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Biomedical subjects

M Conti

Publications and source records attributed to M Conti.

At least 19 recordsLinked to original sources

Density change effects on crystal growth from the melt.

When a crystal grows from its undercooled melt the local density changes, driving a convective flow in the liquid phase. Then, the purely diffusional description of the process ceases to be satisfactory. Moreover, the dynamic pressure associated with the flow field may affect the melting temperature (and the effective undercooling) of the system. Both these effects have been addressed in recent experimental work. In the present study we derive a thermodynamically consistent phase-field model that accounts for the density change effects in the solidification of a pure substance. Starting from a thermodynamic potential that includes squared gradient terms for both the order parameter and the density, the field equations are derived assuming positive local entropy production. The model is numerically solved in one dimension to show deviations from the classic phase-field description of the same phenomenon.

Journal Article↗

Normal scaling in globally conserved interface-controlled coarsening of fractal clusters.

We find that globally conserved interface-controlled coarsening of diffusion-limited aggregates exhibits dynamic scale invariance (DSI) and normal scaling. This is demonstrated by a numerical solution of the Ginzburg-Landau equation with a global conservation law. The general sharp-interface limit of this equation is introduced and reduced to volume preserving motion by mean curvature. A simple example of globally conserved interface-controlled coarsening system: the sublimation/deposition dynamics of a solid and its vapor in a small closed vessel, is presented in detail. The results of the numerical simulations show that the scaled form of the correlation function has a power-law tail accommodating the fractal initial condition. The coarsening length exhibits normal dynamic scaling. A decrease of the cluster radius with time, predicted by DSI, is observed. The difference between global and local conservation is discussed.

Journal Article↗

Long-term hormonal regulation of the cAMP-specific phosphodiesterases in cultured FRTL-5 thyroid cells.

Thyrotropin (TSH) and pharmacological agents that elevate intracellular cAMP concentrations potentiate the mitogenic response of FRTL-5 thyroid cells to insulin-like growth factor-I (IGF-I). This study was undertaken to determine the role of cAMP phosphodiesterases (PDEs) in this TSH-dependent regulation. Incubation of FRTL-5 cells with TSH, forskolin, or dibutyryl cAMP gradually induced the PDE activity, and treatment for 24 h produced a marked increase in type 4 high affinity cAMP PDEs. Under basal conditions, transcripts corresponding to PDE4A, PDE4B, PDE4C, and PDE4D were present. Stimulation for 24 h by TSH, forskolin or dibutyryl cAMP induced an increase in mRNA levels of PDE4B, PDE4D, and PDE4C. To understand the role of this cAMP-dependent PDE regulation in the potentiation of the mitogenic response to IGF-I, thymidine incorporation into DNA in response to IGF-I and TSH was measured in the absence or presence of PDE inhibitors. Exposure of the cells to 3-isobutyl-1-methylxanthine (IBMX) or RO 20-1724 had opposing effects on thymidine incorporation into DNA, depending on the stimulus applied. When IGF-I was used alone, both IBMX and RO 20-1724 potentiated IGF-I-stimulated thymidine incorporation. However, when IGF-I and TSH at high concentrations were used in combination, these PDE inhibitors blocked thymidine incorporation into DNA. In addition, these inhibitors depressed the synergistic increase in cyclin D1 and cyclin D- or cyclin E-associated cyclin-dependent kinase (CDK) activity that is induced by TSH and IGF-I. Increased CDK activities have been shown to play a crucial role in progression through the G(1)/S phase of the cell cycle. These data demonstrate that TSH produces marked changes in the cAMP degradative pathway of FRTL-5 cells by regulating the expression of cAMP PDEs. The regulation of the intracellular cAMP levels by this mechanism may contribute to the TSH- and IGF-I-dependent control of the entry into the S phase of the cell cycle through changes in the cyclin/CDK system in FRTL-5 cells.

1-Methyl-3-isobutylxanthine↗

Incomplete penetrance with normal MRI in a woman with germline mutation of the DCX gene.

X-linked isolated lissencephaly sequence (ILS) and subcortical band heterotopia are allelic human disorders associated with mutations of the DCX gene in both familial and sporadic forms. The authors describe a large Sardinian family in which three brothers with ILS have a missense mutation of the DCX gene. Their mother, a nonmosaic carrier, has a normal phenotype and cranial MRI. Skewed X-inactivation in the lymphocytes was also ruled out. This is the first report of an asymptomatic carrier of a DCX mutation likely due to apparent nonpenetrance.

Adolescent↗

Identification and functional analysis of splice variants of the germ cell soluble adenylyl cyclase.

In mammalian germ cells, cAMP signaling is dependent on two forms of adenylyl cyclase, the conventional membrane-bound ACIII and a soluble form of adenylyl cyclase (sAC). Recent elucidation of the sAC sequence indicates that this enzyme is phylogenetically distinct from the membrane-bound AC, does not interact with G proteins, and its activity is regulated by bicarbonate ions. Here we have investigated the properties and regulation of this enzyme during spermatogenesis. Two different transcripts encoding a full-length and truncated sAC were identified by reverse transcriptase-polymerase chain reaction and RNase protection analysis. The truncated sAC transcript lacks exon 11 with a premature termination of the open reading frame after the catalytic domain. Reverse transcriptase-polymerase chain reaction with testis RNA from adult mouse and rat of different ages, as well as RNase protection, showed that both transcripts are absent at 11 days of age, appear between 20 and 30 days of age, and are retained in the adult testis. The presence of corresponding proteins in testis, germ cells, and spermatozoa was demonstrated by fast protein liquid chromatography and differential immunoprecipitation with full-length sAC-specific antibodies. Bicarbonate ions activated both sAC forms and increased cAMP levels in germ cells isolated from 25- and 50-day-old rats and adult rats in a concentration-dependent manner. These findings provide evidence that full-length and truncated sAC are generated by alternate splicing. Both forms are active in spermatids, and the bicarbonate present in the seminiferous tubule may be a signal that regulates cAMP levels in these cells.

Adenylyl Cyclases↗

Area-preserving dynamics of a long slender finger by curvature: a test case for globally conserved phase ordering.

A long and slender finger can serve as a simple "test bed" for different phase-ordering models. In this work, the globally conserved, interface-controlled dynamics of a long finger is investigated, analytically and numerically, in two dimensions. An important limit is considered when the finger dynamics is reducible to area-preserving motion by curvature. A free boundary problem for the finger shape is formulated. An asymptotic perturbation theory is developed that uses the finger aspect ratio as a small parameter. The leading-order approximation is a modification of the Mullins finger (a well-known analytic solution) whose width is allowed to slowly vary with time. This time dependence is described, in the leading order, by an exponential law with the characteristic time proportional to the (constant) finger area. The subleading terms of the asymptotic theory are also calculated. Finally, the finger dynamics is investigated numerically, employing the Ginzburg-Landau equation with a global conservation law. The theory is in very good agreement with the numerical solution.

Journal Article↗

Phosphodiesterase 4D and protein kinase a type II constitute a signaling unit in the centrosomal area.

The mediation of cAMP effects by specific pools of protein kinase A (PKA) targeted to distinct subcellular domains raises the question of how inactivation of the cAMP signal is achieved locally and whether similar targeting of phosphodiesterases (PDEs) to sites of cAMP/PKA action could be observed. Here, we demonstrate that Sertoli cells of the testis contain an insoluble PDE4D3 isoform, which is shown by immunofluorescence to target to centrosomes. Staining of PDE4D and PKA shows co-localization of PDE4D with PKA-RIIalpha and RIIbeta in the centrosomal region. Co-precipitation of RII subunits and PDE4D3 from cytoskeletal extracts indicates a physical association of the two proteins. Distribution of PDE4D overlaps with that of the centrosomal PKA-anchoring protein, AKAP450, and AKAP450, PDE4D3, and PKA-RIIalpha co-immunoprecipitate. Finally, both PDE4D3 and PKA co-precipitate with a soluble fragment of AKAP450 encompassing amino acids 1710 to 2872 when co-expressed in 293T cells. Thus, a centrosomal complex that includes PDE4D and PKA constitutes a novel signaling unit that may provide accurate spatio-temporal modulation of cAMP signals.

3',5'-Cyclic-AMP Phosphodiesterases↗

Curvature effects in rapid alloy solidification.

The growth of a cylindrical or spherical crystal into its undercooled melt is a process whose description is complicated by the lack of a stationary regime. A simple approach to the problem, justified for low growth rates and widely used in the past for both pure substances and alloy solidification, is based on a quasistatic approximation which assumes an instantaneous adaptation of the diffusional field to the interface configuration. For alloy solidification, assuming isothermal conditions and local interface equilibrium, this simplified model predicts a diffusion controlled growth, with the radius of the crystal increasing asymptotically as ~t(1/2). However, as pointed out by recent investigations, thermal diffusion and nonequilibrium effects enter as essential ingredients in rapid alloy solidification. In the present paper we use the phase-field model to simulate the cylindrical and spherical growth of a solid germ into a supersaturated alloy melt. The problem is treated in its full time-dependent characteristics, accounting for nonequilibrium effects as well as for the rejection of both heat and solute away from the advancing front. We observe a complex behavior and a rich variety of dynamic regimes: in different regions of parameter space the growth rate is limited by diffusion (either thermal or chemical) or is kinetic controlled. Traversing the boundaries which limit these regions, the process undergoes sharp transitions which leave a trace in the solidified alloy. For realistic values of the Lewis number, thermal effects drive the process into a a diffusive regime, in which the rate limiting mechanism is the rejection of solute.

Journal Article↗

Cauda equina syndrome secondary to idiopathic spinal epidural lipomatosis.

STUDY DESIGN: Three cases of idiopathic epidural lipomatosis are reported. OBJECTIVES: Description of the relationship between spinal pathologic overgrowth of fat tissue and neurologic symptoms. SUMMARY OF BACKGROUND DATA: Idiopathic epidural lipomatosis is a very rare condition; it is usually secondary to chronic steroid therapy or endocrinopathic diseases. METHODS: Three men with a mean age of 58.5 years, who experienced intermittent claudication, bilateral radicular pain in both legs, and urinary dysfunction with hypoesthesia in the perineal region, were evaluated by plain radiography and magnetic resonance imaging, the results of which demonstrated a pathologic overgrowth of fat tissue in the spinal canal with a marked impingement of the dural sac. Obesity, endocrinopathic diseases, and chronic steroid therapy were excluded for all patients. Surgical treatment was performed by wide multilevel laminectomies, fat debulking, and instrumented posterolateral fusion. RESULTS: After surgery there was a gradual improvement in symptoms and signs so that 2 years later the patients returned to daily activities and were neurologically normal. CONCLUSIONS: Spinal epidural lipomatosis can be a cause of back pain but rarely radicular impingement. Magnetic resonance imaging is the procedure of choice. The treatment must be performed early by wide surgical decompression.

Aged↗

Randomized controlled trial of a synthetic sealant for preventing alveolar air leaks after lobectomy.

BACKGROUND: The intraoperative application of synthetic surgical lung sealant (SLS) to surfaces leaking air or at risk of air leaks has been advocated to reduce alveolar air leaks (AAL) after lobectomy. METHODS: This study was designed to investigate the effectiveness of SLS in reducing AAL in patients considered intraoperatively to have moderate to severe AAL, after all conventional measures to reduce such leaks had been used. Over 17 months, 124 patients undergoing standard lobectomy were randomized to standard closure of parenchymal surgical sites, with or without SLS. RESULTS: In treated patients, the mean numbers of intraoperative AAL after application of SLS were significantly smaller than in untreated patients (38.5 mL versus 59.9 mL, p = 0.0401). Postoperatively, the mean time to last observable AAL was shorter in the treated group (33.7 hours versus 63.2 hours, p = 0.0134) and the mean percentage of patients free of AAL at days 3 and 4 was smaller (87% versus 58.5%, p = 0.002). However, the occurrence of incomplete lung expansion after drain removal, and the length of the postoperative hospital stay due to prolonged AAL, were not different. In the treatment group, 4 patients developed localized empyema and incomplete lung expansion without bronchopleural fistula 7, 12, 15, and 20 days, respectively, after operation. In these 4 patients, inserted chest tubes drained infected sealant. CONCLUSIONS: Surgical lung sealant may be a useful adjunct to conventional techniques for reducing moderate and severe AAL after lobectomy, but its use seems to increase the risk of postoperative empyema.

Acrylates↗

High glucose and hyperinsulinemia stimulate connective tissue growth factor expression: a potential mechanism involved in progression to fibrosis in nonalcoholic steatohepatitis.

Nonalcoholic steatohepatitis (NASH) may progress to liver fibrosis and cirrhosis. Mechanisms directly involved in the development of fibrosis have been poorly investigated. Because connective tissue growth factor (CTGF) is an intermediate key molecule involved in the pathogenesis of fibrosing chronic liver diseases and is potentially induced by hyperglycemia, the aims of this study were to (1) study the expression of CTGF in vivo both in human liver biopsy specimens of patients with NASH and in an experimental model of obesity and type II diabetes (Zucker rats); and (2) analyze the effects of hyperglycemia and insulin in vitro on hepatic stellate cells. In vivo, CTGF overexpression was observed in the liver tissue of all of the 16 patients with NASH. CTGF immunostaining was mild in 7 cases (44%) and moderate or strong in 9 cases (56%). Staining was mainly detected in the liver extracellular matrix in parallel with the amount of liver fibrosis. Liver from fa/fa rats also showed CTGF overexpression by comparison with Fa/fa rats both at the messenger RNA (mRNA) level (3-fold increase) and protein level. In vitro, both CTGF mRNA and protein were significantly increased when hepatic stellate cells were incubated with either glucose or insulin. A slight increase in type I procollagen mRNA level was also observed in hepatic stellate cells incubated with glucose. In conclusion, this study suggests that hyperglycemia and insulin are key-factors in the progression of fibrosis in patients with NASH through the up-regulation of CTGF.

Adult↗

Increased absorption rate of diclofenac from fast acting formulations containing its potassium salt.

Diclofenac (CAS 15307-86-5) is a non-steroidal anti-inflammatory drug largely used, mainly to relief pain of various origin. Diclofenac is present on the market as free acid, as sodium salt (CAS 15307-79-6) and as potassium salt (CAS 15307-81-0). The last salification form has shown a prompter absorption rate and a faster onset of analgesic activity than the acid form and sodium salt. This paper extensively reviews three trials carried out on healthy volunteers, where potassium salt of diclofenac present in three fast-acting formulations, namely sachets (Trial 1), tablets (Trial 2) and oral drops (Trial 3), were compared to reference tablet formulations from the market. A very fast absorption rate was encountered with the three test formulations, with the peak reached in one case 5 min and in most cases within 10-15 min after dosing. The quick absorption rate of test formulations was attributed to the special combination of the salt of diclofenac with a dynamic buffering agent, namely bicarbonate, present in the test formulations and covered by an international patent. The prompt absorption of diclofenac from the new fast-acting formulations was accompanied by the presence of only one peak, whereas the reference formulations produced in most cases two peaks, as widely described in literature. This finding suggested the hypothesis that the absorption of test formulations should occur in a shorter tract of the gut. The faster absorption of diclofenac from the three fast-acting formulations is expected to produce a faster onset of analgesic action, which highlights these new formulations as particularly indicated to relief pain of any origin.

Adult↗

Self-expandable metal stents in the treatment of antro-pyloric and/or duodenal strictures.

PURPOSE: To assess the usefulness of self-expandable metal stents in the recanalization of antro-pyloric and/or duodenal strictures. MATERIAL AND METHODS: We report our experience of 15 patients with inoperable antro-pyloric and/or duodenal strictures treated by implantation of 21 self-expandable metal stents (18 uncovered and 3 covered) inserted perorally under fluoroscopic guidance. The patients were 11 men and 4 women, mean age 65.3 years. Fourteen of 15 patients were affected by a malignant stricture of the antro-pyloric region and/or duodenum either primary or secondary in 10 and 4 cases, respectively. Only in 1 case there was a benign stricture from postoperative scarring. Stricture length and diameter varied from 3 to 9 cm (mean 5.4 cm) and from 0 to 4 mm (mean 1.27 mm), respectively. RESULTS: Twenty-one stents were placed in 15 patients: Technical success was achieved in all cases while clinical improvement was obtained in 14 cases. No short-term complications were observed. A mean 4.3-month follow-up was obtained. Two patients had emesis secondary to peritoneal dissemination of the tumor after 1 and 2 months, respectively. Two other patients showed tumor overgrowth of the oral edge of the prosthesis after 3 and 2 months, respectively, and required another coaxial stent to bridge the new stenosis. The patient treated for a benign stricture had jaundice after 3 months and percutaneous internal-external biliary drainage was necessary. CONCLUSION: Self-expandable metal stents are a safe and effective treatment of antro-pyloric and duodenal strictures; therefore, they should be considered an alternative to palliative resection in cases of advanced stage disease or poor general physical condition.

Adult↗

Surgically induced cryptorchidism-related degenerative changes in spermatogonia are associated with loss of cyclic adenosine monophosphate-dependent phosphodiesterases type 4 in abdominal testes of rats.

The present study was undertaken to investigate the role of phosphodiesterase type 4 (PDE4) enzymes in cryptorchidism-induced apoptosis of the germ cells. Regulation of expression of PDE4 enzymes was studied in the abdominal and scrotal testes of surgically induced cryptorchid rats for 10, 20, and 30 days. In some cases orchidopexy was performed after 30 days of cryptorchidism, and rats were allowed to recover for an additional 50 days. Upon histological examination, marked degenerative changes in the epithelial lining of the seminiferous tubules within abdominal testes were observed compared with contralateral control or age-matched sham-operated rats. These changes included degeneration of some spermatogonia, apoptosis of the secondary spermatocytes, incomplete spermatogenesis, and lack of spermatozoa in the lumen. In contrast, contralateral scrotal testes exhibited normal histology. Significant improvement in the regeneration of spermatogonia was observed in rats after 50 days of recovery following orchidopexy. Immunocytochemical examination suggested the presence of PDE4A in germ cells while PDE4B was predominantly expressed on somatic cells. Western blotting using PDE4 subtype-selective antibodies showed the presence of two PDE4A variants (a 109-kDa PDE4A8 and a previously uncharacterized 88-kDa PDE4A variant) and two PDE4B (78-kDa PDE4B2 and 66-kDa PDE4B variant) bands. In unilaterally cryptorchid animals, the abdominal testis showed a time-dependent decrease in both PDE4A8 and 88-kDa PDE4A variants. In contrast, the expression of 66-kDa PDE4B was markedly increased in a time-dependent fashion in abdominal testes of cryptorchid rats. Animals surgically corrected for cryptorchidism and allowed to recover for 50 days exhibited normal expression of both PDE4A and PDE4B variants compared with aged-matched, sham-operated controls. In conclusion, this study suggests that down-regulation of PDE4A variants in cryptorchid testes may play an important role in the degeneration of spermatogonia and increased apoptotic activity in the germ cells.

3',5'-Cyclic-AMP Phosphodiesterases↗

Cloning and characterization of the cyclic guanosine monophosphate-inhibited phosphodiesterase PDE3A expressed in mouse oocyte.

In the preovulatory follicle, oocyte meiotic resumption occurs soon after the LH surge and is associated with a decrease in cAMP. Inhibition of cAMP degradation blocks germinal vesicle breakdown as well as activation of meiotic promoting factor, both hallmarks of reentry into the cell cycle. In situ and pharmacological analysis of rodent ovaries suggested the presence of a phosphodiesterase 3 (PDE3) in the germ cell but not the somatic cell compartment. Here we have investigated the structure and properties of the PDE form expressed in mouse oocytes. Polymerase chain reactions using a mouse oocyte cDNA library as a template, and primers based on the conserved sequence of rat and human PDE3As, yielded partial fragments corresponding to mouse PDE3A. Further screening of the mouse oocyte cDNA library and subsequent ligation of individual cDNA clones yielded PDE3A cDNA containing the entire coding region of mouse PDE3A. To determine the kinetic properties of this PDE, the cDNAs encoding the full-length PDE3A and NH(2)-truncation forms Delta 1 (Delta346aa) and Delta 2 (Delta608aa) were expressed in mouse Leydig tumor cells. Whereas the full-length recombinant protein was always found in the particulate fraction, the Delta 1 and Delta 2 truncated PDE3As were recovered mostly in the soluble fraction. The Michaelis constant values for hydrolysis of cAMP of PDE3A Delta 1 and PDE3A Delta 2 were similar to those of intact full-length PDE3A or oocyte PDE (0.2-0.5 microM). More importantly, there was good correlation between the rank of potency of selective and nonselective compounds in inhibiting recombinant PDE3A or PDE activity derived from cumulus-oocyte complexes and in blocking resumption of meiosis. These data provide evidence that the PDE expressed in the oocyte is a soluble form of PDE3A and that activity of this enzyme is involved in the control of resumption of meiosis.

3',5'-Cyclic-AMP Phosphodiesterases↗

Role of phosphodiesterase type 3A in rat oocyte maturation.

It is generally accepted that cyclic nucleotides are key signaling molecules in the control of oocyte meiotic resumption. Given the role of phosphodiesterases (PDEs) in cyclic nucleotide degradation, this study was undertaken to investigate the properties and regulation of PDEs expressed in rat oocytes. Cilostamide-sensitive PDE3 was the major activity detected in denuded oocytes, whereas no PDE3 activity could be detected in cumulus cells. Moreover, comparable levels of PDE3 activity were measured in cumulus-oocyte complexes (COCs) and in denuded oocytes. The oocyte PDE was recovered in the soluble fraction of the homogenate and immunoprecipitated with a specific PDE3A antibody. A significant and transient increase (P < 0.05) in PDE3 activity was measured in the oocytes after 30 min of culture (70 min after isolation) compared with immediately after collection (10 min after isolation). Conversely, no changes in activity were observed when denuded oocytes or cumulus cells were incubated for up to 130 min. Evaluation of oocyte maturation indicated that only 10% of oocytes had resumed meiosis at the peak of the PDE3 activity. A significant increase (P < 0.05) in PDE3 activity was measured in COCs when follicle-enclosed oocytes were cultured in the presence of hCG. Again, this increase preceded oocyte maturation. In conclusion, these data demonstrate that PDE3A is the major PDE form expressed in mammalian oocytes. PDE3A activity increases prior to resumption of meiosis in both spontaneous and gonadotropin-stimulated maturation. These findings strongly support the hypothesis that an increase in oocyte PDE3A activity is one of the intraoocyte mechanisms controlling resumption of meiosis in rat oocytes, at least in vitro.

3',5'-Cyclic-AMP Phosphodiesterases↗

Relevant cAMP-specific phosphodiesterase isoforms in human pituitary: effect of Gs(alpha) mutations.

Both cAMP production by adenylyl cyclase and cAMP degradation by phosphodiesterases account for intracellular cAMP levels. We previously demonstrated an increased phosphodiesterase activity in GH-secreting adenomas bearing the gsp oncogene. Here we characterize both the activity and the expression of cAMP-specific phosphodiesterase genes in the human pituitary and in gsp+ and gsp- GH-secreting adenomas and analyze the impact of this intracellular feedback mechanism on the levels of cAMP-responsive element-binding protein phosphorylation. Normal pituitary and gsp- GH-secreting adenomas showed similar phosphodiesterase activities, and 7-fold higher levels were observed in gsp+ tumors. In these tumors the increased activity was mainly owing to isobutyl-methyl-xanthine-sensitive phosphodiesterase 4 and to isobutyl-methyl-xanthine-insensitive isoforms. By semiquantitative RT-PCR, all phosphodiesterase 4 transcripts were expressed in the normal and tumoral pituitary. However, the levels of phosphodiesterase 4C and 4D messenger RNAs were significantly higher in gsp+ than in gsp- GH-secreting adenomas and normal pituitary. Expression of the thyroid-specific isobutyl-methyl-xanthine-insensitive phosphodiesterase 8B was absent in the normal pituitary but detectable in almost all GH-secreting adenomas and higher in gsp+ (P < 0.02). Therefore, this study provides a characterization of phosphodiesterase expression in human pituitary and demonstrates a dramatic induction of the cAMP-specific phosphodiesterases 4C and phosphodiesterases 4D and phosphodiesterases 8B in gsp+ GH-secreting adenomas. Similar levels of cAMP-responsive element-binding protein phosphorylation were observed in gsp- and gsp+ GH-secreting adenomas; however, phosphodiesterase blockade caused an increase in cAMP-responsive element-binding protein phosphorylation that was significantly higher in gsp+ than in gsp- adenomas. Because cAMP-responsive element-binding protein represents the principal end point of the cAMP pathway, these results suggest that the enhanced phosphodiesterase activity may have a significant impact on the phenotypic expression of gsp mutations.

1-Methyl-3-isobutylxanthine↗