Search

Axial chordoma represents approximately 1% of malignant bone tumors. This tumor expresses cytokeratins, specifically cytokeratin 19, and commonly S100. More recently brachyury, a transcription factor important in mesodermal differentiation, including notochord development, has been detected by immunohistochemistry in axial chordomas and hemangioblastomas but not chondrosarcomas or other neoplasms. In this report, we describe 10 cases (6 men, 4 women: age 18 to 68 y; mean 44.6) of extra-axial tumors, 8 in bone and 2 in soft tissue, with morphologic and immunohistochemical features identical to those of axial chordoma. Imaging excluded metastases from axial chordoma. Three tumors occurred in the tibia, the others in the rib, metatarsal, ulna, femur, pubis: 2 intracortical, 6 intramedullary. Both soft tissue brachyury-positive tumors, one involving the thumb the other the wrist, were sited in the juxta-articular region. Seven of the tumors were widely excised and these patients are disease-free but of the 3 tumors that recurred, 1 was curetted, 1 was marginally excised, and 1 had a pathologic fracture on presentation. Metastases have not occurred in any of the patients. We also confirm the expression of brachyury in hemangioblastomas, and for the first time demonstrates its expression in spermatogonia and testicular germ cell tumors by immunohistochemistry. Brachyury was not detected in a wide range of tumors including carcinomas, lymphomas, and sarcomas. In conclusion, we describe the first series of extra-axial skeletal chordomas bringing the total number of such cases reported in the literature to 11, and present the first report of 2 soft tissue chordomas as defined by brachyury expression.

Recent work has highlighted the involvement of a dopamine derivative, 5-S-cysteinyl-dopamine (CysDA), in neurodegeneration and apoptotic cell death. In this paper we study in further detail the apoptotic process activated by this catechol-thioether derivative of dopamine in SH-SY5Y neuroblastoma cells. CysDA activates a cascade of events by an initial perturbation of Calcium homeostasis in the cell. Cell treatment with the catechol-thioether induces an immediate rise in intracellular Ca(2+) concentration, as demonstrated by a shift in the indo-1 dye emission spectrum, and a sustained high calcium concentration at long times of incubation. Fluorescence microscopy data show that the treatment of cells induces mitochondrial transmembrane potential depolarization, a clear evidence of the onset of apoptotic process. Programmed cell death activation is also demonstrated by cytochrome c release from the mitochondria, by an increased activity of both caspase-8 and -9 and by the poly(ADP-ribose)polymerase (PARP-1) cleavage, yielding the typical 86 kDa fragment due to caspase-3 activity. Overall, our data support the hypothesis that CysDA may induce apoptotic death in neuronal cells, via an initial perturbation of calcium homeostasis in the cytosol., ((c) 2007 Wiley-Liss, Inc.)

Conus is a genus of predatory marine gastropods that poison the prey with a complex mixture of compounds active on muscle and nerve cells. An individual cone snail's venom contains a mixture of pharmacological agents, mostly short, structurally constrained peptides. This study is focused on the composition of the venom employed by Conus ventricosus Gmelin, 1791, a worm-hunting cone snail living in the Mediterranean Sea. For this purpose, LC coupled to MS techniques has been successfully used to establish qualitative and quantitative differences in conopeptides from minute amounts of venom ducts. We were able to prove variability in the venom conopeptide complement, possibly related to different trophic habits of the species in the Mediterranean Sea. Moreover, the information-rich MS techniques enabled us to identify two novel C. ventricosus peptides, here named Conotoxin-Vn and -Conotoxin-Vn. On the basis of the structural data collected so far, we suggest that Conotoxin-Vn is a conopeptide belonging to the -family that recognizes calcium channels through a specific pharmacophore. Similarly, molecular modeling data suggest that -Conotoxin-Vn should represent a competitive antagonist of neuronal nicotinic acetylcholine receptors (nAChRs).

Valdecoxib is an NSAID that is selective for COX-2 (commonly named coxibs). It exhibits anti-inflammatory, analgesic and antipyretic properties in animal models and humans due to inhibition of prostanoid synthesis primarily by affecting COX-2. In this review, the clinical results of cardiovascular effects of valdecoxib and its prodrug parecoxib were analyzed and the information from animal models and clinical pharmacology was exploited, that is, pharmacodynamic and pharmacokinetic data, to give a mechanistic interpretation. Similarly to other coxibs and some traditional (t)NSAIDs less selective for COX-2, such as diclofenac, valdecoxib may increase the risk of thrombotic events through a prostacyclin-based mechanism. The rapid and elevated thrombotic risk detected in two coronary artery bypass graft surgery trials with parecoxib and valdecoxib is coherent with almost complete suppression of COX-2 by supratherapeutic doses (particularly parecoxib), which plausibly translates into a deep suppression of prostacyclin. Drug potency, that is, the degree of suppression of COX-2-dependent prostacyclin, is proposed to represent a strong determinant in the increased incidence of thrombotic events associated with the use of COX-2 inhibitors and some tNSAIDs.

We compared the variability in degree and recovery from steady-state inhibition of cyclooxygenase (COX)-1 and COX-2 ex vivo and in vivo and platelet aggregation by naproxen sodium at 220 versus 440 mg b.i.d. and low-dose aspirin in healthy subjects. Six healthy subjects received consecutively naproxen sodium (220 and 440 mg b.i.d.) and aspirin (100 mg daily) for 6 days, separated by washout periods of 2 weeks. COX-1 and COX-2 inhibition was determined using ex vivo and in vivo indices of enzymatic activity: 1) the measurement of serum thromboxane (TX)B(2) levels and whole-blood lipopolysaccharide-stimulated prostaglandin (PG)E(2) levels, markers of COX-1 in platelets and COX-2 in monocytes, respectively; 2) the measurement of urinary 11-dehydro-TXB(2) and 2,3-dinor-6-keto-PGF(1alpha) levels, markers of systemic TXA(2) biosynthesis (mostly COX-1-derived) and prostacyclin biosynthesis (mostly COX-2-derived), respectively. Arachidonic acid (AA)-induced platelet aggregation was also studied. The maximal inhibition of platelet COX-1 (95.9 +/- 5.1 and 99.2 +/- 0.4%) and AA-induced platelet aggregation (92 +/- 3.5 and 93.7 +/- 1.5%) obtained at 2 h after dosing with naproxen sodium at 220 and 440 mg b.i.d., respectively, was indistinguishable from aspirin, but at 12 and 24 h after dosing, we detected marked variability, which was higher with naproxen sodium at 220 mg than at 440 mg b.i.d. Assessment of the ratio of inhibition of urinary 11-dehydro-TXB(2) versus 2,3-dinor-6-keto-PGF(1alpha) showed that the treatments caused a more profound inhibition of TXA(2) than prostacyclin biosynthesis in vivo throughout dosing interval. In conclusion, neither of the two naproxen doses mimed the persistent and complete inhibition of platelet COX-1 activity obtained by aspirin, but marked heterogeneity was mitigated by the higher dose of the drug.

We provide comprehensive knowledge on the differential regulation of expression and catalysis of cyclooxygenase (COX)-1 and COX-2 in health and disease which represents an essential requirement to read out the clinical consequences of selective and nonselective inhibition of COX-isozymes in humans. Furthermore, we describe the pharmacodynamic and pharmacokinetic characteristics of major traditional nonsteroidal anti-inflammatory drugs (tNSAIDs) and coxibs (selective COX-2 inhibitors) which play a prime role in their efficacy and toxicity. Important information derived from our pharmacological studies has clarified that nonselective COX inhibitors should be considered the tNSAIDs with a balanced inhibitory effect on both COX-isozymes (exemplified by ibuprofen and naproxen). In contrast, the tNSAIDs meloxicam, nimesulide and diclofenac (which are from 18- to 29-fold more potent towards COX-2 in vitro) and coxibs (i.e. celecoxib, valdecoxib, rofecoxib, etoricoxib and lumiracoxib, which are from 30- to 433-fold more potent towards COX-2 in vitro) should be comprised into the cluster of COX-2 inhibitors. However, the dose and frequency of administration together with individual responses will drive the degree of COX-2 inhibition and selectivity achieved in vivo. The results of clinical pharmacology of COX inhibitors support the concept that the inhibition of platelet COX-1 may translate into an increased incidence of serious upper gastrointestinal bleeding but this effect on platelet COX-1 may mitigate the cardiovascular hazard associated with the profound inhibition of COX-2-dependent prostacyclin (PGI2).

In this study we report the effect of AZT on the whole protein expression profile both in the control and the AZT-treated K562 cells, evidenced by two-dimensional gel electrophoresis and peptide mass fingerprinting analysis. Two-dimensional gels computer digital image analysis showed two spots that appeared up-regulated in AZT-treated cells and one spot present only in the drug exposed samples. Upon extraction and analysis by peptide mass fingerprinting, the first two spots were identified as PDI-A3 and stathmin, while the third one was proved to be NDPK-A. Conversely, two protein spots were present only in the untreated K562 cells, and were identified as SOD1 and HSP-60, respectively.

Aspirin affords cardioprotection through the acetylation of serine529 in human cyclooxygenase-1 (COX-1) of anucleated platelets, inducing a permanent defect in thromboxane A2 (TXA2)-dependent platelet function. However, heterogeneity of COX-1 suppression by aspirin has been detected in cardiovascular disease and may contribute to failure to prevent clinical events. The recent recognized capacity of platelets to make proteins de novo paves the way to identify new mechanisms involved in the variable response to aspirin. We found that in washed human platelets, the complete suppression of TXA2 biosynthesis by aspirin, in vitro, recovered in response to thrombin and fibrinogen in a time-dependent fashion (at 0.5 and 24 hours, TXB2 averaged 0.1+/-0.03 and 3+/-0.8 ng/mL; in the presence of arachidonic acid [10 micromol/L], it was 2+/-0.7 and 25+/-7 ng/mL, respectively), and it was blocked by translational inhibitors, by rapamycin, and by inhibitors of phosphatidylinositol 3-kinase. The results that COX-1 mRNA was readily detected in resting platelets and that [35S]-methionine was incorporated into COX-1 protein after stimulation strongly support the occurrence of de novo COX-1 synthesis in platelets. This process may interfere with the complete and persistent suppression of TXA2 biosynthesis by aspirin necessary for cardioprotection.

Purpose: Although the inpatient setting has served as the predominant educational site of internal medicine training programs, many changes and factors are currently affecting education in this setting. As a result, many educational organizations are calling for reforms in inpatient training. This report reviews the available literature on specific internal medicine inpatient educational interventions and proposes recommendations for improving internal medicine training in this setting., Method: We searched Medline for articles published between 1966 and August 2004 which focused on internal medicine training interventions in the inpatient setting; bibliographies of Medline-identified articles, as well as articles suggested by experts in the field provided additional citations. We then reviewed, classified, and abstracted only articles where an assessment of learner outcomes was included., Results: Thirteen studies of inpatient internal medicine educational interventions were found that included an outcome assessment. All were single institution studies. The majority of these studies was of poor methodological quality and focused on specific content areas of internal medicine. None assessed the effectiveness or impact of internal medicine core inpatient experiences or curriculum., Conclusion: This review identifies significant gaps in our understanding of what constitutes effective inpatient education. The paucity of high quality research in the internal medicine inpatient setting highlights the urgent need to formally define and study what constitutes an effective "core" inpatient curriculum.

Background: Sexual difficulties may arise after stroke. This study was aimed at evaluating and quantifying sexual changes 1 year after stroke., Method: Sixty-eight stroke patients, consecutively admitted to our rehabilitation unit, were enrolled in the study. Sixty-two patients were available for response after 1 year--46 men and 16 women with a mean age of 64 years (SD = 9.2). Time interval between stroke event and admission to rehabilitation unit was 15 days. None of the patients presented with lack of comprehension. Methods of data collection at admission were clinical examination, computed tomographic scan or magnetic resonance imaging, the Cumulative Illness Rating Scale, laboratory data, and data collection on sexual life from patients and, separately, from their partners. After 1 year, they were interviewed again to assess sexual performance; the Center for Epidemiologic Studies-Depression scale, Structured Clinical Interview for DSM-IV, and Functional Independence Measure were also performed. A questionnaire designed for this study was also administered for collecting data on patients' private lives., Results: Sexual decline was common in the post-stroke period. Age (p =.009) and disability (p =.0059) were significant variables. There was no correlation between sexual decline and gender, nor injured hemisphere. There was also no correlation to marriage duration, education duration, or depression. Evaluation and analysis of the questionnaires revealed, however, that patients' partners played a substantial role in the decline of sexual activity. Many partners experienced fear of relapse, anguish, lack of excitation, or even horror, which withheld them from encouraging sexual activities., Conclusion: Data from this study demonstrated that sexual decline in the aftermath of stroke is pronounced. Patients suffer from their sexual impairment, but do not conceal that problem. Psychological, rather than medical, aspects account for discontinuity of sexual activity in stroke survivors, and proper counseling is clearly warranted.

A checkerboard methodology, based on standardized methods proposed by the National Committee for Clinical Laboratory Standards for broth microdilution antifungal susceptibility testing, was applied to study the in vitro interactions of flucytosine (FC) and posaconazole (SCH 56592) (FC-SCH) against 15 isolates of Cryptococcus neoformans. Synergy, defined as a fractional inhibitory concentration (FIC) index of <0.50, was observed for 33% of the isolates tested. When synergy was not achieved, there was still a decrease in the MIC of one or both drugs when they were used in combination. Antagonism, defined as a FIC of >4.0, was not observed. The in vitro efficacy of combined therapy was confirmed by quantitative determination of the CFU of C. neoformans 486, an isolate against which the FC-SCH association yielded a synergistic interaction. To investigate the potential beneficial effects of this combination therapy in vivo, we established two experimental murine models of cryptococcosis by intracranial or intravenous injection of cells of C. neoformans 486. At 1 day postinfection, the mice were randomized into different treatment groups. One group each received each drug alone, and one group received the drugs in combination. While combination therapy was not found to be significantly more effective than each single drug in terms of survival, tissue burden experiments confirmed the potentiation of antifungal activity with the combination. Our study demonstrates that SCH and FC combined are significantly more active than either drug alone against C. neoformans in vitro as well in vivo. These findings suggest that this therapeutic approach could be useful in the treatment of cryptococcal infections.

The interaction of amphotericin B (AmB) and azole antifungal agents in the treatment of fungal infections is still a controversial issue. A checkerboard titration broth microdilution-based method that adhered to the recommendations of the National Committee for Clinical Laboratory Standards was applied to study the in vitro interactions of AmB with fluconazole (FLC), itraconazole (ITC), and the new investigational triazole SCH 56592 (SCH) against 15 clinical isolates of Cryptococcus neoformans. Synergy, defined as a fractional inhibitory concentration (FIC) index of < or =0.50, was observed for 7% of the isolates in studies of the interactions of both FLC-AmB and ITC-AmB and for 33% of the isolates in studies of the SCH-AmB interactions; additivism (FICs, >0.50 to 1.0) was observed for 67, 73, and 53% of the isolates in studies of the FLC-AmB, ITC-AmB, and SCH-AmB interactions, respectively; indifference (FICs, >1.0 to < or =2.0) was observed for 26, 20, and 14% of the isolates in studies of the FLC-AmB, ITC-AmB, and SCH-AmB interactions, respectively. Antagonism (FIC >2.0) was not observed. When synergy was not achieved, there was still a decrease, although not as dramatic, in the MIC of one or both drugs when they were used in combination. To investigate the effects of FLC-AmB combination therapy in vivo, we established an experimental model of systemic cryptococcosis in BALB/c mice by intravenous injection of cells of C. neoformans 2337, a clinical isolate belonging to serotype D against which the combination of FLC and AmB yielded an additive interaction in vitro. Both survival and tissue burden studies showed that combination therapy was more effective than FLC alone and that combination therapy was at least as effective as AmB given as a single drug. On the other hand, when cells of C. neoformans 2337 were grown in FLC-containing medium, a pronounced increase in resistance to subsequent exposures to AmB was observed. In particular, killing experiments conducted with nonreplicating cells showed that preexposure to FLC abolished the fungicidal activity of the polyene. However, this apparent antagonism was not observed in vivo. Rather, when the two drugs were used sequentially for the treatment of systemic murine cryptococcosis, a reciprocal potentiation was often observed. Our study shows that (i) the combination of triazoles and AmB is significantly more active than either drug alone against C. neoformans in vitro and (ii) the concomitant or sequential use of FLC and AmB for the treatment of systemic murine cryptococcosis results in a positive interaction.

Candida tropicalis is less commonly isolated from clinical specimens than Candida albicans. Unlike C. albicans, which can be occasionally found as a commensal, C. tropicalis is almost always associated with the development of fungal infections. In addition, C. tropicalis has been reported to be resistant to fluconazole (FLC). To analyze the development of FLC resistance in C. tropicalis, an FLC-susceptible strain (ATCC 750) (MIC = 1.0 microg/ml) was cultured in liquid medium containing increasing FLC concentrations from 8.0 to 128 microg/ml. The strain developed variable degrees of FLC resistance which paralleled the concentrations of FLC used in the medium. The highest MICs of FLC were 16, 256, and 512 microg/ml for strains grown in medium with 8.0, 32, and 128 microg of FLC per ml, respectively. Development of resistance was rapid and could be observed already after a single subculture in azole-containing medium. The resistant strains were cross-resistant to itraconazole (MIC > 1.0 microg/ml) and terbinafine (MIC > 512 microg/ml) but not to amphotericin B. Isolates grown in FLC at concentrations of 8.0 and 32 microg/ml reverted to low MICs (1.0 microg/ml) after 12 and 11 passages in FLC-free medium, respectively. The MIC for one isolate grown in FLC (128 microg/ml) (128 R) reverted to 16 microg/ml but remained stable over 60 passages in FLC-free medium. Azole-resistant isolates revealed upregulation of two different multidrug efflux transporter genes: the major facilitators gene MDR1 and the ATP-binding cassette transporter CDR1. The development of FLC resistance in vitro correlated well with the results obtained in an experimental model of disseminated candidiasis. While FLC given at 10 mg/kg of body weight/day was effective in reducing the fungal burden of mice infected with the parent strain, the same dosing regimen was ineffective in mice infected with strain 128 R. Finally, the acquisition of in vitro FLC resistance in strain 128 R was related to a loss of virulence. The results of our study elucidate important characteristics and potential mechanisms of FLC resistance in C. tropicalis.

A broth microdilution method performed in accordance with the National Committee for Clinical Laboratory Standards guidelines was used to compare the in vitro activity of the new antifungal triazole SCH 56592 (SCH) to that of fluconazole (FLC), itraconazole (ITC), and ketoconazole (KETO) against 257 clinical yeast isolates. They included 220 isolates belonging to 12 different species of Candida, 15 isolates each of Cryptococcus neoformans and Saccharomyces cerevisiae, and seven isolates of Rhodotorula rubra. The MICs of SCH at which 50% (MIC(50)) and 90% (MIC(90)) of the isolates were inhibited were 0.06 and 2.0 microg/ml, respectively. In general, SCH was considerably more active than FLC (MIC(50) and MIC(90) of 1.0 and 64 microg/ml, respectively) and slightly more active than either ITC (MIC(50) and MIC(90) of 0.25 and 2.0 microg/ml, respectively) and KETO (MIC(50) and MIC(90) of 0.125 and 4.0 microg/ml, respectively). Our in vitro data suggest that SCH has significant potential for clinical development.

To determine if vessel wall overstretching during coronary stenting is associated with a higher restenosis rate, the intravascular ultrasound morphological evaluation was performed following ultrasound criteria. A total of 468 lesions with successful coronary Palmaz-Schatz stenting guided by intravascular ultrasound were classified into the no overstretching group (n = 295) and the overstretching group (n = 147). There were 26 lesions not classifiable due to the poststent morphology. Balloon-to-vessel ratio was 1.12 +/- 0.17 in the no focal overstretching group and 1. 20 +/- 0.20 in the overstretching group (P < 0.0002). Follow-up angiogram was performed in 77% of no focal overstretching group and in 75% of the focal overstretching group. The restenosis rate of the no focal overstretching group was 19.8% and 20.9% in the focal overstretching group, respectively (P = 0.65). Focal overstretching was more frequent following balloon oversizing. No increase in restenosis rate, found in focal overstretched stented lesions, leads us to the hypothesis of a regulation of smooth-muscle-cell proliferation activated by the normalization of blood flow and of shear stress, when stent implantation succeeds in optimally improving the lumen. Cathet. Cardiovasc. Intervent. 48:24-30, 1999., (Copyright 1999 Wiley-Liss, Inc.)

We evaluated the in vitro activity of fluconazole, itraconazole, ketoconazole, 5-fluorocytosine and amphotericin B against 30 clinical isolates of Saccharomyces cerevisiae by a broth microdilution method, following the NCCLS recommendation. Testing was performed either in RPMI-1640 or yeast nitrogen base (YNB). YNB supported the growth of all isolates tested, while results in RPMI-1640 were not obtained for six isolates (20%). The MIC of all three azoles in YNB were one or two dilutions higher than those obtained in RPMI-1640 (P=0.0001 for fluconazole and itraconazole, P=0.03 for ketoconazole). Elevated MICs were observed for all three azoles, while all the isolates were susceptible to 5-fluorocytosine and amphotericin B. All MIC values were confirmed by spectrophotometric reading. Six strains of S. cerevisiae isolated from the faeces and consecutive blood cultures from an AIDS patient over a 7-month period were typed by electrophoretic karyotyping (EK). EK showed the maintenance of the same karyotype over time suggesting that the faecal isolate changed from a colonizing to infection-causing strain. The relative resistance of S. cerevisiae to azole drugs as well as its ability to cause widespread infections may promote the emergence of this species as a pathogen in immunosuppressed patients.

Objectives: Stent implantation reduces restenosis in vessels > or =3 mm compared with balloon angioplasty, but few data are available for stents implanted in vessels <3 mm. The aim of this study was to evaluate immediate and follow-up patient outcomes after stent implantation in vessels <3 mm compared to stent implantation in vessels > or =3 mm., Methods: Between March 1993 and May 1996, a total of 1,298 consecutive patients (1,673 lesions) underwent coronary stenting. The study population was divided into two groups based on angiographic vessel diameter. In case of multivessel stenting, patients were randomly assigned only one lesion. Group I included 696 patients (696 lesions) in whom stents were implanted in vessels > or =3 mm, and group II included 602 patients (602 lesions) in whom stents were implanted in vessels <3 mm., Results: There was no difference in procedural success (95.4% in group I and 95.9% in group II), or subsequent subacute stent thrombosis (1.5% in group I and 1.4% in group II, p=NS). The postprocedure residual diameter stenosis was 3.31+/-12.4% in group I and -2.45+/-16.2% in group II. Angiographic follow-up was performed in 75% of patients, restenosis occurred in 19.9% of patients in group I and 32.6% in group II (p <0.0001). Absolute lumen gain was significantly higher in group I compared to group II, but absolute late lumen loss was similar in the two groups (1.05+/-0.91 mm in group I vs. 1.11+/-0.85 mm in group II, p=NS). Subsequently, the loss index was more favorable in group I (0.45 vs. 0.56; p=0.0006). Independent predictors of freedom from restenosis by multivariate logistic regression in the total population were: larger baseline reference diameter (odds ratio 2.032 p=0.006), larger postprocedure minimal stent cross-sectional area (odds ratio 1.190, p=0.0001) and shorter lesions (odds ratio 1.037, p=0.01). At long-term clinical follow-up, patients with small vessels had a lower rate of event-free survival (63% vs. 71.3%, p=0.007)., Conclusions: Coronary stenting can be performed in small vessels with a high success rate and low incidence of stent thrombosis. However, the long-term angiographic and clinical outcome of patients undergoing stent implantation in small vessels is less favorable than that of patients with large vessels.

OBJECTIVE: This pilot study was performed to evaluate the feasibility and safety of intrawall delivery of long acting steroids before stent implantation, testing the efficacy of this treatment in reducing intimal hyperplasia in lesions at high risk for restenosis. METHODS: In 24 patients (40 lesions) local intrawall drug delivery of methylprednisolone acetate, using a new catheter device, was attempted before elective stent implantation. Treated lesions were compared to a matched control group. RESULTS: Lesions were classified AHA/ACC type C in 47% of cases, had a mean lesion length of 13.6 +/- 9.1 mm, and a mean vessel diameter of 2.85 +/- 0.44 mm. In 9 cases (25%) chronic total occlusions were treated. Methylprednisolone acetate (mean 60 +/- 23 mg) was delivered in 36 lesions (21 patients; delivery success 90%) in the remaining 4 lesions (10%) the delivery device did not cross the lesion. After drug delivery 46 stents were implanted (1.2 stent/lesion; stented segment length 30.1 +/- 18.8 mm) using high pressure for stent expansion (mean 16.4 +/- 3.1 atm). Intracoronary ultrasound guidance was used in 64% of cases. Procedural and in-hospital complications were: Two non Q wave myocardial infarctions (8%) and one (3%) subacute stent thrombosis. Angiographic follow-up was obtained in all 36 treated lesions (100%) and the angiographic restenosis rate (³ 50% diameter stenosis) was 39%. A reduction of the incidence of restenosis compared to the matched control group was not observed. CONCLUSIONS: Long acting steroids could be delivered locally with high success and low complication rates. The restenosis rate remained high in a subset of unfavorable lesions with high risk for restenosis.

We report a patient with systemic sclerosis having implantation of a 35 mm beStent with immediate success but developing angina at follow-up. A focal stent collapse with focal hyperplasia in and outside the stent was documented by ultrasound after 2 mos. A 14mm Palmaz-Schatz stent was successfully deployed into the collapsed beStent, with good 6-mo angiographic result. The stent collapse was probably due to unequal distribution of radial forces and possibly reactive hyperplasia in this unique patient with systemic sclerosis.

Background: Randomized clinical trials have shown that multivessel coronary angioplasty is feasible and provides similar long-term survival as bypass surgery in selected patients. However, the higher need for repeat intervention, in particular, coronary artery bypass graft surgery, remains a problem. The objective of this study was to test the hypothesis that multivessel stenting is safe and effective in reducing the need for repeat interventions, in particular, the need for bypass surgery., Methods and Results: Between March 1993 and June 1995, 100 consecutive patients (243 lesions) had multivessel coronary stenting. High-pressure stent optimization was used in all patients. Procedural success was achieved in 97% of lesions; 2 patients (2%) required emergency bypass surgery. Angiographic follow-up was obtained in 89% of patients at 5.2+/-2.5 months. Angiographic restenosis occurred in 22% of the lesions, but 37% of patients had > or = 1 lesion with restenosis. Clinical follow-up was obtained in all patients at 21 +/- 10 months: target lesion revascularization was needed in 30 patients (30%), repeat angioplasty in 28 patients (28%) and coronary bypass surgery in 2 patients (2%); the overall survival rate was 96% (2% noncardiac death)., Conclusions: Multivessel coronary stenting can be performed with high success rate and low need for emergency bypass surgery. Compared with historical results with multivessel percutaneous transluminal coronary angioplasty, patients who undergo multivessel stenting need less repeat interventions, in particular, less coronary bypass surgery and have similar long-term survival.

The aim of the present study was to investigate the implication of protein kinase C (PKC) in the mouse egg activation process. We used OAG (1-oleoyl-2-acetyl-sn-glycerol) as a PKC activator, calphostin C as a specific PKC inhibitor, and the calcium ionophore A23187 as a standard parthenogenetic agent. The exposure of zona-free eggs to 150 microM or 50 microM OAG for 10 min resulted in meiosis II completion in approximately 80% of instances. By contrast, at a lower concentration (25 microM), the PKC stimulator was ineffective as parthenogenetic agent. Shortly after the application of 150 microM OAG, the cytosolic Ca2+ concentration ([Ca2+]i) increased transiently in all the eggs examined, whereas after the addition of 50 microM OAG, [Ca2+]i remained unchanged for at least 20 min. During this period, the activity of M-phase promoting factor (MPF) dramatically decreased and most of the eggs entered anaphase except when the PKC was inhibited by calphostin C. Similarly, MPF inactivation and meiosis resumption were prevented in calphostin C-loaded eggs following treatment with A23187, even though the ionophore-induced Ca2+ signalling was not affected. Taken together, our results indicate that stimulation of PKC is a sufficient and necessary event to induce meiosis resumption in mouse eggs and strongly suggest that, in this species, the mechanism by which a transient calcium burst triggers MPF inactivation involves a PKC-dependent pathway.

Background: In humans with coronary artery disease, ACE inhibition attenuates coronary sympathetic vasoconstriction. Whether this is due to removal of angiotensin (Ang) II production or to a reduced bradykinin breakdown, however, is unknown., Methods and Results: In eight normotensive patients with angiographic evidence of mild left coronary artery lesions (< or = 50%), mean arterial pressure (MAP, intra-arterial catheter), heart rate (HR, ECG lead), coronary sinus blood flow (CBF, thermodilution method), and coronary vascular resistance (CVR, ratio between MAP and CBF) were measured before and during a 15-minute left intracoronary infusion of Ang II at a dose that had no direct coronary or systemic vasomotor effects. The same measurements were made before and during a 15-minute infusion of saline. A 2-minute cold pressor test (CPT) and a 45-second diving were performed at the end of either infusion period. These maneuvers were used because their coronary vasomotor effects are abolished by phentolamine and thus depend on sympathetic activation. During saline infusion, both CPT and diving caused a marked increase in MAP. HR increased with CPT and fell with diving. CBF increased in parallel to the MAP increase, with little change in CVR. The MAP and HR responses were similar during Ang II infusion, which, however, caused either no change or a reduction in CBF with a consequent marked increase in CVR with both CPT and diving. In four additional patients, the diameter of the stenotic vessels remained unchanged during the CPT performed under saline and Ang II infusion., Conclusions: Ang II markedly enhances sympathetic influences on coronary circulation in humans, presumably by acting at the arteriolar level. This may explain the blunting effect of ACE inhibition on sympathetic coronary vasoconstriction in patients with coronary artery disease.

The new generation quantitative angiographic systems apply the interpolated technique to calculate the reference diameter at the site of the stenosis by integrating measurements of the segments proximal and distal to the stenosis. After stent implantation these measurements can be misleading as the treated segment, which is frequently larger than the adjacent not stented segments, is included in the measurements. The consequence is an overestimation of the reference diameter and the residual diameter stenosis. The present study was performed to compare this conventional technique of measurement with a new method which excludes the stented segment for the calculation of the reference diameter. Fifty-two lesions treated with poorly radiopaque stents (56% Palmaz-Schatz, 28% NIR, 10% Gianturco-Roubin, 6% Wallstent) expanded at high pressure (> = or 16 atm) were analyzed according to the conventional and stent excluded method. After stent implantation the reference diameter was 3.39 +/- 0.48 mm with conventional measurements and 3.02 +/- 0.45 mm with the stent excluded method (P < 0.05). The corresponding % diameter stenosis was 13 +/- 9 for the conventional technique and 1 +/- 13 for the stent excluded analysis (P < 0.05). The new approach to quantitative coronary analysis after stenting provides higher accuracy in reference diameter calculations and allows a more appropriate matching of stented segments with adjacent normal segments.

Objective: To analyze the results of implantation of six different intracoronary stents without the use of prolonged anticoagulation., Material and Methods: Between Mar. 30, 1993, and Jun. 30, 1995, 889 patients with 1,194 coronary or vein graft lesions underwent implantation of one of six types of stents-Palmaz-Schatz, Gianturco-Roubin, Wiktor, Micro, Cordis, or Wallstent. The patients were classified into seven groups on the basis of the type of stent that was implanted, including one group with combined use of two or more types of stents. Among the 851 patients with successful stent delivery and without major complications, 801 received only antiplatelet therapy, and 50 received a standard anticoagulation regimen. One-month clinical followup data were obtained in all patients, and clinical events were investigated., Results: The mean number of stents was 1.8 per lesion and 2.4 per patient. Procedural success was achieved in 93% of the lesions. The clinical success rate at 1 month was 90%. Intravascular ultrasound assessment was performed in 90% of the lesions. The final minimal luminal cross-sectional area of the stent increased from 6.8 to 7.8 mm2 after intravascular ultrasound-guided optimization. Within 1 month, 16 stent thrombosis events (1.9%) occurred. No significant differences were noted in stent thrombosis rates among the various stent cohorts. Multivariate logistic regression analysis revealed that the final stent minimal luminal diameter measured by intravascular ultrasonography was the only variable associated with stent thrombosis., Conclusion: This study showed that six different stents could possibly be inserted without subsequent anticoagulation if optimal stent expansion and total lesion coverage were achieved.

Coronary stent implantation has become an accepted treatment for selected patients but the long-term outcome after stent implantation has not been investigated in a large unselected population. This study reports clinical and angiographic results after coronary stent implantation in a consecutive group of 1000 patients treated between November 1989 and June 1994. A total of 2012 stents were implanted in 1216 lesions. The anticoagulation regimen after successful stenting was abolished in the last 499 patients, treated with aspirin and ticlopidina. Complex lesions (type B2, type C lesions and chronic total occlusions) were less frequent in the group with anticoagulation (67%, 355/529 lesions) than in the group without anticoagulation (76%, 485/641; p < 0.01). Vessel size was also significantly smaller in the group with no anticoagulation (3.21 +/- 0.50 mm vs 2.89 +/- 0.70 mm; p < 0.001). Procedural success was achieved in 96% of cases. Major complications occurred in 49 pts, including death (1%), emergency bypass surgery (2.7%), and myocardial infarction (2.1%). Subacute stent thrombosis, observed in 2.6% of the patients treated with anticoagulation, decreased to 1% in the last 499 patients receiving only antiplatelet drugs after high pressure stent expansion controlled with intravascular ultrasound (p < 0.05). Angiographic follow-up was obtained at a mean interval of 6.6 +/- 3.0 months in 718 patients (75% of the eligible candidates). The global restenosis rate (greater than 50% diameter stenosis) was 22%, with no statistically significant difference in patients with and without anticoagulation after stenting. Clinical follow-up was available in 890 patients at a mean interval of 14 +/- 17 months. Late cardiac events occurred in 251 patients and included: 159 repeat coronary angioplasties for restenosis (20%), 114 coronary angioplasties for de novo lesions (10%), 33 elective by-pass surgery operations (3.7%) and 42 late deaths (4.7%). At the most recent follow-up there were 719 pts (81%) free of anginal symptoms, including patients having repeat angioplasty for restenosis or disease progression. Coronary stenting has a low incidence of procedural complications and high-pressure ultrasound guided implantation has drastically reduced the risk of subacute thrombosis. Repeat procedures due to restenosis remain the most important limitation of the technique and do not seem to be affected by evolving technique and operator experience.

Clinical trials have shown that stents are superior to other catheter-based coronary interventions in terms of reduced complications and improved long-term efficacy. With utilization of high-pressure balloon inflation and intravascular ultrasound (IVUS) guidance, stent implantation can now be performed safely without anticoagulation (i.e., with lower rates of stent thrombosis and vascular complications). In 2 recent prospective clinical trials, stent thrombosis occurred in 3.5% of cases despite anticoagulant therapy, which resulted in an average of 7% vascular and bleeding complications. Initial use of IVUS during traditional stent deployment showed that 80% of stents were underexpanded and led to the hypothesis that stent thrombosis might be decreased as a result of optimal stent placement under IVUS guidance without the need for anticoagulation. In a prospective clinical trial to test this hypothesis, three factors were found to reduce stent thrombosis: full stent expansion, complete apposition to the vessel wall, and full lesion coverage. Predictors of thrombotic risk in this era of high-pressure stent deployment without anticoagulation include low ejection fraction, residual dissections, slow flow, multiple stents per lesion, and smaller postprocedure stent luminal diameter. To optimize stent expansion, stent dilation should be performed using a mean inflation pressure of 18 atm with a noncompliant or minimally compliant balloon sized to the vessel being treated (B/V ratio = 1.1). Controversy still remains about the best poststent antiplatelet regimen, and results of a recent trial should indicate whether heparin coating provides additional protection from stent thrombosis.

Plasma endothelin-1 (ET-1) and von Willebrand factor (vWF) levels are elevated in the presence of either macro- or microvascular lesions. Since an increased risk to develop hypertension-related vascular damage has been suggested in human sensitive hypertension, we evaluated both substances in plasma samples from 20 non-diabetic, non obese essential hypertensive men (mean age 49 +/- 4 years). Patients were divided in salt sensitive (n = 9) or salt resistant (n = 11) groups, according to the individual response to both high- and low-sodium diets. Plasma ET-1 levels were also assessed after an oral glucose tolerance test (75 g). Both ET-1 and vWF were higher in salt sensitive than salt resistant patients (ET-1 p < 0.01; vWF p < 0.03). Furthermore, after oral glucose administration, plasma ET-1 concentrations increased very mildly but significantly only in salt sensitive patients (p < 0.05 at 90 min). In conclusion, human salt sensitive hypertension is combined to increased levels of two markers of endothelial damage, and by an augmented ET-1 response to glucose leading, suggesting it is characterized by an increased risk to develop hypertension-related vascular complications.

We report a successfully percutaneous closure of a brisk coronary artery rupture with a custom-made "vein graft stent," a Palmaz-Schatz stent covered with a vein graft. This method is an elegant and effective alternative to the traditional surgical approach and should be considered whenever technically and clinically feasible.

Endothelial injury in vivo induced by Rickettsia Conorii, the etiologic agent of Mediterranean Spotted Fever (MSF) has been recently demonstrated. We sought to determine whether platelet and/or coagulative activation in vivo can be demonstrated in the acute phase of MSF, through measurements of a major metabolite of thromboxane (TX) in the urine (11-dehydro-TXB2) and of plasma prothrombin fragment 1 + 2, whose levels reflect activation of prothrombin to thrombin. Moreover, we measured plasma endothelin-1 as marker of endothelial dysfunction. Our results provide biochemical evidence for the occurrence of TXA2-dependent platelet activation and thrombin generation in vivo, together with endothelial dysfunction. These phenomena could account for clinical manifestations of MSF, such as vasculitis and focal microthrombus formation. These results could also provide a rationale for testing the efficacy of aspirin or heparin in reducing the prothrombotic status of Rickettsiae diseases.

Pig brain cytosolic sialidase purified to homogeneity, showed a single protein band on SDS-PAGE under non-reducing conditions, and three bands using reducing conditions, suggesting a complex of different units. The sialidase complex (molecular mass, M(r), 180 kDa) was resolved into a catalytic unit (M(r) 30 kDa), active but very liable upon storage at 4 degrees C and freezing and thawing, and two protective units (66 kDa and 42 kDa), inactive, but capable to stabilize the catalytic unit. Recombination of the catalytic and protective units (optimal ratio, 1:1, by weight) gave rise to a stable active complex. Using GD1a as substrate, the catalytic unit showed a Michaelis-Menten kinetics, and the complex a sigmoid-shaped kinetics, whereas a Michaelis-Menten kinetics was exhibited with MU-NeuAc in both cases. The apparent Vmax and Km values of the catalytic unit for MU-NeuAc and GD1a were 105.1 and 110.0 mU/mg protein, and 4.2 x 10(-5) and 1.6 x 10(-5) M, respectively. The model we propose for cytosolic sialidase complex is one of each protective units and 2-3 catalytic units. The sialidase complex and protective units did not display any beta-D-galactosidase, beta-D-N- acetylglucosaminidase, alpha-L-fucosidase, alpha-D-glucosidase and carboxypeptidase activities.

Objective: To measure the levels of two adhesion molecules (AM), soluble intercellular adhesion molecule 1 (sICAM-1) and soluble E-selectin (sE-selectin), in serum and synovial fluid (SF) of patients with juvenile chronic arthritis (JCA)., Methods: Both soluble AM levels were tested, in serum and synovial fluid (SF) samples, with an enzyme-linked immunosorbent assay (ELISA) method., Results: Serum levels of sICAM-1 and sE-selectin in JCA patients were not significantly different from those of a control group. Synovial fluid levels of sICAM-1, but not of sE-selectin, assayed significantly higher (p < 0.05) in JCA patients than in controls. Moreover SF levels of both molecules correlated negatively with disease duration, being higher in the earliest phases. No significant correlations were found between JCA sICAM-1 and sE-selectin levels and leukocyte count or ESR., Conclusions: These observations may signify a more important role of ICAM-1 than E-selectin in the migration of inflammatory cells into JCA SF. The negative correlation of both AMSF levels in JCA patients with disease duration could reflect a higher expression of ICAM-1 and E-selectin during the earliest phases of the disease.

Some predictive markers for NaCl sensitivity, related to the red blood cell membrane or to circulating proteins, have already been described in human essential hypertension. The present study was planned to investigate whether or not some hormones produced by the kidney or acting at the kidney level could be used as new markers for NaCl sensitivity. The study was conducted in 28 not previously treated outpatients affected by uncomplicated mild to moderate essential hypertension. After 15 days on a normal NaCl diet, plasma renin activity (PRA), plasma atrial natriuretic peptide (ANP), and the urinary excretion of active kallikrein were evaluated. The sensitivity of blood pressure to changes in NaCl intake was then assessed in all patients, according to a randomized double blind cross-over design. Each patient was assigned to a high (240 mmol of NaCl/day for 15 days) or low (40 mmol of NaCl/day for 15 days) NaCl intake. During the assessment of NaCl sensitivity, the double blindness was achieved by the use of capsules containing either NaCl or placebo. Fifteen patients (11 males and 4 females) resulted as NaCl-sensitive, while 13 patients (8 males and 5 females) were classified as NaCl-resistant. Our results indicate that PRA levels were significantly lower in the NaCl-sensitive group than in the NaCl-resistant one (0.108 +/- 0.05 ng/L/s vs 0.247 +/- 0.16 ng/L/s, p < 0.007), in the presence of raised levels of plasma ANP in NaCl-sensitive hypertensives (18.08 +/- 4.61 fmol/mL vs 12.45 +/- 3.77 fmol/mL, p < 0.006).(ABSTRACT TRUNCATED AT 250 WORDS)

Treatment of human erythrocyte ghosts with phosphatidylinositol-phospholipase C (PIPLC) from Bacillus cereus liberated the ghost-linked sialidase. Maximal release of sialidase (about 70% of total) was achieved by incubating ghosts at 37 degrees C for 60 min, at pH 6.0, with PIPLC (PIPLC total units/ghost protein ratio, 4.5 each time) added at the beginning of incubation and every 15 min (four subsequent additions). Liberated sialidase was fully resistant to at least four cycles of rapid freezing and thawing and to storage at 4 degrees C for at least 48 h. The liberated enzyme had an optimal activity at pH 4.2, degraded ganglioside GD1a better than methylumbelliferyl N-acetylneuraminic acid (about fourfold), and gave a Km value of 2.56 x 10(-4) M and an apparent Vmax of 2.22 mU per mg protein on GD1a. Treatment of intact erythrocytes with PIPLC (PIPLC total units/erythrocyte protein ratio, 8), under conditions where haemolysis was practically negligible, caused liberation of 10-12% of membrane linked sialidase, indicating that the enzyme is, at least in part, located on the outer surface of the erythrocyte membrane. It is concluded that the erythrocyte membrane sialidase is anchored by a glycosylphosphatidylinositol structure sensitive to PIPLC action, and is partly located on the outer surface.

In order to verify the influence of salt sensitivity on the blood pressure response to orally administered kallikrein, we evaluated the efficacy of glandular kallikrein (derived from porcine pancreas) in 28 essential hypertensives (21 males and 9 females) aged between 40 and 62 years. After a placebo run-in period, the patients were assigned to receive oral kallikrein therapy (150 IU 3 times a day; n = 18 patients) or placebo (n = 10 patients) over a period of 8 days in a random double-blind fashion. In the salt-resistant patients (n = 8), kallikrein administration did not modify blood pressure levels. In the same group, natriuresis increased significantly after the treatment [from 94.51 +/- 10.76 to 111.65 +/- 23.19 mEq/24 h (mmol/24 h), p < 0.039]. In the salt-sensitive patients (n = 10), blood pressure decreased with the kallikrein therapy (systolic: from 158.50 +/- 9.20 to 144.50 +/- 10.12 mm Hg, p < 0.005; diastolic: from 99.50 +/- 2.16 to 90.0 +/- 3.67 mm Hg, p < 0.024). In the same patients, urinary Na+ excretion increased considerably after the kallikrein treatment (from 101.07 +/- 18.36 to 134.34 +/- 18.27 mEq/24 h, p < 0.0001). Therefore, our data indicate that the oral kallikrein administration reduces blood pressure levels only in the salt-sensitive hypertensives. In both the salt-sensitive and the salt-resistant groups a marked increase in the 24-hour urinary excretion of sodium was observed after the kallikrein treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Light and heavy lysosomes of mouse forebrain were separated from each other by centrifugation on a Percoll gradient. Light lysosomes were then freed from mitochondria and membranes by sucrose density gradient centrifugation and further purified by floatation-centrifugation on a sucrose gradient. The final preparations of light and heavy lysosomes, fairly homogenous, carried sialidase activity, assayed on MU-NeuAc. The optimal pH was 4.0 and 4.2, the apparent Km value 2.8 x 10(-5) M and 4.2 x 10(-5) M and the apparent Vmax value 0.11 and 0.47 mU.mg-1 protein, for the light and heavy lysosome sialidase, respectively. From 4 days to adulthood the specific activity of the light and heavy lysosome sialidase increased 3-fold and 1.7-fold, respectively.