PubMed Health⌕ Search

Biomedical subjects

G Salviati

Publications and source records attributed to G Salviati.

At least 19 recordsLinked to original sources

Antiphase disorder in GaAs/Ge heterostructures for solar cells

Antiphase disorder in metal organic vapour phase epitaxy grown GaAs/(100)Ge heterostructures has been studied both in as-grown materials and in GaAs solar cells by chemical etching, transmission electron microscopy, and cathodoluminescence. All the samples are single domains at the surface due to the self-annihilation of antiphase domains whose size decreases as the misorientation angle increases. Completely antiphase domain-free epitaxy has been achieved for substrate miscuts greater than 3 degrees off towards [111]. A reversal in sublattice location has been found in the GaAs layers varying the misorientation angle and the growth temperature. A model to explain this result has been proposed based on the role of surface steps in the nucleation process. Strong interaction between antiphase boundaries and misfit dislocations has been found in all the heterostructures. In solar cells antiphase domains have been observed in high densities in the initial layer of GaAs deposited on Ge. The successful realisation of high efficiency solar cells is due to the overgrowth of these domains by single phase material over most of the wafer area.

Journal Article↗

Low-temperature spectrally resolved cathodoluminescence study of degradation in opto-electronic and microelectronic devices

This study reports on the microcharacterization of devices for optoelectronic and for microelectronic applications using low temperature (T = 5 and 77 K) spectrally resolved cathodoluminescence (SCL). The mechanisms leading to compositional inhomogeneities in the regrowth regions of InP-based butt-coupled laser-waveguide devices for semiconducting optical amplifiers (SOAs) and for defect generation in the active and cladding layers of GaAs based pump lasers for erbium-doped optical fibre amplifiers (EDFAs) were studied. Beryllium outdiffusion in the base regions of GaAs-based heterojunction bipolar transistors (HBTs) after bias ageing was also studied. By comparing the CL results with TEM, SIMS and HRXRD studies and with the existing literature, the observed growth and operation induced defects were attributed, respectively, to the following mechanisms: recombination-enhanced defect glide (REDG) in the pump lasers, recombination enhanced impurity diffusion (REID) in the HBTs and electrostatically induced growth flux instabilities in the butt-coupled laser-waveguide devices.

Journal Article↗

Ecto-ATPase activity of alpha-sarcoglycan (adhalin).

alpha-Sarcoglycan is a component of the sarcoglycan complex of dystrophin-associated proteins. Mutations of any of the sarcoglycan genes cause specific forms of muscular dystrophies, collectively termed sarcoglycanopathies. Importantly, a deficiency of any specific sarcoglycan affects the expression of the others. Thus, it appears that the lack of sarcoglycans deprives the muscle cell of an essential, yet unknown function. In the present study, we provide evidence for an ecto-ATPase activity of alpha-sarcoglycan. alpha-Sarcoglycan binds ATP in a Mg2+-dependent and Ca2+-independent manner. The binding is inhibited by 3'-O-(4-benzoyl)benzoyl ATP and ADP. Sequence analysis reveals the existence of a consensus site for nucleotide binding in the extracellular domain of the protein. An antibody against this sequence inhibits the binding of ATP. A dystrophin.dystrophin-associated protein preparation demonstrates a Mg-ATPase activity that is inhibited by the antibody but not by inhibitors of endo-ATPases. In addition, we demonstrate the presence in the sarcolemmal membrane of a P2X-type purinergic receptor. These data suggest that alpha-sarcoglycan may modulate the activity of P2X receptors by buffering the extracellular ATP concentration. The absence of alpha-sarcoglycan in sarcoglycanopathies leaves elevated the concentration of extracellular ATP and the persistent activation of P2X receptors, leading to intracellular Ca2+ overload and muscle fiber death.

Adenosine Triphosphatases↗

Gender- and thyroid hormone-related transitions of essential myosin light chain isoform expression in rat soleus muscle during ageing.

In this brief review, the modulatory influence of essential myosin light chain (MLC) isoforms on muscle cell contractility is discussed. Specific interest is focused on the expression of the MLC1Sa and MLC1Sb isoforms in the slow-twitch soleus muscle in male and female rats, during ageing and after thyroid hormone treatment. According to two-dimensional gel electrophoresis analysis, the MLC1Sa/MLC1SB ratio increased during ageing in both males and females in parallel with the age-related decrease in shortening velocity reported in muscle fibres expressing the slow (type 1) myosin heavy chain (MHC) isoform. However, the MLC1Sa and MLC1Sb isoform expression responded to thyroid hormone treatment in a complex manner which did not parallel the age-related changes in shortening velocity reported in hyperthyroid animals. Thus, if MLC1Sa and MLC1Sb isoforms modulate shortening velocity in type 1 fibres, then other modulators of shortening velocity are not regulated by thyroid hormone in co-ordination with these essential MLCs.

Aging↗

A novel muscle protein located inside the terminal cisternae of the sarcoplasmic reticulum.

An immunofluorescence study of adult rat muscle tissues with a polyclonal antibody against the RGD-directed fibronectin receptor of Friend's erythroleukemia cells (alpha5beta1-integrin) unexpectedly revealed a pattern of intracellular antigen distribution. Western blotting analysis of rat and rabbit membrane fractions indicated that the antibody recognizes a 167-kDa protein expressed both in heart and in skeletal muscle (relative abundance: heart > slow muscle > fast muscle), but not in liver and kidney. The 167-kDa protein did not show altered electrophoretic mobility upon reduction and failed to bind several lectins, including wheat germ agglutinin. A study of its subcellular distribution in rabbit skeletal muscle revealed that the 167-kDa protein is mostly associated with the terminal cisternae of the sarcoplasmic reticulum (SR) and, to a smaller extent, with the sarcolemma, while it is absent in the longitudinal tubules of the SR. The 167-kDa protein is not an integral membrane protein since it can be extracted at pH >/=10. This protein can be proteolytically cleaved only in the presence of detergent, indicating that it resides on the luminal side of the SR. The 167-kDa protein could be resolved from the closely spaced sarcalumenin and histidine-rich protein by column chromatography followed by detergent dialysis and two-dimensional gel electrophoresis. The N terminus and the internal sequences did not match any known sequence in protein and DNA data bases, indicating that the 167-kDa protein is a novel muscle protein selectively localized to the SR. Integrins from rat kidney fibroblasts were not recognized by either (i) a polyclonal antiserum against the purified 167-kDa protein or (ii) the anti-alpha5beta1-integrin antiserum after affinity purification onto the 167-kDa protein. These data indicate that the 167-kDa protein is not immunologically cross-reactive with integrins, despite its reaction with a polyclonal anti-integrin antibody.

Animals↗

Sphingosylphosphocholine modulates the ryanodine receptor/calcium-release channel of cardiac sarcoplasmic reticulum membranes.

Sphingosylphosphocholine (SPC) modulates Ca2+ release from isolated cardiac sarcoplasmic reticulum membranes; 50 microM SPC induces the release of 70 80% of the accumulated calcium. SPC release calcium from cardiac sarcoplasmic reticulum through the ryanodine receptor, since the release is inhibited by the ryanodine receptor channel antagonists ryanodine. Ruthenium Red and sphingosine. In intact cardiac myocytes, even in the absence of extracellular calcium. SPC causes a rise in diastolic Ca2+, which is greatly reduced when the sarcoplasmic reticulum is depleted of Ca2+ by prior thapsigargin treatment. SPC action on the ryanodine receptor is Ca(2+)-dependent. SPC shifts to the left the Ca(2+)-dependence of [3H]ryanodine binding, but only at high pCa values, suggesting that SPC might increase the sensitivity to calcium of the Ca(2+)-induced Ca(2+)-release mechanism. At high calcium concentrations (pCa 4.0 or lower), where [3H]ryanodine binding is maximally stimulated, no effect of SPC is observed. We conclude that SPC releases calcium from cardiac sarcoplasmic reticulum membranes by activating the ryanodine receptor and possibly another intracellular Ca(2+)-release channel, the sphingolipid Ca(2+)-release-mediating protein of endoplasmic reticulum (SCaMPER) [Mao, Kim, Almenoff, Rudner, Kearney and Kindman (1996) Proc.Natl.Acad.Sci. U.S.A 93, 1993-1996], which we have identified for the first time in cardiac tissue.

Animals↗

Thyroid hormone effects on contractility and myosin composition of soleus muscle and single fibres from young and old rats.

1. Young (3-6 months) and old (20-24 months) male Wistar rat soleus muscles were examined for myosin isoform composition, fibre type, contractility and sarcoplasmic reticulum (SR) Ca2+ release properties either in control rats or in rats treated with thyroid hormone (3,5,3'-triiodothyronine, T3) for 4 weeks. 2. T3 treatment had a strong impact on myosin heavy chain (MyHC) and light chain (MyLC) isoform composition in both young and old rats. That is, all single fibres co-expressed type I and IIA (type I/IIA fibres) or type I, IIA and IIX MyHCs (type I/IIAX fibres) after treatment. Slow and fast MyLC isoforms, i.e. MyLC1s, MyLC1f, MyLC2s, MyLC2f and MyLC3, co-existed in each of the type I/IIA and I/IIAX fibres in variable proportions. 3. In old rats the maximum velocity of unloaded shortening (V0) was related to MyHC isoform composition: V0 for type I fibres was less than that for type I/IIA fibres which was less than that for type I/IIAX fibres. In young rats, on the other hand, V0 did not differ between pure type I fibres from controls and those co-expressing type I and type II MyHC isoforms from T3-treated rats. 4. Contraction and half-relaxation times of the isometric twitch were significantly longer in old than in young controls. This was paralleled by an age-related decrease in the caffeine threshold of the SR. Four weeks of T3 treatment eliminated the age-related differences in both speed of twitch contraction and caffeine thresholds. V0, on the other hand, was slower in old than in young animals, both control and T3-treated, when cells with a similar MyHC composition were compared. 5. In conclusion, thyroid hormone can substantially reverse at least some of the changes that occur in ageing muscle. Further, the age-related decline in V0 in soleus fibres from control and hyperthyroid rats suggests that: (1) the identification of beta/slow myosin isoforms is incomplete; or (2) the molecular characteristics of MyHC differ between young and old age; or (3) MyHC is not the only determinant of V0.

Aging↗

Increased expression of dystrophin, beta-dystroglycan and adhalin in denervated rat muscles.

To evaluate a potential regulatory role of the nerve, the distribution and expression of dystrophin, of beta-dystroglycan (43DAG) and adhalin (50DAG), two of the dystrophin-associated proteins and utrophin (dystrophin related protein or DRP) were studied in rat muscles after 2 weeks of denervation. We found that dystrophin, beta-dystroglycan and adhalin were overexpressed in denervated muscle, whereas utrophin did not increase and was found only in the post-synaptic membrane. The study of the distribution of dystrophin in the sarcolemma of single muscle fibres indicates that the molecular organization of dystrophin was maintained after denervation. Dystrophin in addition of forming a scaffold around the fibre was found around the clusters of AChR that reappeared in the extra-synaptic membrane after denervation. Also beta-dystroglycan colocalises at these clusters. These results suggest that the increase in dystrophin, beta-dystroglycan and adhalin is correlated with the reappearance of AChRs in the extra synaptic membrane.

Animals↗

Involvement of ryanodine receptors in sphingosylphosphorylcholine-induced calcium release from brain microsomes.

Sphingosylphosphorylcholine (SPC) releases Ca2+ from brain microsomes. SPC-induced CA2+ release differs from IP3-induced Ca2+ release in that it is more extensive in the cerebrum than in the cerebellum. SPC has little effect on [3H] IP3 binding but enhances [3H] ryanodine binding, as expected for an activator of ryanodine receptors. SPC-induced Ca2+ release is inhibited by ryanodine receptor blockers but not by selective blockers of IP3 receptors. We conclude that SPC releases Ca2+ from brain microsomes by activating ryanodine receptors rather than IP3 receptors. Activation of an additional SPC-sensitive pathway for releasing Ca2+ is not precluded.

Animals↗

Phosphorylation of dystrophin:effects on actin binding.

Dystrophin is phosphorylated by several protein kinases. In this work, we have studied the effects of dystrophin phosphorylation on the binding to actin. Purified dystrophin was phosphorylated in vitro by the catalytic subunit of cAMP-dependent protein kinase (PKA), casein kinase II (CK-II), and protein kinase c (PKC). The results demonstrate that phosphorylation of dystrophin by PKA phosphorylation caused a three fold increase in dystrophin binding to actin. In contrast, phosphorylation by CK-II or PKC inhibited the binding to actin. These results indicate that phosphorylation of dystrophin modulates its interaction with the actin cytoskeleton. It is suggested that phosphorylation may be one mechanism for regulating protein turnover in muscle membrane-skeleton.

Actins↗

Effects of age on sarcoplasmic reticulum properties and histochemical composition of fast- and slow-twitch rat muscles.

Calcium release activity of sarcoplasmic reticulum and enzyme-histochemical properties were investigated in extensor digitorum longus (e.d.l.) and soleus muscles in young (4 months and old (24 months) male rats. With age, the caffeine threshold concentration for calcium release from the sarcoplasmic reticulum of soleus skinned muscle fibres showed only minor modifications. On the other hand, in e.d.l. skinned muscle fibres, the caffeine threshold concentration decreased significantly (P < 0.05). The histochemical fibre type composition changed with age both in soleus and in e.d.l. muscles, showing a common transformation toward a more oxidative histochemical profile. In fact, in aged soleus, a significant (P < 0.05) increase was observed of type 1 fibres to represent almost the totality of the muscle fibres (more than 98%), while types 2C and 2A were reduced in proportion. In aged e.d.l. the percentage of type 1 (P < 0.05), 2A and 2X (a recently identified fourth component of the fast-twitch muscle types) fibres increased, with a reduction of type 2B (P < 0.01) fibres. The present results suggest that the changes in contractile properties of aged muscles may be related to the changes not only in fibre composition but also in the mechanism of calcium release from sarcoplasmic reticulum.

Aging↗

Effects of thyroid hormone on fast- and slow-twitch skeletal muscles in young and old rats.

1. The effects of 4 weeks of thyroid hormone treatment on contractile, enzyme-histochemical and morphometric properties and on the myosin isoform composition were compared in the slow-twitch soleus and the fast-twitch extensor digitorum longus (EDL) muscle in young (3-6 months) and old (20-24 months) male rats. 2. In the soleus of untreated controls, contraction and half-relaxation times of the isometric twitch increased by 19-32% with age. The change in contractile properties was paralleled by an age-related increase in the proportions of type I fibres and type I myosin heavy chain (MHC) and slow myosin light chain (MLC) isoforms. 3. In the EDL of controls, contraction and half-relaxation times were significantly prolonged (21-38%) in the post-tetanus twitch in the old animals. No significant age-related changes were observed in enzyme-histochemical fibre-type proportions, although the number of fibres expressing both type IIA and IIB MHCs and of fibres expressing slow MLC isoforms was increased in the old animals. 4. Serum 3,5,3',5'-tetraiodothyronine (T4) levels were lower (34%) in the old animals, but the primary byproduct of T4, 3,5,3'-triiodothyronine (T3), did not differ between young and old animals. 5. The effects of 4 weeks of thyroid hormone treatment were highly muscle specific, and were more pronounced in soleus than in EDL, irrespective of animal age. In the soleus, this treatment shortened the contraction and half-relaxation times by 35-57% and decreased the number of type I fibres by 66-77% in both young and old animals. In EDL, thyroid hormone treatment significantly shortened the contraction time by 24%, but the change was restricted to the old animals. 6. In conclusion, the ability of skeletal muscle to respond to thyroid hormone treatment was not impaired in old age and the age-related changes in speed of contraction and enzyme-histochemical properties and myosin isoform compositions were diminished after thyroid hormone treatment in both the soleus and EDL.

Aging↗

Modulation of cardiac sarcoplasmic reticulum ryanodine receptor by sphingosine.

Excitation contraction (EC) coupling in muscle cells involves the movement of calcium through the calcium release channel of the sarcoplasmic reticulum (SR) membrane known as the ryanodine receptor. We have recently shown that the novel second messenger, sphingosine, can block calcium release from skinned skeletal muscle fibers and from isolated skeletal muscle SR membranes (Sabbadini et al., J Biol Chem 267: 15475-15484, 1992). In this report, we demonstrate that sphingosine also inhibits calcium release from isolated canine cardiac SR membranes containing the ryanodine receptor when release is induced by caffeine, doxorubicin or by calcium. Sphingosine also prevents the augmentation of [3H]-ryanodine binding normally produced by caffeine and doxorubicin and exerts noncompetitive inhibition with regard to both releasing agents. Sphingosine significantly reduces in a dose-dependent manner [3H]-ryanodine binding to the high affinity site of the receptor and increases by several-fold the Kd for binding, which is consistent with a blocking action of sphingosine on the ryanodine receptor calcium channel. Sphingosine inhibits the extent of calcium-induced calcium release (CICR) and significantly shifts the threshold for CICR so that a higher level of trigger calcium is required to initiate CICR. The sphingosine inhibition of CICR is consistent with the near abolition of calcium dependent [3H]-ryanodine binding. HPLC analysis of cardiac sphingosine content indicates that sphingosine is present in the cardiac cell at moderately high levels (29.4 nmol/g wet wt for the entire cell and approximately 0.4 microM for the cytosol) which are sufficient to produce significant inhibition by sphingosine on calcium release and ryanodine binding. The data suggest that sphingosine acts on the cardiac ryanodine receptor by opposing the physiological stimulus (e.g. trigger calcium entering via the dihydropyridine receptor). We propose that sphingosine is produced by the T-tubule membranes and that sphingosine is released into the protected intracellular environment of the T-tubule/SR junction to negatively modulate calcium release. Consequently, it is possible that sphingosine is a physiologically relevant regulator of calcium levels in the heart.

Animals↗

Reducing body myopathy and desmin storage in skeletal muscle: morphological and biochemical findings.

We describe clinical, morphological and biochemical findings of a patient with reducing body myopathy (RBM). This 15-year-old patient was affected by severe limb-girdle progressive myopathy with asymmetric distribution. Muscle biopsy showed many fibers with cytoplasmic polymorphic masses, which stained dark purple with modified Gomori's trichrome, associated with proliferation of cytoplasmic bodies. Cytoplasmic polymorphic masses showed marked reducing activity with menadione-nitro blue tetrazolium reaction. Ultrastructurally, there was great amount of highly electron-dense tubular-filamentous structures of 16-17 nm in diameter. Immunohistochemistry showed that many fibers were positive for desmin. Sodium dodecyl sulfate-electrophoresis disclosed an increase in two bands of approximately 53 and 70 kDa, and Western blot demonstrated that the 53-kDa band was desmin. It was not possible to characterize the 70-kDa protein further.

Adolescent↗

Control of cardiac Ca2+ levels. Inhibitory actions of sphingosine on Ca2+ transients and L-type Ca2+ channel conductance.

The naturally occurring second messenger sphingosine (SPH) was examined for its ability to influence cardiac myocyte Ca2+ regulation. SPH inhibited intracellular Ca2+ transients in adult and neonatal rat ventricular myocytes. The inhibition was steeply dose dependent, with complete blockage of the Ca2+ transients occurring in the 20- to 25-mumol/L range. Whole-cell patch clamping revealed substantial inhibition of the L-type Ca2+ channel current (ICa) by SPH. The ability of SPH to block both the Ca2+ transients and ICa was not dependent on protein kinases, since the general protein kinase inhibitor H7 failed to prevent the actions of SPH. The specificity of the effect of SPH was determined in experiments showing that SPH analogues did not produce comparable effects. Neither the naturally occurring ceramide, N-stearoyl SPH, nor the cell-permeant ceramide, N-acetyl SPH, had SPH-like actions on the Ca2+ transients or L-type channel conductances. Caffeine-induced Ca2+ transients were also inhibited by the actions of SPH on cardiac sarcoplamic reticulum Ca2+ release, and the threshold for caffeine-induced Ca2+ release was raised. We conclude that SPH inhibits excitation-contraction coupling in cardiac myocytes by reducing the amount of entering "trigger Ca2+" for Ca(2+)-induced Ca2+ release and by simultaneously raising the threshold of the ryanodine receptor for Ca(2+)-induced Ca2+ release. Consequently, we propose that sphingolipids produced by the sphingomyelin signal transduction pathway could be physiologically relevant regulators of cardiac [Ca2+]i and therefore cardiac contractility.

Animals↗

Localization of protein kinase C in skeletal muscle T-tubule membranes.

Membrane fractions enriched in transverse tubules, either predominantly free or junctional, sarcoplasmic reticulum subfractions and purified sarcolemmal preparations have been isolated from rabbit skeletal muscle and examined for their contents of protein kinase C. Using activity measurements and immunoblotting methods, we have been able to detect substantial amounts of endogenous protein kinase C in T-tubules membranes and to a lesser extent, in muscle sarcolemma. Protein kinase C was found to be highest in junctional T-tubules and to be virtually absent from sarcoplasmic reticulum-derived membrane fractions. Immunofluorescence staining of muscle fibers is consistent with a T-tubule localization of the kinase. The T-tubule-associated protein kinase C enzyme phosphorylates several potentially important membrane proteins.

Animals↗