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Biomedical subjects

S Lindskog

Publications and source records attributed to S Lindskog.

At least 37 records · Page 2Linked to original sources
PubMed1998

Crystal structure of carbonic anhydrase from Neisseria gonorrhoeae and its complex with the inhibitor acetazolamide.

The crystal structure of carbonic anhydrase from Neisseria gonorrhoeae has been solved to a resolution of 1.78 A by molecular replacement using human carbonic anhydrase II as a template. After refinement the R factor was 17.8% (Rfree=23.2%). There are two molecules per asymmetric unit (space group P21), but they have essentially identical structures. The fold of the N. gonorrhoeae enzyme is very similar to that of human isozyme II; 192 residues, 74 of which are identical in the two enzymes, have equivalent positions in the three-dimensional structures. This corresponds to 85% of the entire polypeptide chain of the bacterial enzyme. The only two cysteine residues in the bacterial enzyme, which has a periplasmic location in the cell, are connected by a disulfide bond. Most of the secondary structure elements present in human isozyme II are retained in N. gonorrhoeae carbonic anhydrase, but there are also differences, particularly in the few helical regions. Long deletions in the bacterial enzyme relative to human isozyme II have resulted in a considerable shortening of three surface loops. One of these deletions, corresponding to residues 128 to 139 in the human enzyme, leads to a widening of the entrance to the hydrophobic part of the active site cavity. Practically all the amino acid residues in the active site of human isozyme II are conserved in the N. gonorrhoeae enzyme and have similar structural positions. However, the imidazole ring of a histidine residue, which has been shown to function as a proton shuttle in the catalytic mechanism of the human enzyme, interacts with an extraneous entity, which has tentatively been identified as a 2-mercaptoethanol molecule from the crystallization medium. When this entity is removed by soaking the crystal in a different medium, the side-chain of His66 becomes quite mobile. The structure of a complex with the sulfonamide inhibitor, acetazolamide, has also been determined. Its position in the active site is very similar to that observed in human carbonic anhydrase II.

Acetazolamide↗
PubMed1998

Effect of 3-O-methylglucose on insulin secretion in the perfused dog and rat pancreas.

We recently reported that the nonmetabolizable glucose analogue, 3-O-methylglucose, stimulates somatostatin secretion in the perfused dog pancreas. In this study, we report that 3-O-methylglucose also stimulates insulin secretion in the dog pancreas. The effect was present at 5.5 mM glucose (p < 0.001) but not at O or 2.7 mM glucose. The inhibitor of glucose metabolism, mannoheptulose, blocked the insulinotropic action of 3-O-methylglucose. In contrast, 3-O-methylglucose had no effect on insulin secretion in the perfused rat pancreas. We conclude that 3-O-methylglucose stimulates insulin secretion in the dog and that the effect requires the presence of stimulatory concentrations of D-glucose.

3-O-Methylglucose↗
PubMed1998

Chlorhexidine as a root canal medicament for treating inflammatory lesions in the periodontal space.

The purpose of the present study was to investigate the therapeutic effect of intra-canal application of chlorhexidine on inflammatory root resorption. Dental pulps from monkey incisors were infected and resealed prior to extraction of the teeth. Root dentin was mechanically exposed and the teeth were replanted under aseptic conditions. After 4 weeks, the experimental teeth were dressed with chlorhexidine gel and resealed. Animals were sacrificed 4 weeks later, and their jaws prepared for histologic examination. Results showed that both marginal and apical periodontal inflammation and resorption were reduced in the chlorhexidine-treated teeth. It was concluded that the use of intra-canal chlorhexidine may be a useful adjunct in the treatment of inflammatory root resorption, but further human trials need to be undertaken before its clinical use can be recommended.

Animals↗
PubMed1998

Cervical root resorption associated with guided tissue regeneration: a case report.

Root surface resorption, ankylosis (replacement resorption) and alveolar bone resorption are not uncommon sequelae to periodontal healing in both animal and human trials whether the treatment objective is regenerative, preventive, or conservative. The present report describes a case with progressive cervical root resorption in a patient who received periodontal regenerative treatment with guided tissue regeneration (GTR). A 46-year-old woman was referred for treatment of severe periodontitis. Remaining radiographic attachment was less than 50%. Following a period of 18 months, during which non-surgical and surgical therapies were performed, angular defects were diagnosed on radiographs and recurrent bleeding periodontal pockets (6 mm) were found in the proximal areas of 24 and 25. Root caries was not present. Periodontal surgery with GTR was performed in this area. No immediate postsurgical complications were noted. Two years later, clinical and radiographic examinations revealed gingival recession with bleeding periodontal pockets (6 mm) which had partly uncovered severe proximal cervical resorptions in 25. Root surface caries was not present. Following surgical inspection, the root of 25 was removed. The root was subsequently prepared for histological analysis. Resorption cavities covered almost the entire cervical proximal surface of the root above intact infracrestal cementum and were covered by numerous CD68+, both mononuclear and multinucleated cells. In a central area as indicated on the radiographs, the cavities penetrated into the root canal. There was no evidence of root caries.

Antigens, CD↗
PubMed1997

The complete sequence, expression in Escherichia coli, purification and some properties of carbonic anhydrase from Neisseria gonorrhoeae.

The complete nucleotide sequence of the carbonic anhydrase gene from Neisseria gonorrhoeae has been determined. The gene encodes a 252-residue polypeptide with a molecular mass of 28085 Da. The gene has been cloned and overexpressed in Escherichia coli, and the enzyme has been purified. A 26-residue signal peptide is cleaved off by the E. coli processing machinery. Thus, the isolated enzyme contains 226 amino acid residues with a molecular mass of 25314 Da. Most of the enzyme seems to be produced as a soluble protein located in the periplasm of E. coli. The enzyme is homologous to carbonic anhydrases from the animal kingdom; it is an alpha-carbonic anhydrase. A comparison with the amino acid sequences of human carbonic anhydrases I and II suggests that the secondary structures are essentially intact in the bacterial enzyme but that several loops are much shorter than in the human forms. Most of the active-site residues are identical to those found in the high-activity human isozyme II. The bacterial enzyme has a high CO2 hydration activity with a k(cat) of 1.1 x 10(6) s(-1) and Km of 20 mM at pH 9 and 25 degrees C. The enzyme also catalyzes the hydrolysis of 4-nitrophenyl acetate. The pH/rate profile can be described as a titration curve with pKa of 6.7 and a maximal value of the catalytic second-order rate constant, k(enz), of 130 M(-1) x s(-1).

Amino Acid Sequence↗
PubMed1997

Cellular expression of neurotrophin mRNAs during tooth development.

Target-derived neurotrophins support and sustain peripheral sensory neurons during development. In addition, it has been suggested that these growth factors could have developmental functions in non-neuronal tissues. To further elucidate the possible roles of neurotrophins in tooth morphogenesis and innervation, we have used in-situ hybridization to determine the specific sites of neurotrophin gene activity in pre- and postnatal rat jaws from E16 to P7. All four neurotrophins were expressed during tooth development with specific temporospatial patterns. Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) mRNAs were mainly detected in the dental papilla/pulp in postnatal animals, and the pattern of expression correlated with the onset of dental innervation. In contrast, neurotrophin 3 (NT3) and neurotrophin 4 (NT4) mRNA expression patterns were predominantly epithelial and were strongest during early developmental stages when teeth are not yet innervated. Dental papilla NGF-mRNA expression was first seen in both epithelium and mesenchyme and later shifted to the odontoblast layer and the subodontoblast zone. BDNF-mRNA labeling was present in low levels in the early dental organ, but increased in the pulp and in the odontoblast cell layer of the developing teeth at later developmental stages. Both NT3 and NT4 mRNA were observed in the prenatal oral epithelium and the inner dental epithelium. NT3-mRNA labeling was seen mainly in the cervical loop region, fissure system depressions and cuspal tops, while NT4 mRNA was more evenly distributed in the dental epithelium. At P7, NT3-mRNA labeling was below detection level and NT4 mRNA expression was lower than at prior stages. Complementary to reports on the presence of low-affinity neurotrophin receptor (LANR), trkB and trkC mRNA in the developing teeth, our results suggest that neurotrophins may have multiple functions during tooth morphogenesis. Neurotrophins might participate in epithelial-mesenchymal interactions in early tooth morphogenetic events such as proliferation and differentiation of epithelial and mesenchymal cells. In addition, based on mRNA localization in postnatal animals, we also suggest that NGF and BDNF (beside glial cell line-derived neurotrophic factor) might participate in establishing and maintaining the innervation of the teeth, thus acting as classical neurotrophic factors.

Animals↗
PubMed1997

Structure and mechanism of carbonic anhydrase.

Carbonic anhydrase (CA; carbonate hydro-lyase, EC 4.2.1.1) is a zinc-containing enzyme that catalyzes the reversible hydration of carbon dioxide: CO2+ H2O<-->HCO3(-)+H+. The enzyme is the target for drugs, such as acetazolamide, methazolamide, and dichlorphenamide, for the treatment of glaucoma. There are three evolutionarily unrelated CA families, designated alpha, beta, and gamma. All known CAs from the animal kingdom are of the alpha type. There are seven mammalian CA isozymes with different tissue distributions and intracellular locations, CA I-VII. Crystal structures of human CA I and II, bovine CA III, and murine CA V have been determined. All of them have the same tertiary fold, with a central 10-stranded beta-sheet as the dominating secondary structure element. The zinc ion is located in a cone-shaped cavity and coordinated to three histidyl residues and a solvent molecule. Inhibitors bind at or near the metal center guided by a hydrogen-bonded system comprising Glu-106 and Thr-199. The catalytic mechanism of CA II has been studied in particular detail. It involves an attack of zinc-bound OH- on a CO2 molecule loosely bound in a hydrophobic pocket. The resulting zinc-coordinated HCO3- ion is displaced from the metal ion by H2O. The rate-limiting step is an intramolecular proton transfer from the zinc-bound water molecule to His-64, which serves as a proton shuttle between the metal center and buffer molecules in the reaction medium.

Animals↗
PubMed1997

Effect of different concentrations of EDTA on smear removal and collagen exposure in periodontitis-affected root surfaces.

The purpose of the present study was to explore the possibility of obtaining an acceptable smear-removing and collagen-exposing effect following EDTA etching with concentrations lower than supersaturation (24%). A flat dentin surface was created on human teeth extracted due to severe periodontitis. The teeth were etched with the following concentrations of EDTA for 2 min: 1.5% (group I), 5% (group II), 15% (group III) and 24% (group IV), and evaluated with scanning electron microscopy with respect to smear layer removal and exposure of root-associated collagen fibers. It was found that neither 1.5% nor 5% EDTA etching was sufficient to dissolve the smear layer to any higher degree. Etching with 15% EDTA dissolved the smear more effectively than the lower concentrations and collagen fibers were sparsely found. Following etching with 24% EDTA (group IV), significantly less smear was seen on the dentin surfaces than in any of the other groups and collagen fibers were seen covering the entire intertubular dentin surfaces. Based on these findings, the concentration of EDTA should be somewhere between 15 to 24% in order to obtain an acceptable smear removing and collagen-exposing effect within a clinically acceptable time period.

Acid Etching, Dental↗
PubMed1997

Galanin exerts dual action on inositol-specific phospholipase C activity in isolated pancreatic islets.

The intracellular mechanism whereby the neuropeptide galanin inhibits insulin secretion is not establish, since the peptide affects several signal pathways, including intracellular messengers such as calcium and cyclic AMP. In this study, we have assessed the effect of galanin on the inositol-specific phospholipase C (iPLC) activity in isolated rat pancreatic islets. The iPLC activity was measured as the generation of inositol 1,4,5-trisphosphate and its metabolite inositol 1,3,4-trisphosphate from the hydrolysis of polyphosphoinositides. Inositol phosphates were measured by anion-exchange fast protein liquid chromatography (FPLC) analysis of extracts from islets prelabelled with myo-3H-inositol. Galanin (1 to 100 nM) significantly increased the glucose-induced (12 mM) accumulation of inositol 1,4,5-trisphosphate after 2 min, but this stimulation of iPLC activity was followed by a significant suppression after 15 min. In the absence of extracellular calcium, both the stimulatory and inhibitory effects of galanin on the iPLC activity vanished. We therefore conclude that galanin initially stimulates iPLC in a calcium-dependent manner, followed by a secondary inhibitory effect. The secondary inhibition of iPLC activity might contribute to the insulinostatic action of the neuropeptide.

Animals↗
PubMed1997

Periodontal healing in horizontal and vertical defects following surgical or non-surgical therapy.

The aim of the present study was to investigate any relationship between the level of oral hygiene and probing pocket depth reduction over time after periodontal treatment in sites with either vertical or horizontal destructions. The investigation was conducted as a retrospective study on a 3-year consecutive referral population of periodontitis-prone patients based on full-mouth oral radiographic examinations, probing pocket depth registrations and plaque scores. The analyses were performed on a final sample of 3064 sites in 107 patients with regression analysis after adjusting for dependence within the patient. Probing pocket depth was significantly less reduced over time in sites with vertical destructions compared to sites with horizontal destructions following non-surgical treatment. Furthermore, the difference in probing pocket depth reduction between vertical and horizontal defects following non-surgical treatment increased over time in sites with plaque compared to sites without plaque, thus reflecting the importance of the patient's plaque control, especially in sites with vertical destructions. However, the difference in probing pocket depth reduction between vertical and horizontal defects did not increase over time for surgically treated teeth, a finding which probably can be attributed to a more thorough debridement of vertical defects during surgery and/or osteoplasty/osteoectomy limiting the surface area upon which a long junctional epithelium can form, which may facilitate recurrence of a periodontal pocket.

Adult↗
PubMed1997

Prognosis and mortality of root-resected molars.

The purpose of the present study was to compare tooth mortality of root-resected molars with that of root-filled, single-rooted teeth. Survival rates were 68% for root-resected molars and 77% for root-filled single-rooted teeth over a 10-year period. This difference was not statistically significant. Ten-year survival of root-resected molars in patients with radiographic attachment loss in single-rooted teeth of greater than 6 mm was 56% while survival was as high as 89% for root-resected molar patients with radiographic attachment loss in single-rooted teeth less than or equal to 6 mm. In conclusion, the prognosis of root-resection is not poorer than the prognosis of single-rooted teeth with an equal susceptibility to periodontitis, if endodontic conditions and maintenance care are optimal.

Adult↗
PubMed1997

Ultrasonic subgingival root planing and EDTA etching in a one-step procedure.

The purpose of the present study was to investigate if ultrasonic debridement and EDTA etching could be combined in a ore-step procedure. Tap water was exchanged for a 24% EDTA solution as irrigation agent during root planing with a piezo scaler. 24 human teeth extracted due to severe periodontal disease were used and the results were evaluated with scanning electron microscopy. Results indicate that synergistic effects (smear removal and exposure of collagen fibers) were negligible with conventional application of the irrigation fluid to the working area of the scaling tip while a marked effect was evident when the EDTA solution was applied directly to the working area of the piezoelectric scaler through a customized tip.

Acid Etching, Dental↗
PubMed1996

Two point mutations convert a catalytically inactive carbonic anhydrase-related protein (CARP) to an active enzyme.

A murine carbonic anhydrase-related protein (CARP) has been expressed in Escherichia coli and purified to near homogeneity. The polypeptide chain consists of 290 amino acid residues and has a calculated molecular mass of 32,950 Da. By introducing two mutations, Arg117 --> His and Glu115 --> Gln, we created a metal-binding center homologous to that in the carbonic anhydrases from the animal kingdom. In contrast to unmodified CARP, this double mutant was isolated as a 1:1 zinc-protein complex. While unmodified CARP is catalytically inactive, the mutant catalyzes CO2 hydration with a significantly higher efficiency than the mammalian low-activity carbonic anhydrase isozyme III. The activity is strongly inhibited by the powerful and selective carbonic anhydrase inhibitor, acetazolamide.

Acetazolamide↗
PubMed1996

Stimulation of somatostatin secretion by 3-O-methylglucose in the perfused dog pancreas.

CONCLUSION: 3-O-methylglucose stimulates somatostatin secretion from the dog pancreas by a glucose-dependent and glucose-like effect. Therefore, it is possible that 3-O-methylglucose-stimulated somatostatin secretion is dependent on glucose metabolism. BACKGROUND: Somatostatin secretion from the endocrine pancreas is stimulated by glucose, glyceraldehyde, and dihydroxyacetone but not affected by fructose, galactose, or ribose. Whether the nonmetabolizable glucose analog, 3-O-methylglucose affects somatostatin secretion is, however, not known. METHOD: We therefore, examined whether the glucose analog affects somatostatin secretion in the perfused dog pancreas. RESULTS: We found that when added to a medium containing 2.7 mM or 5.5 mM D-glucose, 3-O-methylglucose (10 mM) stimulated somatostatin secretion to the same extent as did an equivalent dose of D-glucose. The same stimulation was observed also with arginine at 2.5 mM in the perfusion medium. In contrast, 3-O-methylglucose did not stimulate somatostatin secretion in the absence of glucose in the perfusion medium. Mannoheptulose (5 mM), which inhibits glucose metabolism, completely blocked the secretion to both hexoses.

3-O-Methylglucose↗
PubMed1996

Cellular expression of GDNF mRNA suggests multiple functions inside and outside the nervous system.

Glial-cell-line-derived neurotrophic factor (GDNF) is a distant member of the transforming growth factor-beta family and has potent neurotrophic effects on several classes of neurons including dopamine neurons and motoneurons. Here, we have used in situ hybridization to describe the development of the cellular expression of GDNF mRNA pre- and postnatally. Consistent with dopaminotrophic activity, GDNF mRNA is expressed in the developing basal ganglia and the olfactory tubercle. It is also found in a thalamic nucleus, in neurons of the substantia innominata, in the developing Purkinje neurons and the developing locus coeruleus area, and in trigeminal brainstem nuclei. In the spinal cord, neuronal expression is found in Clarke's column. GDNF mRNA is also expressed in the dorsal horns during development. Additional GDNF mRNA expression in the head region includes the carotid body, the retina, the vibrissae, the inner ear, the ear canal, and epithelium in the nasal cavity. Prominent expression is also found in the developing teeth. The widespread expression of GDNF in developing skeletal muscle is consistent with trophic activity on alpha-motoneurons. The smooth muscle layers of the gastrointestinal tract are also strongly positive. A very strong signal is found in the outer mesenchyme of the developing metanephric kidney. We conclude that GDNF mRNA is expressed in many different cellular systems inside and outside the central nervous system during development, suggesting multiple functions of GDNF in the developing organism.

Animals↗
PubMed1996

Healing of the root surface-associated periodontium: an immunohistochemical study of orthodontic root resorption in man.

The purpose of the present investigation was to study resorption and regeneration of periodontal tissues incident to orthodontic tooth movement, in particular cells resorbing the root surface and the subsequent regeneration of the periodontal epithelial network and forming reparative cementum. The study was carried out using a select number of immunohistochemical markers on extracted human teeth which had been treated orthodontically. The most striking finding in the resorbing areas was the presence of what appeared to be two populations of KP 1+ mononuclear cells located at a distance of 50-100 microns from the root surface and multinucleated cells in resorption lacunae in close contact with the root surface. KP 1+ has previously not been reported for odontoclasts. The mononuclear KP 1+ cells in the periodontal ligament may represent either precursors to odontoclasts or phagocytic scavenger cells of the macrophage lineage. The subsequent healing of the resorption lacunae was characterized by re-establishment of nervous, vascular and epithelial tissues as evidenced by S-100+ filamentous delicate structures, factor VIII+ vessels and cytokeratin+ clusters of cells, respectively. However, cytokeratin+ single cells in close contact with the unresorbed cementum did not re-appear within the healing period. Although the present results are not quantitative in nature, cementoblasts located in the vicinity of resorption lacunae, especially healing ones, appeared to show an up-regulation of epidermal growth factor (EGF) receptors. It may be suggested the intense positive staining for EGF receptors may be an expression of an auto- or paracrine stimulatory pathway increasing the rate of reparative cementum formation.

Adolescent↗
PubMed1996

Tissue response to space closure in monkeys: a comparison of orthodontic magnets and superelastic coil springs.

Interest in using magnets for generating orthodontic forces started with the widespread availability of rare earth magnetic alloys. In vivo studies have indicated that a static magnetic field and/or corrosion products from the magnetic materials may induce biological effects when in close contact with cells or tissues. In the clinical situation, orthodontic magnets are often situated some distance away from the gingiva and bone. Consequently, the previously observed biological effects may not be found in an experimental situation mimicking the clinical setting. Thus, the present experimental study was undertaken to test this hypothesis using commercially available cobalt-samarium magnets for orthodontic treatment in comparison to treatment with Sentalloy closed coil springs with respect to possible side effects on alveolar bone growth, gingival epithelial thickness as well as rate of space closure. Corrosion of the uncovered areas of the magnets was already evident after 6 weeks. No statistical differences were found between the magnet and coil spring specimens with respect to rate of space closure, bone formation or epithelial thickness. The only two variables that differed significantly between magnet and coil spring specimens was that there were more resorption and more tetracycline labelled osteocyte lacunae under the magnets. In conclusion, although some marginal statistical differences were found between the magnet and coil spring specimens with respect to cell and tissue reactions, the near lack of cell and tissue effects of the magnets in the present clinical experimental situation compared to previous studies in which the magnets were positioned in close contact with the tissue under study, indicate limited adverse clinical effects.

Alveolar Process↗