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Biomedical subjects

M Pascual

Publications and source records attributed to M Pascual.

At least 109 records · Page 6Linked to original sources
PubMed1996

Expression of NGF and NT3 mRNAs in hippocampal interneurons innervated by the GABAergic septohippocampal pathway.

We used in situ hybridization for the detection of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin 3 (NT3) mRNAs combined with immunocytochemistry against the calcium-binding proteins parvalbumin (PARV), calbindin 28k (CALB), and calretinin (CALR) to determine the expression of neurotrophins in functionally distinct subsets of hippocampal interneurons. Most PARV-immunoreactive neurons in the hippocampus were NGF mRNA-positive (82%), which corresponds to 71% of NGF-positive neurons in the hippocampus proper and in the dentate gyrus (excluding granule cells). In contrast, only a subset of CALB- and CALR-immunoreactive interneurons (24% and 23%, respectively) displayed hybridization signals for NGF. Small subsets of PARV- and CALR-positive cells expressed NT3 mRNA, but we did not find hippocampal interneurons expressing BDNF mRNA. These results show that NGF and NT3 genes are differentially regulated in distinct subsets of GABAergic cells, and these interneurons are a major source of NGF production in the hippocampus. We also addressed whether hippocampal interneurons expressing neurotrophins were targets of the GABAergic septohippocampal pathway. We developed a triple-labeling method that combines anterograde tracing of this pathway by means of Phaseolus vulgaris leucoagglutinin injections, with in situ hybridization for the detection of neurotrophins, and immunocytochemistry for calcium-binding proteins. Virtually every PARV-positive neuron innervated by GABAergic septohippocampal baskets expressed NGF mRNA (86%), whereas 39-59% of CALR- and CALB-positive interneurons that were contacted by GABAergic septohippocampal axons showed NGF gene expression. A small subset of NT3 mRNA-expressing interneurons was also innervated by septohippocampal baskets. These findings show that the GABAergic septohippocampal pathway preferentially terminates on interneurons expressing NGF mRNA, suggesting that this neurotrophic factor might be involved in the specification of this connection and in its maintenance and normal function in the adult brain.

Afferent Pathways↗
PubMed1996

New synthetic sulfated oligosaccharides prolong survival of cardiac xenografts by inhibiting release of heparan sulfate from endothelial cells.

Binding of recipient natural antibodies to the endothelium of the graft, complement activation, endothelial cell activation, and microvascular thrombosis are major events in the hyperacute rejection of organ xenografts. The aim of this study was to investigate the effects of two new synthetic sulfated oligosaccharides (A and B) on the survival of discordant cardiac xenografts in the guinea pig-to-rat model. In untreated recipients, hyperacute rejection occurred in 5 min (median; range, 4-6 min) and immunohistological analysis of all the grafts revealed deposition of IgM and C3 along the endothelium. Administration of oligosaccharides A and B prior to revascularization prolonged the survival of xenografts in a dose-dependent manner, up to 113 min (median; range, 42-145 min) and 86 min (median; range, 35-108 min), respectively, when doses of 20 mg/kg were used. There were no bleeding complications. Histological examination of the rejected grafts showed a picture of hyperacute rejection, with no difference in IgM and C3 deposition as compared with the untreated animals. In cell culture experiments, the release of heparan sulfate from guinea pig cardiac endothelial cells induced by rat serum was inhibited by both saccharides in a dose-dependent manner. The results indicate that these new synthetic sulfated oligosaccharides are effective for prolongation of discordant xenograft survival, possibly by interfering with endothelial cell activation. Such substances may be of value in other xenotransplant combinations.

Acute Disease↗
PubMed1996

Preliminary description of focal segmental glomerulosclerosis in patients with renovascular disease.

BACKGROUND: Primary and secondary forms of focal segmental glomerulosclerosis (FSGS) are common causes of glomerular proteinuria. Secondary forms of FSGS seem to be the result of adaptive changes that follow a reduction in renal mass. We saw an elderly patient with severe bilateral renal vascular disease (RVD) who had FSGS on percutaneous biopsy. To find out whether elderly patients with atherosclerotic RVD are predisposed to the development of FSGS, we reviewed all cases of FSGS at our institution between 1990 and 1995. METHODS: We identified 59 cases of biopsy-proven FSGS and examined clinical, histological, and radiographic records. FINDINGS: Of the 59 patients, 24 were older than 50 years; eight of these had RVD. No patient under the age of 50 had RVD. Seven of the eight patients with RVD and FSGS had substantial proteinuria at presentation. All had typical glomerular lesions with focal segmental tuft collapse and synechiae; other glomeruli were hypertrophic. All patients showed further decline in renal function on follow-up. INTERPRETATION: The association of FSGS and RVD may represent an under-recognised aetiology of significant proteinuria in elderly patients.

Aged↗
PubMed1996

Evaluation of recombinant human soluble dimeric tumor necrosis factor receptor for prevention of OKT3-associated acute clinical syndrome.

Tumor necrosis factor alpha (TNFa) has been shown to be the primary cytokine responsible for the OKT3-induced acute clinical syndrome (OKT3-ACS). Recombinant human soluble tumor necrosis factor receptor (TNFR:Fc) is a dimer of the p80 TNF receptor, which binds both TNFa and lymphotoxin (LT). Renal allograft recipients undergoing OKT3 therapy for steroid-resistant rejection were randomized to receive OKT3 alone or in combination with TNFR:Fc to determine its safety and efficacy in decreasing the severity of OKT3-ACS and in restoring renal function. Six of 12 patients were given TNFR:Fc prior to each of the first two injections of OKT3. All patients were monitored for manifestations of OKT3-ACS and changes in renal function. In addition, serial serum samples were assayed for TNFa and TNFR:Fc levels (ELISA) and TNFa bioactivity (L929). No adverse side effects were identified in patients receiving TNFR:Fc. Patients treated with TNFR:Fc had significantly fewer symptoms by day 2 of OKT3, and had a lower overall incidence of chills and arthralgias. Renal dysfunction reversed within 24 hr in the TNFR:Fc-treated group in contrast to the 48-72-hr delay in the control group. Antigenic TNFa levels increased in the control group from < 10 pg/ml pre OKT3 to a mean peak level of 30 +/- 13 pg/ml on day 1 and decreased to pretreatment levels by day 2. TNFR:Fc-treated patients had a mean peak TNFa level of 235 +/- 135 pg/ml, suggesting a carrier effect of TNFR:Fc. In contrast, bioactivity was barely detectable (mean 20 +/- 14 pg/ml) in the day 1 samples from TNFR:Fc-treated patients, whereas significant bioactivity (peak mean 60 +/- 35 pg/ml) was detected in sera from control patients. TNF receptor levels reached 600 ng/ml in treated patients and remained elevated for up to 18 days confirming the long half-life of TNFR:Fc. This phase 1 trial demonstrates that TNFR:Fc is well tolerated and may limit the severity of OKT3-ACS. The most significant observation was a more rapid improvement in renal function in the TNFR:Fc-treated patients. The absence of TNFa bioactivity indicates that TNFR:Fc functions as a TNF antagonist. Further evaluation of higher doses of TNFR:Fc in OKT3-treated patients is currently in progress.

Acute Disease↗
PubMed1996

Molecular basis of hereditary C1q deficiency associated with SLE and IgA nephropathy in a Turkish family.

Two siblings (case 1 and case 2) with homozygous C1q deficiency are described. Both presented with a photosensitive rash, and during follow-up case one developed SLE with nephrotic range proteinuria. Case 2 had microscopic hematuria with a past history of macroscopic hematuria. Renal biopsies revealed mesangioproliferative glomerulonephritis in case 1 and IgA nephropathy in case 2, a new finding in association with C1q deficiency. Since the classical pathway of complement plays a role in the development of antibody responses, the family was also evaluated for the immune response to hepatitis B vaccine. Antibody response to hepatitis B vaccine was normal in both affected members and the rest of the family. The A-, B- and C- chain genes of C1q were amplified by PCR and directly sequenced. A homozygous C to T point mutation was identified in genomic DNA isolated from the patients at codon 186 in the A chain that resulted in a premature stop codon. This mutation was present in both parents and both unaffected sibs in the heterozygous state. This mutation was identical to that previously described in a Slovakian family with C1q deficiency. Because of this finding, a series of 92 genomic DNA samples was screened from ethnically distinct patient groups with SLE to test the hypothesis that this mutation of C1q may be a widespread disease susceptibility gene. No further examples of this mutation were found.

Adolescent↗
PubMed1996

Complement depletion during haemofiltration with polyacrilonitrile membranes.

BACKGROUND: Polyacrylonitrile (PAN, AN69) dialysis membranes have been shown to improve the outcome of critically ill patients. Factor D is an essential enzyme of the alternative pathway of complement and is increased during renal failure. On the other hand the contact of blood with biomaterials activates the complement cascade through the alternative pathway. PAN filters adsorb factor D which looses its enzymatic activity whilst bound to the membrane [1]; the complement alternative pathway function of serum exposed to PAN filters is greatly diminished and restored after addition of purified factor D [1]. The aim of our study was to measure the time course of factor D adsorption and its blood concentration during CVVH in critically ill patients with acute renal failure. METHODS: We studied seven critically ill patients with ARF before, during and after continuous veno-venous haemofiltration (CVVH) with AN69. RESULTS: There was a rapid decrease of factor D levels to 62(+/-6%) of the pre-CVVH value during the first 2 h, which continued to 51(+/-7.3%) after 12 h; at 24 h there was a slight rise to 62 +/- 12%. Sequential use of Polyacrylonitrile (AN69) filters lowered factor D levels below the normal plasma concentration in three patients, thus producing a state of factor D depletion. CONCLUSION: The significant reduction of factor D levels during CVVH with PAN filters suggests that frequent changes of PAN filters may reduce alternative pathway function by lowering factor D levels. CVVH (as opposed to intermittent dialysis) with PAN membranes may further improve the outcome of critically ill patients.

Acrylic Resins↗
PubMed1995

Insulin-like growth factor I (IGF-I) affects plasma lipid profile and inhibits the lipolytic action of growth hormone (GH) in isolated adipocytes.

The actions of insulin-like growth factor I (IGF-I) and growth hormone (GH) on lipid metabolism have been investigated in hormonally intact rats. The subcutaneous injection of IGF-I (100 micrograms) and GH (100 micrograms) lowered total cholesterol plasma levels. The in vitro assays on isolated rat adipocytes showed an antilipolytic effect of IGF-I on GH induced lipolysis. All these data support an important role for IGF-I and GH on the regulation of lipid metabolism.

Adipocytes↗
PubMed1995

Another function of erythrocytes: transport of circulating immune complexes.

OBJECTIVE: To review the main characteristics of the 'immune complex (IC) transport system of erythrocytes', and its possible relevance for transfusion medicine. DATA SOURCES: Literature search, and results Of Studies performed in the laboratory of Dr J. Schifferli from 1986 to 1995. SELECTION CRITERIA: Peer review journals and work relevant to the topic. RESULTS: When antigen/antibody IC form in the presence of complement. C3b binds covalently to the complexes. Such opsonized complexes attach to cells bearing complement receptor 1 (CR1), which is mostly found on erythrocytes in the circulation. This allows IC to be transported through the circulation to the fixed macrophage system of the liver and spleen. In addition, C3b bound to the complexes is catabolized by factor 1 (with CR1 as a cofactor) so that the complement-activating properties of the complexes are reduced. CONCLUSIONS: Complement and erythrocyte CR1 contribute to the safe and effective elimination of IC in humans. This physiologic system prevents IC deposition in the vessel walls.

Antigen-Antibody Complex↗
PubMed1995

Organ-specific effects and interactions of acute insulin-like growth factor I (IGF-I) and growth hormone (GH).

The acute organ-specific and metabolic actions of Insulin-like Growth Factor-I (IGF-I) and Growth Hormone (GH) were investigated in normal rats. Rats received a single subcutaneous injection of IGF-I (100 micrograms), GH (100 micrograms) or a combined treatment with IGF-I (100 micrograms) plus GH (100 micrograms) following a 2(2) factorial design. The acute treatment with IGF-I produced an increase in its plasma concentration along with a reduction in insulin levels. Plasma glucose and total cholesterol decreased in all treated groups as well as triglycerides in IGF-I treated animals as compared to controls. The rates of protein synthesis, measured by amino acid incorporation were not affected by any of these treatments in muscle or liver. However, GH treatment raised the rate of protein synthesis in the jejunum, while IGF-I treatment produced an increase in tibia protein synthesis rate. Negative interactions between GH and IGF were noted concerning tibia and jejunum protein formation. Thus, GH appeared to inhibit the response to IGF-I in bone, while IGF-I inhibited the response of the jejunum to GH administration. Also, liver cathepsin activity was reduced, while bone alkaline phosphase was increased by the GH treatment. Therefore, these results have demonstrated some acute organ-specific anabolic effects and interactions of IGF-I and GH on different aspects of metabolism in normal rats.

Acid Phosphatase↗
PubMed1994

Identification of membrane-bound CR1 (CD35) in human urine: evidence for its release by glomerular podocytes.

Complement receptor 1 (CR1) is present on erythrocytes (E-CR1), various leucocytes, and renal glomerular epithelial cells (podocytes). In addition, plasma contains a soluble form of CR1 (sCR1). By using a specific ELISA, CR1 was detected in the urine (uCR1) of normal individuals (excretion rate in 12 subjects, 3.12 +/- 1.15 micrograms/24 h). Contrary to sCR1, uCR1 was pelleted by centrifugation at 200,000 g for 60 min. Analysis by sucrose density gradient ultracentrifugation showed that uCR1 was sedimenting in fractions larger than 19 S, whereas sCR1 was found as expected in fractions smaller than 19 S. The addition of detergents reduced the apparent size of uCR1 to that of sCR1. After gel filtration on Sephacryl-300 of normal urine, the fractions containing uCR1 were found to be enriched in cholesterol and phospholipids. The membrane-association of uCR1 was demonstrated by analyzing immunoaffinity purified uCR1 by electron microscopy which revealed membrane-bound vesicles. The apparent molecular mass of uCR1 was 15 kD larger than E-CR1 and sCR1 when assessed by SDS-PAGE and immunoblotting. This difference in size could not be explained on the basis of glycosylation only, since pretreatment with N-glycosidase F reduced the size of all forms of CR1; however, the difference in regular molecular mass was not abrogated. The structural alleles described for E-CR1 were also found for uCR1. The urine of patients who had undergone renal transplantation contained alleles of uCR1 which were discordant with E-CR1 in 7 of 11 individuals, indicating that uCR1 originated from the kidney. uCR1 was shown to bind C3b-coated immune complexes, suggesting that the function of CR1 was not destroyed in urine. A decrease in uCR1 excretion was observed in 3 of 10 patients with systemic lupus erythematosus, corresponding to the three who had severe proliferative nephritis, and in three of three patients with focal sclerosis, but not in six other patients with proteinuria. Taken together, these data suggest that glomerular podocytes release CR1-coated vesicles into the urine. The function of this release remains to be defined, but it may be used as a marker for podocyte injury.

Adult↗
PubMed1994

Acute effects of insulin-like growth factor-I (IGF-I) on bone protein synthesis in rats.

The acute systemic effects of IGF-I on bone protein metabolism from hormonally intact rats have been investigated by measuring the fractional rate of protein synthesis. The animals received a single subcutaneous injection of rhIGF-I (100 micrograms). Treatment with IGF-I increased the rate of tibia protein synthesis (Ks) while plasma glucose and insulin concentrations were decreased. These results are in favor of an endocrine role of IGF in stimulating bone growth.

Animals↗