Cripto in tumors and embryo development.
Explore the source record for details and available documents.
Amino Acid Sequence↗
Biomedical subjects
S Parisi
Publications and source records attributed to S Parisi.
Sensitivity of mitochondrial respiration to different inhibitors in Venturia inaequalis.
The sensitivity of Venturia inaequalis field isolates to inhibitors of the cytochrome bc1 complex at the Qo site (QoIs) was characterised at the molecular, biochemical and physiological level, and compared to other respiration inhibitors. Comparison of a sensitive and a QoI-resistant isolate revealed very high resistance factors both in mycelium growth and spore germination assays. Cross-resistance was observed among QoIs such as trifloxystrobin, azoxystrobin, famoxadone, strobilurin B and myxothiazol. In the mycelium growth assay, antimycin A, an inhibitor of the cytochrome bc1 complex at the Qi site, was less active against the QoI-resistant than against the sensitive isolate. The mixture of QoIs with salicylhydroxamic acid (SHAM), an inhibitor of the alternative oxidase, exerted synergistic effects in the spore germination but not in the mycelium growth assay. Thus, the cytochrome and the alternative respiration pathways are assumed to play different roles, depending on the developmental stage of the fungus. Induction of alternative oxidase (AOX) by trifloxystrobin was observed in mycelium cells at the molecular level for the sensitive but not the resistant isolate. Following QoI treatment, respiration parameters such as oxygen consumption, ATP level, membrane potential and succinate dehydrogenase activity were only slightly reduced in Qo-resistant mycelium cells, and remained at much higher levels than in sensitive cells. In contrast, no difference was observed between sensitive and resistant isolates when NADH consumption was measured. Comparison of the cytochrome b (cyt b) gene of the sensitive and resistant isolates did not reveal any point mutations as is known to occur in resistant isolates of other plant pathogens. It is assumed that QoI resistance in V inaequalis may be based on a compensation of the energy deficiency following QoI application upstream of the NADH dehydrogenase of the respiratory chain.
Adenosine Diphosphate↗
Characterization of rec7, an early meiotic recombination gene in Schizosaccharomyces pombe.
rec7 is involved in intra- and intergenic meiotic recombination in all tested regions of the genome of the fission yeast Schizosaccharomyces pombe. Segregational analysis in a rec7 gene disruption mutant revealed frequent occurrence of two-spored asci. Spores giving rise to diploid colonies were shown to derive from skipping of the second meiotic division. Nondisjunction of homologous chromosomes at the first meiotic division was also frequent. The cytological structures and processes, such as formation of linear elements, pairing of homologous chromosomes, and clustering of telomeres and centromeres, are regular in the mutant. Northern blot experiments revealed meiosis-specific expression of rec7. Screening of a meiotic cDNA library also identified transcripts from the opposite strand in the rec7 region. A Rec7-GFP fusion protein was localized in the nucleus of whole cells before karyogamy, during prophase, and after meiosis I. On spreads of prophase nuclei approximately 50 foci of Rec7-GFP were counted. Some of the observed phenotypes of the disruption mutant and the N-terminal sequence homology suggest that Rec7p is a functional homolog of Rec114p of Saccharomyces cerevisiae. The observed phenotypes of the disruption and the appearance of Rec7-GFP in mating haploid cells and after meiosis I are consistent with Rec7p functions before, during, and after meiotic prophase.
Blotting, Northern↗
Structure-function analysis of the EGF-CFC family member Cripto identifies residues essential for nodal signalling.
Cripto is the founding member of the family of EGF-CFC genes, a class of extracellular factors essential for early vertebrate development. In this study we show that injection of Cripto recombinant protein in mid to late zebrafish Maternal-Zygotic one-eyed pinhead (MZoep) blastulae was able to fully rescue the mutant phenotype, thus providing the first direct evidence that Cripto activity can be added extracellularly to recover oep-encoded function in zebrafish early embryos. Moreover, 15 point mutations and two deletion mutants were generated to assess in vivo their functional relevance by comparing the ability of cripto wild-type and mutant RNAs to rescue the zebrafish MZoep mutant. From this study we concluded that the EGF-CFC domain is sufficient for Cripto biological activity and identified ten point mutations with a functional defective phenotype, two of which, located in the EGF-like domain, correspond to loss-of-function mutations. Finally, we have developed a three-dimensional structural model of Cripto protein and used it as a guide to predict amino acid residues potentially implicated in protein-protein interaction.
Amino Acid Sequence↗
Analysis of patient flows via data mining.
The paper presents DoMiner, a data mining tool for the analysis of patient flows among public hospitals and care units. DoMiner is based on the theory of rough sets, and allows for the extraction of association rules from data base tables. The paper describes both the clustering and rule extraction algorithm of DoMiner, and illustrates an introductory analysis example.
Algorithms↗
Membrane-anchorage of Cripto protein by glycosylphosphatidylinositol and its distribution during early mouse development.
cripto is the original member of the family of EGF-CFC genes, recently recognized as novel extracellular factors essential for vertebrate development. During the early stages of mouse gastrulation, cripto mRNA is detected in mesodermal cells; later, cripto mRNA is detected only in the truncus arteriosus of the developing heart. Here we describe the in vivo distribution of Cripto protein throughout mouse embryo development and show that cripto mRNA and protein colocalize. By means of immunofluorescence analysis and biochemical characterization, we show that Cripto is a membrane-bound protein anchored to the lipid bilayer by a glycosylphosphatidylinositol (GPI) moiety. We suggest that presentation of Cripto on the cell surface via a GPI-linkage is important in determining the spatial specificity of cell-cell interactions that play a critical role in the early patterning of the embryo.
Animals↗
[Primary tumors of the central nervous system: histogenetic classification, grade of malignity, clinico-pathologic features, and therapeutic criteria].
We review the current classification of the central nervous system neoplasms focusing on grading and new clinicopathological entities. Moreover a brief synopsis on up to date treatments in different groups of neoplasms is given.
Astrocytoma↗
The msh2 gene of Schizosaccharomyces pombe is involved in mismatch repair, mating-type switching, and meiotic chromosome organization.
We have identified in the fission yeast Schizosaccharomyces pombe a MutS homolog that shows highest homology to the Msh2 subgroup. msh2 disruption gives rise to increased mitotic mutation rates and increased levels of postmeiotic segregation of genetic markers. In bandshift assays performed with msh2Delta cell extracts, a general mismatch-binding activity is absent. By complementation assays, we showed that S. pombe msh2 is allelic with the previously identified swi8 and mut3 genes, which are involved in mating-type switching. The swi8-137 mutant has a mutation in the msh2 gene which causes a truncated Msh2 peptide lacking a putative DNA-binding domain. Cytological analysis revealed that during meiotic prophase of msh2-defective cells, chromosomal structures were frequently formed; such structures are rarely found in the wild type. Our data show that besides having a function in mismatch repair, S. pombe msh2 is required for correct termination of copy synthesis during mating-type switching as well as for proper organization of chromosomes during meiosis.
Alleles↗
Rec8p, a meiotic recombination and sister chromatid cohesion phosphoprotein of the Rad21p family conserved from fission yeast to humans.
Our work and that of others defined mitosis-specific (Rad21 subfamily) and meiosis-specific (Rec8 subfamily) proteins involved in sister chromatid cohesion in several eukaryotes, including humans. Mutation of the fission yeast Schizosaccharomyces pombe rec8 gene was previously shown to confer a number of meiotic phenotypes, including strong reduction of recombination frequencies in the central region of chromosome III, absence of linear element polymerization, reduced pairing of homologous chromosomes, reduced sister chromatid cohesion, aberrant chromosome segregation, defects in spore formation, and reduced spore viability. Here we extend the description of recombination reduction to the central regions of chromosomes I and II. We show at the protein level that expression of rec8 is meiosis specific and that Rec8p localizes to approximately 100 foci per prophase nucleus. Rec8p was present in an unphosphorylated form early in meiotic prophase but was phosphorylated prior to meiosis I, as demonstrated by analysis of the mei4 mutant blocked before meiosis I. Evidence for the persistence of Rec8p beyond meiosis I was obtained by analysis of the mutant mes1 blocked before meiosis II. A human gene, which we designate hrec8, showed significant primary sequence similarity to rec8 and was mapped to chromosome 14. High mRNA expression of mouse and human rec8 genes was found only in germ line cells, specifically in testes and, interestingly, in spermatids. hrec8 was also expressed at a low level in the thymus. Sequence similarity and testis-specific expression indicate evolutionarily conserved functions of Rec8p in meiosis. Possible roles of Rec8p in the integration of different meiotic events are discussed.
Amino Acid Sequence↗
[Radiotherapy alone in endometrial neoplasms. The authors' own experience].
INTRODUCTION: Adenocarcinoma of the endometrium is the most common invasive genital malignancy in women and the majority of the cases are in stage I (80-85%) at the time of diagnosis. Total abdominal hysterectomy with bilateral salpingo-oophorectomy is the treatment of choice for most patients with uterine carcinoma. However, a number of women with endometrial cancer cannot undergo surgical treatment because of poor medical conditions. In these patients, who cannot tolerate surgery, radiation therapy is the only effective alternative. It is performed either as external treatment plus intracavitary brachytherapy or as brachytherapy alone. This retrospective study reports our data on survival, complications and local control in a consecutive series of patients treated with curative radiotherapy. MATERIAL AND METHODS: From January, 1985, to December 1995, at the Radiotherapy Department of "Casa Sollievo della Sofferenza" Hospital in San Giovanni Rotondo (Foggia, Italy), 60 patients were treated with combined external beam radiation therapy (ERT) and high-dose-rate intracavitary brachytherapy (HDR-BRT) or with high dose rate intracavitary brachytherapy alone. The average age of patients was 69 years (range 50-90). FIGO stage distribution was: 41 patients in stage I, 11 in stage II and 8 in stage III. The ERT was given by means of 6-8 MV linear accelerator, with conventional technique (with two opposed AP-PA pelvic fields in 15 patients, with the four fields-box technique in 41 patients) and a daily fraction to a total dose of 45-50 Gy. HDR-BRT was delivered by means of an HDR remote afterloading unit, containing a linear source of 192-Iridium (370 Gbq). The dose was specified to Point A in 32 patients and to uterine outline in 26. 2-3 intracavitary insertions (mean dose 6-8 Gy per fraction) were performed with weekly intervals. RESULTS: At the time of the analysis, all the patients were available for follow-up. Median follow-up was 25 months. 60% of patients were alive and well with no evidence of disease; 3.3% were alive with disease; 20% had died of this and 16.7% of other diseases. Five-year actuarial specific survival, obtained with the Kaplan and Meier method, was 77.7% in stage I, 90% in stage II, and 75% in stage III. Local relapses were observed in 14 patients. Complications (grade 2-3) scored with the French-Italian Glossary, were gastrointestinal in 10% of cases and genital in 6.6%. CONCLUSIONS: Radical radiotherapy achieved acceptable specific survival, local control and complications rates in patients with medically or surgically inoperable uterine carcinomas. Complications and survival rates, in our experience, are consistent with the literature data. The treatment is comfortable for the patients, because there is no need for long immobilization and it can frequently be performed on an outpatient basis. Besides, the completely standardized procedure was carried out easily with remote control allowing maximal radiation protection.
Aged↗
Mechanism of the persisting TxA2 receptor antagonism by picotamide.
Picotamide is a dual TxA2 receptor antagonist/TxA2 synthetase inhibitor. As the picotamide effect is maximum only after 5-10 min of incubation, aim of the present work was to study whether the effect of picotamide could be observed even after wash out of the drug from the external buffer. Platelet aggregation, serotonin release and changes in intracellular calcium were analyzed in platelets treated with picotamide and then gel-filtered, in comparison with gel-filtered platelets treated with picotamide. To exclude the possibility that the inhibitory effect of picotamide is due to its intracytosolic storage, serotonin release in digitonin-permeabilized platelets was measured. Platelet aggregation and serotonin release in response either to U46619, a synthetic agonist of the thromboxane A2 receptor, or arachidonic acid or collagen, as well as the changes in intracellular calcium concentration after U46619 or arachidonic acid stimulation, were inhibited by picotamide even when it had been washed out by gel-filtration. Both inhibitions were very similar to that observed in experiments in which picotamide was present in the medium. As the serotonin release in digitonin permeabilized platelets presented the same inhibition it may be excluded that picotamide effect is consequent upon the cytosolic storage of the drug. Since our results clearly indicate that the action of picotamide persists even after washing out of the drug from the medium, the idea that this antagonistic effect may be dependent on its binding to platelet plasma membrane rather than on its cytosolic concentration, is strongly substantiated.
Blood Platelets↗
[Preliminary results of preoperative radiotherapy schedule in the treatment of rectal tumors].
From March, 1988, to October, 1993, fifty-six consecutive patients with rectal adenocarcinoma in clinical stage T3NxM0, underwent preoperative radiation therapy at the "Casa Sollievo della Sofferenza" Hospital of San Giovanni Rotondo (Italy). The patients were treated with the four-field technique with 6 to 8 MV X photons on the pelvis. The dose given was 36 Gy in 12 fractions of 3 Gy each. Surgery was performed 2-3 weeks after completion of radiotherapy. Six patients were excluded from this study for metastatic involvement of the liver found at surgery. 48% of 50 assessable patients underwent abdominoperineal resection and 52% anterior resection. 68% of patients were in pathologic stage pT0-3 pN0 and 32% in pT0-3 pN1-2. Metastatic nodes were found in 16 patients (32%) (11 pN1 and 5 pN2). 4% of patients achieved a complete response. The follow-up ranged 24 to 91 months (mean: 46 months). None of the 50 patients died during the postoperative period and the specific morbidity was 26%. Side-effects, requiring surgery, were found in 4% of patients (1 retroperitoneal fibrosis and 1 small bowel occlusion). The incidence of local relapse was 8%. The overall survival at 5 years, in all stages, calculated with the Kaplan and Meyer method, was 76.5%. The disease-free survival rate was 81.1% in all stages: 94.1% in pT0-3 pN0 patients and 54.1% in pT0-3 pN1-2 patients. The disease-free survival rate related to nodal involvement was 72.7% in pN1 patients and 20% in pN2 patients. Our experience confirms the effectiveness of preoperative radiation therapy to improve local control in rectal cancer patients. In the future, it will be useful to assess the impact on prognosis of the schedules using chemotherapy, different fractionation of radiotherapy, delayed surgery and biological predictors of response to irradiation.
Adenocarcinoma↗
Studies on in vitro effect of picotamide on human platelet aggregation in platelet-rich plasma and whole blood.
Picotamide is a new antiaggregating agent influencing the platelet prostaglandin pathway through an inhibition of thromboxane A2 (TXA2) synthesis and a competitive antagonism of platelet TXA2 receptors. In the present study, we investigated the in vitro effect of this drug on human platelet aggregation induced by different agents (adenosine 5'-diphosphate [ADP], collagen, Na arachidonate) both in platelet-rich plasma (PRP; Born's method) and whole blood (WB; impedance method). For each aggregating agent, ED50 value (agonist concentration necessary to induce a maximal aggregation of 50%) was determined in control samples and following addition of different picotamide concentrations on the basis of dose-response curves. Picotamide decreased the response to each aggregating agent in both WB and PRP samples. In WB, 25 microM picotamide was able to induce a highly significant enhancement of ED50 values for ADP (from 6.6 +/- 1 microM to 12.7 +/- 1.7 microM, p < 0.01), Na arachidonate (from 740 +/- 240 microM to 1,080 +/- 280 microM, p < 0.01) and collagen (from 2.4 +/- 0.3 micrograms/ml to 3.8 +/- 0.15 micrograms/ml, p < 0.01). In PRP, the same picotamide concentration significantly enhanced ED50 for each aggregating agent (from 2.0 +/- 0.1 microM to 3.1 +/- 0.3 microM for ADP, p < 0.01; from 960 +/- 80 microM to 1,850 +/- 260 microM for Na arachidonate, p < 0.001; from 3.0 +/- 0.3 microgram/ml to 5.0 +/- 0.8 micrograms/ml for collagen, p < 0.01). Present results show that picotamide effect on platelet response is present also in WB. Data might support the use of picotamide as antiaggregating agent in vascular diseases.
Adenosine Diphosphate↗
Effect of picotamide on the calcium mobilization and phospholipase C activation in human platelets.
The effect of picotamide (G137 or N,N'-bis-3-picolyl-4-methoxyisophthalamide), a dual thromboxane A2 (TxA2) synthetase inhibitor/TxA2 endoperoxide receptor antagonist, on the phospholipase C (PLC) activation and calcium mobilization in human platelets stimulated by arachidonic acid (AA) and TxA2 receptor synthetic agonist U46619, has been studied. Preincubation with picotamide (10(-4) M) for 1 and 3 min significantly reduced (p < 0.03 and p < 0.005 respectively) the calcium concentration changes induced in gel-filtered platelets (GFPs) by U46619 125 nM and 250 nM. Picotamide also reduced the calcium concentration changes induced in GFPs by AA 75 and 150 microM and by ADP 5 and 10 microM. In thrombin degranulated platelets picotamide inhibited the effect of U46619 up to 500 nM. The PLC activation, as indicated by inositol-1,3,4 P3 (Ins 1,3,4 P3) formation in response to U46619 250 nM and AA 150 microM was also inhibited by picotamide. These results may suggest a dual effect of picotamide on the receptor/effector systems through which TxA2 mediates platelet activation.
15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
Chest pain self-management training for patients with coronary artery disease.
This study was designed to evaluate the efficacy of a brief, structured pain management program to improve control over chest pain episodes in patients diagnosed with coronary artery disease. Twenty-six male veterans who attended the three-session program were compared with twenty-six matched controls. Results indicated significant short-term reductions in self-report of number of chest pain episodes in treated subjects. Self-report of pretreatment daily physical activity level moderated treatment outcome, as individuals reporting lower levels of physical activity derived greater benefit than their high-activity counterparts. These results suggest the potential utility of incorporating chest pain control strategies into comprehensive cardiac rehabilitation programs.
Adult↗
Bone formation markers and pain palliation in bone metastases treated with strontium-89.
In several bone disorders, including those with metastatic involvement, changes in procollagen type I C-terminal and type III N-terminal peptides are detected, as indications of altered bone metabolism. Assessment of bone turnover could play a role in the evaluation of response to Strontium-89 used as palliative treatment in symptomatic bone metastases from various primary tumors. A correlation between bone formation rate markers procollagen I and III and efficacy of ionic Strontium-89 was shown in a group of 13 patients who underwent treatment with 4 mCi of Strontium-89 for painful bone metastases: 5 from breast, 7 from prostate, and 1 from lung carcinoid cancer. Assessed as a modification of analgesic intake, pain, and ambulation, there were 6 complete remissions, 3 partial remissions, and 4 nonresponders. The duration of the response was from 2 to 11 months. Procollagen I and III levels were found to be highly abnormal in those with no benefit from Strontium-89 administration but were in the normal range or only slightly elevated in those achieving complete or partial pain control, thus correlating with the clinical response.
Adult↗
Analgesic rebound headache: experience in a community hospital.
In the past, chronic daily headaches have been attributed to many possible causes, including "tension," excessive contraction of pericranial muscles, and depression. A careful history reveals that many patients start out with fairly typical intermittent migraine attacks that evolve over the years into a pattern of daily or almost daily head pain. Overuse of analgesics is usually a part of this clinical picture, which has been termed "analgesic rebound headache." Fifty-three of the first 100 patients admitted to a community hospital's inpatient headache treatment program had chronic daily headaches and were overusing analgesics or other headache relief medications. We report our retrospective review of these patients and describe treatment of this difficult and sometimes unrecognized syndrome, primarily with intravenous dihydroergotamine (DHE). Results at hospital discharge and at follow-up several months later showed significant reduction of pain.
Adult↗
Determination of picotamide in human plasma and urine by high-performance liquid chromatography.
A high-performance liquid chromatographic method for the determination of picotamide in human plasma and urine is described. After addition of an internal standard (bamifylline), the plasma and urine samples were subjected to liquid-liquid extraction and clean-up procedures. The final extracts were evaporated to dryness and the resulting residues were reconstituted in 100 microliters of methanol-water (50:50, v/v) and chromatographed on a LiChrosorb RP-SELECT B reversed-phase column coupled to an ultraviolet detector monitored at 230 nm. Chromatographic analysis takes about 10 min per sample. The assay was linear over a wide range and has a limit of detection of 0.005 and 0.1 micrograms/ml in plasma and urine, respectively. It was selective for picotamide, accurate and robust and thus suitable for routine assays after therapeutic doses of picotamide.
Chromatography, High Pressure Liquid↗