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Biomedical subjects

P Dupuy

Publications and source records attributed to P Dupuy.

At least 19 recordsLinked to original sources

Evidence that human kidney produces a protein similar to lithostathine, the pancreatic inhibitor of CaCO3 crystal growth.

Pancreatic juice is supersaturated in calcium carbonate. CaCO3 crystal growth is controlled by lithostathine, a secretory protein synthesized by pancreatic acinar cells, first described as a constituent of pancreatic stones. It was recently reported that, in the thin descending limb of the Henle's loop, urine was supersaturated in CaCO3 (Coe FL, Parks JH: Defenses of an unstable compromise: crystallization inhibitors and the kidney's role in mineral regulation. Kidney Int. 1990: 38, 625-631. This observation suggested the presence in kidney of a similar inhibitor. In this study, we show that a protein immunologically related to lithostathine is actually present in urine of healthy subjects and in renal stones. Immunocytochemistry of kidney sections localized the protein to cells of the proximal tubules and thick ascending limbs of the Henle's loops. Protein extracts of renal stones inhibited CaCO3 crystal growth in vitro and this inhibition was significantly lifted by incubating the extracts with antibodies to lithostathine. The protein is not immunologically related to nephrocalcin. Because of its structural and functional similarities with pancreatic lithostathine, it was called renal lithostathine.

Calcium Carbonate

[The metastatic liver: a rare cause of hepatocellular insufficiency].

Liver metastases are frequent, often large and sometimes very late in skin or choroid metastatic melanoma. Hepatic failure is a rarely described complication in metastatic liver melanoma. We report a case of liver failure in a metastatic liver melanoma in a woman 76 years old, treated 16 years before for a choroid melanoma. About this case, we discuss the frequency, the location of primitive tumors and mechanism of liver failure in metastatic liver.

Aged

Preliminary treatment of urinary proteins improves electrophoretic analysis and immunodetection.

Analysis of urinary protein composition is an important tool in studies on renal physiology and physiopathology. Urine is, however, a complex mixture containing, besides protein, a variety of compounds such as salts, peptides, oligosaccharides, and glycosaminoglycans. Some of these compounds interfere with the electrophoretic migration of protein in sodium dodecyl sulfate-polyacrylamide gels and prevent correct analysis of the protein pattern. We describe a simple method for extracting urinary proteins that considerably improves their electrophoretic migration and subsequent immunodetection. This treatment involves ammonium sulfate fractionations (for precipitating proteins), EDTA (for inhibiting protein aggregation), and HCl hydrolysis (for removing glycosylaminoglycans). Recovery during extraction was found to be almost quantitative for total protein and three representative proteins: albumin, alpha 1-glycoprotein acid, and beta 2-microglobulin.

Albuminuria

Change in bile salt dependent lipase in human breast milk during extended lactation.

Two hundred eighty-one milk samples collected from Zaïrian nonprivileged, undernourished mothers, in series of nine groups from 1 month to 18 months after parturition, and 66 milk samples collected from French privileged mothers in series of four groups from 2 days to 16 months postpartum, were analyzed for their lactose, lipid and protein contents. In addition, the activity of bile salt-dependent lipase (esterase), which may play an important role in the newborn infant's lipids digestion, was measured. After the first month postpartum, independent of the nutritional state of the mother, sugar and protein concentrations were identical. Lipid content of French mothers' milk was lower in transitional milk, but appeared constant in mature milk with an average value of 29.1 +/- 5.8 mg/mL of milk. In Zaïrian mothers' milk, the lipid content of mature milk plateaued at around 50-55 mg/mL independent of the stage of lactation. Bile salt-dependent lipase showed constant esterase activity within the lactation stage in privileged mothers' milk, but decreased by almost 80-90% during the first four months of lactation in undernourished mothers. The data suggest that milk from nonprivileged mothers may lose some of its ability to hydrolyze milk lipid esters, which could also be of consequence to the infant's normal growth in view of its effect on the esters of the lipid-soluble vitamins A, E and D.

Adult

Cyclosporin A inhibits the antigen-presenting functions of freshly isolated human Langerhans cells in vitro.

Cyclosporin A (CsA) is a strong inhibitor of skin allograft rejection. It has been also reported to act, not only on helper T cells, but also on the antigen-presenting functions of mouse epidermal cells (EC) enriched in Langerhans cells (LC). We tested the effects of CsA on the human allogeneic mixed epidermal cell-lymphocyte reaction (MECLR) using whole EC and freshly isolated LC as stimulator cells. Results were as follows. 1) CsA inhibited the lymphocyte proliferative response in a dose-dependent fashion, by about 80% for a CsA concentration of 10(-7) M. To evaluate the effects of the drug on the two cell populations involved in MECLR, stimulator EC and responder PBMC were separately pulsed for 2 h with CsA, washed, and combined to form MECLR. Inhibition by CsA of the alloantigen-dependent lymphocyte proliferation appeared to be multifactorial, because CsA-pulsed EC and CsA-pulsed effector PBMC led to identical reductions of 40% each in proliferation. 2) The nature of EC sensitivity to CsA during MECLR was then analyzed after freshly separating highly purified CD1-positive LC (greater than 95%) and LC-depleted EC (mainly keratinocytes), using an immunomagnetic particle technique. When responder PBMC were cultured with CsA-pulsed LC, a highly significant reduction of lymphocyte proliferation was observed, indicating that CsA has direct effects on LC. 3) Some of the possible mechanisms by which CsA might act on LC were studied. Substantial IL-1 activity and PGE2 amounts were induced during MECLR by LC and keratinocytes, but CsA did not act via these factors. Neither did it significantly modify HLA-DR, DQ, or DP antigen expressions on EC. In conclusion, CsA directly inhibits antigen presentation by human LC. This inhibition may partly explain the beneficial effects of CsA on skin allografts and certain cutaneous immune reactions.

Adolescent

Changes in pancreatic exocrine secretion with age: pancreatic exocrine secretion does decrease in the elderly.

Pancreatic exocrine secretion was estimated in 180 normal control patients, free of abdominal and pancreatic disease, aged from 16 to 83 years. Duodenal juice was collected in two 15-min fractions after a single intravenous injection of 1 U/kg secretin + 3 U/kg CCK. Volume, maximal concentration and output of bicarbonate, lipase, phospholipase and chymotrypsin were estimated as well as minimal concentration and output of chloride and calcium. Each parameter was plotted against age, either individually or after separation into two age groups. Volume linearly increased up to the 3rd decade, and thereafter linearly decreased. Bicarbonate secretion paralleled fluid secretion and also decreased after the 3rd decade. The changes in chloride and calcium concentrations were different: concentrations linearly increased after the 3rd decade. Calcium concentration linearly increased with age (p less than 0.02) while chloride output was unchanged. The three enzymes that were studied linearly decreased in concentration as well as in output with age from the 3rd decade (p less than 0.02). Protein secretion decreased before water and bicarbonate secretion. One can conclude that pancreatic secretion changes in humans with age. Aging alters pancreatic secretion, through a decrease in flow rate, bicarbonate and enzyme secretion while calcium concentration is enhanced. Although not requiring substitutive therapy in the whole population, individual cases of pancreatic exocrine insufficiency might be explained by aging, without malnutrition.

Adult

[Antropyloric lithiasic obstruction. A variant of Bouveret's syndrome].

We report about one case of cholecystoduodenal fistula complicated by antropyloric lithiasic obstruction, which was treated surgically with gastrotomy and extraction of the calculus, in an 82-year-old woman. This case represents an anatomic variant of Bouveret's syndrome, which is classically defined as a duodenal lithiasic obstruction. On the basis of this case, the authors discuss the diagnostic and possibly therapeutic merits of digestive endoscopy and define the main clinical, anatomical and evolutive characteristics of this unfrequent complication of biliary lithiasis.

Aged

Characterization of a monoclonal antibody against a mucin-type glycoprotein in human sweat.

We report the preparation and characterization of an IgG2 monoclonal antibody (MAb), HSMA, prepared against a human pooled sweat extract (HPSE). The major component of HPSE was a mucin-type molecule, as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) stained with periodic acid-Schiff (PAS) reagent. By immunoblotting, HSMA revealed a smear in the high molecular weight range, typical of mucins. In enzyme-linked immunosorbent assay (ELISA), HSMA failed to react with HPSE fractions isolated after anionic exchange gel chromatography. Similarly, radio-immunobinding assays demonstrated no reactivity between HSMA and A, B, H, and Lewis blood group-related structures. The immunohistological labeling on normal skin showed that HSMA reacted with the cells of eccrine sweat glands, and to a lesser extent, with sebaceous glands and epidermal cells. Periodate treatment in situ abolished these reactions, thus suggesting the carbohydrate structure of the HSMA-epitope. In indirect immunofluorescence (IF) studies, HSMA also reacted with other exocrine glands, e.g. mammary glands, sublingual glands, mixed sero-mucous glands of the trachea, and in the pancreas. Sparse positive cells were also observed in the testis, kidney, thyroid and digestive tract.

Animals

T-cell receptor-gamma/delta bearing lymphocytes in normal and inflammatory human skin.

Murine dendritic epidermal T cells (DETC) were recently reported to express T-cell receptor (TCR)-gamma/delta chains. In a search for the human equivalent of these cells, we tested normal and lesional skin with MoAb which react with the TCR-gamma/delta heterodimer. We performed indirect immunofluorescence (IF) on epidermal sheets, and alkaline-phosphatase-anti-alkaline-phosphatase complex (APAAP) on epidermal cell smears. Frozen skin sections from normal skin and various cutaneous lymphocyte infiltrates were also studied. A few CD3+ T lymphocytes were consistently found in normal epidermis. Most of these cells appeared to be TCR-alpha/beta +, and some CD4+ or CD8+. On epidermal sheets and cell smears, only a very small TCR-gamma/delta + cell population was visualized (less than 0.1% of the total). On normal skin sections, we observed 0 to 3 gamma/delta + cells per section. When present, they were often located in the epidermal basal layer, and were round or dendritic. Double immunolabeling revealed that gamma/delta + cells differed from CD1+ Langerhans cells, and that they had a similar phenotypic pattern as gamma/delta + peripheral lymphocytes (PBL): CD2+, CD3+, CD4-, and CD8-. Immunostaining from various inflammatory skin lesions showed that the dermal infiltrates included CD3+ T lymphocytes but virtually no gamma/delta + cells. Only a few gamma/delta + cells were found in some end-evolutive infiltrates. Taken together, these results strongly suggest that normal human epidermis occasionally harbors TCR-gamma/delta complex bearing lymphocytes, which constitute a small fraction of the CD3+ cutaneous T lymphocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal

[Postoperative enoximone in coronary surgery. Systemic and coronary hemodynamics and regional systolic function].

The effects of Enoximone (50 mg in 20 minutes) on systemic, pulmonary and coronary haemodynamics and on regional systolic function were examined after surgical myocardial revascularisation in 5 patients without overt cardiac failure. The flow in mammary artery grafts and the study of regional systolic function were assessed by Doppler probes implanted during the operation. Enoximone increased Qc and Qco without changing blood pressure or cardiac output, due to arterial vasodilatation and inotropic stimulation. Pulmonary vasodilatation resulted in hypoxemia which must be prevented. The increase in VO2 was unrelated to oxygen transport suggesting a specific drug related effect.

Cardiotonic Agents

Portal and hepatic arterial blood flow measurements of human transplanted liver by implanted Doppler probes: interest for early complications and nutrition.

Hepatic artery thrombosis and early acute rejection are severe complications of orthotopic liver transplantation (OLT). Their rapid detection is most desirable. The purpose of this study was to assess the usefulness of monitoring hepatic artery (HABF) and portal vein (PVBF) blood flows during the first week after OLT. At the end of operation, microprobes were sutured to the vessels, and their connecting tubes were externalized and connected to a pulsed Doppler flowmeter operating at 8 MHz. In 10 patients (ages ranged from 2 to 54 years) of 106, measurements of HABF and PVBF were done during alternative clamping of both vessels and before and after abdominal closure, every 12 hours during 7 days, and at day 7, before and after a 150 gm carbohydrate meal. At day 7 the probes were pulled out by gentle traction without complication, and all patients were allowed to go home. Reciprocal increase of flow during selective clamping was only observed for HABF (+45.8% +/- 47.6%; p less than 0.01). Abdominal closure decreased both HABF and PVBF by 13.8%, p less than 0.01, and 26%, p less than 0.05, respectively. In seven cases no significant variation of HABF and PVBF was observed during 7 days. In two patients with histologically confirmed early acute rejection, a marked decrease of diastolic HABF, without modification in PVBF, was the first manifestation and was rapidly corrected by boluses of steroids. In one patient disappearance of systolic and diastolic HABF led us to diagnose an arterial obliteration caused by a plicature, which was successfully surgically treated in the emergency department. In all patients, after oral ingestion of the carbohydrate meal, and only after this type of diet, a significant and deep decrease (-87%, p less than 0.001) of HABF was observed between 7 and 120 minutes without any change in PVBF. Such an effect was not observed in control patients. We conclude that this Doppler flowmetric technique with implantable microprobes is useful for rapid diagnosis of and strategy in treating early complications and is a new tool for pathophysiologic study of OLT consequences.

Dietary Carbohydrates

Synthetic retinoids inhibit the antigen presenting properties of epidermal cells in vitro.

The clinical efficacy of retinoids in benign and malignant skin diseases involving immune mechanisms suggests that they affect the immunologic functions of the epidermis. However, these effects have yet to be demonstrated. The action of vitamin A (retinol) and the synthetic retinoids, isotretinoin, etretinate, acitretin, and arotinoid-free acid have been studied on the lymphocyte proliferation induced by phytohemagglutinin (PHA), by the mixed lymphocyte reaction (MLR), and the mixed epidermal cell-lymphocyte reaction (MECLR). The results for PHA-induced proliferations were highly variable for all the retinoids. However, in MECLR, the synthetic retinoids consistently reduced the proliferation by 20%-30%. This occurred at therapeutic drug concentrations of about 10(-7)M. In MLR, a minor decrease of 10%-15% was only found for higher concentrations (10(-5)M). Retinol induced no effect in either reaction. Further analysis of acitretin on MECLR showed that it reduced lymphocyte proliferation in a dose-dependent fashion. This reduction was combined with a decrease in cytotoxic T-lymphocyte induction (CTL). Addition of 10(-6)M acitretin at various times also revealed that its presence at cell culture initiation was necessary to inhibit proliferation significantly. Furthermore, cell treatments prior to MECLR showed that exposure of epidermal cells to acitretin was essential to produce this inhibition, suggesting that it acts directly on epidermal cells. Consequently, it is suggested that the specific inhibitory effect of synthetic retinoids on lymphocyte activation in MECLR may partly account for their therapeutic action on the skin.

Acitretin

Pancreatic stone protein: quantification in pancreatic juice by enzyme-linked immunosorbent assay and comparison with other methods.

To quantitate pancreatic stone protein (PSP), a competitive radioimmunoassay using monoclonal antibodies to PSP extracted from pancreatic stones and a sandwich enzyme-linked immunosorbent assay (ELISA) using monospecific polyclonal antibodies to the secretory forms of PSP (PSP S) were established. When PSP concentrations were measured in pancreatic juice by radioimmunoassay, no difference could be found between patients suffering from chronic calcifying pancreatitis and other diagnostic groups. Yet, with the ELISA technique involving polyclonal antibodies, decreased concentrations were found in chronic calcifying pancreatitis patients when compared to controls (p less than 0.001), chronic alcoholics without pancreatic symptoms, or obstructive pancreatitis patients. These discrepancies are discussed. The monoclonal antibodies recognizing the C-terminal part of PSS S (PSP S1), results from the radioimmunoassay indicate that the concentration of that polypeptide is identical in the juice of controls and patients. Results from the ELISA obtained with polyclonal antibodies raised against PSP S2-5 molecules, i.e., recognizing the PSP S1 part and the N-terminal portion of the molecule, indicate that the differences observed reflect differences in the juice concentration of that N-terminal peptide.

Antibodies