PubMed Health⌕ Search

Biomedical subjects

T J Sims

Publications and source records attributed to T J Sims.

At least 55 records · Page 3Linked to original sources
PubMed1995

Non-enzymic glycation of fibrous collagen: reaction products of glucose and ribose.

Non-enzymic glycation of collagen involves a series of complex reactions ultimately leading to the formation of intermolecular cross-links resulting in changes in its physical properties. During analysis for the fluorescent cross-link pentosidine we identified the presence of an additional component (Cmpd K) in both glucose and ribose incubations. Cmpd K was formed more quickly than pentosidine in glucose incubations and more slowly than pentosidine in ribose incubations. Cmpd K represented 45% of the total fluorescence compared with 15% for pentosidine in glucose incubations and 25% of the total fluorescence compared with 30% for pentosidine in the ribose incubations. Cmpd K is not an artefact of in vitro incubations, as it was shown to be present in dermal tissue from diabetic patients. Subsequent high-resolution h.p.l.c. analysis of glucose-incubated collagen revealed Cmpd K comprise two components (K1 and K2). Further, a similar analysis of Cmpd K from the ribose incubations revealed two different components (K3 and K4). These differences indicate alternative mechanisms for the reactions of glucose and ribose with collagen. The amounts of these fluorescent components and the pentosidine cross-link determined for both glucose and ribose glycation were found to be far too low (about one pentosidine molecules per 200 collagen molecules after 6 months incubation with glucose) to account for the extensive cross-linking responsible for the changes in physical properties, suggesting that a further additional series of cross-links are formed. We have analysed the non-fluorescent high-molecular-mass components and identified a new component that increases with time of in vitro incubation and is present in the skin of diabetic patients. This component is present in sufficient quantities (estimated at one cross-link per two collagen molecules) to account for the changes in physical properties occurring in vitro.

Adolescent↗
PubMed1995

Abnormal collagen cross-linking in the cartilage of a diastrophic dysplasia patient.

The abnormal organization of the cartilage collagen in diastrophic dysplasia is not generally reflected in the levels of the major stabilizing cross-link, hydroxylysyl-pyridinoline. However, in one case there was a marked decrease in the pyridinoline concomitant with the appearance of two unknown components in the cross-link region of the chromatogram. A decrease in cross-linking of this magnitude could lead to mechanically weakened cartilage. Insufficient material was available to characterize these unknown components. The disorganization of cartilage in some cases of diastrophic dysplasia could therefore be due to post-translational modifications, including defective cross-linking.

Amino Acids↗
PubMed1994

Regrowth of dorsal root axons into a radiation-induced glial-deficient environment in the spinal cord.

Exposure of the lumbosacral spinal cord of early postnatal rats to X-rays reduces the glial populations within the irradiated region. The present study examines the ability of axons of a dorsal root subjected to a crush-freeze lesion to grow back into this glial-deficient spinal cord environment, in contrast to the non-irradiated rat. Ultrastructural examination of the dorsal root entry zone (DREZ) 60 days after root injury revealed a well-formed astrocytic scar in this zone and adjacent regions of spinal cord in non-irradiated rats. In contrast, scar formation did not occur in irradiated root-lesioned animals in which the astrocytic response was quite limited. Axons were present in the DREZ and underlying spinal cord in irradiated root-lesioned rats at this time but were absent from these regions in the non-irradiated lesioned controls. These ultrastructural findings are highly suggestive that axons are capable of regrowth into the irradiated spinal cord. Axonal regrowth was assessed further by tracing techniques after application of a combination of peroxidase-labeled wheat germ agglutinin and horseradish peroxidase to the cut end of the root distal to the previously injured site. Labeled axons were readily identified within the spinal gray matter in irradiated lesioned but not in the non-irradiated lesioned rats. These data, together with the ultrastructural observations, are supportive of regrowth of the dorsal root axons into the spinal cord. The radiation-induced changes in the glial populations are discussed with regard to conversion of a normally non-permissive environment into one conducive for axonal regrowth.

Animals↗
PubMed1994

Glial development in primary cultures established from normal and X-irradiated neonatal spinal cord.

The glial population of the lumbosacral spinal cord of the rat can be markedly depleted by exposure to ionizing radiation during the first postnatal week. Identification of specific cell populations which survive the exposure to radiation is difficult in situ; therefore, the present investigation used in vitro approaches to address issues related to specific phenotypes and maturational states of glia in cultures derived from non-irradiated (control) and irradiated (experimental) lumbosacral spinal cords of 3-day-old rats. Cultures were established from the spinal cords 2 to 4 hours following irradiation and were compared to cultures from non-irradiated, littermate controls. By 4 days in vitro (DIV) the numbers of cells in experimental cultures were profoundly reduced when compared to controls, and this reduction persisted through the termination of the study (8 DIV). In addition to reduction in numbers, astrocyte phenotypes were altered in experimental cultures, with greater proportions of the astrocyte population being constituted by the flat angular, large angular, and pancake types and a lesser proportion by stellate cells. The non-astrocytic cell types were dramatically reduced as evidenced by the paucity of oligodendrocytes immunoreactive for galactocerebroside and of small, non-process bearing cells binding the lectin, Griffonia (Bandeiraea) simplicifolia, a marker for microglia. Experimental cultures contained an increased incidence of binucleate astrocytes, an increase not restricted to a particular astrocyte phenotype. This study established the feasibility of utilizing this combined in vivo/in vitro approach in assessment of glial populations in immature spinal cords, and further investigations are in progress using this model.

Animals↗
PubMed1994

Regeneration of dorsal root axons into experimentally altered glial environments in the rat spinal cord.

Exposure of the lumbar spinal cord of rats to X-rays 3 days after birth results in changes in the composition of central glia. Shortly after irradiation, there is both retardation of central myelin formation and a loss of integrity of the astrocyte-derived glia limitans on the dorsal surface of the cord. Subsequently, Schwann cells invade, undergo division and myelinate axons in the dorsal funiculi in the irradiated region of the cord, creating there an environment similar to that of peripheral nerve. The present study was undertaken to compare the ability of lesioned dorsal root axons to grow back into the altered glial environments that exist within the spinal cord after irradiation. This regrowth was assessed by injecting Fluoro-Gold into the spinal cord and subsequently examining neurons in the dorsal root ganglia (DRG) for the presence of this label. Numbers of retrogradely labeled neurons were counted in the DRG in both injured and contralateral non-injured sides. Non-irradiated control rats had almost no labeled DRG neurons on the injured side, whereas Fluoro-Gold labeled neurons were observed in substantial numbers in the DRG on the injured side of irradiated rats. There was a definite trend in the data, indicating that the longer the interval between irradiation and root injury, the greater the number of labeled neurons. Since the Fluoro-Gold labeling technique does not allow for visualization of the labeled axons within the spinal cord, a few animals were used to assess anterograde labeling with wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP/HRP) from the dorsal root into the spinal cord.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

The continuous elongation technique for severe Dupuytren's disease. A biochemical mechanism.

Continuous extension of Dupuytren's contracture prior to fasciectomy results in a softening of the tissue, allowing straightening of the fingers. The observed change in cross-link profile indicates an increase in newly synthesised collagen due to increased turnover. This was confirmed by demonstration of the increases in levels of the degradative enzymes, the neutral metalloproteinases, collagenase and gelatinase and the acidic cathepsins B and L. Both types of enzyme effectively depolymerize the collagen fibres, albeit by different mechanisms, leading initially to loss of tensile strength and ultimately to solubilization. We suggest that the increase in enzyme activity is generated by tension on the fibroblasts of this metabolically active tissue produced during the continuous extension of the retracted fingers. The weakening of the fibres by degradation and the increase in newly synthesized collagen provide an explanation for the extension of the tissue without trauma.

Amino Acids↗
PubMed1993

Chemistry of collagen cross-links: glucose-mediated covalent cross-linking of type-IV collagen in lens capsules.

The incubation of lens capsules with glucose in vitro resulted in changes in the mechanical and thermal properties of type-IV collagen consistent with increased cross-linking. Differential scanning calorimetry (d.s.c.) of fresh lens capsules showed two major peaks at melting temperatures Tm 1 and Tm 2 at approx. 54 degrees C and 90 degrees C, which can be attributed to the denaturation of the triple helix and 7S domains respectively. Glycosylation of lens capsules in vitro for 24 weeks caused an increase in Tm 1 from 54 degrees C to 61 degrees C, while non-glycosylated, control incubated capsules increased to a Tm 1 of 57 degrees C. The higher temperature required to denature the type-IV collagen after incubation in vitro suggested increased intermolecular cross-linking. Glycosylated lens capsules were more brittle than fresh samples, breaking at a maximum strain of 36.8 +/- 1.8% compared with 75.6 +/- 6.3% for the fresh samples. The stress at maximum strain (or 'strength') was dramatically reduced from 12.0 to 4.7 N.mm.mg-1 after glycosylation in vitro. The increased constraints within the system leading to loss of strength and increased brittleness suggested not only the presence of more cross-links but a difference in the location of these cross-links compared with the natural lysyl-aldehyde-derived cross-links. The chemical nature of the fluorescent glucose-derived cross-link following glycosylation was determined as pentosidine, at a concentration of 1 pentosidine molecule per 600 collagen molecules after 24 weeks incubation. Pentosidine was also determined in the lens capsules obtained from uncontrolled diabetics at a level of about 1 per 100 collagen molecules. The concentration of these pentosidine cross-links is far too small to account for the observed changes in the thermal and mechanical properties following incubation in vitro, clearly indicating that another as yet undefined, but apparently more important cross-linking mechanism mediated by glucose is taking place.

Animals↗
PubMed1993

Titer and subclass distribution of serum IgG antibody reactive with Actinobacillus actinomycetemcomitans in localized juvenile periodontitis.

Most patients with localized juvenile periodontitis (LJP) manifest serum IgG antibodies specifically reactive with antigens of Actinobacillus actinomycetemcomitans serotype b (Aa-b). Whether these antibodies are protective, destructive, or irrelevant to the progress of the disease remains unclear. We report results of studies aimed at assessing the subclass IgG responses in 35 LJP patients and 35 periodontally normal control subjects using well-characterized monoclonal antibody subclass reagents in an enzyme-linked immunosorbent assay. Our data show that the mean value for total IgG reactive with antigens of Aa-b was more than sevenfold higher for patients than for normal control sera (2349.6 micrograms/ml for patients vs 332.2 micrograms/ml for controls). Individual patients and control subjects were classified as high- or low-titer, using twice the median value for total anti-Aa-b IgG in control sera as the cutoff. Of 35 patients, 26 (74%) were high-titer, and 9 (26%) were low-titer. This compares to 5 normal control subjects (14%) high-titer and 30 (86%) low-titer. IgG2 accounted for the major quantitative response in both patients and control subjects. Indeed, the mean IgG2 values for both concentration and percentage of total specific IgG were greater than the combined values for specific anti-Aa-b IgG1, IgG3, and IgG4. Of the 26 high-titer sera, IgG2 predominated in 24, with IgG1 and IgG3 predominating in 1 each; IgG2 predominated in only 2 of the low-titer sera.

Actinobacillus Infections↗
PubMed1993

Schwann cell induction in the ventral portion of the spinal cord.

Schwann cell development can be induced in a predictable manner in the dorsal aspect of the lumbosacral spinal cord of the immature rat by exposing that structure to ionizing radiation. This development occurs in essentially all animals and becomes evident between 2 and 3 weeks postirradiation (P-I). Occasionally, intraspinal Schwann cells were observed ventrally at later intervals following irradiation, usually more than 45 days P-I. The present study focused on the development of Schwann cells within the ventral portion of the lumbosacral spinal cord in 53 animals followed for periods up to 7 months P-I. Ventrally located intraspinal Schwann cells developed in approximately 40% of these animals, in contrast to the development dorsally in all animals. The ventrally located aggregates were generally smaller than those dorsally and occurred more frequently in gray matter than in white matter. An interesting feature of the ventrally located Schwann cells was that they were often associated with blood vessels, which raised the possibility that these cells developed from undifferentiated cells of the vascular walls or used the vessels as a pathway for migration.

Animals↗
PubMed1993

Effects of treatment on antibody titer to Porphyromonas gingivalis in gingival crevicular fluid of patients with rapidly progressive periodontitis.

Twenty-eight patients diagnosed as having rapidly progressive periodontitis (RPP) were enrolled in a study in which samples of subgingival microflora were harvested from test teeth and assayed for the presence of Porphyromonas gingivalis, and GCF collected and analyzed by ELISA for specific antibody for P. gingivalis. Clinical conditions were measured and recorded, and treatment by scaling and root planing provided at baseline and at 3, 6, 9, and 12 months. Reduction in pocket depth, stabilization of attachment level, and resolution of inflammation were comparable to previously reported values. By 3 months, mean and median specific antibody concentration had decreased, and continued to decrease through 12 months. The proportion of samples in which specific antibody was not detectable increased from 27% at baseline to 73% at month 12. GCF samples from sites at which P. gingivalis was present had greater than 2-fold higher median specific antibody than samples from P. gingivalis-negative sites. At baseline, specific antibody titer of 30-second GCF samples positively correlated with pocket depth, and GCF volume significantly correlated with antibody titer and concentration, and with pocket depth. In addition, change in specific antibody titer of 30-second samples from baseline to both 6 and 12 months correlated positively with pocket depths. Thus sites infected by P. gingivalis manifested high levels of specific antibody, and levels were related to clinical status. Following treatment, antibody levels decreased significantly as pocket depths decreased, attachment levels stabilized, and inflammation resolved.

Adult↗
PubMed1993

Biochemical changes in the collagen of human osteoporotic bone matrix.

Although it is known that collagen imparts mechanical strength to bone no detailed biochemical analysis has been made of osteoporotic bone collagen. We report for the first time significant changes in the properties of the collagen. Analysis of collagen types revealed little change in the proportion of Type III collagen, but in some cases there was a significant loss of the Type VI. However, the major differences were observed in the post-translational modifications, namely, in the stabilizing cross-links and the hydroxylation of the collagen. These changes indicated a higher turnover in the head region compared to the neck region of the femoral head and are consistent with the susceptibility of the neck region to fracture. Clearly, the collagen is altered in osteoporosis and these changes may play a role in the pathogenesis of the disease.

Aged↗
PubMed1992

Quantitative analysis of collagen and elastin cross-links using a single-column system.

The separation of both the immature and mature cross-links present in collagen together with the stable cross-links of elastin has been achieved on a single ion-exchange column. This technique avoids the current necessity for two different systems, ion-exchange and high-performance liquid chromatography-fluorescence techniques. The value of the method is illustrated by the comparison of the cross-link contents of aging bovine skin, from foetal to old age.

Aging↗
PubMed1992

Post-translational modifications in the collagen of human osteoporotic femoral head.

No detailed biochemical analysis has been made of the possible compositional changes in the collagen relating to the fragility of osteoporotic bone. We report for the first time significant changes in the compositional properties of the collagen. The major differences were observed in the post-translational modifications, namely, in the hydroxylation of lysine residues and the nature of the stabilizing cross-links of the collagen fibre. The increase in hydroxylation was greater in the head region compared to the neck region of the femoral head, whilst the decrease in the intermediate cross-links was greater in the neck region. Clearly, the collagen is altered in osteoporosis and it is important that these changes are recognised in studies of bone metabolism in osteoporosis since they may play a role in the pathogenesis of the disease.

Adult↗
PubMed1992

Glial response to dorsal root lesion in the irradiated spinal cord.

Exposure of the rat lumbar spinal cord to X-rays during the early postnatal period results in a marked reduction in the glial populations within the irradiated region. The present study was undertaken to determine what effects this reduction of glia, particularly astrocytes, has on the pattern and characteristics of the scar formation that follows root injury in the normal spinal cord. Morphological assessments 60 days following injury of the right L4 dorsal root revealed a distinct difference in the extent of the astrocyte response between the irradiated and the nonirradiated rats. In the nonirradiated animals, a thick astrocytic scar composed of multiple layers of astrocyte processes formed over the dorsal horn and adjacent portions of the dorsal surface of the cord. This astrocytic response was not confined to the surface of the spinal cord but extended also into the root, i.e., into regions normally considered as PNS. In irradiated rats, the astrocytes did not form a thick scar nor did they extend into the injured root. Instead, they formed a glia limitans and were at most only one or two layers thick over the region of cord comparable to that occupied by the thick astrocytic scar in the nonirradiated rats. Mechanisms involved in the glial response of the irradiated spinal cord to dorsal root injury are discussed, particularly with regard to the possible positive effect that this reduction in scar formation may have on regrowth of injured dorsal root axons into the spinal cord environment.

Animals↗
PubMed1991

Radial glia give rise to perinodal processes.

Nodes of Ranvier in the central nervous system in mammals are characterized by the presence of perinodal astrocytic processes. This study examines the association between processes of radial glia and the axolemma at nodes of Ranvier in the spinal cord of the mature axolotl, an animal in which radial glia represent a large portion of the total glial population. The radial glial cells have their cell bodies located close to the central canal. Those situated dorsal to the canal send long processes to the dorsal surface of the spinal cord. Along this trajectory these processes coalesce into large fascicles in the midline and form the dorsal median septum. Slender branches rise from the processes in these fascicles and extend into the adjacent white matter to terminate in close apposition to the axolemma at nodes of Ranvier. This arrangement provides an intracellular pathway extending from the perinodal region to the surface of the spinal cord. Radial glia ventral to the central canal give rise to processes that project to the glia limitans adjacent to the ventral spinal artery. These ventrally projecting processes appear to be more irregular in their branching pattern than their dorsal counterparts. Multiple slender processes are seen in close apposition to the nodal axolemma of myelinated axons in the ventral white commissure, again providing an intracellular pathway that runs from the perinodal region to the cord surface. In one instance a radial glial process was observed to occupy a pocket formed by the invagination of the nodal axolemma. The axonal cytoplasm adjacent to the invagination contained a variety of organelles, e.g. multivesicular bodies, vesicles and endoplasmic reticulum, suggesting that this relationship between the radial glial process and the axon is more than a passive interaction. These observations are consistent with the view that processes of radial glial cells may regulate the extracellular environment adjacent to the nodal axolemma, and/or play an active role in the maintenance of the nodal membrane. The existence of perinodally-directed processes of radial glial cells in the salamander indicates that this axo-glial specialization reflects an important functional interaction preserved across a large segment of the phylogenetic scale.

Ambystoma mexicanum↗
PubMed1991

The immunodominant outer membrane antigen of Actinobacillus actinomycetemcomitans is located in the serotype-specific high-molecular-mass carbohydrate moiety of lipopolysaccharide.

Most patients with juvenile periodontitis manifest serum antibodies, sometimes at very high titers, to antigens of Actinobacillus actinomycetemcomitans, but the antigens inducing the immune response have been only partly characterized. We separated A. actinomycetemcomitans serotype b cells into protein, lipopolysaccharide (LPS), and soluble polysaccharide fractions and characterized them. Coomassie blue- and silver-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels were used to detect protein and LPS components, and gas-liquid chromatography was used to determine their carbohydrate and fatty acid composition. Western blots, dot blots, and enzyme-linked immunosorbent assay inhibition with high-titer sera from juvenile periodontitis patients revealed which components were highest in antibody binding activity. These results showed that the major portion of the immunoglobulin G binding activity resides in the purified mannan-free LPS, with lesser amounts in the total protein fraction. Using Sephacryl S-300 chromatography, we separated LPS into high-molecular-mass components with high carbohydrate contents by gas-liquid chromatography and a low-molecular-mass component consisting mainly of lipid A and the inner core sugar heptulose. The results of quantitative dot blot assays and enzyme-linked immunosorbent assay inhibition show that the serotype-specific antibody binding activity is highly concentrated in the high-molecular-mass carbohydrate-rich LPS fraction and is almost completely absent in the low-molecular-weight lipid-rich fraction. Our observations contrast with previous reports that the predominant serotype antigen of A. actinomycetemcomitans resides in a mannan-rich polysaccharide isolated from spent culture medium. These observations support the conclusion that the immunodominant antigen of the outer membrane is the O antigen of the LPS.

Actinobacillus↗
PubMed1991

Antigens of Actinobacillus actinomycetemcomitans recognized by patients with juvenile periodontitis and periodontally normal subjects.

Most juvenile periodontitis patients respond to infection by Actinobacillus actinomycetemcomitans by producing serum antibodies. Specific antigens inducing the humoral immune response have not been identified, nor has the role of the resulting antibodies in disease progression been determined. Adsorbed and unadsorbed sera from juvenile periodontitis patients and normal subjects were analyzed by enzyme-linked immunosorbent assay and Western blots (immunoblots), using digested and undigested bacterial sonicates and French pressure cell fractions to determine the biochemical class, cross-reactivity, and cellular location of the antigens in different A. actinomycetemcomitans serotypes. Antigens detected by using high-titer sera included the following: (i) serotype-specific nonprotein material located on the cell surface, (ii) soluble-fraction proteins showing highly variable antibody binding, (iii) cross-reactive proteins, and (iv) a protein present in soluble and cell wall fractions and immunopositive for all sera tested. In addition, one apparently nonprotein component that was enriched in the cell wall fraction was observed. Sera with high immunoglobulin G titers to one, two, three, or none of the three A. actinomycetemcomitans serotypes were observed. There was a high degree of variation from one patient to another in the humoral immune response to serotype-specific and cross-reactive antigens. As demonstrated by whole-cell adsorption experiments, the serotype-specific surface antigen accounted for approximately 72 to 90% of the total antibody-binding activity for sera with titers greater than 100-fold above background, while cross-reactive antigen accounted for less than 28%. Antibody binding the whole-cell sonicate for high-titer sera was inhibited 90% by lipopolysaccharide from the same serotype, strongly suggesting that lipopolysaccharide is the immunodominant antigen class.

Actinobacillus↗