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Biomedical subjects

J R Swafford

Publications and source records attributed to J R Swafford.

15 recordsLinked to original sources
PubMed2001

Molecular structure of acid-etched dentin smear layers--in situ study.

It is commonly reported that acid etchants remove the smear layer, but to date, there has been no chemical evidence to support these observations. The purpose of this study was to determine the molecular structure of acid-etched carbide- and diamond-bur-created smear layers. This project tested the null hypothesis that such smear layers are totally removed with current etchants. Smear layer/demineralized/mineralized dentin interfaces were analyzed at 1- m intervals by micro-Raman spectroscopy. Features associated with the organic component were substantially broadened with loss of fine structure, and mineral peaks were clearly evident in the spectra of acid-etched smear layers. The organic features in the spectra of the EDTA-treated smear layer showed relative intensity ratios similar to demineralized dentin without contribution from the mineral phase. The disorganized collagen within the smear layer was not removed but was denatured by the acid treatment; the mineral was trapped in this gelatinous matrix and shielded from complete reaction.

Acid Etching, Dental↗
PubMed2000

Morphology of azurophil lysosomes in polymorphonuclear leukocytes from humans with rapidly progressive periodontitis.

Previous investigations have shown that subjects with rapidly progressive periodontitis (RPP), an early-onset aggressive form of periodontitis, have polymorphonuclear leukocytes (PMNs) with increased intracellular levels of beta-glucuronidase, a characteristic enzyme of azurophil lysosomes. The current study attempted to account for that increase. Ten healthy controls and 10 otherwise healthy subjects with RPP participated. PMNs from peripheral blood were separated, fixed and reacted for peroxidase to identify azurophil lysosomes. Using transmission electron microscopy, 20 PMNs per subject were photographed at 10,000x. Photographs were subsequently digitized and analyzed by computer. RPP PMNs had a higher percentage of the area of the cell profile occupied by azurophil lysosomes compared to control subjects' PMNs. The RPP subjects also had greater absolute numbers of azurophil lysosomes per cell. Lysosome shape was assessed visually. There were no differences between RPP and control groups for lysosome shape, with the majority of lysosomes in each group exhibiting a round or oval shape. RPP lysosomes did exhibit a significantly greater mean size.

Acute Disease↗
PubMed2000

Interfacial chemistry of the dentin/adhesive bond.

To date, the dentin/adhesive (d/a) bond has primarily been studied by morphologic analysis in conjunction with bond strength measurement. Although these analyses have enhanced our understanding, numerous questions about the chemistry have not been answered. The purpose of this study was to determine, at the molecular level, quantitative differences in the composition of the d/a interface formed under "wet" bonding conditions. The occlusal one-third of the crown was removed from 10 extracted, unerupted human third molars. The prepared dentin surfaces were treated, per manufacturers' instructions, with either Single Bond (3M) or One-Step adhesive (Bisco). Three-micron-thick sections of the d/a interface were cut and stained with Goldner trichrome for light microscopy. Companion slabs were analyzed with micro-Raman spectroscopy; the sample was placed at the focus of a 100x microscope objective, and spectra were acquired at 1-microm intervals across the d/a interface. Reference spectra were collected on model compounds of type I collagen and adhesive; the relative ratios of the integrated intensities of spectral features from adhesive and collagen were determined and plotted as a function of wt% adhesive. The same ratios were determined for the interface samples; by comparing these ratios with the calibration curve generated from the model compounds, we determined the percent of adhesive as a function of spatial position across the d/a interface. The relative percent of Single Bond adhesive was < 50% throughout more than half of the hybrid layer; One Step adhesive was > or = 50% throughout most of the hybrid. The results from this study provide the first direct chemical evidence of phase separation in a dentin adhesive and its detrimental effect on the dentin/adhesive bond.

Bisphenol A-Glycidyl Methacrylate↗
PubMed1999

Effective laser ablation of bone based on the absorption characteristics of water and proteins.

BACKGROUND: Collateral thermal damage and residual char formation have severely limited the use of conventional lasers in the surgical preparation of bony tissue. Thermal damage from lasers can be minimized by selecting a wavelength that is strongly absorbed and by reducing the laser pulse duration. In contrast to the fixed wavelengths and microsecond pulse intervals of conventional lasers, the Vanderbilt free electron laser (FEL) can be set at wavelengths ranging from 2.1 to 9.8 microm, and the pulse duration can be reduced to a series of 1 to 2 picosecond (ps) micropulses delivered in succession over intervals of 4 microsecond macropulses. The purpose of this study was to compare the morphologic and chemical changes induced in the near-surface region of bone following exposure to the FEL at 3.0, 6.1, and 6.45 microm wavelengths. The selected wavelengths coincide with the vibrational modes of proteins and water within bone. METHODS: Under general anesthesia, laser incisions were made in the tibias of 14 skeletally mature rabbits. Laser parameters included 22.5+/-2.5 mJ/pulse delivered in individual 4 microsecond macropulses at a repetition rate of 30 Hz, focused to 200 microm and 500 microm spot sizes. Laser incisions were made using a computer-assisted surgical program, and control incisions were created with a bone saw. Rabbits were euthanized after the final incision, tibias recovered, and non-decalcified specimens processed for light microscopy. Separate samples were prepared for FTIR (Fourier transform infrared) photoacoustic spectroscopic analysis. RESULTS: The light microscopy sections of the ablation defects created at the differing wavelengths showed similar features, i.e., 2 zones of collateral damage, a zone generally < 10 mm of extensive thermal damage, and a wider zone of empty lacunae. In comparing treated and untreated surfaces, the spectral differences were limited to a relative decrease in intensity of the amide II and III absorption peaks in all laser-treated surfaces. CONCLUSIONS: Spectroscopic and histologic results indicated minimal thermal damage to bone ablated at 3.0, 6.1, and 6.45 microm wavelengths using the FEL (Fourier transform infrared) at the specified parameters. The FTIR photoacoustic spectroscopic results suggest that the char layer is limited to an area less than approximately 6 microm from the surface.

Absorption↗
PubMed1999

Unprotected protein at the dentin-adhesive interface.

OBJECTIVE: With dental bonding systems that require acid etching of dentin, inadequate adhesive penetration can leave exposed collagen at the dentin-adhesive interface. The exposed collagen could be degraded by bacterial proteases, compromising the integrity of the dentin-adhesive bond and, ultimately, the restoration. The purpose of this study was to develop a nondestructive staining technique to identify exposed collagenous protein at the dentin-adhesive interface. METHOD AND MATERIALS: The following adhesives were placed, according to manufacturer's instructions, on dentin cut from 15 human third molars: Scotchbond Multi-Purpose, Scotchbond Multi-Purpose Plus, and 3M Single Bond. Light microscopic sections of native dentin-adhesive interfaces of each tooth were cut and stained with Goldner's trichrome. This reagent stained exposed protein in the sections a distinct red-orange. RESULTS: Exposed protein was identified at the dentin-adhesive interface with each of the adhesives. Corollary scanning electron microscopic examination confirmed the presence of exposed protein, i.e., protein that was removed by sodium hypochlorite, at the interface. Sites of exposed protein that were clearly identified in the light microscopic sections were obscured in the transmission electron microscopic sections. CONCLUSION: In vitro identification of inadequacies in the dentin-adhesive bond is the first step in determining sites that may be vulnerable to premature breakdown under clinical conditions.

Azo Compounds↗
PubMed1992

Serosal mast cells maintain their viability and promote the metabolism of cartilage proteoglycans when cocultured with chondrocytes.

OBJECTIVE: To determine the consequences of mast cell (MC)-chondrocyte interactions. METHODS: Cocultured cells were analyzed histochemically, morphologically, biochemically, and functionally. RESULTS: Cocultured MC adhered to the chondrocytes and remained viable. Chondrocytes cocultured with nonactivated MC produced more proteoglycans than did chondrocytes cultured alone, and these proteoglycans possessed an intact hyaluronic acid-binding region. In contrast, most of the proteoglycans produced by chondrocytes cocultured with activated MC were degraded. CONCLUSION: These studies indicate that a complex interaction occurs in which the nonactivated MC stimulates biosynthesis and the activated MC degrades cartilage proteoglycans.

Animals↗
PubMed1991

Freshly isolated and cultured human monocytes obtained by plasmapheresis kill schistosomula of Schistosoma mansoni.

The efficacy of human peripheral blood monocytes (PBM) in killing of schistosomula is controversial. The purpose of this study was to determine the schistosomulacidal activity of human monocytes isolated by two different techniques. Peripheral blood monocytes were obtained either by venipuncture (PBMv) or plasmapheresis (PBMp), purified on Ficoll-Paque, and cultured briefly. The cells then were incubated with schistosomula (cell parasite ratio of 10(4):1) for 16 to 18 hours with or without interferon-gamma IFN-gamma (600 U/ml) or sera from patients with schistosomiasis as a source of antischistosomal antibodies (HASA). Freshly isolated PBMv treated with IFN-gamma or HASA did not kill schistosomula. Freshly isolated PBMp alone killed 22 +/- 13% (mean +/- standard deviation [SD]; n = 9) of worms over background and after incubation with IFN-gamma and HASA, 30 +/- 17%. PBMp cultured in vitro for 7 days killed 50 +/- 15% (mean +/- SD; n = 12) of the schistosomula. Pretreatment of the cells with IFN-gamma and incubation with HASA did not significantly enhance the parasite killing beyond this level. Electron microscopy showed that freshly isolated PBMp attached to the worms and fused occasionally with the outer tegumental membrane. Granules constituted 1.4% of the cytoplasmic volume. Degranulation onto the parasite surface was not observed. Peripheral blood monocytes obtained by plasmapheresis accumulated glycogen during in vitro culture with the parasite and released threefold more H2O2 than PBMv after exposure to phorbol myristate acetate. Thus plasmapheresis increases the schistosomulacidal activity of PBM, enhances the generation of H2O2 and promotes the accumulation of glycogen.

Animals↗
PubMed1985

Localization of the enantiozymes of 6-hydroxy-nicotine oxidase in Arthrobacter oxidans by electron immunochemistry.

During the course of growth of Arthrobacter oxidans, induction of the enantiozymes 6-hydroxy-D-nicotine oxidase and 6-hydroxy-L-nicotine oxidase occurred in the presence of DL-nicotine. Cryoultramicrotomed sections obtained from cells grown to stationary phase were gold immunolabeled. The results obtained demonstrate that both enzymes are localized in the cytoplasm.

Arthrobacter↗
PubMed1984

Studies on Genetic Male-Sterile Soybeans : III. The Initiation of Monocarpic Senescence.

Soybean (Glycine max [L.] Merr.) germplasm, isogenic except for loci controlling male-sterility (ms(1)) and nodulation (rj(1)) was utilized to investigate the effects of reproductive tissue development and nitrogen source on the initiation of monocarpic senescence. The experimental genotypes (Ms(1)Rj(1), Ms(1)rj(1), ms(1)Rj(1), and ms(1)rj(1), were selected from a cross between N69-2774 and N59-5259, and were inbred to the F(5) generation. Green-house-grown plants were collected during the period of flowering (77 days after transplanting) until maturity (147 days after transplanting). Leaf tissues from the respective genotypes were analyzed at the various harvest dates for RNA, phenolic, and chlorophyll concentrations; acid protease activity; polypeptide banding patterns of chloroplast thylakoids; and chloroplastic ultrastructure.Regardless of nitrogen source, total chlorophyll concentrations declined between 77 and 119 days after transplanting, resulting in a 40% loss of chlorophyll per square centimeter in all genotypes. Leaf chlorophyll levels continued to decline at a constant rate in male-fertile genotypes, but remained at a constant level (26 micrograms chlorophyll per square centimeter) in male-sterile genotypes, for the remainder of the study. With increased leaf age, a gradual disruption of thylakoid structures was observed, particularly in chloroplasts from the male-fertile genotypes. Chloroplasts from the male-sterile genotypes appeared to lose starch grains but increased their number of chloroplastic lipid bodies with leaf aging. These data suggest that monocarpic senescence in soybeans was initiated at or before flowering. Although reproductive tissue development probably augmented the process, the response attributed to seed formation was not apparent until the mid-pod fill stage (119 days after transplanting). All genotypes had similar changes in other cellular components that are recognized as indicators of plant senescence regardless of whether the plants produced seed.

Journal Article↗
PubMed1983

Immunochemical localization of NADP-specific isocitrate dehydrogenase in Escherichia coli.

The intracellular localization of isocitrate dehydrogenase was determined by immunochemical techniques with ultrathin sections of Escherichia coli. The thin sections, which were obtained by ultracryomicrotomy, were incubated first with antiserum specific for the enzyme and then with a protein A-gold complex. Transmission electron microscopy showed that the gold label was dispersed mainly in the cytoplasm.

Escherichia coli↗
PubMed1982

Characterization of Chloroplasts Isolated from Triazine-Susceptible and Triazine-Resistant Biotypes of Brassica campestris L.

Chloroplasts isolated from triazine-susceptible and triazine-resistant biotypes of Brassica campestris L. were analyzed for lipid composition, ultrastructure, and relative quantum requirements of photosynthesis. In general, phospholipids, but not glycolipids in chloroplasts from the triazine-resistant biotype had a higher linolenic acid concentration and lower levels of oleic and linoleic fatty acids, than chloroplasts from triazine-susceptible plants. Chloroplasts from the triazine-resistant biotype had a 1.6-fold higher concentration of t-Delta3-hexadecenoic acid with a concomitantly lower palmitic acid concentration in phosphatidylglycerol. Phosphatidylglycerol previously has been hypothesized to be a boundary lipid for photosystem II. Chloroplasts from the triazine-resistant biotype had a lower chlorophyll a/b ratio and exhibited increased grana stacking. Light-saturation curves revealed that the relative quantum requirement for whole chain electron transport at limiting light intensities was lower for the susceptible biotype than for the triazine-resistant biotype. Although the level of the chlorophyll a/b light-harvesting complex associated with photosystem II was greater in resistant biotypes, the increased levels of the light-harvesting complex did not increase the photosynthetic efficiency enough to overcome the rate limitation that is inherited concomitantly with the modification of the Striazine binding site.

Journal Article↗
PubMed1973

Crystallization of NADP-specific isocitrate dehydrogenase.

The first crystalline preparation of isocitrate dehydrogenase specific for nicotinamide adenine dinucleotide phosphate has been obtained with enzyme isolated from Escherichia coli. Scanning electron microscopy was employed to elucidate the structure of the crystals, which were found to exist as regular octahedrons ranging in size from 5 to 90 micrometers.

Crystallization↗
PubMedDate not supplied

Evidence for eumelanin and pheomelanin producing genotypes in the Arabian horse.

The ultrastructural imaging of melanocytes coupled with analyses to detect sulfur-containing melanosomes by energy-dispersive X-ray spectroscopy were used to test the hypothesis that the yellowish-red and black pigments found in Arabian horses result from pheomelanogenesis and eumelanogenesis, respectively. These procedures detected pheomelanosomes in follicles at the base of hairs in chestnut horses and eumelanosomes in follicles at the base of hairs in black horses. By analyzing tissue obtained by skin biopsy, these procedures also demonstrated that skin melanocytes in a chestnut horse produce eumelanosomes, and follicular melanocytes in the same horse produce pheomelanosomes. It was also shown that the type of follicular melanosome present in light bay horses is correlated with the color of the hair. The results of this study give experimental evidence for the Odriozola-Adalsteinsson hypothesis that the e allele is responsible for the chestnut phenotype; they also give fine structure and chemical confirmation of the action of the A and E loci in the Arabian horse as currently proposed for the mouse and other mammals.

Alleles↗