Mechanisms of cell activation by metal ions: influence of pH.
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Biocompatible Materials↗
Biomedical subjects
S R Line
Publications and source records attributed to S R Line.
Absence of mutations in the homeodomain of the MSX1 gene in patients with hypodontia.
Hypodontia, the congenital absence of one or a few permanent teeth, is one of the most frequent alterations of the human dentition. Although hypodontia does not represent a public health problem, it may cause both speech and masticatory dysfunction and esthetic problems. A missense mutation in the homeodomain of MSX1 gene has been associated with hypodontia of second premolars and third molars in humans. However, another study excluded this gene as causative locus for hypodontia of incisors and premolars. To further investigate the role of the MSX1 gene in human hypodontia, we analyzed the homeobox region of the MSX1 gene in 20 individuals with different patterns of familial or isolated hypodontia. The direct sequencing of PCR products did not show any polymorphisms or mutations in the human MSX1 gene. Our results indicate that inactivation of MSX1 gene in humans must have a highly selective effect on dentition, and other genes must be involved in the cause of hypodontia in humans.
Anodontia↗
Incremental markings of enamel in ectothermal vertebrates.
The deposition of enamel is marked by the formation of growth lines, which reflect incremental growth. Although periodic markings have been observed in enamel of non-mammalian vertebrates, the cross-striation interval and the pattern of enamel deposition have not been formally investigated. Here a structural study was made of the enamel in four non-mammalian vertebrates, with emphasis on periodic markings. Teeth from Rana catesbeiana, Tropidurus torquatus, Caiman crocodilus and a Canadian carnosaur were analysed. Enamel of T. torquatus and R. castebeiana was aprismatic; that of C. crocodilus and the carnosaur was formed by large, prism-like structures. Conspicuous incremental lines were observed in the enamel of the three living species, which presented a cross-striation repeat smaller than the prism cross-striations of mammalian enamel. Incremental lines of carnosaur enamel had a mean repeat interval similar to that of mammalian prism cross-striations. As metabolic activity in ectotherms is influenced by environmental conditions, the analysis of incremental markings of enamel is a potentially valuable source of information in the study of living and fossil reptiles and amphibians.
Alligators and Crocodiles↗
The effect of lead on the eruption rates of incisor teeth in rats.
The effects of lead on the continuously erupting rat incisors under normo-, hyper- and hypofunctional conditions were investigated. Left lower incisors of 20 rats were rendered unimpeded (hypofunctional) by cutting them out of occlusion every 2 days; the right lower incisors of these rats were considered hyperfunctional. Measurements on normally growing teeth (normofunctional) were carried out in a group of ten rats whose teeth were not cut but only marked every 2 days. On day 7 of the experiment, half of the rats from these two groups were given a single intraperitoneal injection of lead acetate (40 mg/kg), and the other half received sodium acetate (22 mg/kg). Another group of 15 rats was used to obtain blood samples for lead determination 1 h, and 10, 20, and 30 days after lead administration. Animals were killed on day 32. Hypofunctional incisors from lead-treated rats erupted more slowly than control ones (P<0.05). These results show a previously unreported toxic effect of heavy metals.
Animals↗
Fluoride effect on the activity of enamel matrix proteinases in vitro.
Dental fluorosis is common in individuals exposed to different sources of fluoride during tooth development. The mechanism causing this enamel defect is still unknown. Enamel matrix proteinases play a central role in the maturation of dental enamel, and inhibition of these enzymes by fluoride has been one explanation for dental fluorosis. We have investigated the effect of fluoride on the activity of enamel matrix proteinases using a colorimetric assay, casein zymography, and an enamel protein degradation assay. Fluoride (625 microM to 10 mM) inhibited neither the enzymatic activity of the crude matrix extract nor the activity of individual enamel enzymes separated by SDS-PAGE. The proposition that fluoride could directly inhibit enzymes was not confirmed in this study.
Analysis of Variance↗
Effect of lead, cadmium and zinc on the activity of enamel matrix proteinases in vitro.
Environmental contamination with heavy metals leads to their uptake by the body. Dental tissues are known to harbor metals in concentrations related to the exposure at the time of dentin or enamel formation. Several enzymes involved in the metabolism of extracellular matrix components have been shown to be inhibited by excess metal ions. Enamel matrix proteinases seem to play a central role in the maturation of dental enamel, and inhibition of these enzymes by metals could interfere with amelogenesis. We have investigated the effect of lead, cadmium and zinc on the activity of enamel matrix proteinases using a colorimetric assay, casein and gelatin zymography, and an assay of enamel protein degradation in vitro. All three metals inhibited the proteolysis, as shown by the three methods employed, in concentrations as low as 110 microM. This study showed that contaminant metals can inhibit proteinolysis when a crude enamel extract containing enamel proteases is tested in vitro. Therefore, the presence of heavy metals in enamel matrix could interfere with amelogenesis due to alteration in normal proteolysis.
Amelogenesis↗
Immunochemical analysis of laminin during postnatal development of the rat submandibular gland.
Anti-laminin serum was used to investigate distribution patterns and chain composition of laminin during postnatal development of rat submandibular gland. Immunohistochemical analysis showed that in glands of newborn rats laminin was not uniformly present around growing acini. Staining was frequently weak or absent in clefts formed between adjacent cells. This irregular staining pattern decreased progressively over the subsequent periods, and in 30-day-old animals immunoreactivity was observed only at the periphery of glands. Immunoblot analysis showed that laminin was composed of bands corresponding to the alpha 1, beta 1 and gamma 1 polypeptides. The correlation between the pattern of laminin expression and gland maturation suggests a role of laminin in the functional maturation of acinar cells.
Age Factors↗
Pulpal lesions in normal and cyclosporin A treated rats.
The objective of this study was to examine the development of pulpal lesions in the lower molar of control and cyclosporin A (CyA) treated rats. The pulps of the first lower molars of 20 normal and 20 CyA treated rats were exposed and left open into the oral cavity. Five animals of each group were killed at 7, 14, 21, and 28 days after the pulp exposure. The specimens were sectioned sagittally at a thickness of 7 microns and stained with hematoxylin and eosin. The pulpal lesions were similar for both normal and CyA treated rats in all studied periods and the differences between both groups were not statistically significant by the Student t test at the 5% (0.05) level of significance, indicating that the immunosuppression did not alter the evolution of the inflammatory process.
Animals↗
HIV prevalence in dental outpatients in Brazil.
A series of dental outpatients in Brazil was anonymously screened for HIV antibodies in whole unstimulated saliva with an immunoglobulin G antibody-capture enzyme-linked immunosorbent assay. Salivary HIV antibodies were detected in 40 patients in the control group who were known to be HIV-seropositive but were not detected in any of a series of 40 known HIV-seronegative patients in the control group, confirming the very high sensitivity and specificity of the immunoglobulin G antibody-capture enzyme-linked immunosorbent assay. Only one patient from 84 consecutive dental outpatients of unknown HIV serostatus who were examined anonymously for HIV by immunoglobulin G antibody-capture enzyme linked immunosorbent assay showed HIV positivity (1.2% of the population).
Adolescent↗
Immunochemical analysis of laminin in duct-ligated submandibular glands of rats.
Immunochemical methods have been used to study the expression of laminin during experimental atrophy of the submandibular gland of rats caused by ductal ligation. In normal submandibular glands, laminin immunoreactivity appeared as continuous linear staining around acini and ducts. In the ligated glands, it exhibited an irregular pattern and intensity. Staining was usually stronger around small ducts and acini, which were most prominent in glands ligated for 30 days. Immunoblot analysis showed that the laminin of the rat submandibular gland contains bands that correspond to the EHS alpha 1, beta 1 and gamma 1 chains, and that the composition of the laminin chains does not change during the atrophy.
Animals↗
Immunohistochemical and biochemical analysis of laminin in neonatal rat first molars.
Rabbit polyclonal antibody against mouse EHS laminin was used to investigate the distribution and composition of laminin in the rat first molar tooth germ. Immunohistochemical analysis showed that laminin is expressed in the inner and outer epithelia of the enamel organ and in small blood vessels in the dental papilla and strellate reticulum. Immunoblots revealed that tooth germ laminin differs from EHS laminin. Tooth germ laminin contains beta chains, while the alpha 1 chain is substituted by a 300-kDa chain. Two-dimensional electrophoresis analysis of tooth germ extract showed that beta chains appeared as four spots with approximate pI values of 6.6, 7.5, 7.8 and 8.5. These results indicate that more than-one type of laminin isoform is present in the first molar tooth germ. Additionally, we have shown that despite the early degradation of tooth germ basement membrane, the laminin molecule is still intact at the time of birth.
Animals↗
Basement membrane associated changes in the rat ventral prostate following castration.
This study focuses on the basement membrane associated modifications that take place after androgen blockade, by studying some of its main components, through histochemical, immuno-histochemical and Western blotting tests, and its ultrastructural aspects. It was demonstrated that laminin and collagen type IV remain associated with a thickened basement membrane and that there is an apparent increase in heparan sulfate content 21 days after castration. Ultrastructurally, basal lamina appeared extensively folded and pleated. It was also observed that detachment of epithelial cells is not dependent of basal lamina degradation and that the free basal lamina surfaces are folded by the action of adjacent cells. We have also observed some aspects of smooth muscle cell degeneration and death, that lead to modifications of the associated basal lamina. In this case, residual basal lamina also shows extensive folding. The results suggested that degradation of excess basement membrane does not occur or is a very slow process within the period examined, and that basement membrane is left re-organized but ultrastructurally and compositionally unaffected.
Androgens↗
Immunisation of Brazilian dentists against hepatitis B.
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Brazil↗
Use of TCA as a decalcifying agent for laminin immunohistochemistry.
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Animals↗
Immunochemical characterization and distribution of laminin in the rat tongue.
Anti-laminin serum was used to investigate the distribution and composition of laminin in the rat tongue. Immunohistochemical analysis showed that laminin is a useful marker to study the microscopic anatomy of the tongue. Laminin is expressed in most tissues that form this organ, such as smooth and striated muscle, epithelium, nerves, and endothelium, allowing the observation of fine details of neuromuscular junctions, taste papillae and microvasculature. Western blot analysis revealed that laminin extracted from the tongue differs from EHS tumor laminin. Tongue laminin contains B chains, while A chain seems to be present only in small amounts.
Animals↗
Laminin and collagen IV distribution and ultrastructure of the basement membrane of the gingiva of the rat incisor.
The continuous growth of the rat incisor is associated with renovation of the junctional epithelium and resorption of the periodontal ligament. The circumdental papilla separates the connective tissue suffering resorption from the rest of the gingiva. Laminin and collagen IV were detected by the immunoperoxidase technique on the basement membrane of all regions of the gingival epithelium of the rat incisor, except the internal basal lamina and the internal surface of the circumdental papilla. The internal basal lamina is formed by a granular electron-dense material, without the organization of a typical basal lamina. Areas of the internal surface of the circumdental papilla, negative for laminin and collagen IV, lack the basal lamina. These data suggest that these molecules are not components of the dento-epithelial junction of the distal surface of the rat incisor. In addition, the basal lamina is absent or fragmented on the internal surface of the circumdental papilla, adjacent to the areas of the connective tissue undergoing resorption.
Animals↗
Molecular analysis of spontaneous nephrotropic anti-laminin antibodies in an autoimmune MRL-lpr/lpr mouse.
To explore the genetic relationship between anti-laminin and anti-DNA autoantibodies (autoAb), VH gene and gene family expression were determined among autoAb derived from an individual 6-mo-old MRL-lpr/lpr mouse. Whereas 85% of the anti-DNA Ig were identified by one of two VH family probes, 7183 and VHJ558, none of the anti-laminin antibodies (Ab) examined were recognized by these probes. Subsequent V region sequence analysis of three of the anti-laminin Ab revealed that they in fact utilized a J558 VH gene (VH50). Furthermore, FR2 and CDR2 oligonucleotide probes complementary to VH50 recognized multiple anti-laminin Ab by Northern blot analysis; the FR2 probe recognized two control anti-DNA Ab, but neither probe recognized anti-DNA Ab from the same mouse. Polymerase chain reaction amplification of MRL-lpr/lpr genomic liver DNA using primers generated from VH50 and Vk50 sequences indicated that all three anti-laminin Ig have a single replacement mutation in both their VH and Vk genes. Search of the nucleic acid databases revealed that both germline VH and Vk genes are expressed unmutated by murine lupus anti-dsDNA autoAb, previously sequenced in other laboratories. Sequence comparisons suggest that differences in anti-DNA and anti-laminin reactivity may be dependent upon somatically generated differences in the CDR3 regions of the H and L chains. The results indicate that lupus anti-laminin Ab can arise from distinct B cell populations but express the same unmutated germline V region genes as lupus anti-dsDNA autoAb. They further raise the possibility that these distinct B cell populations may be activated and expanded either: independently, by distinct Ig receptor ligands such as the Ag, laminin and DNA; or simultaneously, by a common ligand such as an anti-Id recognizing a common V region epitope.
Amino Acid Sequence↗