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Biomedical subjects

E Marchetti

Publications and source records attributed to E Marchetti.

At least 37 records · Page 2Linked to original sources

Acylphosphatase: a potential modulator of heart sarcolemma Na+,K+ pump.

Acylphosphatase, purified from cardiac muscle, catalyzes the hydrolysis of the phosphorylated intermediate of heart sarcolemmal Na+,K(+)-ATPase. This effect was remarkable even using acylphosphatase amounts (100-300 units/mg of membrane protein) near the lower limit of the physiological range; besides the low value of the apparent Km, on the order of 10(-7) M, indicates that the enzyme has a high affinity for this special substrate. The results of a dot-immunobinding assay suggest the possibility of an interaction between acylphosphatase and native, undenaturated Na+,K(+)-ATPase. Moreover, when added to sarcolemmal vesicles, acylphosphatase was found to affect the functional properties of the Na+,K+ pump with regard to the rate of both ATP hydrolysis and cation transport. However, while ATPase activity and Na+ uptake were stimulated, the last at a greater extent, the active K+ transport was inhibited, so that the Na+/K+ ratio, which was calculated as 1.50 without acylphosphatase, rose to 6.68 in the presence of 300 units/mg of vesicle protein of this enzyme. Taken together, the reported results indicate that acylphosphatase, because of its hydrolytic activity on the phosphoenzyme intermediate, induces a sort of "uncoupling" effect on the heart sarcolemmal membrane Na+,K+ pump. Possible mechanisms for such an effect, which suggests a potential role of acylphosphatase in the control of this active transport system, are discussed.

Acid Anhydride Hydrolases↗

Transposable elements are stable structural components of Drosophila melanogaster heterochromatin.

We determined the distribution of 11 different transposable elements on Drosophila melanogaster mitotic chromosomes by using high-resolution fluorescent in situ hybridization (FISH) coupled with charge-coupled device camera analysis. Nine of these transposable elements (copia, gypsy, mdg-1, blood, Doc, I, F, G, and Bari-1) are preferentially clustered into one or more discrete heterochromatic regions in chromosomes of the Oregon-R laboratory stock. Moreover, FISH analysis of geographically distant strains revealed that the locations of these heterochromatic transposable element clusters are highly conserved. The P and hobo elements, which are likely to have invaded the D. melanogaster genome at the beginning of this century, are absent from Oregon-R heterochromatin but clearly exhibit heterochromatic clusters in certain natural populations. Together these data indicate that transposable elements are major structural components of Drosophila heterochromatin, and they change the current views on the role of transposable elements in host genome evolution.

Animals↗

Modulation of cellular chemoresistance in keratinocytes by activation of different oncogenes.

Response to chemotherapeutic agents in malignant tumors depends on many factors, most of which are as yet unknown. We investigated the correlation between the activation of different oncogenes and protein-kinase-C (PKC) modulation, and the cytotoxicity of some of the most widely used anti-cancer drugs. We transformed the murine keratinocyte cell line PAM 212, with different oncogenes (v-H-ras, v-myc and adenovirus E1a) and a mutant p53 suppressor gene (mp53). The cytotoxic effect of cisplatin (CDDP), doxorubicin (DOX) and vincristine (VCR), together with the concomitant action of modulators of PKC, TPA and staurosporine were evaluated by the crystal-violet method, thymidine incorporation and flow cytometry. We report that (a) the oncogene v-H-ras induces resistance to CDDP (> 50%), DOX (> 25%) and VCR (> 20%); (b) the E1a oncogene induces only resistance to VCR (> 40%) and marked sensitivity to CDDP and DOX; (c) the mp53 oncogene induces more resistance to VCR and insignificant resistance to the other drugs; and (d) activation of PKC by TPA increases the resistance to VCR and DOX in cells transformed by the v-H-ras, while it significantly increases the lethality with CDDP of the E1a-transformed cells. Staurosporine increases the cytoxicity of all the drugs, especially in the E1a-transformed keratinocytes. In the flow-cytometry analysis, the percentage of BUdR incorporation was related to sensitivity to anti-cancer drugs.

Animals↗

Influence of eye motion on adaptive modifications of the vestibulo-ocular reflex in the rat.

While sustained retinal slip is assumed to be the basic conditioning stimulus in adaptive modifications of the vestibulo-ocular reflex (VOR) gain, several observations suggest that eye motion-related signals might also be involved. We oscillated pigmented rats over periods of 20 min around the vertical axis, at 0.3 Hz and 20 degrees/s peak velocity, in different retinal slip and/or eye motion conditions in order to modify their VOR gain. The positions of both eyes were recorded by means of a phase-detection coil system with the head restrained. The main findings came from the comparison of two basic conditions--including their respective controls--in which one or both eyes were reversibly immobilised by threads sutured to the eyes. In the first condition the animals were rotated in the light with one eye immobilised and the other eye free to move but covered. Rotation in the light in this open-loop condition immediately elicited high-gain compensatory eye movements of the non-impeded, covered eye. At the end of this training procedure, the VOR gain increased by 43.2%. In the second condition, both eyes were immobilised and one eye was covered. The result was an increase in the VOR gain of 26.3%. These two conditions were similar as to the visuo-vestibular drive during the exposure, but different as to the resulting--and allowed--eye motion, showing that the condition where the larger eye movements occurred yielded the larger VOR gain change. Our data support the idea proposed by Collewijn and Grootendorst (1979, p. 779) and Collewijn (1981, p. 146) that "[retinal] slip and eye movements seem to be relevant signals for the adaptation of the rabbit's visuo-vestibular oculomotor reflexes".(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗

Modifications induced by acylphosphatase in the functional properties of heart sarcolemma Na+,K+ pump.

Acylphosphatase purified from cardiac muscle actively hydrolyzes the phosphoenzyme intermediate of heart sarcolemma Na+,K(+)-ATPase. This effect occurred with acylphosphatase amounts (up to 800 units/mg membrane protein) that fall within the physiological range and the low value of the apparent Km (0.69 x 10(-7) M) indicates a considerable affinity of the enzyme towards this specific substrate. Acylphosphatase addition to purified sarcolemmal vesicles significantly increased the rate of Na+,K(+)-dependent ATP hydrolysis. Maximal stimulation, observed with 800 units/mg protein, resulted in an ATPase activity which was about 2-fold over basal value. The same acylphosphatase amounts significantly stimulated, in a similar and to an even greater extent, the rate of ATP driven Na+ transport into sarcolemmal vesicles. These findings lead to suppose that an accelerated hydrolysis of the phosphoenzyme may result in an enhanced activity of heart sarcolemmal Na+,K+ pump, therefore suggesting a potential role of acylphosphatase in the control of this active transport system.

Acid Anhydride Hydrolases↗

Dermal fibroblasts tumor suppression of ras-transformed keratinocytes is associated with induction of squamous cell differentiation.

We have previously reported that tumor formation of ras-transformed keratinocytes can be suppressed by dermal fibroblasts through production of a diffusible growth inhibitory factor of the transforming growth factor-beta (TGF-beta) family. Keratinocytes transformed by ras and E1a oncogenes or papilloma-derived keratinocytes transformed by a ras oncogene show concomitant resistance to dermal fibroblast tumor suppression and TGF-beta growth inhibition. We report here that dermal fibroblast tumor suppression is associated with a striking induction of squamous cell differentiation and that this effect is blocked in tumors resistant to dermal fibroblast inhibition. This experimental system strongly supports the notion that suppression of tumorigenicity and induction of a differentiated phenotype are closely associated events.

Animals↗

Acylphosphatase induced modifications in the functional properties of erythrocyte membrane sodium pump.

Human red cell acylphosphatase actively hydrolyzes the Na+/K(+)-ATPase phosphoenzyme from erythrocyte membrane. This effect occurred with amounts of acylphosphatase (up to 10 units/mg membrane protein) within the physiological range, and the low value of the apparent Km (0.147 +/- 0.050 microM) indicates that the enzyme has a high affinity for this substrate. When added at the above concentration to inside out vesicles from human erythrocytes, acylphosphatase significantly enhanced the rate of strophantidine-sensitive ATP hydrolysis. The same amounts of acylphosphatase stimulated, although to a lower extent, the rate of ATP-dependent 22Na+ influx (normal efflux). Thus, the calculated stoichiometry for Na+/ATP was 2.68 in the absence of acylphosphatase and 1.06 in the presence of 10 units/mg vesicle protein of the enzyme. Conversely, acylphosphatase addition strongly decreased the rate of ATP-dependent 86Rb+(K+) efflux (normal influx) which, with 10 units/mg vesicle protein, was almost suppressed. As a consequence, the Na+/Rb+ ratio, calculated as 1.52 in the absence of acylphosphatase rose to 72.5 in the presence of 10 units/mg vesicle protein of this enzyme. These results suggest that, because of its hydrolytic activity on the phosphoenzyme intermediate, acylphosphatase 'uncouples' erythrocyte membrane Na+,K+ pump. Possible mechanisms for this effect are discussed.

Acid Anhydride Hydrolases↗

Clinical usefulness of estrogen receptor immunocytochemistry in human breast cancer.

We assessed the reliability of the immunocytochemical assay of estrogen receptor (ER-ICA) as a marker of clinical outcome. Relapse-free interval (RFI) and overall survival (OS) according to ER-ICA status were retrospectively evaluated on a series of 210 patients who had undergone surgery for primary breast cancer between January 1985 and December 1988. ER assay by the dextran-coated charcoal method (DCC) was also performed in 189 tumors. A significant positive correlation was found between the DCC and ER-ICA assays, with an overall agreement of 79%. ER-ICA status showed a prognostic predictive power with respect to OS and RFI in the whole series of patients and in the subset of node-positive patients. It was also a marker of outcome with respect to OS in the subsets of node-negative patients and patients with tumors < or = 2 cm in diameter. Moreover, the predictive value of the ER-ICA assay was higher than that of the DCC assay in the present study. These findings emphasize the clinical usefulness of the ER-ICA assay as a measure for prognosis.

Adult↗

Alterations induced by acylphosphatase in the activity of heart sarcolemma calcium pump.

We studied the effect of muscle acylphosphatase on the Ca2+ pumping ATPase of heart sarcolemma. Acylphosphatase addition to calmodulin-depleted sarcolemmal vesicles produced a significant increase in the rate of Ca(2+)-dependent ATP hydrolysis, even higher than obtained with exogenously added calmodulin. Maximal stimulation (about four fold over basal value) was obtained with 550 units/mg vesicle protein, a concentration that fall within the physiological range. Conversely, similar amounts of acylphosphatase decreased the rate of ATP-dependent Ca2+ transport into the sarcolemmal vesicles. The maximal statistically significant inhibition of Ca2+ uptake was observed with the same acylphosphatase concentration that gave the maximal stimulation of Ca(2+)-ATPase activity. From these findings acylphosphatase appears to reduce the efficiency of heart sarcolemmal Ca2+ pump with an impairment of the coupling between ATP hydrolysis and Ca2+ transport. A possible mechanism of this effect is discussed.

Acid Anhydride Hydrolases↗

Hydrolysis by acylphosphatase of erythrocyte membrane Na+, K(+)-ATPase phosphorylated intermediate.

Acylphosphatase, purified from human erythrocytes, actively hydrolyzes the phosphoenzyme intermediate of human red blood cell membrane Na+, K(+)-ATPase. This effect occurred with acylphosphatase amounts (up to 10 units/mg membrane protein) that fall within the physiological range. Acylphosphatase addition to erythrocyte membranes resulted in a significant increase in the rate of Na+, K(+)-dependent ATP hydrolysis. Maximal stimulation, observed with 10 units/mg membrane protein, was of about 80% over basal value. The same acylphosphatase amount enhanced of about 40% the rate of ATP driven Na+ transport into inside out red cell membrane vesicles. Taken together these findings suggest a potential role of acylphosphatase in the control of the activity of erythrocyte membrane Na,K pump.

Acid Anhydride Hydrolases↗

Immunohistochemical expression of c-erbB-2 in human breast cancer by monoclonal antibody: correlation with lymph node and ER status.

c-erbB-2 Protein expression was investigated in a series of fifty primary breast cancers by means of a specific monoclonal antibody and immunocytochemistry. Specific staining was observed at the plasma membrane level of neoplastic cells, according to the reported localization of c-erbB-2 protein. Sixty-four percent of tumors scored positive, with a variable amount of stained cells. The rate of protein expression was found to exceed the reported gene amplification. No relationship was observed between c-erbB-2 protein staining and age, menopausal status or histologic subtypes. An inverse association was found between c'erbB-2 protein staining and estrogen receptor content of tumors, assayed by immunocytochemistry. A positive relationship was observed between c-erbB-2 protein expression and presence of axillary node metastasis. These findings suggest that c-erbB-2 protein expression is a marker of tumor aggressiveness and that its prognostic power deserves further investigation both in node-positive and node-negative patients.

Adult↗

Assessment of proliferative rate of breast cancer by Ki-67 monoclonal antibody.

The proliferative activity of 163 primary breast cancers was assessed by immunocytochemistry with the mouse monoclonal antibody Ki-67, which recognizes a nuclear antigen expressed in all phases of the cell cycle except for Go. The overall frequency distribution of Ki-67 staining was of exponential type, with percentage of positive staining cells ranging from 0.3 to 88.3%, with a median value of 10.1%. No relationship was observed between Ki-67 values and menopausal status of patients. A significant positive correlation was found between Ki-67 values and tumor grade, especially mitotic grade. Estrogen Receptors (ER) were assayed by immunocytochemistry with ER-ICA method and by dextran-coated charcoal method (DCC) in 129 and 141 tumors, respectively. A negative correlation was found between the ER content by both methods and Ki-67 score. Eighty-nine percent of cases with Ki-67 value less than 10.1% contained more than 10% ER-ICA-positive cells. Progesterone receptors (PgR) were assayed by immunocytochemistry with PgR-ICA method and by DCC in 62 and 141 tumors, respectively. A negative correlation was observed between the PgR content by both methods and Ki-67 score. No correlation was found between Ki-67 score and lymph node involvement by tumor. These findings suggests that Ki-67 score could be used as an independent prognostic marker, useful to distinguish different risk for recurrence within the two clinically heterogeneous groups of N- and N+ patients.

Adult↗

Oculo-manual tracking of visual targets: control learning, coordination control and coordination model.

The processes which develop to coordinate eye and hand movements in response to motion of a visual target were studied in young children and adults. We have shown that functional maturation of the coordination control between eye and hand takes place as a result of training. We observed, in the trained child and in the adult, that when the hand is used either as a target or to track a visual target, the dynamic characteristics of the smooth pursuit system are markedly improved: the eye to target delay is decreased from 150 ms in eye alone tracking to 30 ms, and smooth pursuit maximum velocity is increased by 100%. Coordination signals between arm and eye motor systems may be responsible for smooth pursuit eye movements which occur during self-tracking of hand or finger in darkness. These signals may also account for the higher velocity smooth pursuit eye movements and the shortened tracking delay when the hand is used as a target, as well as for the synkinetic eye-arm motions observed at the early stage of oculo-manual tracking training in children. We propose a model to describe the interaction which develops between two systems involved in the execution of a common sensorimotor task. The model applies to the visuo-oculo-manual tracking system, but it may be generalized to other coordinated systems. According to our definition, coordination control results from the reciprocal transfer of sensory and motor information between two or more systems involved in the execution of single, goal-directed or conjugate actions. This control, originating in one or more highly specialized structures of the central nervous system, combines with the control processes normally operating in each system. Our model relies on two essential notions which describe the dynamic and static aspects of coordination control: timing and mutual coupling.

Adult↗

Expression of p21 ras gene products in breast cancer relates to histological types and to receptor and nodal status.

Expression of the p21 product of the ras gene family was investigated in a series of 142 infiltrating primary breast tumors by two specific ras p21 monoclonal antibodies with an immunocytochemical technique. The majority of tumors demonstrated a varying number of positive cells. A significant association between p21 expression and tumor histotype was found: among ductal carcinomas the comedo variety was always positive; conversely, lobular tumors were more frequently negative. Nodal status was recorded for all patients. A significant difference was found in nodal status with respect to p21 expression: tumors with more than 50% positive cells were more often N+. Estrogen receptor (ER) status was determined in 77 tumors. Tumors with higher levels of p21 contained a high percentage of estrogen receptor positive cells. The present results show that p21 expression in human breast cancer could be a marker of tumor aggressiveness and might thus improve the predictive power of known prognostic factors such as estrogen receptor and nodal status.

Breast Neoplasms↗

Commentary on human mammary preneoplasia. The estrogen receptor-promotion hypothesis.

In the carcinogenic process, promotion is the process whereby an initiated tissue develops focal proliferations which act as proximate precursors. The evidence obtained from the immunocytochemical staining by monoclonal anti-receptor antibodies indicates that the early steps (atypical hyperplasias) in the carcinogenic process of the breast show an increased and homogeneous expression of the estrogen receptor. These observations suggest that the persistent sensitivity to estrogen may be critical in sustaining the growth of mammary preneoplastic changes and their progression to ultimate precursors and to invasive cancer.

Breast Neoplasms↗

[Pituitary prolactin reserve in acute exacerbation of chronic respiratory insufficiency].

The level of pituitary prolactin reserve (PPR) was studied in 56 patients with chronic respiratory failure (CRF) in the acute stage after injections of metoclopramide and Thyrotropin Releasing Hormone (TRH). PPR was low in 90% of the subjects in both groups so that the pathogenic mechanism is more likely to be hypophyseal than hypothalamic and due to the reduced synthesis of PRL by the pituitary gland as a result of inadequate ATP synthesis and/or functional alterations to the receptors or pituitary cells following hypoxaemia and hypercapnia.

Acidosis, Respiratory↗