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Biomedical subjects

R K Clark

Publications and source records attributed to R K Clark.

At least 19 recordsLinked to original sources
PubMed1995

Time-related changes in myeloperoxidase activity and leukotriene B4 receptor binding reflect leukocyte influx in cerebral focal stroke.

In previous studies, we have used histological methods to characterize cellular changes, and validated the use of the myeloperoxidase (MPO) activity assay to quantitate increased neutrophil infiltration in ischemic stroke. We also identified increased leukotriene B4 (LTB4) binding sites as a potential marker for neutrophil infiltration into focal ischemic tissue. However, these studies were conducted at only one time-point, 24 h after ischemia. In the present study, we examined the full time-course of MPO activity and LTB4 receptor binding following middle cerebral artery occlusion (MCAO) made permanently (PMCAO) or transiently (160 min followed by reperfusion; TMCAO) in spontaneously hypertensive rats, and compared the results to previously characterized histologic changes in these models. Ischemic and contralateral (control) cortical tissue samples were assayed for MPO (U/g wet wt) and [3H]LTB4 receptor binding (fmol/mg protein). Following PMCAO, MPO activity significantly increased as early as 12 h and continued to increase over the next 5 d (p < 0.05). Following TMCAO, MPO activity was significantly elevated already after only 6 h of reperfusion and also continued to increase over the next 5 d of reperfusion (p < 0.05). LTB4 receptor binding and MPO activity were highly correlated during periods when both measures increased together (i.e., 0.5-5 d; p <0.01). However, by 15 d post-MCAO, LTB4 receptor binding remained elevated after MPO activity levels had returned to normal. This persistent LTB4 binding was associated with the significant gliosis that was characterized previously to persist in these models. The time-course of increased MPO activity and initially increased LTB4 binding post-MCAO correspond very well to our previous histological data that characterized the time-course for leukocyte infiltration under these conditions. Therefore, the increased MPO activity over time was associated with initial neutrophil and later mononuclear cell infiltration into ischemic tissue in these models. In addition, the present studies utilized histochemical analysis to demonstrate peroxidase activity in macrophages within the cerebral infarct following MCAO, thus validating that MPO activity originates from the later infiltrating mononuclear cells in addition to the early infiltrating neutrophils that had been previously characterized in the same manner. TMCAO produces a significantly larger and earlier increase in ischemic cortex MPO activity and a similar later increase in MPO activity compared to PMCAO treatment.(ABSTRACT TRUNCATED AT 400 WORDS)

Analysis of Variance↗
PubMed1995

Contamination of titanium castings by aluminium oxide blasting.

OBJECTIVES: It is desirable that the surfaces of surgical implants be uncontaminated by foreign materials to avoid untoward tissue reactions, and grit blasting is widely assumed to leave clean metal surfaces. SEM examination and X-ray microanalysis of a recovered 'pure' titanium implant casting that was associated with tissue breakdown revealed embedded particles of alumina. The casting had been cleaned of investment by blasting with alumina grit. METHODS: A variety of treatments of cast titanium plates was used: (a) to establish that the observed aluminium was due to the blasting grit, and (b) to determine whether removal of investment could be achieved effectively by other means. SEM examination and X-ray microanalysis were used. RESULTS: The detected aluminium was associated with embedded fragments identified as coming from the blasting grit. Acid-pickling and mechanical (rotary instrument) trimming produced minimally contaminated surfaces. CONCLUSIONS: Whilst unproven, the presence of the alumina is viewed with great concern as a possible causative agent in the observed tissue breakdown and procedures avoiding alumina blasting are recommended as a precautionary measure.

Aluminum Oxide↗
PubMed1995

Temperature and dimensional changes in the two-stage processing technique for complete dentures.

OBJECTIVES: This study concerned the temperature and linear dimensional change of heat-cured acrylic resin in the two-stage processing technique for complete dentures. METHODS: Thermocouples were incorporated in the acrylic resin for recording temperatures. Measurements between reference marks were made by a high-resolution digital measuring microscope. RESULTS: No increase in temperature associated with the exothermic nature of the polymerization reaction was recorded. The temperature of the resin followed the waterbath temperature closely. The temperatures recorded at various regions were in phase with each other. The total linear shrinkage of the base after two processing cycles was less than 1% and compares favourably with studies on the single-stage processing technique. CONCLUSIONS: Temperature differential is excluded as a reason for the warpage of dentures. The dimensional changes of the denture base resulting from the two-stage processing technique cannot be considered to be of any clinical significance.

Acrylic Resins↗
PubMed1995

The bonding of cold-cured acrylic resin to acrylic denture teeth.

The necessary improvement of the bonding of denture teeth to the base resin has been hampered by inconsistent results and the consequent lack of clear guidance as to the best procedure to use. This state of affairs is in part due to poor experimental design. The tensile strength of notched planar interfaces between acrylic denture tooth material and cold-cured acrylic resin was determined for various treatments under a protocol designed to minimize spurious failures. The tooth material treatment consisting of grinding and exposure to a methyl methacrylate-trichloromethane mixture was, of those treatments, the most reliable in that no interfacial failures were observed. Strength was indistinguishable from that of both tooth and base acrylic under the same conditions.

Acrylic Resins↗
PubMed1995

A reappraisal of indirect retention in removable partial dentures with long bilateral distal-extension saddles.

When only the six anterior teeth remain, it may appear that indirect retention cannot be achieved because the fulcrum line around which the denture may move when the distal extensions lift away from the mucosa is located behind the retentive tips of the clasps on the abutment teeth. This will be the case when the indirect retainer and the tips of the clasps are on the same level; however, if the tips of the clasps are nearer to the gingival margin than the indirect retainer, indirect retentive effects will be achieved.

Dental Clasps↗
PubMed1994

Human laminin produces human platelet aggregation in vitro.

The effects of laminin isoforms on platelet aggregation were compared and characterized in platelet rich plasma (PRP) obtained from 26 healthy human volunteers. In approximately 38% of the individuals tested, human laminin produced a biphasic platelet aggregation response. Human laminin produced only a primary phase in the remaining "non-responsive" individuals. Mouse laminin, rat laminin and human merosin did not cause platelet aggregation in any of the volunteers. The biphasic platelet aggregation response caused by human laminin was concentration-dependent (0.3-30 nM) and was consistently observed upon repeated testing of "responsive" individuals. The secondary phase of aggregation produced by human laminin in "responsive" individuals was abolished by aspirin, SQ 29,548, a selective thromboxane antagonist, and SK&F 106760, an RGD-derived platelet fibrinogen receptor (GPIIb/IIIa) antagonist. Also, the secondary phase of aggregation was not observed in washed platelets. Both the primary and secondary platelet responses produced by human laminin were abolished by a VLA-6 (alpha 6 beta 1) monoclonal antibody, but not by the YIGSR pentapeptide. In conclusion, human laminin causes thromboxane-dependent platelet aggregation, in vitro, in a significant population of human volunteers. The aggregation response was dependent upon the interaction of human laminin with platelet VLA-6 (alpha 6 beta 1). These novel results suggest that in some individuals laminin may play an important role in hemostasis and thrombogenesis.

Adult↗
PubMed1994

Reperfusion following focal stroke hastens inflammation and resolution of ischemic injured tissue.

Previously, we described cellular changes following Permanent Middle Cerebral Artery Occlusion (PMCAO) in spontaneously hypertensive rats. Ischemic changes following PMCAO included a time-related focal pan necrosis, inflammatory cell infiltration, gliosis, and eventual loss of necrotic tissue post PMCAO. We have now characterized changes which occur after Temporary Middle Cerebral Artery Occlusion (TMCAO; 80 or 160 min) followed by reperfusion and compared these changes to those which occur following PMCAO. TMCAO with reperfusion results in cortical infarcts which vary in size in an occlusion-time-dependent manner. After 1 h of reperfusion, ischemic changes were observed histologically, including microhemorrhages and the beginning of a slight inflammatory infiltration in and around the meningeal vasculature. This infiltrate consisted primarily of neutrophils, which by 6 h of reperfusion was significant with infiltration from deep blood vessels into brain tissue, including the presence of some monocytes adhering within blood vessels. Neutrophil infiltration occurred sooner and to a greater extent in reperfused tissues than in permanently occluded tissues, where it only began at 12 h post PMCAO. As occurred following PMCAO, increased Glial Fibrillary Acidic Protein (GFAP) immunoreactivity indicating astrogliosis was first observed at 12 h postTMCAO. Over 1-3 days of reperfusion, a heavy macrophage infiltrate was observed in the reperfused tissues in addition to a continued influx of neutrophils. Following 5 days of reperfusion, the lesion was completely replaced with inflammatory cells, of which macrophages predominated. Unlike PMCAO, which resulted in focal spots of neutrophil accumulation, neutrophils were more distributed throughout the infarcted cortex following TMCAO.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Enhanced leucocyte adhesion to interleukin-1 beta stimulated vascular smooth muscle cells is mainly through intercellular adhesion molecule-1.

OBJECTIVE: The aim was to investigate whether interleukin-1 beta (IL-1 beta) plays a role in modulating the adhesion of monocytes and neutrophils to vascular smooth muscle cells, and to identify what molecules on these cells may be involved in the adhesion. METHODS: Rat aortic smooth muscle cells were challenged with IL-1 beta and tested for adhesion of prelabelled monocytes and neutrophils. Northern analysis, reverse transcription/polymerase chain reaction (RT/PCR), and immunocytochemical staining were used to measure the changes of intercellular adhesion molecule-1 (ICAM-1) and other adhesion molecules in response to IL-1 beta stimulation. Neutralising antibody against ICAM-1 was used to demonstrate a role of ICAM-1 in this IL-1 beta induced adhesion. RESULTS: IL-1 beta induced the adhesion of monocytes and neutrophils to aortic smooth muscle cells in a concentration and time dependent manner. IL-1 beta-induced adhesion was inhibited by preincubation of the cells with an IL-1 receptor antagonist (IL-1ra). Northern analysis and RT/PCR showed that ICAM-1 mRNA represents a predominant adhesion molecule induced by IL-1 beta, and that the expression of ICAM-1 mRNA precedes and parallels the induced adhesion profiles of aortic smooth muscle cells for leucocytes. Immunocytochemical staining confirmed the IL-1 beta induced ICAM-1 expression on the smooth muscle cells. Moreover, a monoclonal anti-rat ICAM-1 antibody produced a concentration dependent inhibition of the IL-1 beta induced adhesion of monocytes and neutrophils to the smooth muscle cells. CONCLUSIONS: IL-1 beta actively regulates functional ICAM-1 expression in vascular smooth muscle cells. The IL-1 beta-induced expression of ICAM-1 on the smooth muscle cells may be an important contributor to the increased adhesion by monocytes and neutrophils to these cells and suggests that IL-1 beta might play a role in the proinflammatory and immune functions of the modified smooth muscle cells during atherosclerosis and restenosis.

Animals↗
PubMed1994

Tumor necrosis factor-alpha expression in ischemic neurons.

BACKGROUND AND PURPOSE: Tumor necrosis factor-alpha (TNF-alpha) is a cytokine with diverse proinflammatory actions, including endothelial leukocyte adhesion molecule expression. Since leukocytes infiltrate into ischemic brain lesions, the present study was conducted to examine whether TNF-alpha messenger RNA (mRNA) and peptide are expressed in the brain after experimental focal stroke and before leukocyte accumulation. METHODS: TNF-alpha mRNA and protein expression were monitored in the ischemic and nonischemic cerebral cortex of rats after focal ischemia produced by permanent middle cerebral artery occlusion. The effect of TNF-alpha administered by microinjection into the brain cortex on leukocyte adherence to brain capillaries was also studied. RESULTS: Induction of TNF-alpha mRNA, normalized to a standard reference rat macrophage TNF-alpha mRNA, was detected as early as 1 hour after middle cerebral artery occlusion. TNF-alpha mRNA was elevated by 3 hours (29 +/- 6% versus 2 +/- 1% in sham-operated rats) only in the ischemic cortex, with peak expression at 12 hours (104 +/- 8%; P < .01). Five days after middle cerebral artery occlusion, TNF-alpha mRNA levels in ischemic cortex were still significantly elevated (38 +/- 5%; P < .05). Also, TNF-alpha mRNA expression was greater in the ischemic cortex of spontaneously hypertensive rats than in normotensive rats (P < .05). Double-labeling, immunohistochemical studies revealed the presence of TNF-alpha protein localized within nerve fibers in the evolving infarct at 6 and 12 hours after ischemia and further expression in the tissues immediately adjacent to the infarct 24 hours after ischemia. After 5 days, the neuronally localized peptide had diminished greatly, but macrophages located within the infarcted tissues were immunoreactive. Cortical microinjections of TNF-alpha (10 ng in 1 microL) produced a significant neutrophil adherence/accumulation in capillaries and small blood vessels 24 hours later. CONCLUSIONS: These results represent the first demonstration that focal cerebral ischemia in rats results in elevated TNF-alpha mRNA and protein in ischemic neurons. The neuronal expression of peptide appears to facilitate the infiltration of inflammatory cells that can further exacerbate tissue damage in cerebral ischemia and might contribute to increased sensitivity and risk in focal stroke.

Animals↗
PubMed1994

Geographic analysis of pathogen exposure in bighorn sheep (Ovis canadensis).

Antibody responses were examined among 998 bighorn sheep (Ovis canadensis) in California (USA) to determine spatial patterns of pathogen exposure. Using a shifting frame analysis, a specific geographic region was delineated that contained bighorn sheep with higher (P < 0.05) levels of multiple exposure (antibodies detected against > or = two pathogens), as well as higher prevalence values for eight of ten individual pathogens. This region in southwestern California encompassed all of the peninsular populations of bighorn sheep recently proposed for listing as endangered by the U.S. Fish and Wildlife Service.

Animals↗
PubMed1994

The edentulous mandible opposed by natural maxillary teeth: a report of six cases treated with implant-retained prostheses.

Dental restoration of the edentulous mandible opposing natural maxillary teeth is often associated with a number of problems, particularly in the occlusion, and an accelerated rate of alveolar bone resorption in the edentulous mandible resulting from occlusal loading. The advent of osseointegrated implants would seem to provide a new means of treating these cases. Accordingly, the cases of six patients for whom either fixed partial dentures or over-dentures supported by Brånemark implants were provided, are reviewed. Three of the mandibles were normal and three had bone grafts. Results so far indicate that this is a viable alternative for treating such cases.

Dental Implantation, Endosseous↗
PubMed1993

Neuron-specific enolase increases in cerebral and systemic circulation following focal ischemia.

Neuron-specific enolase (NSE) is an isoform of the glycolytic enzyme, enolase, and is found in neurons and neuroendocrine cells. We evaluated cerebral immunohistologic and plasma changes in NSE in rats from 2 h to 15 days following permanent or transient middle cerebral artery occlusion (MCAO). At 1-2 days post-MCAO, loss of NSE immunofluorescence from within neurons to the extracellular space was observed in the infarcted areas of all MCAO animals. NSE also was identified intravascularly throughout the brain following MCAO. NSE in plasma was determined by a specific radioimmunoassay. Plasma NSE following permanent or transient MCAO was increased significantly from that observed in controls (2.8 +/- 0.3 ng/ml) beginning at 2 h and persisting for 2.5 days post-MCAO (maximum levels of 8.8 +/- 0.9 to 9.6 +/- 0.5 ng/ml after 6-12 h; P < 0.05, n = 4-9). Quantified contralateral forelimb and hindlimb neurological deficits in these animals were significant and persisted for at least 15 days following MCAO but were not observed following sham surgery. These data suggest that MCAO-induced cortical infarction and neurological dysfunction is associated with neuronal depletion and vascular redistribution of brain NSE resulting in a measurable increase in plasma NSE. Such diffusion of NSE into the cerebral vasculature and systemic circulation from ischemic tissue can be expected to serve as a marker for the incidence of cerebral damage in acute and chronic ischemic brain infarcts.

Animals↗
PubMed1993

A computer-aided study of speaking spaces.

A computer-aided system was devised to investigate the speaking space. Thirty native Cantonese speakers with Class I occlusion were selected and test sentences were designed for speech analysis by a Sona-Graph. The investigation indicated that the sibilant sounds produced the closest speaking space and that the mean and the variability of the closest speaking space in Cantonese speakers were smaller than that in English speakers. The beliefs that the closest speaking space was smaller than that of the freeway space and that the speaking space of m sound was similar to the freeway space were not supported by this study.

Adult↗
PubMed1993

A human lymphoid recombinant cell line with functional human immunodeficiency virus type 1 envelope.

Our goal has been to develop a safe and effective system that would allow us to explore the functions of the human immunodeficiency virus (HIV) envelope. We have generated a human lymphoid cell line (TF228.1.16) that stably expresses functional HIV envelope proteins on its cell surface, and therefore closely mimics the viral envelope and virus-infected cells. The TF228.1.16 line forms syncytia with human cells of the CD4+ phenotype and provides a facile virus-free cell-based assay for examining the mechanism of syncytia formation and for evaluating novel agents that may disrupt this process. The TF228.1.16 cells also provide an opportunity to present the HIV envelope proteins to the immune system in cellular form. In vitro immunization of human peripheral blood mononuclear cells (PBMC) and in vivo immunization of rhesus monkeys with this reagent results in the production of antibodies with neutralizing (anti-syncytia) activities. When the HIV envelope is expressed against the background of human lymphoid cells, it may exhibit immune protection with unique properties that have not yet been explored. Our results indicate that a virus-free cell system can play an important role in exploring the biology and function of HIV-envelope proteins without the interference of other viral components present in infected cells. This paper discusses these results, and examines the potential use of TF228.1.16 as a vaccine.

Animals↗
PubMed1993

Restriction endonuclease analysis of herpesviruses isolated from two peninsular bighorn sheep (Ovis canadensis cremnobates).

In 1989, herpesviruses were isolated from nasal swabs taken from two peninsular bighorn sheep (Ovis canadensis cremnobates) in the Anza-Borrego Desert State Park, San Diego County, California (USA). Using restriction endonuclease analysis (REA) with Pst1 enzyme, each isolate was found to be similar to the Cooper strain of infectious bovine rhinotracheitis virus (IBRV). The REA patterns of the two herpesviruses from bighorn sheep were typical of either field strains or vaccine strains of IBRV commonly associated with cattle in the USA.

Animals↗
PubMed1992

The facial soft tissue profile of the southern Chinese: prosthodontic considerations.

A simple photographic setup was established to produce standardized life-sized black and white prints of Southern Chinese adults. Facial soft tissue profile analysis was performed on 28 men and 31 women aged 19 to 30, dentate Chinese, having class I occlusion and originating from Guangdong province. The facial profile values studied included the profile convexity, the interlabial contour, the nasolabial contour, the columella and upper lip inclination angles to the true horizontal, and the relative positions of upper and lower lips from the Esthetic plane. Data obtained were compared with those from previous Caucasian studies and other accepted empirical values. The lower third of the face presented the greatest ethnic difference: the interlabial contour was more convex and the upper and lower lips were more protrusive among the Southern Chinese. The widely-used standard of a right-angled nasolabial contour proved to be applicable among the Southern Chinese adult males. However, a more obtuse nasolabial angle, almost 100 degrees, occurred among the women.

Adult↗
PubMed1992

Reperfusion increases neutrophils and leukotriene B4 receptor binding in rat focal ischemia.

BACKGROUND AND PURPOSE: Neutrophils are critically involved with ischemia and reperfusion injury in many tissues but have not been studied under conditions of reperfusion after focal cerebral ischemia. The present studies were conducted to confirm our previous observations quantifying neutrophils in rat permanent focal stroke using a myeloperoxidase activity assay and to extend them to transient ischemia with reperfusion. In addition, leukotriene B4 receptor binding in ischemic tissue was evaluated as a potential marker for inflammatory cell infiltration. METHODS: Histological, enzymatic, and receptor binding techniques were used to evaluate neutrophil infiltration and receptor binding in infarcted cortical tissue 24 hours after permanent middle cerebral artery occlusion (n = 25) or temporary occlusion for 80 (n = 12) or 160 (n = 22) minutes followed by reperfusion for 24 hours in spontaneously hypertensive rats. RESULTS: Sham surgery (n = 26) produced no changes in any parameter measured. After permanent middle cerebral artery occlusion, neutrophil accumulation was observed histologically, but the infiltration was moderate and typically within and adjacent to blood vessels bordering the infarcted cortex. After temporary middle cerebral artery occlusion with reperfusion, marked neutrophil infiltration was observed throughout the infarcted cortex. Myeloperoxidase activity was increased (p less than 0.05) after permanent occlusion and to a greater extent after temporary occlusion with reperfusion. Myeloperoxidase activity (units per gram wet weight) in ischemic cortex was increased over that in nonischemic (control) cortex 32.2-fold, 54.6-fold, and 92.1-fold for permanent occlusion and 80 and 160 minutes of temporary occlusion with reperfusion, respectively (p less than 0.05). Sham surgery produced no changes in myeloperoxidase activity. Leukotriene B4 receptor binding also was increased (p less than 0.05) after focal ischemia and paralleled the increases in myeloperoxidase activity. Ischemic cortex-specific receptor binding (femtomoles per milligram protein) was 3.87 +/- 0.63 in sham-operated rats and 4.57 +/- 0.98, 8.98 +/- 1.11, and 11.12 +/- 1.63 for rats subjected to permanent occlusion and 80 and 160 minutes of temporary occlusion with reperfusion, respectively (all p less than 0.05 different from sham-operated). Cortical myeloperoxidase activity was significantly correlated with the degree of cortical leukotriene B4 receptor binding (r = 0.66 and r = 0.79 in two different studies, p less than 0.01). CONCLUSION: These data indicate that neutrophils are involved in focal ischemia and that there is a dramatic accumulation of neutrophils in infarcted tissue during reperfusion that can be quantified using the myeloperoxidase activity assay. Leukotriene B4 receptor binding increases in infarcted tissue in a parallel manner, which suggests that the increased leukotriene B4 binding is to receptors located on the accumulating neutrophils.

Animals↗