Biomedical subjects

H Lepidi

Publications and source records attributed to H Lepidi.

45 records · Page 3Linked to original sources

PubMed1995

Expression of developmentally regulated cytoskeleton and cell surface proteins in childhood spinal muscular atrophies.

Expression of some developmentally regulated cytoskeleton components (desmin, vimentin and myosin heavy chain isoforms) and cell surface proteins (including neural cell adhesion molecule (NCAM), its polysialylated (PSA) isoform and CD24) have been studied by immunohistochemical detection in a series of 23 infantile spinal muscular atrophies (SMA). According to the clinical classification established by Byers and Banker in 1961, 8 cases were type I SMA (Werdnig-Hoffmann's disease), 10 cases were type II (intermediate form), and 5 cases were type III (Kugelberg-Welander's disease). In 15 cases, the percentage of immunoreactive fibers with the various antibodies used has been quantified and the results correlated with clinical data. The aim of the study was to search for variations in the pattern of expression of the proteins to improve the accuracy of diagnosis and prognosis, and to gain an understanding of the pathological processes involved in SMA. The results showed that the pattern of expression of these cytoskeleton and cell surface proteins is abnormal in all types of SMA. However, it was strikingly different in type I and II SMA as opposed to type III. In type I and II SMA, strong NCAM and developmental myosin heavy chain (MHC) expression was observed in atrophic fibers. Numerous atrophic fibers co-expressed desmin and vimentin as well as slow and fast adult MHC. Very few of them expressed PSA NCAM, fetal MHC and CD24. In type III SMA, the number of fibers expressing NCAM, developmental MHC and co-expressing slow and fast adult MHC was low and virtually none of them expressed vimentin or desmin. These findings are in favor of a denervation process occurring very early in life, probably even in utero, in type I and II SMA and leading to a severe impairment of muscle fibers maturation. In contrast, in type III SMA, the process is initiated well after birth and affects mature muscle fibers. In all types of SMA, the ability of muscle fibers to regenerate is low, although some fibers may be reinnervated. Immunohistochemical data was not related to the patients follow-up and thus has no prognostic value.

Adolescent↗

PubMed1995

Morphological polarization of human polymorphonuclear leucocytes in response to three different chemoattractants: an effector response independent of calcium rise and tyrosine kinases.

Chemoattractants such as interleukin-8, C5a and N-formylmethionyl-leucyl-phenylalanine induce a cytosolic calcium rise involved in triggering the secretory functions of human polymorphonuclear leucocytes. We studied the possible role of calcium rise in membrane ruffling, actin polymerization, filamentous actin distribution, and morphological polarization, which are all events contributing to chemotaxis. Membrane ruffling was assessed by right-angle light-scatter changes, the cellular content of polymerized actin by fluorescence of bodipy phallacidin, the intracellular distribution of filamentous actin by fluorescence microscopy and image digitization, and morphological polarization by scanning electron microscopy. Pretreatment of polymorphonuclear leucocytes with 50 microM BAPTA/AM, an intracellular calcium chelator, lowered the basal level in cell calcium and inhibited the transient calcium rise stimulated by 2 nM interleukin-8, 2 nM C5a, and 10 nM N-formylmethionyl-leucyl-phenylalanine. However, BAPTA pretreatment of polymorphonuclear leucocytes did not modify membrane ruffling, actin polymerization, filamentous actin distribution, and morphological polarization stimulated by chemoattractants. Downstream effectors may be protein tyrosine kinases. However, the tyrosine kinase inhibitor tyrphostin did not affect the cytoskeletal characteristics elicited by chemoattractants. Taken together, our results suggest that the transductional pathway leading to cytoskeleton organization and morphological polarization of polymorphonuclear leucocytes is different from that leading to secretion.

Actins↗

PubMed1993

F-actin content and spatial distribution in resting and chemoattractant-stimulated human polymorphonuclear leucocytes. Which role for intracellular free calcium?

Intracellular free calcium concentration ([Ca2+]i) plays a pivotal role for many responses in polymorphonuclear leucocytes (PMNs) stimulated by chemoattractants such as N-formylmethionyl-leucyl-phenylalanine (fMet-Leu-Phe). The importance of [Ca2+]i in the morphological polarization was investigated by using calcium-manipulated PMNs. We loaded human PMNs with BAPTA/AM to buffer or chelate [Ca2+]i in the presence or the absence of extracellular calcium by using fluo-3/AM as calcium indicator. The shape changes of PMNs were determined by microscopic examination, and membrane ruffling by right-angle light-scatter changes. Actin polymerization and F-actin distribution were recorded by staining PMNs with bodipy-phallacidin and quantified by quantitative fluorescence microscopy. We found that calcium-free incubation of PMNs loaded or not with 50 microM BAPTA/AM did not modify morphological polarization, membrane ruffling, actin assembly and F-actin distribution of PMNs stimulated with fMet-Leu-Phe, suggesting that these responses were probably functionally linked. It should be noted that incubation of PMNs in calcium-free conditions resulted in a radial distribution of F-actin and a moderate polymerization of actin, but not in morphological polarization of PMNs. Moreover, both calcium-sensitive and calcium-insensitive mechanisms of actin polymerization were additive, and inhibitable by 5 micrograms/ml cytochalasin B.

Actins↗

PubMed1993

Role of calcium in the shape control of human granulocytes.

The possible role of cytoplasmic calcium in the shape control of human blood granulocytes was explored with two complementary sets of experiments. First, cells were stimulated with N-formyl methionyl leucyl phenylalanine (FMLP) with or without depletion of free intracellular calcium (by incubation with BAPTA/AM in calcium-deprived medium). Control cells displayed a rapid and transient rise of free intracellular calcium (peaked after a few seconds, returned to the basal level after 2-3 minutes), extensive morphological polarization (more than 90% polarized cells after 15 minutes), and reorganization of actin filaments (as assessed by image analysis on cells labeled with a fluorescent phallacidin derivative). Calcium-depleted cells displayed no calcium rise, but both morphological polarization and actin reorganization were indistinguishable from those of controls. In a second set of experiments, individual granulocytes were labeled with fluorescent calcium probes and aspirated in a micropipette on the stage of a confocal laser microscope. About half of the cells displayed a transient calcium rise, which was concomitant with the formation of a protrusion in most cases. However, extensive protrusions were shown by the cells that did not exhibit calcium flashes. Furthermore, if cells were treated with ionomycin, in calcium-containing or calcium-deprived medium, then aspirated, they showed protrusions comparable to those of controls. Finally, when cells were treated with ionomycin during micropipette aspiration, a calcium rise was followed by a moderate increase of the rate of protrusion growth. It is concluded that free calcium alone cannot play a major role in the control of cell shape.

Actin Cytoskeleton↗

PubMed1993

[Gastric adenocarcinoma with an endocrine component. Report of nine cases].

Nine gastric adenocarcinomas containing endocrine cells were retrospectively studied to evaluate anatomo-clinical and immunohistochemical features, and the out-come of the patients. These gastric tumors were bulky with frequently lymph node metastasis. The immunohistochemical analysis had shown that all tumors contain EC cells, two tumors D cells, one tumor PP cells, but neither G cells nor A cells were detected. Five-years survival rate of these gastric tumors was slightly but not significantly increased as compared to usual adenocarcinomas (45.5% vs 36.5%). However, one can raise the question of possible control of local hormonal secretions on tumoral growth, particularly the anti-trophic part of the somatostatin.

Adenocarcinoma↗

PubMed1992

Double localization of F-actin in chemoattractant-stimulated polymorphonuclear leucocytes.

Uniform concentrations of chemoattractants such as formylpeptides induced a morphological polarization of human polymorphonuclear leucocytes (PMNs) and a concentration of F-actin at the cell front. They also induced a transient increase in filamentous actin (F-actin) which preceded the cell shape change. We combined fluorescence microscopy and image analysis to study the localization of F-actin, as revealed by a specific probe (bodipyTM phallacidin) in suspended PMNs stimulated by chemoattractants. F-actin exhibited remarkable concentration in focal points after a 30 s exposure to 10(-8) M formylmethionyl-leucyl-phenylalanine (fMet-Leu-Phe), although no shape change of PMNs was detectable. A 10-min incubation with formylpeptide (10(-6) to 10(-9) M) induced the morphological polarization of PMNs and the appearance of a principal focus of F-actin in the cell head region and a secondary focus in the cell posterior end. The distribution of F-actin-associated fluorescence in 2D images of polarized PMNs might be due to an actual concentration of F-actin in privileged areas, to a local concentration of plasma membrane drawing filamentous actin or to variations in the cell volume. Then, we studied the distribution of a cytoplasmic marker, fluorescein diacetate and a membrane probe, TMA-DPH, in unstimulated rounded PMNs and in spherical and morphologically polarized PMNs stimulated by formylpeptide. The distribution of neither of these probes was correlated with F-actin distribution, especially in rounded PMNs stimulated 30 s with 10(-8) M fMet-Leu-Phe, suggesting that F-actin was concentrated in two foci located in the cell head region and in the cell posterior end. In addition, zymosan-activated serum induced the morphological polarization of PMNs and the appearance of two foci of filamentous actin, demonstrating that binding of formylpeptide to its specific receptor was not required for F-actin reorganization. We conclude that the accumulation of F-actin probably resulted from local filament assembly and put forward the hypothesis that microfilament reorganization in two centres drives the morphological polarization of PMNs.

Actin Cytoskeleton↗

PubMed1991

Lymphadenopathic tumor exhibiting intermingled features of Kaposi's sarcoma, malignant lymphoma, and angiofollicular hyperplasia.

A 56-year-old man presented with an inguinal lymph node enlargement. Histologic study of the tumor revealed three intermingled pathologic lesions: a nodular small cell lymphoma, an angiofollicular hyperplasia of vasculohyaline type, and a vascular neoplasia closely resembling Kaposi's sarcoma. The patient was immunocompetent and denied any homosexual relationships, transfusions, or drug use. The serum was negative for the presence of human immunodeficiency virus antibody. Computed tomographic scan and ultrasound examination revealed no other lymphadenopathies. This case shows that both hyperplastic and neoplastic lymphoid proliferations can occur simultaneously with vascular neoplasia. It thereby suggests that the neoplastic populations might interact to favor the tumor growth, the sequence and the nature of the stimulating events remaining unclear.

Castleman Disease↗

PubMedDate not supplied

Protein tyrosine kinases and TNF alpha secretion in human monocytes.

The role of protein tyrosine kinases (PTK) in TNF alpha secretion by human monocytes was investigated in this report. We showed that an immunomodulator such as Nocardia lysozyme digest (NLD) and a particulate agonist, zymosan, stimulated an increase in tyrosine phosphorylation of several endogenous substrates including 53-56 kDa protein which was the predominant phosphoprotein. In addition, NLD and zymosan induced TNF alpha secretion which was impaired by a PTK inhibitor, tyrphostin. We suggest that a cascade of kinases including PTK is involved in NLD and zymosan signalling.

Humans↗

PubMedDate not supplied

Severe lower limbs lymphedema following breast carcinoma treatment revealing radiation-induced constrictive pericarditis--a case report.

In patients treated for breast carcinoma, unilateral lymphedema of the upper limb is usual. However, to the authors' knowledge, lower limb lymphedema has never been reported as a complication of breast carcinoma therapy. They report here the first case of a radiation-induced constrictive pericarditis revealed by severe lower limbs lymphedema. A 60-year-old woman was treated for left breast carcinoma with quadrantectomy, axillary lymphadenectomy, and combined radio chemotherapy (60 grays). Three and a half years later she suffered from a diffuse and increasing lower limbs lymphedema, which became huge and disabling. Radiation-induced constrictive pericarditis was evidenced by right cardiac cavities catheterization. A dramatic improvement was rapidly obtained after pericardectomy. Histopathologic analysis of the pericardium did not reveal neoplastic cells. Radiation-induced constrictive pericarditis is usually responsible for lower limbs edema, but lymphedema is exceptional. This case highlights the need to search for a constrictive pericarditis also in the case of lower limbs lymphedema, particularly in a patient treated with mediastinal radiotherapy or combined radio chemotherapy.

Antineoplastic Combined Chemotherapy Protocols↗