Biomedical subjects
Publications and source records attributed to A Ferraro.
The case of a patient who developed catatonia one week following Lysergic Acid Diethylamide (LSD) ingestion is presented. The psychosis developed two days after the intake. The catatonic syndrome resolved dramatically following one treatment of electroconvulsive therapy (ECT). This is perhaps the first case report of catatonia following the use of LSD. The need for a diagnostic category of organic catatonia is highlighted.
In 551 patients with osteosarcoma of the extremities treated between 1980 and 1991 in our Institution with surgery only (35 cases), surgery combined with adjuvant chemotherapy (147 cases) or neoadjuvant chemotherapy (369 cases) the relapse patterns were analyzed. Adjuvant chemotherapy was performed according to 2 different protocols and neoadjuvant chemotherapy according to 3 different protocols successively activated. In the 252 patients who relapsed, the interval between initial treatment and first relapse was significantly longer in the group treated with adjuvant and neoadjuvant chemotherapy (18.1 and 21.3 mo) than in the group treated with surgery only (5.4 mo). For patients treated with neoadjuvant chemotherapy, a longer interval was seen in the most effective regimen of neoadjuvant chemotherapy (25 mo). No significant differences were seen among the 3 groups, according to the site of first metastasis, although in patients treated with the most effective neoadjuvant regimen there was a higher incidence of bone metastasis. In patients who relapsed with pulmonary metastases the average number of nodules seen by standard X-rays, as well as CT scans, was significantly higher in patients treated with surgery only (3.6) than in patients treated with adjuvant or neoadjuvant chemotherapy (2.5 and 2.6 nodules). We conclude that these changes in metastatic pattern in patients treated with adjuvant and neoadjuvant chemotherapy are important, because they may encourage the use of salvage therapy with thoracotomy in a larger number of patients. Prolongation of time relapsed after more effective regimens of adjuvant and neoadjuvant chemotherapy should be considered when evaluating the preliminary results of new chemotherapy protocols.
To detect the interactions of DNA with the nuclear matrix proteins, DNA-protein cross-linkages were induced in intact nuclei from chicken liver by the use of cis-diammine dichloroplatinum. Methods have been devised for fast purification both of the proteins and of the DNA fragments involved in the cross-linked complexes. By Southern-Western blotting a number of matrix proteins isolated from the complexes have been shown to recognize specifically DNA sequences present in the cross-linked DNA fragments. This experimental approach not only allows to identify the nuclear matrix-DNA interactions existing in the nucleus before its disruption, but also provides a preparation of matrix proteins enriched in those species which are involved in such interactions and which can therefore be detected with high sensitivity.
DNA in cell nuclei is organized in large loops, which are formed by the binding of DNA to proteins present in a nuclear structure called matrix or scaffold. There is ample evidence that these interactions have not only a structural role, but also a functional one. Studies of these interactions have so far been carried out mainly in vitro. Their relevance, therefore, for the in vivo situation is not proven. We have analyzed the DNA-protein interactions directly in the intact nucleus by means of cross-linking reactions. Cross-linking by UV irradiation and by cis-diamine dichloro platinum (II) have been performed on nuclei from chicken liver and compared. The platinum complex has been found to be more efficient. The proteins complexed to DNA have been isolated and analyzed, and have been found to be enriched in species present in the nuclear scaffold, independently from its method of preparation.
RNA polymerase II purified from wheat germ has been treated with a series of cleavable bifunctional reagents and the resulting crosslinked products have been analyzed by diagonal electrophoresis. The results indicate that the three largest subunits (220, 140, and 42,40 kDa, respectively) form a core around which the smaller subunits are bound. The 220- and 140-kDa subunits can be also crosslinked together in the absence of bifunctional reagents by disulfide bond(s) formation. The 27-, 16.3- and 16-kDa subunits appear to be close to the largest subunit (220 kDa). The 21-kDa subunit is close to the 27- and 25-kDa subunits. The reaction of monofunctional reagents with the enzyme shows that the 42,40-kDa subunit is partially hidden in the interior of the protein molecule. On the basis of these results a model of the quaternary structure of the enzyme is proposed.
The nuclear matrix from chicken liver cells contains a small amount of glycoproteins recognized by Concanavalin A. These proteins are present not only in the peripheral matrix, but also in the internal one. In this latter localization many glycoprotein species appear, by cross-linking experiments, to be placed in the proximity of DNA. The effect of a partial enzymatic deglycosylation of matrix preparations suggests that these proteins contribute to the stabilization of the native matrix structure.
Human immunodeficiency virus type 1 (HIV-1) can infect various cell lines in culture and be maintained in a chronic state of restricted replication. These states of proviral latency are characterized by a predominance of spliced compared to unspliced viral RNA species. The proximate molecular mechanisms leading to restricted HIV-1 replication may differ in various cell lines. Importantly, recent studies have demonstrated that the site of integration is the critical parameter leading to proviral latency in ACH-2 cells. Utilizing murine retroviral shuttle vectors, the HIV-1 Tat protein was demonstrated to dramatically increase HIV-1 expression in the restrictively infected U1 monocytic cell line but not in the ACH-2 T-lymphocytic line. The HIV-1 Rev protein only modestly increased viral expression in both of these cell types. Thus, these data support the hypothesis that the mechanisms which initiate and/or maintain restricted HIV-1 expression may differ in various cell types in cell culture, and possibly in vivo.
The presence of glycoproteins and sugar-binding sites in the nucleus is well ascertained. In order to verify the existence of specific nuclear protein-carbohydrate interactions, oligosaccharides were released from nuclear or non-nuclear glycoproteins by Peptide-N-glycosidase F, labeled by reduction with NaB3H4 and added to whole chicken liver nuclei and to subnuclear insoluble fractions (nucleoli, nuclear matrix and nuclear envelope). The analysis of the binding of the oligosaccharides to the nuclear fractions, performed in the presence or absence of competitor sugars, suggest that some rare oligosaccharides species, present only among the nuclear carbohydrates, are specifically recognized by the nuclear matrix.
Proteins of the internal nuclear matrix from chicken liver were fractionated, by chromatographic procedures, in non denaturing conditions. At least two fractions were present with phosphatidylinositol-specific phospholipase C and three with thiol:protein-disulfide oxidoreductase activity. A 57kDa protein was isolated which copurified with both these activities. Partial amino acid sequences showed a high degree of homology with a cytosolic protein previously identified as a phospholipase C and with a microsomal protein identified as a thiol:protein-disulfide oxidoreductase. Our finding leaves the question still unanswered of the real function of this protein, which for the first time has been isolated from the nuclear matrix.
BACKGROUND: It is known that many patients with osteosarcoma have high serum alkaline phosphatase (SAP) levels. The prognostic significance of this finding, however, is still controversial. METHODS: The pretreatment SAP levels of 656 patients with osteosarcoma of the extremities (107 metastatic and 549 localized at presentation) were examined to evaluate whether the enzyme levels had a clinical value in predicting the course of the disease. RESULTS: The percentage of patients with increased SAP levels was significantly higher in the metastatic group than in the group of patients with localized disease (91.5% versus 61.3%; P < 0.001). In the latter group, treated with adjuvant and neoadjuvant chemotherapy, the relapse rate was significantly higher in patients with elevated pretreatment SAP levels than in those with normal levels (55.1% versus 26.4%; P < 0.001). Among patients with elevated SAP levels, the percentage of relapses was higher in patients with high levels of the enzyme in comparison with patients with moderately elevated values (66.4% versus 47%; P < 0.05). After treatment, SAP levels almost always return to normal values, and a correlation between postoperative levels of the enzyme and prognosis could not be made. CONCLUSIONS: These data demonstrate that in osteosarcoma of the extremities, pretreatment SAP levels have a prognostic value and they should be considered when comparing the results achieved with different therapeutic protocols and in planning new randomized clinical trials.
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Between January 1988 and December 1990, 74 patients with localized Ewing's sarcoma of bone were treated with a new protocol that consisted of an initial 6-week period of chemotherapy with vincristine (VCR), adriamycin (ADM) and cyclophosphamide (EDX) followed by local therapy and additional chemotherapy with the same drugs previously indicated plus ifosfamide and VP-16. The rationale for the addition of ifosfamide and VP-16 to the four drugs of the standard chemotherapy of this tumor was that this drug combination was previously very effective in the treatment of metastases from Ewing's sarcoma even in patients who did not respond to cyclophosphamide. As local treatment all patients were offered surgery, when feasible (70 cases). Forty-three patients accepted and 27 refused. These patients, as the 4 patients in whom surgery was not considered feasible, were treated with radiation therapy alone (50-60 Gy). In the remaining patients amputation was performed in 4 cases, rotationplasty in 3 and resection in 36. Where conservative surgery was marginal or intralesional (30 cases), radiotherapy at lower doses (40-45 Gy) was also delivered. At a mean follow-up of 3.5 years (2-7), 43 patients (58%) remained continuously disease-free and 31 relapsed (29 with metastases and 2 with both metastases and local recurrences). These results do not differ from those obtained at our Institution in 98 patients treated between 1983 and 1988 with a neoadjuvant protocol in which only VCR, ADM, EDX and dactinomycin (DAC) were used (3-year continuously disease-free survival (CDFS) respectively of 54% and 55%). Despite the fact that these results came from a nonrandomized study, the Authors conclude that the addition of ifosfamide and VP-16 to the four-drug standard regimen did not improve the outcome of the patients with Ewing's sarcoma of bone which remains a lethal disease in about 50% of the cases. These findings stress the need to find more effective chemotherapeutic regimens for the associated treatment of this tumor.
It has been observed that double stranded DNA in solution is thermally destabilized by the presence of several organic molecules, among which are alcohols and polyols. This finding, and the fact that sugar moieties are components of some nuclear proteins and DNA binding drugs, may suggest a role for oligosaccharides in DNA recognition and/or interaction. In order to investigate this possibility, the effect of several sugars and other polyols on the thermal stability and on the conformation of DNA has been studied by high sensitivity differential scanning calorimetry and circular dichroism. While addition of small size polyols does not influence appreciably the melting enthalpy, it always results in a decrease in the DNA melting temperature. The magnitude of the destabilizing effect depends on the concentration of additives, reaching 13.5 degrees C in 50% (5.4M) glycerol. It also depends on the nature of the additives, e.g., sorbitol is more efficient than inositol, while dextran has no effect on the DNA melting temperature. According to circular dichroism results, DNA undergoes a significant structural change in the presence of small sugars or polyols: a continuous loss in 'B' character is observed as the glycerol concentration is gradually increased. This conformation change appears to be related to the decrease in thermostability.
In the last few years increasing attention has been given to investigate the role of large arteries in the pathophysiology of arterial hypertension and cardiovascular diseases. Thanks to their elastic properties, these vessels convert pulsatile flow in continuous flow at capillary level, thus significantly affecting the tissue perfusion. Various factors can modify the elastic properties of large arteries (arterial-wall structure alteration, vasoactive substances, elevated transmural pressure) and thus have positive or negative effect on cardiac function, vascular damage and systemic circulation. New ultrasonic techniques allow one to evaluate these vascular properties and to detect early changes in vessel structure and function. These techniques appear to be a promising tool to provide early markers of hypertensive vascular disease and of vascular risk in general, as well as to follow the vascular effects of treatment.
From September 1986 to December 1989, 144 patients with osteosarcoma of the extremities were treated with combined surgery and neoadjuvant chemotherapy. The disease-free survival was 79% for good responders (necrosis greater than 90%) and 72% for poor responders (necrosis less than 90%), and the local recurrence rate was low. Improvement in long-term prognosis and the increase of limb-sparing surgery determine a higher rate of immediate and late complications. Most of the complications were observed in limb-salvage procedures; 63% of these procedures presented one or more complications. In nine rotationsplasties, there were four complications, and in 13 amputations no complications were observed. Therefore, 55% of patients were affected by surgical complications. Twenty-eight complications were considered minor (not requiring surgery), whereas 77 complications were major. Functional results, evaluated according to Enneking's new system, were higher than 50% in two thirds of the limb-salvage procedures. Complications in limb-salvage procedures are more influenced by the type of reconstruction than by the surgical procedure used. Probably the most troublesome consequence of surgical complications in osteosarcoma is the deviation or delay in administering postoperative chemotherapy, which jeopardizes survival.
Investigation of structural features of native chromatin requires the use of intact nuclei, a turbid material which cannot be analyzed by optical methods. Differential scanning calorimetry does not require optically clear samples and has been proved by a number of authors to be a powerful tool in this field of study. By this technique, chicken erythrocyte nuclei were found to undergo at least four thermal transitions, centered at 59, 74, 88 and 98 degrees C. The highest temperature transition is strongly dependent on age and storage conditions of the nuclei. Adequate storage conditions overcame this problem and reproducible scans were obtained over a period of several months. This technical improvement has permitted the reconsideration of the occurrence of the fourth calorimetric transition, previously believed to be displayed only in replicating nuclei. Evidence gathered in the presence of perturbants and possible ligands allows the assignment of the four transitions to a nuclear protein scaffold, histones, nucleosomal DNA and a superstructured form of DNA. Moreover, it suggests that the higher-order structure is stabilized by fibronectin-like proteins.
In order to detect the nuclear matrix proteins involved in DNA binding, avoiding possible artifacts derived from the disruption of nuclei, proteins were crosslinked to DNA by the action of cis-diamminedichloroplatinum on intact chicken liver cells and analyzed by two-dimensional gel electrophoresis. At least eleven species of crosslinked proteins were found to derive from the nuclear matrix prepared from the same cell type, and five of these were found also among the proteins crosslinked to DNA in intact liver cells from ox and pig. This subset of common proteins, conserved in different animal species, is likely to have a fundamental role for the anchorage of DNA to the nuclear matrix.
We compared three methods of coronary stenosis (S) sizing in a multihospital PTCA restenosis trial: visual analysis by independent observers, single point caliper measurements, and quantitative coronary angiography (QCA). Cine films from 50 patients were submitted from the clinical site to the quantitative angiographic center, where visual analysis and computer quantitation were performed. Regression analysis revealed a correlation coefficient of .857 for caliper vs. QCA (p less than .0001) and .876 for visual observation vs. QCA (p less than .0001). No significant differences were found between any of the 3 methods for pre- or post-PTCA stenosis values. However, QCA yielded smaller PTCA changes in stenosis severity than either caliper or visual observation (p less than .05). Both caliper and visual methods correlated well with QCA in assessing stenosis size pre- and post-PTCA. Trained observers can visually assess lesion severity with accuracy approaching QCA. QCA is likely to be less expensive when compared to visual analysis by three experienced observers and should be the method used for estimating the results of PTCA in clinical trials.