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G Salvatori

Publications and source records attributed to G Salvatori.

At least 19 recordsLinked to original sources
PubMed2006

Management of fluid overload in congestive heart failure: learning from a case report.

A case of refractory fluid overload due to congestive heart failure and consequent renal insufficiency is reported. The case was approached multidisciplinarily, at the beginning with conservative and pharmacological therapy, subsequently with extracorporeal fluid removal in which a specific attention was payed to the maintenance of circulating blood volume and achievement of dry weight, and finally with chronic peritoneal dialysis as a maintenance therapy. The case seems to summarize the pathway of many patients seen initially in intensive care and cardiology departments and subsequently in nephrological wards.

Aged↗
PubMed2005

Effect of cyclosporine, mycophenolate mofetil, and their combination with steroids on apoptosis in a human cultured monocytic U937 cell line.

Transplant patient plasma produces an increased rate of mononuclear cell apoptosis despite a normal serum creatinine value. Immunosuppressive medications may be one factor that causes an altered apoptotic pattern. We evaluated the in vitro effects of various doses of cyclosporine, mycophenolate mofetil, and steroids on apoptosis of a cultured human monocytic U937 cell line, using estimates by fluorescence microscopy and annexin V assays. Increasing cyclosporine concentrations (100 to 800 ng/mL) progressively increased apoptosis rates (16% to 32%). The combination of steroid (0.01 microg/mL) and cyclosporine increased the apoptosis rate to 45%. Mycophenolate mofetil alone (0.3 microg/mL) led to an apoptosis rate of 34%. Therapeutic levels of mycophenolate mofetil from 3 to 7 microg/mL led to apoptosis rates from 56% to 67%. The combination of cyclosporine, steroid, and mycophenolate mofetil increased the rate of apoptosis to 95%. Immunosuppressive therapy may contribute to the high rate of apoptosis observed among mononuclear cells of transplanted patients. This effect may alter patient susceptibility to infections and contribute to a unique mechanism of immunosuppression.

Annexin A5↗
PubMed2004

Pulse high volume hemofiltration.

The sepsis syndrome is the most common cause of acute renal failure and multiple organ dysfunction in critically ill subjects and continues to have an alarmingly high mortality. Normal immune homeostasis is interrupted by a complex storm of inflammatory mediators responsible for the deleterious effects. Extracorporeal blood purification techniques can confer benefits in sepsis by proven non-specific removal of these mediators (pro- or anti-inflammatory), and provide a logical and adequate approach to treat this syndrome. High volume hemofiltration (HVHF) has had the most dramatic effect conferring benefits in hemodynamics, reduction in vasopressor doses and improvement in survival. "Pulse HVHF" is the latest approach which may offer the most efficient results: a daily schedule of 6-8 hours followed by standard CVVH. This paper describes the rationale and potential of this technique. Reliability and tolerance of this technique and biological effects are described.

Acute Kidney Injury↗
PubMed2004

First clinical trial for a new CRRT machine: the Prismaflex.

A new CRRT machine has been designed to fulfill the expectations of nephrologists and intensivists operating in the common ground of critical care nephrology. The new equipment is called "Prismaflex" (Gambro-Dasco, Mirandola, Modena) and it is the natural evolution of the Prisma machine that has been utilized worldwide for CRRT in the last decade. We performed a preliminary "alfa trial" to establish usability, flexibility and realiability of the new device. Accuracy was also tested by recording various operational parameters during different intermittent and continuous renal replacement modalities. Forty-one runs were conducted on 13 patients and the difference between delivered and prescribed parameters was always lower than 2%. We concluded that the new Prismaflex is a well designed new machine for CRRT and can be safely and effectively utilized in the critical care nephrology setting.

Acute Kidney Injury↗
PubMed2004

Continuous renal replacement technology: from adaptive devices to flexible multipurpose machines.

OBJECTIVE: To review the evolution of technologies in the development of renal replacement therapies. DATA SOURCES: Articles and published reviews on renal replacement therapies. SUMMARY OF REVIEW: Continuous arterio-venous haemofiltration (CAVH) was the first continuous renal replacement technique capable of overcoming the traditional haemodialysis-related side effects, making possible the treatment of critically ill patients safely and with less physiological instability. The evolution of technology and the progress experienced in intensive care units (ICUs) has made it possible to start renal replacement therapy programs in the absence of a chronic dialysis facility or a trained nephrological team. Initial limitations and draw-backs of CAVH, stimulated the ICU staff to explore new avenues for better therapy. Extracorporeal therapies are today a routine experience in the ICUs: continuous renal replacement therapies are a broadly accepted treatment for acute renal failure. Furthermore, alternative indications for extracorporeal blood circulation (e.g. sepsis, liver failure, congestive heart failure, drug intoxications, hyperthermia, immuno-mediated syndromes) are becoming more and more popular. The ideal machine has yet to be completed, but progress has occurred and has opened a new era for critical care nephrology and the further expansion of blood purification technology in the ICU. CONCLUSIONS: Technical advances in renal replacement therapies have increased their functionality (i.e. used in hepatic failure, sepsis, cardiac failure and immuno-mediated syndromes), are easier to operate and have less side-effects compared with their standard extracorporeal counterparts. Further improvements may see them become a routine part in the management of the critically ill patient.

Journal Article↗
PubMed2001

Growth, slaughter and intra-muscular collagen characteristics in Garganica kids.

Growth performance, ossification of metacarpal growth plate, and intra-muscular collagen characteristics in three muscles of 10 intact male Garganica kids were studied. Each week, starting from the 47th day of age to slaughter (75th day of age), kids were weighed and blood samples taken for plasma non-protein hydroxyproline analysis. At slaughter, metacarpal, metatarsal bones and metacarpal growth plate thickness were measured. The semitendinosus (ST), semimembranosus (SM), and rectus medialis (RM) muscle samples were analysed for collagen concentration and hydroxylysyl pyridinoline (HP) cross-links. ST muscle had a collagen amount higher than SM and RM (P<0.01), and RM muscle had the highest (P<0.01) HP concentration. Tenderness index was the highest (P<0.01) in RM muscle. Animal growth rates slowed, starting from 61st day of age (P<0.01); this trend coincided with a marked reduction in concentration of plasma non-protein hydroxyproline (P<0.01). In addition, plasma non-protein hydroxyproline concentration was positively correlated (r=0.61, P=0.001) with growth rate of animals, suggesting that collagen turnover decreases during a slow growth phase. Based on the metacarpal growth plate width, the rate of bone ossification appears to be slower in goats than sheep.

Journal Article↗
PubMed2001

Oxidative status of plasma and muscle in rabbits supplemented with dietary vitamin E.

Thirty New Zealand white rabbits, mean weight 2 kg, were divided into three equal groups balanced for body weight and randomly assigned to a diet containing 60 (C), 150 (T1) or 375 (T2) mg/kg of all-rac-alpha-tocopheryl acetate. After 29 days, the animals were slaughtered. alpha-Tocopherol was assayed in muscle (longissimus dorsi) and plasma; triglycerides and cholesterol (total, high density lipoprotein, low density lipoprotein) were analysed in plasma; reactive oxygen metabolites (ROMs) were analysed in serum; and thiobarbituric acid-reactive substances (TBARS) were analysed in muscle. There were no body weight and food intake differences between the groups. The plasma vitamin E and vitamin E:lipid ratio were significantly higher in groups T1 and T2 than in C, but increases were not linearly related to dietary levels. Muscle alpha-tocopherol concentrations in the treated groups were significantly higher than in C, and linearly related (R =.67) to the vitamin E:lipid ratio. ROM and vitamin E levels in blood were inversely related (R =.74), with ROMs significantly lower in the treated groups than in C. The 60-mg/kg dose of C recommended by the National Research Council was unable to control ROM production. Lipid oxidation in muscle was significantly lower in T2 than in the other groups, and TBARS correlated significantly with muscle vitamin E (R =.61) and serum ROM (R =.73). These data suggest that vitamin E supplemented at 375 mg/kg diet can effectively control ROM production and improve muscle lipostability. ROM assay provides a useful indirect estimate of the oxidative status of muscle in vivo.

Journal Article↗
PubMed2000

Determination, diversification and multipotency of mammalian myogenic cells.

In amniotes, myogenic commitment appears to be dependent upon signaling from neural tube and dorsal ectoderm, that can be replaced by members of the Wnt family and by Sonic hedgehog. Once committed, myoblasts undergo different fates, in that they can differentiate immediately to form the myotome, or later to give rise to primary and secondary muscle fibers. With fiber maturation, satellite cells are first detected; these cells contribute to fiber growth and regeneration during post-natal life. We will describe recent data, mainly from our laboratory, that suggest a different origin for some of the cells that are incorporated into the muscle fibers during late development. We propose the possibility that these myogenic cells are derived from the vasculature, are multi-potent and become committed to myogenesis by local signaling, when ingressing a differentiating muscle tissue. The implications for fetal and perinatal development of the whole mesoderm will also be discussed.

Animals↗
PubMed1999

Reversible immortalization of human myogenic cells by site-specific excision of a retrovirally transferred oncogene.

Myogenic cells have a limited life span in culture, which prevents expansion at clinically relevant levels, and seriously limits any potential use in cell replacement or ex vivo gene therapy. We developed a strategy for reversibly immortalizing human primary myogenic cells, based on retrovirus-mediated integration of a wild-type SV40 large-T antigen (Tag), excisable by means of the Cre-Lox recombination system. Myogenic cells were transduced with a vector (LTTN-LoxP) expressing the SV40 Tag under the control of an LTR modified by the insertion of a LoxP site in the U3 region. Clonal isolates of Tag-positive cells showed modified growth characteristics and a significantly extended life span, while maintaining a full myogenic potential. Transient expression of Cre recombinase, delivered by transfection or adenoviral vector transduction, allowed excision of the entire provirus with up to >90% efficiency. Cultures of Cre-treated (Tag-) or untreated (Tag+) myogenic cells were genetically labeled with a lacZ retroviral vector, and injected into the regenerating muscle of SCID/bg immunodeficient mice. Tag- cells underwent terminal differentiation in vivo, giving rise to clusters of beta-Gal+ hybrid fibers with an efficiency comparable to that of control untransduced cells. Tag+ cells could not be detected after injection. Neither Tag+ nor Tag- cells formed tumor in this xenotransplantation model. Reversible immortalization by Tag therefore allows the expansion of primary myogenic cells in culture without compromising their ability to differentiate in vivo, and could represent a safe method by which to increase the availability of these cells for clinical application.

3T3 Cells↗
PubMed1999

Effects of intramuscular injection of alpha-tocopheryl acetate on fatty acid profile in lamb liver.

The effects of intramuscularly administrated vitamin E on total lipids, fatty acid profile, and lipid stability to oxidation was investigated in lamb liver. Twenty-four 5-day-old lambs were allotted to 4 groups of 6 each and given respectively 0 (control), 125, 200, 300 mg dl-alpha-tocopheryl acetate weekly from day 5 to 33. alpha-Tocopherol stored in lamb liver at the end of experiment showed linear correlation with the level of injected vitamin E. No effect on total lipids was found. A decrease in the level of liver thiobarbituric-acid reactive substances (TBARS), significantly correlated with liver alpha-tocopherol content, was found in vitamin E groups. The amount of linoleic and linolenic acids significantly increased in the vitamin E groups as compared to control group, and were correlated with the liver alpha-tocopherol content. TBARS were negatively correlated with the concentration of unsaturated fatty acids. Finally, in the liver of the treated groups, vitamin E concentrations in the range 30-50 micrograms/g showed adequate for an efficient protection from peroxidation of membrane lipids, and determined an increase in the unsaturated to saturated fatty acid ratio.

Animals↗
PubMed1999

Influence of dietary vitamin E supplementation on "heavy" pig carcass characteristics, meat quality, and vitamin E status.

Crossbred "heavy" pigs (average weight 120 kg, slaughter weight above 160 kg) were supplemented with all-rac-alpha-tocopheryl acetate during the last 60 d of late finishing at doses of 25 (control), 50, 100, 200, or 300 mg/kg of diet. At the end of this period, the pigs were slaughtered. Carcass characteristics and the meat quality of pork chops from longissimus muscle (LM) at the last lumbar vertebra were evaluated on eight barrows from each dosage group. Alpha-tocopherol levels were determined in plasma during supplementation and in LM muscle after slaughter. Thiobarbituric acid reactive substances (TBARS) and drip loss were also evaluated in meat. Plasma alpha-tocopherol levels increased (P < .005) during supplementation in treated animals compared to controls, with a peak at 40 d. Alpha-tocopherol levels were higher (P < .05) in LM from pigs treated with 300 mg/kg than in controls (8.4 vs 5.6 microg/g). Dressing percentages correlated (P < .05) with the ratio of plasma alpha-tocopherol levels to the sum of cholesterol and triglycerides. Inhibition of TBARS during storage was related (P < .005) to vitamin E supplementation level, but drip losses in chops were not related to supplementation levels. We concluded that dietary vitamin E supplementation to heavy pigs during the last 60 d of finishing improves dressing percentage, reduces lipid oxidation, and increases the alpha-tocopherol concentration of tissues.

Animal Feed↗
PubMed1999

Influence of multiple injections of vitamin E on intramuscular collagen and bone characteristics in suckling lambs.

We studied the influence of vitamin E on intramuscular collagen characteristics and on metacarpal growth plate evolution in suckling lambs. Twenty-four 5-d-old Ile de France suckling male lambs were divided into four equal weight groups, and weekly i.m. injections of DL-alpha-tocopheryl acetate (Control group, 0 IU; Group 1, 625 IU; Group 2, 1,000 IU; and Group 3, 1,500 IU) were given until the lambs were 33 d old. Blood samples were withdrawn for plasma alpha-tocopherol, cholesterol, and triglyceride analyses when the lambs were slaughtered at 40 d of age. Hot carcass weight, metacarpal and metatarsal bone characteristics, and metacarpal growth plate width were measured. After 24 h at 2 to 4 degrees C, semitendinosus muscles were removed for intramuscular collagen analyses. Weight was not influenced by treatment, but lambs in Group 1 had a higher (P < .01) intramuscular collagen content than the other groups. A positive quadratic correlation was found between total collagen and vitamin E doses (r = .511; P < .05). Amount and percentage of soluble collagen increased in muscles of all three vitamin E-treated groups compared with the control group. However, hydroxylysyl pyridinoline concentration decreased with vitamin E injection, but the difference compared to control lambs was only significant for the group injected with 1,000 IU. Amount of soluble collagen was strongly correlated to total collagen concentration (r = .879; P < .0001). It was also related to the alpha-tocopherol/(cholesterol + triglycerides) ratio in plasma (r = .431; P < .04). However, the correlation between soluble collagen percentage and alpha-tocopherol concentration in plasma was not significant (r = .369; P < .08). Bone length, weight, and diameter did not change, but the thickness of metacarpal growth plate increased as the DL-alpha-tocopheryl acetate dose increased; in Groups 2 and 3, the growth plate was significantly wider than in Group 1 and the controls. Growth plate width correlated with alpha-tocopherol/(cholesterol + triglycerides) ratio in plasma (r = .481; P < .02).

Animals↗
PubMed1998

High efficiency myogenic conversion of human fibroblasts by adenoviral vector-mediated MyoD gene transfer. An alternative strategy for ex vivo gene therapy of primary myopathies.

Ex vivo gene therapy of primary myopathies, based on autologous transplantation of genetically modified myogenic cells, is seriously limited by the number of primary myogenic cells that can be isolated, expanded, transduced, and reimplanted into the patient's muscles. We explored the possibility of using the MyoD gene to induce myogenic conversion of nonmuscle, primary cells in a quantitatively relevant fashion. Primary human and murine fibroblasts from skin, muscle, or bone marrow were infected by an E1-deleted adenoviral vector carrying a retroviral long terminal repeat-promoted MyoD cDNA. Expression of MyoD caused irreversible withdrawal from the cell cycle and myogenic differentiation in the majority (from 60 to 90%) of cultured fibroblasts, as defined by activation of muscle-specific genes, fusion into contractile myotubes, and appearance of ultrastructurally normal sarcomagenesis in culture. 24 h after adenoviral exposure, MyoD-converted cultures were injected into regenerating muscle of immunodeficient (severe combined immunodeficiency/beige) mice, where they gave rise to beta-galactosidase positive, centrally nucleated fibers expressing human myosin heavy chains. Fibers originating from converted fibroblasts were indistinguishable from those obtained by injection of control cultures of lacZ-transduced satellite cells. MyoD-converted murine fibroblasts participated to muscle regeneration also in immunocompetent, syngeneic mice. Although antibodies from these mice bound to adenoviral infected cells in vitro, no inflammatory infiltrate was present in the graft site throughout the 3-wk study period. These data support the feasibility of an alternative approach to gene therapy of primary myopathies, based on implantation of large numbers of genetically modified primary fibroblasts massively converted to myogenesis by adenoviral delivery of MyoD ex vivo.

Adenoviridae↗
PubMed1997

Vitamin E nutritional status and serum lipid pattern in normal weanling rabbits.

In a 6-wk postweaning trial (Trial 1), 30 male New Zealand White rabbits were fed a starter diet with three different levels of all-rac-alpha-tocopheryl acetate (60, 160, or 260 mg/kg diet). Plasma concentrations of alpha-tocopherol and serum concentrations of total cholesterol, high density lipoprotein (HDL) cholesterol, and triglycerides were measured at 2, 4, and 6 wk. In a 20-wk postweaning trial (Trial 2), 48 male New Zealand White rabbits were divided into two groups; one group was fed a vitamin E-free diet, and the other was fed the same diet supplemented with 60 mg/kg of all-rac-alpha-tocopheryl acetate. Plasma concentrations of alpha-tocopherol and serum total cholesterol, HDL cholesterol, and triglycerides were measured at 2, 4, 6, and 20 wk. In Trial 1, split-plot ANOVA showed that dietary all-rac-alpha-tocopheryl acetate increased plasma alpha-tocopherol (P < .01); in addition, serum total cholesterol and triglycerides declined more rapidly (P < .01) and HDL cholesterol increased more rapidly (P < .005) in rabbits fed supplemental all-rac-alpha-tocopheryl acetate than in controls. The all-rac-alpha-tocopheryl acetate intake was significantly correlated with serum total cholesterol reduction and HDL cholesterol increase only when plasma alpha-tocopherol was relatively low (< 23.5 mumol/ L). In trial 2, the vitamin E-free diet caused an increase (P < .05) in total serum cholesterol and a reduction in HDL cholesterol (P < .05 at 6 wk and P < .01 at 20 wk). The overall results of this study suggest that vitamin E plays an important role in the regulation of serum concentrations of cholesterol and lipoproteins in weanling rabbits. Consequently, the maintenance of an adequate nutritional status of vitamin E in the postweaning period is important to avoid alterations of serum lipid pattern.

Aging↗
PubMed1997

A temperature-conditional mutant of simian virus 40 large T antigen requires serum to inhibit myogenesis and does not induce DNA synthesis in myotubes.

The temperature-conditional mutant tsA58 of SV40 large T antigen (Tag) increases the proliferation rate and the number of cell divisions in primary murine and human myogenic cells when expressed under permissive conditions (i.e., at 33 degrees C in medium containing high levels of serum). Under these conditions, Tag also prevents terminal differentiation. Under nonpermissive conditions (i.e., at 39 degrees C in medium containing low levels of serum) in which Tag is largely inactive, proliferation is arrested, and differentiation occurs. However, even at a permissive temperature, the removal of serum induced myosin expression and the fusion of myogenic cells, which continued to express functional Tag. Although Tag was complexed with pRb, as expected from a functional protein, proliferation was nevertheless arrested, and differentiation was induced. Consistent with these findings, the exposure of Tag-expressing differentiated myotubes to serum at 33 degrees C did not reinduce DNA synthesis in these cells. Thus, in myogenic cells, temperature-conditional mutants of Tag stimulate proliferation in the presence of serum but neither prevent terminal differentiation in the absence of serum nor induce DNA synthesis once complete withdrawal from the cycle has occurred.

Animals↗
PubMed1996

Terminally differentiated skeletal myotubes are not confined to G0 but can enter G1 upon growth factor stimulation.

Terminally differentiated cells are specialized cells unable to proliferate that constitute most of the mammalian body. Despite their abundance, little information exists on the characteristics of cell cycle control in these cells and the molecular mechanisms that prevent their proliferation. They are generally believed to be irreversibly restricted to the G0 state. In this report, we define some features of a paradigmatic terminally differentiated system, the skeletal muscle, by studying its responses to various mitogenic stimuli. We show that forced expression of a number of cell cycle-regulatory genes, including erbB-2, v-ras, v-myc, B-myb, ld-1, and E2F-1, alone or in combinations, cannot induce terminally differentiated skeletal muscle cells (myotubes) to synthesize DNA. However, serum-stimulated myotubes display a typical immediate-early response, including the up-regulation of c-fos, c-jun, c-myc, and ld-1. They also elevate the expression of cyclin D1 after 4 hours of serum treatment. All these events take place in myotubes in a way that is indistinguishable from that of quiescent, undifferentiated myoblasts reactivated by serum. Moreover, pretreatment with serum shortens the time required by E1A to induce DNA synthesis, confirming that myotubes can partially traverse G1. Serum growth factors do not activate late-G1 genes in myotubes, suggesting that the block that prevents terminally differentiated cells from proliferating acts in mid-G1. Our results show that terminally differentiated cells are not confined to G0 but can partially reenter G1 in response to growth factors; they contribute to a much-needed definition of terminal differentiation. The important differences in the control of the cell cycle between terminally differentiated and senescent cells are discussed.

Adenovirus E1A Proteins↗
PubMed1995

A retroviral vector containing a muscle-specific enhancer drives gene expression only in differentiated muscle fibers.

Genetically modified myogenic cells have a number of potentially relevant applications for gene therapy of genetic defects. Retroviral vectors proved to be a safe and efficient tool to transfer and express genes into satellite cells and their differentiated progeny, although muscle-specific regulation of the transferred gene is very difficult to achieve in a conventional vector framework. We modified a Moloney murine leukemia virus (MoMLV)-derived retroviral vector containing a bacterial beta-galactosidase (beta-Gal) reporter gene by inserting a muscle creatinine kinase (MCK) enhancer element into the U3 region of the viral long terminal repeat (LTR). The resulting vector (mLBSN) was transferred into cells of different histological origin, including undifferentiated murine and human myogenic cells, which were unable to express the transgene at detectable levels. Instead, gene expression from the modified LTR was obtained in a mouse myogenic cell line and in human primary satellite cells upon induction of differentiation into myotubes in culture, and correlated with the activation of the muscle differentiation program. beta-Gal-negative, mLBSN-transduced human satellite cells were also transplanted into the quadricep muscle of immunodeficient mice, where activation of the transgene expression was observed in vivo after differentiation and fusion into muscle fibers. These results show that retroviral vectors carrying LTRs modified in the enhancer sequences may be used to target tissue- and differentiation-specific gene expression into the muscle. For practical purposes, satellite cells engineered by muscle-specific retroviral vectors might represent an effective tool to deliver expression of a given gene product specifically into the muscle tissue, avoiding undesired protein accumulation in mononucleated cells. More generally, this type of vector might be useful whenever regulated expression of a transferred gene is necessary in a target cell or tissue.

Animals↗