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Biomedical subjects

F Pinet

Publications and source records attributed to F Pinet.

At least 37 records · Page 2Linked to original sources

Immunocytochemical and biochemical evidence of renin in human lactotrophic cell cultures.

Primary cell cultures from human prolactin (PRL)-secreting adenomas were used to test the ability of human lactotrophs to synthesize renin in vitro. The renin content of the culture medium and of cellular extracts was measured by enzyme-linked immunosorbent assay. The level of PRL release in the culture medium and the amount of PRL in a cellular extract were determined by radioimmunoassay. Morphologic studies included indirect immunofluorescence, pre-embedding immunoelectron microscopy using a three-layer peroxidase-antiperoxidase method and postembedding immunoelectron microscopy using protein A-gold complexes. Renin was detected in cellular extracts and was found to be absent in the culture medium, whereas PRL was extracellularly secreted. PRL and renin immunoreactivity was observed in all the cultures studied by immunofluorescence. The subcellular localization of renin was found to be similar to that of PRL and was observed in the rough endoplasmic reticulum, the Golgi apparatus, and cytoplasmic secretory granules. The results suggest that, in vitro, renin may be synthesized and intracellularly metabolized in human adenomatous lactotrophic cells rather than secreted. Cell cultures may be a useful model to further the understanding of the role of a local renin-angiotensin system in PRL secretion.

Female

Renin and cathepsin B in human pituitary lactotroph cells. An ultrastructural study.

Renin, prorenin and cathepsin B were localized in human lactotrophs using immunoelectron microscopic techniques. Renin and prorenin were found in numerous cytoplasmic granules. Cathepsin B, a lysosomal enzyme known to be able to activate prorenin into renin, was also present in cytoplasmic granules of lactotrophs. The co-localization of renin and prolactin in the same secretory granules was demonstrated by double immunolabelling. Renin and cathepsin B were co-localized in some granules by the same technique. These results suggest a local activation of renin in the secretory granules of lactotrophs and support the hypothesis of a possible autocrine action of the renin-angiotensin system on prolactin release.

Adenoma

[Rapidly developing benign teratoma of the mediastinum in the adult].

Benign mediastinal teratomas are rare entities, representing 8 to 18% of all mediastinal tumors. Two cases of benign teratomas with a rapid growth suggesting malignancy are reported. CT and MR allowed precise characterization of the multi-tissue composition of the tumor (various CT densities were noted corresponding to adipose, cystic, calcified and hemorrhagic components), and definition of the extension of the process before surgical removal. The per-operative pathological analysis in one patient confirmed the benign nature of the lesion based on the absence of immature tissue, allowing complete surgical removal. The final pathological examination showing mucus secretion by bronchial glandular tissue and intra-tumoral hemorrhage may explain the rapid growth of this kind of lesion. The post-operative follow-up of these benign teratomas is favourable.

Adult

Isolation and characterization of renin-producing human chorionic cells in culture.

Chorionic tissue is one of the major extrarenal sites of renin production, and as such, cultured chorionic cells are a potential model for in vitro studies of renin biosynthesis and regulation. Human chorionic cells were isolated from four chorions and maintained in tissue culture for a total of eight subcultures. Total renin production was considerable in the primary cultures, but fell gradually with successive passages. The cells could be frozen and thawed without losing their ability to divide or produce renin. Both the primary cultures and the subcultures contained a single type of elongated cell containing abundant rough endoplasmic reticulum and myofibrils, but no renin granules, suggesting that the cells had smooth muscle-like features. Immunocytochemistry indicated that they contained both renin and prorenin. The renin produced by the chorionic cells was not stored within the cells, but was released rapidly into the medium. More than 95% of the renin produced was prorenin, which, after activation, had biochemical and immunological properties similar to those of pure human renin. The cells contained a renin mRNA that had the same size as that for renal renin (1.6 kilobases), confirming the synthesis of renin by these cells. The cells were also examined for the presence of other components of the renin-angiotensin system. Angiotensinogen and angiotensin I were not detected, but angiotensin-converting enzyme was present in extracts of primary and secondary cultured cells. beta hCG and progesterone were also found in the medium of primary culture. However, the production of beta hCG and progesterone fell after the primary culture, and beta hCG and progesterone were indetectable in secondary and tertiary cultures, respectively. These experiments suggest that these two hormones do not influence renin synthesis or vice versa. Thus, these cultures of human chorionic cells synthesized considerable quantities of prorenin and can provide a permanent source of nonrenal prorenin-producing cells.

Cell Separation

[New imaging technics in dissection of the thoracic aorta].

The multiplicity of imaging for investigation of the thoracic aorta requires that the physician choose a strategy adapted to the disorder involved: this is the case, for example with dissection of the aorta. A critical study of the new methods available which include digital angiography; echocardiography; CT scan; magnetic resonance imaging (MRI), should be compared to "the" reference technique of conventional methods: aortography and two-dimensional cineangiography. In addition, the decision must take into account specific characteristics of the disorder: natural history of the disease and anatomopathological form which are useful for the surgeon in case surgery is decided upon. The strategy proposed remains faithful to two-dimensional cineangiography during the first 72 hours of acute dissection because this disorder is a true surgical emergency at this stage if the ascending aorta is involved. Later, or if the ascending aorta does not appear to be involved, MRI has now proven this efficacy as a first-line diagnostic procedure. The judiciousness of preoperative aortography is still the subject of debate.

Aortic Dissection

[Digital arteriography of the lower limbs by sequential, automatic table translation. Feasibility. Preliminary results].

Digital subtraction angiography (DSA) as a method of evaluating lower limb arteries required examination of one limb at a time up to now. Moreover repeated injections of contrast material were necessary. This problem was mainly due to the relative small image intensifier field available and to the difficulty of repositioning the patient between mask and image acquisition. The authors report the preliminary results obtained with 24 patients studied on angiographic unit equipped with: A DSA system (DG 300-CGR) using 512 X 512 matrix size, an angiographic table (Angix M 200) allowing an automatic sequential translation, a removable image intensifier using a 16 inch field below the table. Thus, all those characteristics allow DSA of lower limb arteries. Only one injection of contrast material (Ioxaglate: 80 ml at 8-10 ml/second) was used per procedure. This, usually, allows good quality examination of lower limb arteries.

Angiography

Detection and localization of renin messenger RNA in human pathologic tissues using in situ hybridization.

In order to investigate the synthesis of renin in human pathologic tissues, the authors used in situ hybridization to detect and localize renin messenger RNA (mRNA). The probe was a 35S-radiolabeled 1.1-kb length complementary DNA of human renal renin. To compare the synthesis with the presence and the storage of renin, renin antigen was assessed by immunohistochemistry in the same tissues. The human pathologic tissues were as follows: two ischemic kidneys related to renovascular hypertension; two renal juxtaglomerular cell tumors; one extrarenal renin-secreting epithelioid sarcoma of soft tissues. In ischemic kidneys, the cells containing both renin mRNA and renin protein were found in numerous juxtaglomerular apparatus and in the wall of arterioles, shown by combined in situ hybridization and immunohistochemistry. Most of the tumor cells in the juxtaglomerular cell tumors and scarce tumor cells in the epithelioid sarcoma of soft tissues were positive by in situ hybridization and immunohistochemistry. These findings demonstrate that the presence of renin in these tissues is associated with local cellular production of renin. In particular, smooth muscle cells of the wall of arterioles are definitely capable of synthesizing renin. Moreover, in these tissues, gene expression (renin synthesis) and renin storage are concordant.

Adult

Embolization of the systemic arteries of the lung.

Embolization of the systemic arteries of the lung (described by Remy and colleagues in 1973) is now a useful method for the treatment of hemoptysis or hemorrhagic lesions of the lung prior to surgical treatment, or for local treatment of hemoptysis when surgery is contraindicated or unnecessary. The technique is based on the anatomy of the different divisions of the systemic circulation (bronchial and extrabronchial), which for various physiologic reasons may develop hypervascularization. The results, complications, and contraindications of systemic embolization have previously been described and the technique is now commonly practiced.

Embolization, Therapeutic

Renin secretion from malignant pulmonary metastatic tumour cells of vascular origin.

1. A pulmonary chemodectoma/glomangiosarcoma that had metastasized from the thigh was studied after removal from a 22 year old Algerian patient with hypertension, high plasma prorenin and signs of secondary aldosteronism. 2. Renin and renin mRNA were localized in sections of the tumour tissue using monoclonal anti-human renin antibody and human renin cDNA probe, respectively. 3. The cells grew prolifically in culture, but, even though their renin content was similar to that of transfected human juxtaglomerular cell tumour cells (approximately 1 pg/microgram DNA), their rate of secretion of renin was much lower (0.05-0.15 cf. 0.5-1.5 pg/h per microgram DNA). 4. Forskolin (10 mumol/l for 24 h) increased secretion of renin from 1.9 +/- 0.36 to 4.1 +/- 0.64 pg/ml per h of culture (P less than 0.001, n = 11), consistent with cAMP being a second messenger in the secretory mechanism. 5. The cells should provide valuable information about intracellular mechanisms for the regulation of renin synthesis and secretion.

Adolescent

Regulation of prorenin secretion in cultured human transfected juxtaglomerular cells.

The regulation of renin secretion was studied in continuous culture of human juxtaglomerular cells (JGC), which provided a permanent source of human renin production (Pinet, F., M. T. Corvol, F. Dench, J. Bourguignon, J. Feunteun, J. Ménard, and P. Corvol, 1985, Proc. Natl. Acad. Sci. USA, 82:8503-8507). 95% of the renin species secreted was prorenin, and therefore this study concerned primarily prorenin secretion. Renin production was stable, since the cells had been maintained in culture for more than two years. In culture, these human cells formed colonies of smooth musclelike cells, and electron microscopy showed the presence of cytoskeleton structures including myofibrils and attachment bodies. This human model was used to investigate the control of prorenin secretion in vitro at cellular level. Various pharmacological agents known to stimulate or inhibit renin secretion were tested in the cell cultures. The variations in prorenin secretion were measured in the supernatant. Forskolin, an independent receptor activator of adenylate cyclase, stimulated prorenin secretion in a dose-dependent manner and this stimulation was mediated by 3',5' cyclic-AMP (cAMP). Angiotensin II (AII) was found to inhibit prorenin secretion directly in a dose-dependent manner and atrial natriuretic factor (ANF), whose effects on human JGC were characterized for the first time, was also shown to exert such inhibition. When the effects of this inhibition by AII and ANF were tested on forskolin-mediated stimulation of prorenin secretion, the latter was inhibited and no change occurred in cAMP release. When JGC were treated with histamine, bradykinin, or one or two bradykinin analogues, the responses suggested that in these cells, H2-histamine receptors and kinin receptors are dependent on adenylate cyclase. One peptide, substance P, had an inhibitory effect on prorenin secretion but it was less important than AII and ANF. The present results demonstrate that the adenylate cyclase system of human JGC remains intact during culture and supports the hypothesis that cAMP is the second messenger and Cai2+, the final messenger involved in renin secretion. The cell system used here permits the evaluation of cellular responses and intracellular events in granulated cells in a human model.

Angiotensin II

Role of cyclic AMP in renin secretion by human transfected juxtaglomerular cells.

A human juxtaglomerular (JG) cell tumour was used for immortalization of renin secreting cells with three SV40 mutants. These transformed cells retained the same light and electron microscopic morphology as the human renal JG cells throughout subculture. Immunocytochemical staining showed the presence of renin in the elongated cells containing myofilaments and secretory granules. The renin produced was not stored within the cells but was released rapidly into the medium as prorenin. This permanent source of renin-producing cells was used for the study of renin regulation in vitro. Agents known to induce renin release such as dibutyryl cyclic AMP (cAMP), forskolin, isoproterenol and histamine were tested in cell culture. Forskolin induced renin secretion in a dose-dependent manner, as did dibutyryl cAMP. The JG cells responded to beta-agonists and to histamine by an H2 receptor. These results offer direct support for the hypothesis that cAMP is the second messenger in stimulation of renin secretion from human juxtaglomerular cells.

Bucladesine

[Radiology in renovascular hypertension from i.v. urography to digital subtraction angiography. A critical evaluation].

For the last 20 years, the value of radiology in renovascular hypertension (RVH) is well established from diagnostic and therapeutic points of view. However things are changing now due to a better appraisal of the incidence of RVH, new antihypertensive drugs, and new radiological procedures (digital angiography, percutaneous transluminal angioplasty). The approach now takes in account the mode of imaging for a better evaluation of the renal artery stenosis and costs of the procedures as well. Finally a new strategy has been set up for RVH patients management.

Angiography