes in rheumatoid arthritis.

In twelve rheumatoid arthritis (RA) patients, receiving only nonsteroid anti-inflammatory therapy, superoxide anion (02-) generation by polymorphonuclear leucocytes (PMNs) was assessed by Cytochrome C reduction. The 02- production by non-activated PMNs in RA patients was significantly higher than in healthy controls. The 02- production by PMNs activated by zymosan and phorbol myristate acetate was significantly lower than in controls. PMNs from controls preincubated with rheumatoid sera generated higher 02- levels than the same PMNs incubated in normal sera.

Adult↗
PubMed1989

Specific interactions between DNA left-handed supercoils and actinomycin D.

The interactions between the natural cyclopentapeptide antibiotic actinomycin D (ACT) and circular pBR322 DNA have been studied by freezing the topological state of the DNA in the complex by topoisomerase I reaction. Both supercoiled and relaxed DNAs, in the complexes at low antibiotic/DNA base-pair ratios, showed a dramatic decrease in linking number that cannot be explained by taking into account only the generally accepted unwinding of 28 degrees for each ACT molecule bound. Recent results derived from the crystallographic analysis of the complex between GpC and ACT suggest that ACT could mediate non-covalent cross-links between distant sections of DNA. Bridges between ACT and different sections of the pBR322 double helix could also explain our results. Two-dimensional gel electrophoresis of ACT-relaxed pBR322 DNA complexes reveals that all supercoils induced by ACT are negative. Two models of the complexes which correspond to the stabilization of DNA crossing by one or two molecules of ACT are proposed. In both cases the ability of ACT to stabilize only DNA left-handed supercoils is derived from the chirality of ACT, when it interacts with DNA.

Animals↗