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Biomedical subjects

P Volpe

Publications and source records attributed to P Volpe.

At least 73 records · Page 4Linked to original sources
PubMed1994

[Propafenone and verapamil poisoning. Report of 2 clinical cases].

We described the self-poisoning of two young adolescents who took improper doses of two major cardiovascular drugs: propafenone and verapamil. The young girls developed markedly different clinical patterns: ECG abnormalities without clinical consequences were found in one case progressively ingravescent ECG abnormalities leading to cardiac arrest in the other. These differences are probably due to varying doses taken and metabolic states. Conventional detoxication and resuscitation techniques proved successful in both cases.

Adolescent↗
PubMed1994

[Radical prostatectomy after complete androgen blockade in stage B2 and C cancer].

Despite recent advances in imaging techniques, the staging process is still unsatisfactory in cancer of the prostate. The underestimation for stage B2 and C tumours in about 50%. We present our findings in a retrospective study analyzing the clinical and biological effects of complete androgen blockade before radical prostatectomy in patients with advanced stage localized tumours. We treated 21 patients from 1989 to 1993. All received preoperative homonotherapy by complete androgen block for at least 3 months before node dissection preceding suprapubic radical prostatectomy. Only 20 prostatectomies were performed as metastasis was found in the extemporaneous examination in 1 patient. The volume of the prostate gland had diminished in all patients after the hormonotherapy (27.8%) as did PSA (95%). When evaluated, the tumour stage of the surgical specimen was always more advanced than the needle biopsy. Only 1 tumour was strictly limited to the intracapsule and all the others had either invaded the capsule, reached the margins or had invaded the seminal vessels or lymph nodes. With a mean follow up of 45 months, recurrence rate is 50%, mainly due to tumours with positive margins or seminal invasion in patients who were not given adjuvant treatment. Our results are in agreement with those in the literature showing that although the volume of the prostate is reduced and PSA declines, no improvement in pathology staging is observed.

Aged↗
PubMed1993

Expression of the calsequestrin gene in chicken cerebellum Purkinje neurons.

Intracellular rapidly exchanging Ca2+ stores are identified and defined in terms of intralumenal low-affinity, high-capacity Ca(2+)-binding proteins, of which calsequestrin (CS) is the prototype in striated muscles. In chicken striated muscles, there is a single gene for CS [Choi and Clegg (1990) Dev. Biol. 142, 169-177]. In the chicken brain, the gene for CS was found to be selectively expressed in Purkinje neurons, as judged by Northern blotting, in situ hybridization and immunocytochemistry. The synthetic machinery for CS was found to be restricted to the cell body, i.e. excluded from dendrites and axon.

Animals↗
PubMed1993

5-Methylcytosine in genes with methylation-dependent regulation.

An asymmetric distribution of deoxy-5-methylcytidylic acid-inhibiting restriction sites (dcm-sites) takes place in ten human genes regulated by 5-methylcytosine. These genes are dcm-site enriched upstream and dcm-site poor downstream. Along them, there is a scattering of hypermethylatable introns and hypomethylatable exons with a common code: the 5mCpG dinucleotides characterize promoters; Gp5mCs characterize introns; Tp5mCs and Cp5mCs are in small concentrations in exons. Housekeeping genes contain more dcm-sites when compared with tissue-specific genes. This depends on the higher number of dcm-sites in their promoters and introns. In exons, the relatively lower number of dcm-sites is almost the same in both housekeeping and tissue-specific genes. Going from 5' to 3', the average frequency of occurrence of these sites per nucleotide units decreases in introns and increases in exons. This difference is highly discriminated for tissue-specific and less discriminated for housekeeping genes.

5-Methylcytosine↗
PubMed1993

Intracellular Ca2+ stores of rat cerebellum: heterogeneity within and distinction from endoplasmic reticulum.

Rat cerebellum microsomes were subfractionated on isopycnic linear sucrose (20-42%)-density gradients. The distribution of endoplasmic reticulum (ER) markers (RNA, signal-sequence receptor alpha, calnexin, calreticulin, the immunoglobulin-binding protein Bip) and markers of intracellular rapidly exchanging Ca2+ stores [Ca2+ channels sensitive to either Ins(1,4,5)P3 or ryanodine) was investigated biochemically and immunologically. The comparison indicates that: (a) vesicles bearing the InsP3 receptor were separated from those bearing the ryanodine receptor; (b) ER markers, i.e. Bip, calnexin, signal-sequence receptor alpha, RNA, did not sediment as either InsP3 or ryanodine receptors did; (c) calreticulin, an intralumenal low-affinity high-capacity Ca(2+)-binding protein, had a widespread distribution, similar to that of Bip and calnexin, and was present in Purkinje, granule, Golgi and stellate neurons, as indicated by immunofluorescent labelling of cerebellum cortex cryosections. The present results show that the ER is not a homogeneous entity, and that Ca2+ stores are heterogeneous insofar as InsP3 receptors and ryanodine receptors are segregated, either to discrete intracellular organelles or to specialized ER subcompartments.

Animals↗
PubMed1993

Ontogenesis of chick iris intrinsic muscles: evidence for a smooth-to-striated muscle transition.

The iris sphincter muscle consists of striated muscle fibers in the adult chicken. The ontogenetic development of this muscle has been studied by immunocytochemistry, from Embryonic Day 7 to the time of hatching. The time course of expression of specific markers of either smooth or striated muscle, i.e., smooth and skeletal muscle myosin heavy chains along with the inositol 1,4,5-trisphosphate (IP3) receptor, the intracellular Ca2+ release channel gated by the second messenger IP3, was investigated. We observed the parallel and transient expression, from about Embryonic Day 8 until hatching, of smooth muscle myosin heavy chain and IP3 receptor in early differentiating cells at the pupillary margin, as well as in migrating cells at subsequent stages; the sequential expression of sarcomeric myosin heavy chain around the end of the second embryonic week; and its spreading throughout the iris sphincter by the time of hatching. The present findings are consistent with a smooth muscle differentiation stage as an intermediate stage in the ontogenic development of the iris sphincter muscle of the chicken.

Animals↗
PubMed1993

The endoplasmic reticulum-sarcoplasmic reticulum connection. II. Postnatal differentiation of the sarcoplasmic reticulum in skeletal muscle fibers.

The postnatal differentiation of sarcoplasmic reticulum (SR) of rabbit skeletal muscles (the slow-twitch soleus and the fast-twitch adductor muscles) was monitored between Days 1 and 12 by following on Western blots the expression and accumulation of molecular markers specific not only for the muscle endomembrane system, i.e., calsequestrin (CS) and the ryanodine-sensitive Ca2+ release channel, but also for the endoplasmic reticulum (ER) at large, i.e., BiP, calnexin (CN) and calreticulin. Our results demonstrate that SR development, documented by the increase of the SR fractional volume, terminal cisternae proliferation, and reorientation of triads, is accompanied by the accumulation of the SR-specific proteins and also of CN, with no change of the other ER general markers. Moreover, the distribution of two of the markers, BiP and CS, was investigated by immunocytochemistry at both the light and the electron microscope level. At Day 1 CS was found to be concentrated both within the few recognizable triad terminal cisternae and within the lumen of numerous, apparently discrete cisternae and tubules, widely scattered throughout both the contractile and the subplasmalemmal areas of the cytoplasm. These structures remain evident until Day 12, when most triad junctions have acquired proper configuration, composition and orientation. BiP, on the other hand, appears widely distributed within the ER/SR of the fibers. From the early stages of postnatal development it does colocalize with the Ca2+ binding protein in the lumen of the CS-rich structures and appears also within the longitudinal SR and the conventional ER cisternae.

Animals↗
PubMed1993

Postnatal development of rabbit fast-twitch skeletal muscle: accumulation, isoform transition and fibre distribution of calsequestrin.

The time-course of disappearance of slow-cardiac calsequestrin (CS) and that of appearance of the skeletal CS isoform were investigated in developing fast-twitch skeletal muscle of the rabbit between postnatal days 1 and 60, along with changes in density of the ryanodine receptor (RyR)/Ca2+ release channel. Western blot data on skeletal muscle membranes, purification of two CS isoforms by phenyl-Sepharose chromatography, and their immunolocalization in muscle fibres, all show that both CS isoforms are coexpressed in neonatal muscles. Our results, at the protein level, indicate that the turning off of synthesis of cardiac CS and its total disappearance from fast-twitch fibres take place at critical periods between two and four weeks postnatally, i.e. past changes in the respective mRNA. In contrast, the accumulation in muscle membranes of both the RyR and the skeletal CS isoform proceeds steadily up to one month, to reach adult values at about two months of age. These findings seem to argue that myogenic factors, in addition to the morphogenetic influence on the sarcoplasmic reticulum from the neural input to the muscle, may be involved in the developmental transition of CS isoforms in mammalian fast-twitch muscle fibres.

Animals↗
PubMed1993

Ryanodine receptors: how many, where and why?

Ryanodine receptors are intracellular Ca2+ channels that have been known for more than a decade to have a role in releasing Ca2+ from the sarcoplasmic reticulum to regulate contraction in skeletal and cardiac muscle fibres. Vincenzo Sorrentino and Pompeo Volpe review some recent developments: the ryanodine receptor channels have now been found to be expressed in the central nervous system, and the cloning of a third ryanodine receptor gene (RYR3) has revealed that this new isoform is widely expressed in several tissues and cells. In consequence, the view of ryanodine receptors as Ca2+ channels of muscle cells is rapidly changing, and these channels seem set to take a more central position on the stage of intracellular Ca2+ signalling.

Animals↗
PubMed1993

Multiple/heterogeneous Ca2+ stores in cerebellum Purkinje neurons.

1. The rapid and transient redistribution of Ca2+ from intracellular membrane-bound compartments (stores) is a key event of cell activation. 2. The cytological nature and molecular composition of such Ca2+ stores have been the object of intense investigation in recent years. 3. Here we review: (a) the current knowledge on intracellular Ca2+ stores of Purkinje neurons at the functional, biochemical, molecular, morphological and ultrastructural level; and discuss: (b) the relationship between Ca2+ stores and the endoplasmic reticulum, and (c) the occurrence of multiple/heterogeneous Ca2+ stores.

Animals↗
PubMed1993

Inositol 1,4,5-trisphosphate receptor in heart: evidence for its concentration in Purkinje myocytes of the conduction system.

Inositol 1,4,5-trisphosphate (IP3) is one of the second messengers capable of releasing Ca2+ from sarcoplasmic reticulum/ER subcompartments. The mRNA encoding the intracellular IP3 receptor (Ca2+ channel) has been detected in low amounts in the heart of various species by Northern blot analysis. The myocardium, however, is a heterogeneous tissue composed of working myocytes and conduction system cells, i.e., myocytes specialized for the beat generation and stimulus propagation. In the present study, the cellular distribution of the heart IP3 receptor has been investigated. [3H]IP3 binding experiments, Western blot analysis and immunofluorescence, with anti-peptide antibodies specific for the IP3 receptor, indicated that the majority of Purkinje myocytes (the ventricular conduction system) express much higher IP3 receptor levels than atrial and ventricular myocardium. Heterogeneous distribution of IP3 receptor immunoreactivity was detected both at the cellular and subcellular levels. In situ hybridization to a riboprobe generated from the brain type 1 IP3 receptor cDNA, showed increased accumulation of IP3 receptor mRNA in the heart conduction system. Evidence for IP3-sensitive Ca2+ stores in Purkinje myocytes was obtained by double immunolabeling experiments for IP3 receptor and cardiac calsequestrin, the sarcoplasmic reticulum intralumenal calcium binding protein. The present findings provide a molecular basis for the hypothesis that Ca2+ release from IP3-sensitive Ca2+ stores evoked by alpha 1-adrenergic stimulation is responsible for the increase in automaticity of Purkinje myocytes (del Balzo, U., M. R. Rosen, G. Malfatto, L. M. Kaplan, and S. F. Steinberg. 1990. Circ. Res. 67:1535-1551), and open new perspectives in the hormonal modulation of chronotropism, and generation of arrhythmias.

Animals↗
PubMed1993

The endoplasmic-sarcoplasmic reticulum of smooth muscle: immunocytochemistry of vas deferens fibers reveals specialized subcompartments differently equipped for the control of Ca2+ homeostasis.

Cryosection immunofluorescence and immunogold labeling with antibodies against specific markers were used in rat vas deferens smooth muscle fibers to reveal the molecular arrangement of the endomembrane system (referred to variously in the text as ER or sarcoplasmic reticulum [SR]; S-ER or ER/SR) known to participate in the control of Ca2+ homeostasis. The lumenal ER chaperon, immunoglobulin binding protein (BiP), as well as protein disulfide isomerase, and calreticulin, a Ca2+ binding protein expressed by most eukaryotic cells, appeared to be evenly distributed throughout the entire system (i.e., within [a] the nuclear envelope and the few rough-surfaced cisternae clustered near the nucleus; [b] single elements scattered around in the contractile cytoplasm; and [c] numerous, heterogeneous, mainly smooth-surfaced elements concentrated in the peripheral cytoplasm, part of which is in close apposition to the plasmalemma). All other structures, including nuclei, mitochondria, Golgi complex, and surface caveolae were unlabeled. An even distribution throughout the endomembrane system appeared also for the proteins recognized by anti-ER membrane antibodies. In contrast, calsequestrin (the protein that in striated muscles is believed to be the main actor of the rapidly exchanging Ca2+ storage within the lumen of the sarcoplasmic reticulum) was found preferentially clustered at discrete lumenal sites, most often within peripheral smooth-surfaced elements of moderate electron density. Within these elements dual labeling revealed intermixing of calsequestrin with the other lumenal ER proteins. Moreover, the calsequestrin-rich elements were enriched also in the receptor for inositol 1,4,5-trisphosphate, the second messenger that induces Ca2+ release from intracellular stores. These results document the previously hypothesized molecular heterogeneity of the smooth muscle endomembrane system, particularly in relation to the rapid storage and release of Ca2+.

Animals↗
PubMed1993

Intracellular magnesium and inositol 1,4,5-trisphosphate receptor: molecular mechanisms of interaction, physiology and pharmacology.

This review focuses on the action of intracellular magnesium ion on the inositol 1,4,5-trisphosphate (IP3) receptor, the almost ubiquitous membrane-bound Ca2+ channel gated by the intracellular second messenger IP3. Experimental data have shown that Mg2+ is a non-competitive inhibitor of the IP3-gated Ca2+ channel and of [3H]-IP3 binding. The relevance of the Mg2+ effect is discussed in relation to: (a) the physiological role of the IP3-induced release of Ca2+ from intracellular Ca2+ stores, i.e., Ca2+ homeostasis of activated cells, in particular cardiac myocytes; (b) the modulation exerted by changes of [Mg2+]o and [Mg2+]i on basic cardiac functions, e.g., inotropism, chronotropism and automaticity; and (c) the pathogenesis of automatic arrhythmias, caused by either early or delayed afterdepolarizations, in which IP3-induced release of Ca2+ may be involved.

Animals↗
PubMed1992

The endoplasmic reticulum-sarcoplasmic reticulum connection: distribution of endoplasmic reticulum markers in the sarcoplasmic reticulum of skeletal muscle fibers.

The skeletal muscle sarcoplasmic reticulum (SR) was investigated for the presence of well-known endoplasmic reticulum (ER) markers: the lumenal protein BiP and a group of membrane proteins recognized by an antibody raised against ER membrane vesicles. Western blots of SR fractions revealed the presence of BiP in fast- and slow-twitch muscles of the rabbit as well as in rat and chicken muscles. Analyses of purified SR subfractions, together with cryosection immunofluorescence and immunogold labeling, revealed BiP evenly distributed within the longitudinal SR and the terminal cisternae. Within the terminal cisternae BiP appeared not to be mixed with calsequestrin but to be distributed around the aggregates of the latter Ca2+ binding protein. Of the various membrane markers only calnexin (91 kDa) was found to be distributed within both SR subfractions, whereas the other markers (apparent molecular masses of 64 kDa and 58 kDa and a doublet around 28 kDa) were concentrated in the terminal cisternae. These results suggest that the SR is a specialized ER subcompartment in which general markers, such as the ones we have investigated, coexist with the major SR proteins specifically responsible for Ca2+ uptake, storage, and release. The differential distribution of the ER markers reveals new aspects of the SR molecular structure that might be of importance for the functioning of the endomembrane system.

Animals↗
PubMed1992

Coexistence of two calsequestrin isoforms in rabbit slow-twitch skeletal muscle fibers.

The cardiac and skeletal muscle isoforms of calsequestrin (CS), the low affinity, high capacity Ca2+ binding protein localized in the lumen of sarcoplasmic reticulum, are the products of two different genes (Fliegel, L., Leberer, E., Green, N.M. and MacLennan, D.H. (1982) FEBS Lett. 242, 297-300), and can be both purified from slow-twitch skeletal muscle of the rabbit (Damiani, E., Volpe, P. and Margreth, A. (1990) J. Muscle Res. Cell Motil. 11, 522-530). Here we show that both CS isoforms coexist in slow-twitch muscle fibers as indicated by indirect immunofluorescent staining of cryosections with affinity-purified antibodies specific for each CS isoform.

Animals↗
PubMed1992

Ryanodine receptor autoantibodies in myasthenia gravis patients with a thymoma.

Sera from patients with myasthenia gravis were examined by Western blot for the presence of antibodies to proteins of the sarcoplasmic reticulum from rabbit skeletal muscle. Fourteen of 30 patients with myasthenia gravis and a thymoma had IgG autoantibodies to the calcium release channel of the sarcoplasmic reticulum (the ryanodine receptor), which plays a crucial role in the mechanism of excitation-contraction coupling in striated muscle. Ryanodine receptor autoantibodies were not detected in any of the 45 sera from patients with myasthenia gravis without a thymoma. Ryanodine receptor autoantibodies may have pathogenetic relevance in thymoma-associated myasthenia gravis.

Autoantibodies↗