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Biomedical subjects

E Lopez

Publications and source records attributed to E Lopez.

At least 19 recordsLinked to original sources

Nacre initiates biomineralization by human osteoblasts maintained in vitro.

When nacreous shell produced by the marine oyster Pinctada maxima, used as a biomaterial in oral surgery, is implanted in human bone, new bone formation occurs, resulting in a tight welding of the bone to the nacre [16]. These findings are consistent with the possibility that nacre adjacent to bone can locally stimulate osteogenic activity. To test this hypothesis, we have evaluated the effect of the simultaneous presence of bone and nacre on human osteoblasts in vitro. Nacre chips (1 mm3) were placed at approximately 1 mm distance from a similarly sized bone chip on a layer of first passage human osteoblasts. None of the chemical inducers generally required to obtain bone mineralization in vitro (in particular, beta-glycerophosphate) was added to the cultures. Mineralized sections of the cultures were evaluated by light and electron microscopy, contact microradiography, and Laser Raman Spectroscopy. The results demonstrated that nacre has strong osteogenic effects on human osteoblasts when placed in proximity to bone in vitro. New bone formation occurred by both appositional growth on the existing bone and by the formation of mineralized nodules within the matrix adjacent to the bone explant. Electron microscopic evaluation of these sites demonstrated findings typical of those described in the course of bone formation in vivo, and no evidence of toxicity was observed. In addition, under the conditions of culture used, nacre can also promote the formation by osteoblasts of a structure with characteristics similar to nacre (e.g., lamellar organic matrix mineralized with aragonite, as demonstrated by Laser Raman Spectroscopy).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Demonstration of the capacity of nacre to induce bone formation by human osteoblasts maintained in vitro.

Nacre implanted in vivo in bone is osteogenic suggesting that it may possess factor(s) which stimulate bone formation. The present study was undertaken to test the hypothesis that nacre can induce mineralization by human osteoblasts in vitro. Nacre chips were placed on a layer of first passage human osteoblasts. None of the chemical inducers generally required to obtain bone formation in vitro was added to the cultures. Osteoblasts proliferated and were clearly attracted by nacre chips to which they attached. Induction of mineralization appeared preferentially in bundles of osteoblasts surrounding the nacre chips. Three-dimensional nodules were formed by a dense osteoid matrix with cuboidal osteoblasts at the periphery and osteocytic-like cells in the center. These nodules contained foci with features of mineralized structures and bone-like structures, both radiodense to X-ray. Active osteoblasts (e.m.) with abundant rough endoplasmic reticulum, extrusion of collagen fibrils and budding of vesicles were observed. Matrix vesicles induced mineral deposition. Extracellular collagen fibrils appeared cross-banded and electrodense indicating mineralization. These results demonstrate that a complete sequence of bone formation is reproduced when human osteoblasts are cultured in the presence of nacre. This model provides a new approach to study the steps of osteoblastic differentiation and the mechanisms of induction of mineralization.

Animals

Europium(III) cryptate: a fluorescent label for the detection of DNA hybrids on solid support.

We report here a new detection method for DNA hybrids on dot blots. The process utilizes DNA or oligonucleotide probes labeled with biotin, followed by recognition with a conjugate of streptavidin and europium cryptate, a time-resolved fluorescent label. Unlike the other lanthanide chelates, this label is an organic molecule embedding a europium ion into an intramolecular cavity. This structure has a better stability in diluted assay media, a good sensitivity even on solid support, and an elevated fluorescence lifetime which allows elimination of most of the background generated by other species present in the assay medium. This procedure is quantitative and detects down to 2 amol of a model DNA, which is similar to other nonisotopic (especially colorimetric) methods. The main advantages of this method are easy automation, quantitation, and rapidity of measurements.

Bacterial Proteins

Giant liposomes: a model system in which to obtain patch-clamp recordings of ionic channels.

Cell-size, giant liposomes have been formed by submitting a mixture of asolectin lipid vesicles and native membranes from Torpedo, highly enriched in acetylcholine receptor (AcChR), to a partial dehydration/rehydration cycle [Criado, M., & Keller, B. U. (1987) FEBS Lett. 224, 172-176]. Giant liposomes can be prepared in bulk quantities, in the absence of potentially damaging detergents or organic solvents, and their formation is mediated by membrane fusion phenomena. In fact, fluorescence microscopy and freeze-fracture data indicate that protein and lipid components of the initial membranes and lipid vesicles are homogenously distributed in the resulting liposomes. Giant liposomes containing AcChR have been used as a model to evaluate whether this system can be used to monitor the activity of ionic channels by using high-resolution, patch-clamp techniques. Excised liposome patches in an "inside-out" configuration have been used in this work. We find that the most frequent pattern of electrical activity in response to the presence of acetylcholine in the patch pipet corresponds to a cation-specific channel exhibiting a dominant conductance level and a sublevel of approximately 78 and 25 pS, respectively. Such channel activity exhibits the pharmacological specificity, ion channel activation, ion selectivity, and desensitization properties expected from native Torpedo AcChR. Thus, it appears that the giant liposome technique offers a distinct advantage over other reconstitution procedures in that it provides a unique opportunity to undertake simultaneous biochemical, morphological, and electrophysiological studies of the incorporated ionic channel proteins.

Animals

[Hormonal stimulation, in vivo, of the ovary of European eel in the yellow stage].

The immature ovary of yellow eel was stimulated by a long-term treatment with a gonadotropin II rich extract of carp pituitary. Ovarian follicles of treated yellow eels showed an accumulation of lipidic vacuoles in the oocyte cytoplasm and a thickened follicular envelope (in this regard, they resembled the ovarian follicles of previously studied silver eels). The treatment also resulted in a significant but limited increase in the gonadosomatic ratio and mean follicle size (these values were much lower than those measured in silver eels). No change in pigmentation or general morphology was observed. Thus, under our experimental conditions, a gonadotropic stimulation of the yellow eel leads to some but not the totality of the ovarian modifications which occur physiologically during silvering.

Anguilla

Partial purification of parathyrin from the corpuscles of Stannius (PCS) of the eel (Anguilla anguilla, L.).

It has been previously shown that the eel corpuscles of Stannius (CS) synthesize and secrete a substance (PCS) which is functionally and immunologically related to the mammalian parathyrin family. Purification of PCS, including anion-exchange chromatography, ODS C-18 reverse-phase HPLC, and affinity chromatography, showed that a biologically active peak, eluted in 32% acetonitrile, contains a 32- to 34-kDa protein which is 600-fold more potent than the crude extract is a test involving the hypocalcemic response in the CS-deprived eel. Specific immunoprecipitation of protein encoded by mRNA extracted from eel CS indicates that a 45-kDa precursor is involved in PCS synthesis. The hypothetical significance of a "large" parathyrin-like molecule in fish is suggested in relation to what is known about mammalian parathyrin gene expression.

Alkaline Phosphatase

[Soldering without transition (osteoassimilation) between the human maxillary bone and a compact dental implant in natural calcite from marine invertebrate].

We present evidence for the formation of woven bone between human maxillary lamellar bone (the target site) and a coral implant (Corallium johnsoni Gray). This newly formed bone is characterized by numerous osteocytes located in spherical lacunae and having many ramified processes directed towards the coral; some of these processes continue into the implant surface. Healthy osteocytes were detected within the superficial part of the coral, next to the newly formed bone. Neither osteoclast nor surface of resorption has been noticed. All these observations indicate tight welding or osteoassimilation without any transition between the maxillary bone and the graft. We have obtained analogous results by using grafts of molluscan mother-of-pearl. Our results differ from those reported by other authors, probably due to the compactness of the biomaterial used here.

Animals

Stimulation of gonadotropin release and of ovarian development, by the administration of a gonadoliberin agonist and of dopamine antagonists, in female silver eel pretreated with estradiol.

In freshwater or seawater female silver eel, the release of gonadotropin (GTH) accumulated in the pituitary under estradiol (E2) influence could be stimulated by a conjugated treatment with a mammalian gonadoliberin agonist (GnRH-A = des-Gly10, (D-Ala6)-LH-RH ethylamide) and a blocker of dopamine receptor (pimozide). Furthermore, despite the GTH release, no reduction or even a significant increase in pituitary GTH levels were noted, indicating a stimulation of GTH synthesis. In consequence of the endogenous GTH release, a stimulation of ovarian development was induced, as demonstrated by the gonadosomatic index and histological study. Similar results were obtained with a combined treatment with GnRH-A and an inhibitor of catecholamine synthesis (L-alpha-methyl-3,4-dihydroxyphenylalanine). In contrast, no effect was produced by GnRH-A, pimozide, or L-alpha-methyl-DOPA, given alone. The results suggest that a double neuroendocrine mechanism (a lack of GnRH production and a dopaminergic inhibition of GnRH action) is involved in the prepubertal blockage of eel gonadotropic function before the reproductive migration.

Anguilla

Calcium-dependent metabolism of 25-hydroxycholecalciferol in silver eel tissues.

We investigated the in vitro metabolism of [26,27-3H]-25-(OH)D3 in different eel tissues. After incubation with [3H]-25-(OH)D3, tissues were extracted with methanol-chloroform and chromatographed on Sephadex LH 20 columns. Two derivatives less polar than 25-(OH)D3 were detected, the first one being sensitive to KOH treatment. Three peaks more polar than 25-(OH)D3 were also found: peak I migrated close to the 24,25-(OH)2D3 area and was quantitatively the most important, but the presence of 24,25-(OH)2D3 could not be demonstrated; peak II migrated in the 1,25-(OH)2D3 region; and peak III had an elution position twice that of peak II. After 6-h incubation of tissues isolated from control eels, peak I was found in all tissues including intestine and gills. It was highest in pituitary gland and brain and lowest in ovaries and muscle. It was not significantly modified 20 days after ablation of the corpuscules of Stannius. In contrast, in vivo daily calcium chloride injection was followed 24 hr later by a significant increase in the [3H]-25-(OH)D3 conversion into peak I in gills, intestine, and the spinal cord and by an inhibition of this conversion in pituitary gland, skin, and muscle. The inhibition was found in all tissues after five daily calcium injections. Calcium injection had no effect on the in vitro metabolite synthesis by the corpuscules of Stannius. These results suggest that vitamin D is not metabolized in the same way in eel as in mammals and that this metabolism could in part be calcium dependent.

Animals

Anticytotoxin-neutralizing antibodies in immune globulin preparations: potential use in hemolytic-uremic syndrome.

The pathogenesis of primary (classic) hemolytic-uremic syndrome (HUS) is thought to be related to cytotoxin-producing enteric pathogens such as Shigella dysenteriae serotype 1 and Escherichia coli serotypes O157:H7 and 026:H11. The relevant cytotoxins include Shiga toxin and the closely related Shiga-like toxins (SLTs) produced by some E. coli strains. Intravenously administered immune globulin (IVIG) therapy has been reported to be beneficial in a few children with HUS. We therefore examined commercially available immune globulin preparations for the presence of anticytotoxin-neutralizing antibodies. Cytotoxicity and neutralization of the HUS-associated cytotoxins were quantitatively determined by means of a (3H)thymidine-labeled HeLa cell assay. The immune globulin preparations tested almost completely neutralized Shiga toxin (produced by S. dysenteriae 1) and SLT-I (produced by E. coli serotype 026:H11). Twofold dilutions of the preparations showed significant (p less than 0.01) neutralizing titers of 1:64 to 1:128. No significant neutralization (greater than 20%) of SLT-II (produced by E. coli strain C600 (933W] was noted. The IVIG preparation lost its inhibitory activity when passed through a protein A-Sepharose column, which bound immune globulin, indicating that its neutralizing effect is related to the antibody content. We also examined sera from 30 children without diarrhea or HUS; only one child had neutralizing titers against Shiga toxin (1:64) and SLT-I (1:128). Immune globulin preparations contain anticytotoxin-neutralizing antibodies, a finding that warrants further investigation of the therapeutic role of these preparations in early treatment of children with HUS related to Shiga toxin and SLT-I.

Adolescent

Perineal urethrocutaneous fistula complicating ischiectomy.

The role of ischiectomy for the repair and/or prevention of ischial ulcers is controversial. We report a rare complication of a perineal urethrocutaneous fistula following ischiectomy and flap closure to repair an ischial ulcer. This complication and its management are reviewed.

Adult

A method for quantifying the area of closed pressure sores by sinography and digitometry.

Neurologically impaired and elderly patients who remain in the same position for prolonged periods risk developing pressure sores. The most insidious of these are closed pressure sores. Conservative treatment of closed pressure sores frequently is the only treatment option. However, no systematic method to assess the effectiveness of conservative treatment modalities exists. The purposes of this study were to develop a method for quantifying the area of closed pressure sores by means of sinography and digitometry and to determine the utility of this method for monitoring healing. A convenience sample of 40 veterans with closed pressure sores was selected. Four individuals independently traced 116 regions from 30 sinograms of closed pressure sores from a subsample of ten patients. A fifth individual calculated areas of closed pressure sores in units, using digitometry. Findings and nursing implications will be presented.

Adult

Correlations among measures of cognitive ability, creativity, and academic achievement for gifted minority children.

This study explored the correlations among nonverbal reasoning ability, creativity, and academic achievement in gifted minority children, 89 girls and 71 boys in Grades 1 through 8 in a program for gifted. A random half of students from all grade levels were tested at the beginning of the year and the remaining half after 7 mo. with Raven Progressive Matrices, Torrance Tests of Creative Thinking, and the California Achievement Test. Pearson correlations reflected limited relations among these variables except for a significant positive value between creativity and reading achievement. Suggestions for further study and implications for identification procedures and program development were provided.

Achievement

Induction of spermatogenesis and spermiation by a single injection of human chorionic gonadotropin in intact and hypophysectomized immature European eel (Anguilla anguilla L.).

Intact and hypophysectomized male silver eels (Anguilla anguilla) in fresh water received a single injection of human chorionic gonadotropin (hCG) (250 C) or solvent (0.15 M NaCl). No effect of solvent was observed. Spermatogonia proliferated in testis of hCG-treated intact or hypophysectomized eels. One month after the injection, primary and secondary spermatocytes were found. After 3 months, numerous spermatozoa were present. In hypophysectomized eels, hCG was also effective even though maturing germ cells were less numerous and spermiation was less frequent than in intact animals. Within 1 week after hCG injection, plasma levels of free and glucuroconjugated androgens (testosterone and 11-oxotestosterone) rose significantly in intact and hypophysectomized fish. The highest values were observed within 1 month, and then plasma levels decreased to pretreatment values. The most important changes were observed in the case of free 11-oxotestosterone. The long-term effects of hCG can be explained partly by the long half-life of this hormone. The effects of hypophysectomy on the response of testis to hCG caused us to think that some endogenous pituitary secretions must interfere in the intact fish so that maximal effects of hCG, especially on the induction of spermiation, are obtained.

Anguilla

Placental abnormalities in systemic lupus erythematosus: in situ deposition of antinuclear antibodies.

We studied 5 placentae from patients with systemic lupus erythematosus (SLE) for the presence of immunopathologic changes. We found antinuclear antibodies (ANA) deposited along the villi, trophoblast and amnion producing a lupus band-like pattern. The ANA were IgG class and had C3 fixing activity. Of the T cell subsets found in the placenta, OKT8 was the predominant type in patients with SLE and controls. All the patients were ANA positive, and 1 was also positive for anti-Ro(SSA). One newborn, positive for ANA and anti-Ro(SSA), developed neonatal lupus, 1 infant with a cardiac septal defect was ANA negative, and the other infants were normal. Our study demonstrates that in pregnant women with SLE, ANA deposition occurs in the placenta in situ.

Antibodies, Antinuclear