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Biomedical subjects

S Gomez

Publications and source records attributed to S Gomez.

At least 37 records · Page 2Linked to original sources

Matrix vesicles and focal proteoglycan aggregates are the nucleation sites revealed by the lanthanum incubation method: a correlated study on the hypertrophic zone of the rat epiphyseal cartilage.

Correlated studies were performed with light and electron microscopy, and backscattered electron image in conjunction with X-ray microanalysis, of lanthanum-incubated epiphyseal cartilage of the young rat. The hall-mark of this procedure is the appearance of LaP electron-dense deposits (not present in control sections) in precise sites of the hypertrophic zone. The ultrastructural study revealed a dual nature of these sites: "dense matrix vesicles" and "focal filament aggregates". The dense matrix vesicles are a specific type of matrix vesicle with the intrinsic capacity of precipitating LaP mineral, as soon as they originate from the hypertrophic chondrocytes. Furthermore, the matrix vesicles were found to be heterogeneous because lanthanum-devoid, "light matrix vesicles" were also present. The focal filament aggregates, which were not recognized in unstained sections and in controls, are apparently focal concentrations of proteoglycans with high lanthanum binding capacity, although the presence in them of other components (e.g., type X collagen, C-propeptide of type II collagen) cannot be excluded. The were in close connection with the light matrix vesicles in the upper hypertrophic zone, and were loaded with a variable quantity of LaP irregular electron-dense deposits in the lower hypertrophic zone. These irregular deposits are similar to, but distinct from, calcification nodules. The lanthanum incubation method indirectly detects the matrix Ca-binding components (which bind La ions), and the calcification initiation sites (which precipitate a LaP-mineral phase). A sequence is proposed of successive steps of LaP nucleation within the focal filament aggregates, which possibly mimics calcium phosphate deposition. Such a sequence seems to require the participation not only of dense matrix vesicles, but also of the filamentous components of the focal aggregates, possibly together with the activity of alkaline phosphatase.

Animals↗

The GPI-anchored adhesion molecule F3 induces tyrosine phosphorylation: involvement of the FNIII repeats.

The glycosyl-phosphatidylinositol (GPI)-anchored F3 molecule, a member of the Ig superfamily made up of Ig and FNIII-like domains, is involved in cell-cell adhesion, neuronal pathfinding and fasciculation. Little is known about the mechanism(s) that governs the F3-mediated cell-cell recognition. In particular, it is not known whether F3 transduces signals across the membrane. Here we show that in F3-transfected CHO cells (1A cells) an increase in tyrosine phosphorylation occurs during F3-mediated aggregation. Moreover, under aggregation conditions F3 immunoprecipitated from 32P-metabolically labeled 1A cells associated with three major phosphorylated proteins. Interestingly, genistein inhibited the F3-mediated aggregation. Increased tyrosine phosphorylation was also observed using antibody-mediated F3-cross-linking. Furthermore, F3 expressed both in 1A cells and in post-natal mouse cerebellum forms non-covalent soluble complexes with protein tyrosine kinase(s). In cerebellum the F3-associated kinase was identified as fyn. By contrast, a truncated F3 protein, expressed in CHO cells, from which all the FN type III repeats have been deleted, does not associate with a kinase. Cross-linking of the F3-truncated form does not induce modulation of tyrosine phosphorylation. Taken together these data demonstrate that F3 is a molecule that transduces signals through both association with protein tyrosine kinase and modulation of protein tyrosine phosphorylation. The presence of FN type III domains is essential for the activation of the intracellular signaling pathway.

Animals↗

The use of caffeine versus prophylactic naps in sustained performance.

Previous studies have shown that performance during sleep loss is improved by prophylactic naps as a function of varying nap length. Based on single-dose caffeine studies, a similar dose-response effect has been hypothesized on performance, alertness and mood during sleep loss. The present study compared the effects of repeated versus single-dose administration of caffeine and varying amounts of sleep taken prior to sleep loss on performance, mood and physiological measures during 2 nights and days of sleep loss. A total of 140 normal, young adult males participated at one of two study sites. Ninety-eight subjects at one site were randomly assigned to one of four nap conditions (0, 2, 4 or 8 hours) and 42 subjects at the second site were assigned to one of four caffeine conditions. After a normal baseline night of sleep and morning baseline tests of performance, mood and nap latency, subjects in the nap groups returned to bed at noon, 1600 hours, 1800 hours or not at all. Bedtimes were varied so that all naps ended at 2000 hours. Subjects in the caffeine groups received either a single 400-mg dose of caffeine at 0130 hours each night or repeated doses of 150 or 300 mg every 6 hours starting at 0130 hours on the 1st night of sleep loss. A placebo control group (no nap and placebo administered every 6 hours on the repeated caffeine schedule) was run at both sites.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Social work in pediatric oncology: a family needs assessment.

When a child is diagnosed with cancer, the family is confronted with meeting both the physical and psychosocial needs of the child and maintaining normal family functioning. This study assessed the perceived psychosocial needs of 77 families who have a child diagnosed with cancer. Preliminary results suggest practical application for social work interventions in specific areas such as the development of an informal support network, enhancement of communication within families concerning the disease, the need for adequate information at various stages of the disease, and continued supportive services for the family.

Adult↗

Correlated alkaline phosphatase histochemistry and quantitative backscattered electron imaging in the study of rat incisor ameloblasts and enamel mineralization.

The different functional conditions of pre-ameloblasts, secretory ameloblasts, and maturation ameloblasts in 9 day rat incisors were recognized using high resolution light microscopic alkaline phosphatase histochemistry: Digital backscattered electron imaging was performed using the block surfaces from which thin sections were taken for histochemical study. It was possible to correlate exact locations in histochemical sections with positions in the block face at all stages of enamel mineralization from early secretion through late maturation. The first steep increase in the rate of mineralization of completed enamel matrix occurs after the first transition from smooth ended ameloblasts to ruffle ended ameloblasts. In the 9 day rat incisors used for this purpose, there are only two smooth to ruffle cyclical transitions, and the width of successional smooth ended bands of ameloblasts in the maturation cycling process is always narrow. Nevertheless, there seems to be a good correlation between mineralization increase and the acquisition of the high alkaline phosphatase activity in the deeply enfolded distal cytoplasm of the ruffle-ended maturation stage ameloblasts.

Alkaline Phosphatase↗

Different domains of the F3 neuronal adhesion molecule are involved in adhesion and neurite outgrowth promotion.

The mouse F3 cell surface protein is preferentially expressed on axons of subpopulations of neurons and is anchored to the membrane by a glycosyl-phosphatidylinositol group. It consists of six immunoglobulin-like domains and four fibronectin type III homologous repeats, and can be found both in membrane-anchored and soluble forms. We have previously established that F3 fulfills the operational criteria of a cell adhesion molecule when anchored to the plasma membrane and that its soluble form stimulates neurite initiation and neurite outgrowth. To further characterize F3-mediated adhesion and to investigate whether adhesion and neurite outgrowth promoting activities are displayed by different parts of the molecule, we (i) selected F3 transfected CHO cells expressing increasing levels of F3 at their surface and (ii) prepared transfectants expressing an F3 molecule with its fibronectin type III repeats deleted. We show that the F3 molecule mediates divalent-cation-independent, temperature-dependent binding. The levels of aggregation of F3 transfectants are proportional to the level of F3 expression. Transfectants expressing F3 deleted of the fibronectin type III repeats lose their adhesive properties; conversely, cells expressing wild-type F3 and treated with collagenase, specifically removing the immunoglobulin-like domains, are still able to aggregate. Therefore, in this model adhesion site(s) mapped to the fibronectin type III repeats. By contrast, transfectants expressing deleted F3, as well as the soluble forms of this F3 deleted molecule, were able to stimulate neurite outgrowth of sensory neurons similarly to wild-type F3. Our data indicate that F3 is a multifunctional molecule and that adhesion and neurite outgrowth promoting properties are expressed by distinct and independent domains.

Animals↗

Anti-immunoglobulin antibodies in children with Schönlein-Henoch syndrome. Absence of serum anti-IgA antibodies.

Circulating immune complexes (CIC) that simultaneously contain IgG and IgA are frequently found in IgA nephropathy (IgA-N) and the Schönlein-Henoch syndrome (SHS). The presence of anti-immunoglobulin antibodies (IgA anti-IgG and IgG anti-IgA) was studied by ELISA in the serum of 39 children with SHS and compared to 30 normal children. The mean level of IgG anti-IgA antibodies (240 +/- 104 u/ml) in SHS patients was similar to control values (251 +/- 85 u/ml); the IgA anti-IgG antibodies were increased, although only the antibodies against Fc fraction of IgG were elevated (185 +/- 71 u/ml in patients vs 127 +/- 24 mu/ml in controls, P < 0.0001) without a significant increase of IgA anti-IgGFab antibodies (141 +/- 54 mu/ml vs. 137 +/- 25 u/ml); 16/39 (41%) of the patients had increased levels of IgA anti-IgGFc and 6 of these had also high IgA anti-IgGFab. None of these patients had high IgA anti-IgGFab antibodies without simultaneous augmentation of IgA anti-IgGFc. Only 3/39 (7.7%) of SHS patients showed high levels of IgG anti-IgA antibodies. The correlation of IgA anti-IgGFc antibodies with IgA anti-IgGFab was very strong (P < 0.0001) but lower with IgG anti-IgA antibodies (P < 0.002). In addition, 8/39 children had renal involvement, nevertheless in these patients the findings were quite similar, with a non-significant elevation of IgA anti-IgGF ab antibodies. These results show that the IgA anti-IgG antibodies are more frequently increased than IgG anti-IgA antibodies in the SHS; moreover they are mainly directed against Fc fraction and are IgA-FR isotype. Our findings suggest that the CIC in SHS are likely formed by the reaction of IgA anti-bodies against IgG and not vice versa.

Antibodies, Anti-Idiotypic↗

Antineutrophil cytoplasmic antibodies (ANCA) in idiopathic pulmonary hemosiderosis.

Four children were diagnosed with idiopathic pulmonary hemosiderosis (IPH), over a period of 4 years. Retrospectively, antineurtrophil cytoplasmic antibodies (ANCA) were studied by indirect immunofluorescence (IIF) and ELISA in 18 sera from these patients, stored at -20 degrees C. ANCA-positive sera, from 1/20 to 1/1, 200 dilution, were found in 3/4 of the patients, by IIF. The patient with the highest titre of ANCA died 3 months later during an acute crisis, the other two patients need a minimal dose of steroids. In one case only, a patient who is still without treatment, had no ANCA. The antibodies anti-myeloperoxidase and anti-proteinase-3 were negative or at border line levels. Rheumatoid factor, antinuclear (Hep-2), anti-endomysial, anti-reticulin and antibasement membrane antibodies were negative in all sera. The surviving patients were followed-up for more than 10 years with no systemic or renal disease appearances. The presence of serum ANCA may help to classify children with pulmonary haemorrhage and may have a prognostic value.

Antibodies, Antineutrophil Cytoplasmic↗

Interaction between band 3 and ankyrin begins in early compartments of the secretory pathway and is essential for band 3 processing.

In many cell types, membrane proteins are specifically segregated to particular areas of the cell surface. It is known that this segregation is stabilized by anchorage proteins which interact with the cytoskeleton. However, the mechanism by which the interactions with anchorage proteins occur, as well as whether they may also play a role in the process of sorting, is not known. Using differentiated murine erythroleukemic cells, we have investigated the association between band 3 (a major transmembrane anion exchanger), and ankyrin (a cytoplasmic protein that links band 3 to the cytoskeleton). Our data demonstrate that the association between band 3 and ankyrin occurs in the endoplasmic reticulum or the first Golgi compartment. These data support a model in which the band 3-ankyrin complex is inserted as a "cassette" at the plasma membrane into the cytoskeletal network. Biosynthetic studies on cotransfected 293 cells with cDNAs encoding band 3 and the band 3 binding fragment of ankyrin (AnK-90), suggest that ankyrin is not only responsible for the anchorage of band 3 to the cytoskeleton but is also involved in the exit of band 3 out of the endoplasmic reticulum.

Animals↗

Rhabdomyosarcomas developing in association with mediastinal germ cell tumours.

Two mediastinal rhabdomyosarcomas that arose in association with germ cell tumours are reported. One presented as a small component of a mixed germ cell tumour with yolk sac and immature teratomatous elements. The other appeared as a large mass 4 months after diagnosis of a yolk sac carcinoma that had been treated with chemotherapy. The first patient was alive and free of disease 7 years later, whereas the second died of tumour 3 months post-operatively. The proportion of rhabdomyosarcoma within the germ cell tumours appears to have influenced the prognosis of these patients. This observation emphasizes the necessity of performing a thorough search for sarcomatous elements and quantifying their relative proportion in germ cell tumours of the mediastinum.

Adult↗

Detection of Mycobacterium paratuberculosis antigen with colloidal immunogold in naturally infected sheep.

The anti-Mycobacterium paratuberculosis polyclonal serum is proved useful for labelling Mycobacterium paratuberculosis in glutaraldehyde-osmium-fixed and epon-embedded intestinal samples from sheep with clinical symptoms of paratuberculosis. M. paratuberculosis marked with antibody-coated colloidal gold stain was seen in macrophages, epithelioid cells, giant cells and neutrophils throughout intestinal mucosa. In large macrophages with a low lysosomal content, a great number of intact mycobacteria was seen within phagosomes. In macrophages with average lysosomal content, very few intact mycobacteria or mycobacterial debris were present and lysosome-phagosome fusions were observed. Mycobacteria within neutrophils were scanty. These results show the usefulness of colloidal immunogold techniques for studies of the pathogenesis of paratuberculosis.

Animals↗

Prosomatostatin is processed in the Golgi apparatus of rat neural cells.

Proteolytic processing of somatostatin precursor produces several peptides including somatostatin-14 (S-14), somatostatin-28 (S-28), and somatostatin-28 (1-12) (S-28(1-12)). The subcellular sites at which these cleavages occur were identified by quantitative evaluation of these products in enriched fractions of the biosynthetic secretory apparatus of rat cortical or hypothalamic cells. Each of the major cellular compartments was obtained by discontinuous gradient centrifugation and was characterized both by specific enzyme markers and electron microscopy. The prosomatostatin-derived fragments were measured by radioimmunoassay after chromatographic separation. Two specific antibodies were used, allowing the identification of either S-28(1-12) or S-14 which results from peptide bond hydrolysis at a monobasic (arginine) and a dibasic (Arg-Lys) cleavage site, respectively. These antibodies also revealed prosomatostatin-derived forms containing at their COOH terminus the corresponding dodeca- and tetradecapeptide sequences. Whereas the reticulum-enriched fractions contained the highest levels of prosomatostatin, the proportion of precursor was significantly lower in the Golgi apparatus. In the latter fraction, other processed forms were also present, i.e. S-14 and S-28(1-12) together with the NH2-terminal domain (1-76) of prosomatostatin (pro-S(1-76). Inhibition of the intracellular transport either by monensin or by preincubation at reduced temperature resulted in an increase of prosomatostatin-derived peptides in the Golgi-enriched fractions. Finally, immunogold labeling using antibodies raised against S-28(1-12) and S-14 epitopes revealed the presence of these forms almost exclusively in the Golgi-enriched fraction mainly at the surface of saccules and vesicles. Together these data demonstrate that in rat neural cells, prosomatostatin proteolytic processing at both monobasic and dibasic sites is initiated at the level of the Golgi apparatus.

Animals↗

Transient elevation of serum lactic dehydrogenase following autologous bone marrow transplantation.

Transient elevated levels of serum lactic dehydrogenase (LDH) were reported immediately following autologous bone marrow transplantation consecutively in 15 patients. Isoenzyme 2 was the predominant elevated isoenzyme of LDH, suggesting that the source of LDH is either hemolysis of RBCs, which occurs during freezing and thawing of bone marrow in the bags and in vivo following infusion of residual dimethyl sulfoxide.

Adolescent↗

Site-specific mutagenesis identifies amino acid residues critical in prohormone processing.

Peptide hormones are generally synthesized as inactive higher mol. wt precursors. Processing of the prohormone into biologically active peptides by specific proteolytic cleavages occurs most often at pairs of basic amino acids but also at single arginine residues. To study the role of protein secondary structure in this process, we used site-directed mutagenesis to modify the predicted secondary structure around the cleavage sites of human prosomatostatin and monitored the processing of the precursor after introduction of the mutated cDNAs in Neuro2A cells. Amino acid substitutions were introduced that affected the possibility of forming beta-turn structures in the immediate vicinity of the somatostatin-28 (S-28) and somatostatin-14 (S-14) cleavage sites. Infection of Neuro2A cells with a retrovirus carrying a human somatostatin cDNA resulted in the expression of prosomatostatin and its processing into S-28 and S-14, indicating that these cells have the necessary enzymes to process prohormone at both single and paired amino acid residues. Disruption of the different beta-turns had various effects on prosomatostatin processing: substitution of Ala for Pro-5 drastically decreased prosomatostatin processing and replacement of Pro-9 by Ala led to the accumulation of the intermediate maturation product [Arg-2Lys-1]-S-14. In contrast, substitution of Ala for Asn-12, Gly+2 and Cys+3 respectively had only very little effect on the proteolytic processing of prosomatostatin. Our results show that amino acids other than the basic amino acid residues are required to define the cleavage sites for prohormone proteolytic processing and suggest that higher orders of protein structure are involved in substrate recognition by the endoproteases.

Acyltransferases↗

Cytological, immunocytochemical and ultrastructural study of the adenohypophyseal pars distalis of the kid (Capra hircus): the TSH cell.

The structure and ultrastructure of the adenohypophyseal pars distalis in kids of the Murciano-granadiana breed (18 males and 12 females), aged 2-3 months and with an average carcass weight of 8 kg has been studied. Techniques of staining (Tetrachrome Herlant's, and Cleveland-Wolfe) and histochemistry (PAS, PAS-OG and BA-PAS-OG) contrasted with immunolabelling (PAP) have been used. In addition an ultrastructural study has been made and nucleus and cytoplasm, secretory granules, mitochondria and lysosomes have been measured with a semiautomatic image analyzer (IBAS-1). TSH cells are found in sagittal section in the anterior area and in an antero-caudodorsal band, and transversally in the ventral and medial region; they are 6% and their average size is 14.15 microns. Ultrastructurally they are a single cellular type with spherical granules whose size is 195 nm in males and 149 in females; these granules are characterized by a clear halo and a variable electronic density. The rough endoplasmic reticulum is found as slightly enlarged vesicles with a homogeneous and moderately electro-dense content or in concentric stratifications.

Animals↗