PubMed HealthSearch

Biomedical subjects

M Mattioli

Publications and source records attributed to M Mattioli.

At least 19 recordsLinked to original sources

Inhibition of Ca2+ and K+ currents by "antifreeze" proteins.

For the last two decades, the research on fish "antifreeze" proteins has focused exclusively on their ability to depress noncolligatively blood plasma freezing points, presumably by binding to ice crystals. We report evidence that antifreeze polypeptides from the winter flounder (Pseudopleuronectes americanus) have another special property, the ability to block ion channels. In experiments with porcine granulosa cells we show, using the patch-clamp technique in the whole cell configuration, that these proteins suppress effectively calcium and potassium currents. The results of dose-response studies indicate a protein-protein interaction mechanism.

Animals

[Oral contrast media in the study of the esophagus with magnetic resonance. Proposal of a new preparation].

The major problem in MRI of the esophagus is the lack of a reliable oral contrast agent. To determine the value of Gd-DTPA as an oral contrast medium for the esophagus as a part of phase III clinical trial, 17 patients (10 esophageal neoplasms, 4 neoplasms of the pharyngo-laryngeal tract, 3 Zenker's diverticula) underwent MRI. The oral contrast medium is proposed in a new preparation: 5 cc of oral Gd-DTPA were emulsionated with 30 g of a low-density barium paste for esophageal CT (3% p/v). High signal intensity in the esophageal lumen was observed in all patients and in all sequences. In neoplastic lesions, the c.m. improved the definition of both the level of stenosis and the longitudinal extent of the lesion. In diverticula, the real and the false lumen could be demonstrated. In one patient the exam could not be completed. No adverse reactions were observed.

Administration, Oral

[Magnetic resonance angiography in the study of abdominal neoplasms. Preliminary experience].

This study was designed to evaluate MRA imaging as a potential non-invasive method to study vascular infiltration in patients with abdominal neoplasms. Forty-three patients with abdominal tumors proven by CT and/or MRI were examined with MRA and subsequently with angiography. Of 15 cases of liver tumor, MRA allowed poor intratumoral vascularization to be demonstrated in 1 patient only, but it always provided a characteristic sign--i.e., the complete absence of flow in the hepatic segments infiltrated by the tumor and the evidence of pericapsular neovascularization. In 8 cases portal vein involvement was observed (4 cases of compression/dislocation, 2 cases of infiltration and 2 of thrombosis). In all cases MRA allowed the relationship between tumor and venous structures to be evaluated. It also demonstrated collateral vessel formations in 8 patients with cancrocirrhosis. Two cases of thrombosis and 3 infiltrations of the vena cava were demonstrated. Neither hepatic artery nor intrahepatic arterial vascularization could be correctly evaluated. Of 10 cases of renal tumors and in 3 of adrenal tumors, renal vein infiltration was seen in 4 cases and compression in 3, with only 1 false-positive finding. In these patients and in those with retroperitoneal masses, MRA provided valuable information on the relationship between tumor and vascular structures and was useful in detecting collateral vessels. In our experience, MRA is to be considered as a complementary technique to be performed after CT or MRI when additional information is needed as to the relationship between tumor and vascular structures. In the preoperative evaluation of abdominal neoplasm, angiography remains the gold standard, in spite of its invasiveness.

Abdominal Neoplasms

Follicle somatic cells influence pig oocyte penetrability and cortical granule distribution.

The influence of somatic cells on oocyte penetrability was studied during in vitro maturation. Four experiments were carried out. In the first, pig oocytes were fertilized in vitro immediately after collection (immature oocytes) or after being cultured for 44 hr with cumulus cells connected to the whole wall of the extroverted follicle (follicle oocytes) or without cumulus cells (denuded oocytes) (Mattioli et al.: Gamete Res 20:177-183, 1988a). In follicle and immature oocytes, 12 hr after insemination, the sperm were equally distributed between zona and ooplasm; in denuded oocytes, the majority (90.5%) of the sperm were located in the zona. In the second experiment, zonae pellucidae of follicle and denuded oocytes, obtained after 44 hr of culture, were incubated with a sperm suspension for 2 hr at 39 degrees C to evaluate the zona binding. The different number of sperm found on the zona pellucida of follicle or denuded oocytes (49.3 +/- 2.97 vs. 37.8 +/- 1.77) did not achieve statistical significance (P greater than 0.05). In the third experiment, follicle and denuded oocytes were denuded of their zonae after maturation and then fertilized in vitro. The number of sperm recorded in the ooplasm of zona-free follicle oocytes was significantly higher than that recorded in zona-free denuded oocytes (9.2 +/- 0.93 vs 1.19 +/- 0.28) (P less than 0.01). In the last experiment the influence of somatic cells on the distribution of cortical granules was evaluated. Pig oocytes were denuded at different stages of culture and, after completion of maturation, processed for electron microscopy.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Changes in maturation-promoting activity in the cytoplasm of pig oocytes throughout maturation.

Maturation-promoting factor (MPF) was examined in maturing pig oocytes by electrofusing them with germinal vesicle (GV) oocytes. Oocytes containing high levels of MPF (MI or MII stages) induced the breakdown of the GV introduced by fusion and the formation of the metaphase plate in 1 hr. A similar effect was seen when two or three GV oocytes were fused with a MII oocyte and then incubated for 1 hr in the presence of cycloheximide (a specific protein synthesis inhibitor), indicating that high levels of preformed MPF are present at the metaphase stage. During the maturation in vitro of cumulus-enclosed oocytes, a first sharp rise in MPF was seen between 26 and 29 hr of culture (MI stage); MPF declined after 2 hr (AI-TI stages) and again reached high levels at 35 hr, where it remained for the rest of maturation. Denuded oocytes showed a similar behavior, but MPF appeared 9 hr earlier and the rise, due to the asynchronous maturation of these oocytes, was not as sharp as in cumulus enclosed oocytes. Cycloheximide was used to study protein synthesis requirements for oocyte maturation. Intact GV were observed after 44 hr of culture when cycloheximide was added at 26 hr or earlier, and chromosome decondensation and pronuclear formation were observed when the drug was added at 32 hr. Transcriptional requirements were investigated by treating the oocytes with alpha-amanitin, an RNA polymerase inhibitor. This drug could completely inhibit the maturation of cumulus-enclosed oocytes, but this was a somatic cell-mediated effect since denuded oocytes were insensitive to this treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Luteinizing hormone inhibits potassium outward currents in swine granulosa cells by intracellular calcium mobilization.

Potassium currents of swine granulosa cells were studied using the patch clamp technique in the whole cell configuration. Granulosa cells stepped to positive potentials (+60 mV) from -40 mV holding potential exhibit a slowly activating, noninactivating outward potassium current. Tail current reverse potential (between -90 and -100 mV) and the current inhibition brought about by the replacement of KCl with CsCl in the pipette solution indicate that this current is carried by K ions. LH was found to significantly reduce the amplitude of this current. The effect was dose and time dependent. Similar inhibition (20-30% of the initial current) was reached with doses of 1-50 micrograms/ml, but in times proportionally shorter as the dose increased (50% inhibition was reached in 170-180 and 30-40 sec with 1 and 50 micrograms LH/ml, respectively). Much longer and variable times (3-10 min) were required with lower doses (0.2 microgram/ml). The effect of LH was independent of extracellular Ca, while preexposure of cells to TMB-8, an inhibitor of intracellular Ca mobilization, completely prevented the effect of LH. Outward currents after LH treatment could be completely restored by perfusing the cells with ionomycin in Ca-free medium to facilitate calcium efflux from the cells. The present studies indicate that LH modifies the bioelectrical properties of swine granulosa cells. This effect is mediated by an elevation of intracellular calcium, probably mobilized from intracellular stores. The induced changes in K conductance may play a specific role in the transduction mechanisms for LH.

Animals

The effect of antifreeze glycopeptides on membrane potential changes at hypothermic temperatures.

The research on antifreeze glycopeptides (AFGPs) from Antarctic and Arctic fishes has focused primarily on their interaction with ice crystals. This study reports results of experiments in which pig oocytes, known to be sensitive to hypothermic temperatures, were exposed to 4 degrees C for various periods of time, in solutions of different molecular weight AFGPs from Antarctic nototheniid fishes. The membrane potential was measured across the oolemma following hypothermic exposure. The results show that a physiological combination of the different molecular weight AFGPs protects the structural integrity of the oolemma and inhibits ion leakage across the oolemma at hypothermic temperatures. The results also show that the hypothermic protection is nonlinearly dependent on concentration and that separately, the different molecular weight glycopeptides do not stop ion leakage even at very high concentration. The protection of membranes at hypothermic temperatures is a new property of AFGPs which was not known prior to our work.

Animals

Maturation of pig oocytes: observations on membrane potential.

The membrane-potential changes of pig oocytes during maturation are described. Cumulus-enclosed oocytes have a resting potential of -41.81 +/- 0.60 mV; the removal of cumulus cells caused this potential to drop to -30.95 +/- 0.43 mV. Adding LH to the culture medium did not influence the potential of denuded oocytes but depolarized the potential of cumulus-enclosed oocytes to -32.90 +/- 0.43 mV. FSH did not affect the membrane potential of denuded or cumulus-enclosed oocytes, but significantly reduced the amplitude of the depolarization induced by LH. The effect of gonadotropins on cultured granulosa cells was also investigated. Plated granulosa cells have a resting potential of -45.21 +/- 0.72 mV, similar to that of cumulus-enclosed oocytes. As recorded in cumulus-enclosed oocytes, LH depolarized granulosa cell membrane potential (-30.33 +/- 0.69 mV) and FSH reduced this effect. To evaluate if oocyte maturation in vivo is accompanied by membrane-potential depolarization, follicular growth and oocyte maturation were induced in 6 prepubertal gilts by using an eCG-hCG treatment. Twenty hours after the beginning of oocyte maturation in vivo (induced by hCG), the membrane potential of the oocyte was depolarized to -28.84 +/- 1.01 mV, a value similar to that observed in vitro. These data indicate that both LH and FSH can influence the membrane potential of follicular somatic cells and, consequently, that of the oocyte. The electrical coupling between somatic cell and oocyte may represent a means by which the gonadotropin message is passed to the germinal cell by the somatic compartment.

Animals

Effect of PGF-2 alpha on progesterone production in swine luteal cells at different stages of the luteal phase.

Suspensions of luteal cells were prepared by enzymatic dispersion of pig corpora lutea obtained at specific times during the estrous cycle. Luteal cells from early corpora lutea produced more progesterone (4.73 +/- 0.84 nmol/10(6) cells, day 3) than those from late diestrus (0.73 +/- 0.04 nmol/10(6) cells, day 15); (P less than 0.05). Bovine LH enhanced progesterone production in a dose dependent manner particularly in cells from 9 to 15 day corpora lutea. Also PGF-2 alpha enhanced progesterone output in cells from mid-late corpora lutea. PGF-2 alpha did not exert any antigonadotropic effect since it further increased the progesterone production induced by LH. Luteal cells produced PGF-2 alpha with levels ranging between 1.6 and 2.7 pmol/10(6) cells throughout the whole luteal phase. The cellular content of cAMP was markedly increased by LH (556 +/- 60%) while it was not affected by PGF-2 alpha. Plasma membrane receptors for PGF-2 alpha were not detected in the analyzed tissue.

Animals

Kidney 15-hydroxy-prostaglandin-dehydrogenase activity during the development of experimental hypertension in the rat.

15-hydroxy-prostaglandin-dehydrogenase (PGDH) activity was studied in rat kidney homogenates during the development of hypertension, within 20 days after left renal artery constriction by a solid silver clip. In the ischemic kidney PGDH activity increased at day 6, reached maximum at day 10, then progressively at day 15 and returned to normal levels at day 20. No difference was found between contralateral kidneys and kidneys of normotensive control rats. Variations of PGDH activity did not seem to be related to either renal perfusion pressure or renin production. Increased PGDH activity may be a consequence of an enzyme induction following increased PG-synthetase activity, or it could be viewed as a defence mechanism, according to the hypothesis of a prohypertensive effect of PG in the rat.

Animals

Ferritin distribution and synthesis in sex-linked anemia.

The ferritin concentration of duodenum, liver, and spleen and the incorporation of L-leucine-3H into immunoprecipitated duodenal and liver ferritin was measured in genotypically normal (+/Y) mice and mice with sex-linked anemia (sla/Y), an X-linked recessive trait determined by a defect in intestinal iron absorption. Liver and splenic ferritin concentration was lower in sla/Y animals than in +/Y animals. Parenteral iron administration produced an increase in the duodenal, liver, and splenic ferritin concentration in both sla/Y and +/Y animals that was most striking in the case of the liver. Duodenal ferritin synthesis, both in vivo and in vitro, was increased in iron-deficient sla/Y animals and decreased in iron-deficient +/Y animals. In contrast, liver ferritin synthesis was decreased in both sla/Y and +/Y iron-deficient animals. In sla/Y animals fed an iron-deficient diet, duodenal ferritin synthesis decreased to near normal levels. These results indicating a high level of duodenal ferritin synthesis in standard-fed mice with sex-linked anemia suggest that the primary genetic defect is more likely a disorder of intramucosal iron transport than a primary disturbance of ferritin metabolism.

Anemia

Antinuclear antibodies (ANA): immunologic and clinical significance.

The methods currently used for the detection of ANA have been analyzed, with emphasis on their practical application to the diagnosis of the CTD. The use of the indirect IF-ANA test was recommended as a screening procedure to detect ANA. The need to standardize the technique using a single substrate and fluorescent conjugates with uniform F/P ratios was stressed. Most importantly, the value of titrating ANA for the diagnosis of the CTD was discussed. ANA titers higher than 1/500 are usually very significant clinically, often found in spontaneous or drug-induced SLE and few other CTD. The immunologic aspects of ANA and their potential value as aids in the diagnosis and management of the CTD were discussed. Anti-nDNA antibodies have been found to have a high degree of specificity for SLE and high titers of these antibodies correlate well with low levels of serum complement and severity of kidney involvement. The spectrum of ANA in the sera from patients with SLE has been expanded with the finding of anti-Sm antibodies which, when detected by gel precipitation with prototype serum, have been found so far only in SLE. Some of these antibodies have been found to have prognostic significance. Patients with MCTD and a group of patients with SLE have high titers of serum ANA with specificity for an RNase-sensitive component of ENA. The group of SLE patients defined by the presence of these antibodies (anti-Mo) have a better prognosis and in general develop only mild nephritis or have no kidney involvement at all. High titers of pure antinucleolar antibodies probably are found almost exclusively in the sera of patients with scleroderma. Some ANA have organ specificity, and GS-ANA have been found in all patients with Felty's syndrome and in a large proportion of patients with RA. One of the great advances in the field has been the recognition that ANA can be induced in the human and in experimental animals by the use of a number of therapeutic agents. Some of these agents can also induce a clinical picture resembling spontaneous SLE, though kidney involvement does not occur or is extremely mild. It is interesting that the whole spectrum of ANA can be found in drug-induced LE except anti-nDNA antibodies which have been associated to the pathogenesis of immune complex nephritis in spontaneous SLE. There is no doubt that research on ANA has contributed a great deal to the understanding of the CTD and will continue to be a valuable tool for the clinician and the investigator.

Animals