Effect of welding parameters of the Nd:YAG laser on the penetration depth of cobalt chromium alloys.

The aim of the investigation was to study the effect of the laser welding parameters of energy and spot diameter on the penetration depth of the weld of cast Co-Cr alloy when a single weld was performed. Within the limitations of the study as voltage increased and the spot diameter decreased, penetration depth increased. However, SEM investigation showed more defects in the welded area under these circumstances. The clinical significance is that during selection of the welding parameters the thickness of the components to be welded should be considered to achieve an extended welded area without the induction of micro-structural defects.

Flexural strength and degree of polymerization of a proprietary denture base acrylic resin designed to be cured using long or short cycles.

The flexural strength and degree of polymerization of Diamond D acrylic resin prepared with a long cure monomer and a short cure monomer were investigated using Trevalon as a control. Flexural strength and degree of polymerization of Diamond D acrylic resin were not affected by either using a long cure monomer or the short cure monomer. There was no significant difference in the glass transition temperature Tg between the long and slow cure Diamond D. Provided the manufacturer's instructions are followed the flexural strength, degree of polymerization and glass transition temperature are comparable with more traditional products.

The retentive forces of the locator attachment system at different angulations.

The change of retentive force of three types of Locator inserts when the implant analogue was positioned perpendicular to horizontal, 5 degrees and 10 degrees to perpendicular after 4,200 cycles in vitro was measured using an EnduraTech fatigue testing machine lubricated with artificial saliva. The more rapid decrease in retention occurred up to three months and stabilized after one year of simulated use. The Locator inserts provided more retention when the analogue was placed at 5 degrees to perpendicular compared to 0 degrees and 10 degrees after 9 months of simulated clinical use. After 2 years of simulated clinical use, there was a reduction in retention for all the three inserts of between 70% and 80%.

The effect of monomer/polymer mixing ratio, time between mixing and packing of heat cured acrylic resin denture base material and bond assisting agents on the bond strength to acrylic resin denture teeth.

The aim of the study was to investigate the effect of varying the monomer/polymer mixing ratio, the time from mixing to packing heat cured acrylic resin and the effect of two bond assisting agents on the strength of the bond between denture base acrylic resin and acrylic resin denture teeth. Statistical differences were found in bond strength with monomer/polymer ratio and time between mixing and packing with one of the heat cured resins investigated. The benefit of using the bonding agents was not demonstrated.

Effect of curing cycles on the mechanical properties of heat cured acrylic resins.

Traditionally long curing cycles have been recommended for heat cured acrylic resin denture base materials. Recently manufacturers have produced materials for which they recommend short curing cycles. Specimens conforming to British Standards Specification were made using three different brands of heat cured acrylic resin denture base material. Each material was processed in three batches using either the manufacturer's recommended short curing cycle, an arbitrary medium curing cycle or a traditional long cycle. Specimens were subjected to a three point bending test. With one exception using the arbitrary medium curing cycle, all specimens achieved the British Standard suggesting that the manufacturers' recommended cycles should be followed.

Immunohistochemical analysis of antiserum from rhesus monkeys immunized with human colon carcinoma.

In an attempt to generate antibodies which recognize novel tumor-associated antigens we have immunized Rhesus monkeys (Macaca mulatta) with human colon carcinoma cells prepared from freshly excised tumors. Immunohistochemical characterization of polyclonal antisera from one monkey (DF6) revealed preferential reactivity with primary and metastatic colon carcinoma tissue, and a general lack of recognition of nonneoplastic mucosa. Immunoreactivity was localized to the luminal contents of glandular structures and to the apical surfaces of cells lining these glands. Immunoreactivity was not observed with any normal tissue examined. Examination of neoplastic tissues revealed reactivity with two gastric carcinoma specimens (n = 2) and one breast carcinoma (n = 7). In reactive colon carcinoma tissues, the pattern of staining with DF6 was similar to that of several other antibodies including anti-carcinoembryonic antigen, B72.3, anti-Le(x) and anti-Le(y). However, the panel of tissues recognized by these antibodies and DF6 differed significantly, suggesting that the DF6-reactive epitopes are unique. Human colon carcinoma cell lines maintained in vitro also expressed antigens recognized by DF6 in a pattern similar to that of surgically excised tissue. This preliminary characterization of DF6 antiserum suggests that immunization of Rhesus monkeys is a potentially useful protocol for identifying antigens preferentially expressed by human colon carcinoma.

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.

Immunoblotting of keratin polypeptides extracted from tissues preserved in standard histologic fixatives.

Cytoskeletal polypeptides from fresh placental tissue, tissue stored at -30 degrees C, and tissue fixed in 10% buffered formalin, Bouin's solution, and Carnoy's solution were extracted, separated by electrophoresis, and immunoblotted using monoclonal antibodies immunoreactive with keratin polypeptides. Storage of the placental tissue at -30 degrees C, or fixation in Carnoy's solution did not alter the extractability, migration pattern, or immunoreactivity of the keratin polypeptides. Keratin polypeptides could not be adequately demonstrated in extracts prepared from formalin- or Bouin's solution-fixed tissues. Several unmasking procedures used on tissues before extraction and on nitrocellulose blots before application of primary antibodies failed to unmask keratin polypeptides, either in Coomassie blue-stained gels or in immunoblots reacted with anti-keratin antibodies. These data indicate that Carnoy's solution is the fixative of choice for tissues in which electrophoretic and immunoblotting analyses of keratin polypeptides might be required.

Suppression of nonspecific binding of avidin-biotin complex (ABC) to proteins electroblotted to nitrocellulose paper.

Nitrocellulose blots of cell extracts reacted in sequence with biotinylated lectins and horseradish peroxidase-labeled avidin-biotin complex (ABC) often show considerable nonspecific staining of protein bands. Experiments were performed to determine which of the components of the ABC were responsible for this and whether or not the nature and ionic strength of the buffer used could alter this binding. Furthermore, as powdered non-fat milk has been proposed as a possible blocking agent for nonspecific binding of ABC, we sought to determine if it would adequately block that binding in our system. The initial experiments showed that nonspecific binding of ABC to proteins transferred to nitrocellulose membranes was due to the avidin component of the ABC; little, if any, binding was seen if biotin alone was incubated with these blots. The spurious binding was shown to be primarily due to the high affinity of avidin to proteins electroblotted to nitrocellulose, when incubated in low-salt buffers. Low-fat milk added to the buffer reduced overall nonspecific reactivity but produced additional artifacts in the form of bands that were not seen in other preparations. Nonspecific avidin binding to proteins transferred to nitrocellulose can therefore be effectively reduced by adding extra salt to buffers, whereas the addition of non-fat dry milk does not seem suitable for this purpose.

Mixed glycosidase pretreatment reduces nonspecific binding of antibodies to frozen tissue sections.

Indirect immunohistochemical studies of frozen mouse tissues with mouse monoclonal antibodies yield, in general, suboptimal results primarily because of indiscriminate binding of secondary antibody to all mouse immunoglobulins, i.e., to the monoclonal reagent and to endogenous immunoglobulin nonspecifically trapped in the tissue. To reduce this nonspecific staining, frozen sections of mouse kidney were treated enzymatically. Optimal results were obtained following a 2 hr treatment with 20 mg/ml of mixed glycosidases (MG). This treatment reduced the nonspecific background staining of the interstitial spaces and blood vessels, but did not affect the reactivity of structurally bound immunoglobulin G (IgG) in the glomeruli or alter the reactivity of mouse renal tissue to the monoclonal antibody that recognizes an oligosaccharide antigenic determinant (SSEA-1). Eluates from enzyme-treated frozen tissue sections contained normally immunoreactive IgG in the form of dimers. These data indicate that MG treatment of frozen sections could be safely used to reduce the content of nonstructurally bound immunoglobulins in frozen tissues and thus improve the visualization of specific monoclonal antibody binding.

Antiserum to a novel peptide sequence reacts selectively with epithelial subpopulations.

A synthetic peptide corresponding to a novel protein sequence isolated from bovine kidney was used to immunize rabbits. When applied to Western blots of bovine kidney extracts, antiserum to this peptide recognizes proteins with molecular weights of 23 and 18 KD. Immunohistochemical examination of a variety of bovine and rat tissues with this antiserum revealed a unique distribution of immunoreactivity with the intermediate layers of a variety of stratified epithelia, in addition to renal glomeruli. The pattern of reactivity differed from previously described epithelial markers such as cytokeratins. These results indicate that this antiserum may be useful as a tool for the identification of cells of the intermediate layer of stratified epithelia and, as such, may aid in the study of this differentiating/proliferating tissue compartment.

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.

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.

Quantitative comparison of magnetic resonance imaging (MRI) and histologic analyses of focal ischemic damage in the rat.

Hemispheric swelling and area of infarction, two parameters of cerebral focal ischemic damage, were identified and quantified from T2-weighted magnetic resonance imaging (MRI) two days after occlusion of the middle cerebral artery (MCAO) in spontaneously hypertensive rat (SHR) brains. Results were compared with these measures quantified from 2,3,5-triphenyltetrazolium hydrochloride (TTC)- and hematoxylin and eosin (H&E)-stained histologic sections in the same brains. The degree of hemispheric swelling and infarct size determined by MRI were highly correlated to the measurements as determined in the TTC- and H&E-stained tissues. These results demonstrate that the focal ischemic damaged area and associated tissue swelling identified by MRI is quantitatively similar to, and thus, is representative of actual tissue damage/changes that can be identified by gross or histologic examination.

Development of tissue damage, inflammation and resolution following stroke: an immunohistochemical and quantitative planimetric study.

Development and resolution of the lesion produced by occlusion of the middle cerebral artery (MCAO) was studied through quantitative planimetry and histologic/immunohistochemical techniques. MCAO, performed in spontaneously hypertensive rats (SHR), initially (1-3 days) produced large, consistent cerebral cortical infarctions and an increase in ipsilateral hemispheric size (i.e., swelling) quantitated by planimetry on 2,3,5-triphenyltetrazolium chloride (TTC)-stained gross tissue sections. These initial changes correlated well with changes identified from 2 h to 3 days using hematoxylin and eosin stained histologic tissue sections and immunohistochemical techniques including: the progressive development of a cortical area of pan necrosis, infiltration of neutrophils into infarcted tissues, and activation of astroglia. During the initial 2 days following MCAO, glial fibrillary acidic protein immunoreactive cells increased in number and became larger and more intensely fluorescent medial to the cortical infarct. At 5 to 15 days, both the infarct and the ipsilateral hemisphere decreased in size. These changes correlated with the presence of abundant macrophages, and cavitation of the lesion along its medial border. Also during this period, a loose connective tissue matrix formed along the superficial aspect of the infarct. This connective tissue contained fibroblasts, extracellular matrix immunoreactive for laminin and collagen, capillary buds indicating neovascularization, and abundant macrophages. By the final timepoint (30 days), necrotic tissue could no longer be detected in either gross or histologic tissue sections, the inflammatory infiltrate had resolved, and the connective tissue was removed.(ABSTRACT TRUNCATED AT 250 WORDS)

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)

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.

Pressure and temperature changes in heat-cured acrylic resin during processing.

OBJECTIVES: The aims of this study were to measure the pressure and temperature changes of acrylic resin during processing, to record the highest temperature reached when fast cured in boiling water and to determine the elevated boiling point of monomer under high pressure. METHODS: A subminiature pressure transducer (temperature compensated to 94 degrees C) and a thermocouple were placed on the palate of a standardized maxillary complete denture base. A heat-cured resin (Trevalon C) was polymerized by a long heating cycle (72 degrees C for 6.5 h and 92 degrees C for 1.5 h). Recordings of pressure and temperature (n=6) were made at initial clamping of denture flasks and throughout the processing cycles of resin. The temperature of the resin was also monitored during a fast cycle, which was accomplished by placing the flask directly into boiling water for 40 min. RESULTS: The pressure of acrylic dough inside the clamped flask was initially 11.5 atm (SD=3.2) and reached a peak of 22.0 atm (SD=3.5) during the long heating cycle. The elevated boiling point of monomer at increased pressure was calculated to be about 193 degrees C (at 11.5 atm) and 228 degrees C (at 22.0 atm). These elevated boiling points are higher than the maximum temperature 131 degrees C (SD=6.6) reached during the fast curing cycle. No porosity was observed even in the denture bases heat-cured by the fast cycle. SIGNIFICANCE: The highest temperature reached by heating of resin during processing is well below the elevated boiling point of monomer. Monomer therefore does not boil in clamped denture flasks under sufficient pressure. Thus adequate clamp pressure prevents gaseous porosity irrespective of curing cycle used.

Reproducibility of natural head position.

The orientation of the head, when the natural head position was adopted, was measured relative to the true vertical on standardized black and white profile photographs. Two methods of obtaining the natural head position were compared and their reproducibility tested. No statistically significant difference was found between the two different methods or at different sittings.

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.