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1.
J Biomed Mater Res ; 62(1): 106-18, 2002 Oct.
Article in English | MEDLINE | ID: mdl-12124792

ABSTRACT

Retainers were collected from private, university, and dental labs. After viewing these corroded and control appliances using scanning electron microscopy, corroded maxillary and mandibular retainers were selected along with a control stainless-steel retainer for in-depth chemical analysis. Using electron spectroscopy for chemical analysis, monochromated Al x-rays were rastered over areas 1.5 x 0.3 mm. After survey spectra were acquired, high-resolution multiplex scans were obtained and binding energy shifts were noted. Using Auger electron spectroscopy, a spot size of approximately 30 nm was analyzed. Photos, survey scans, and depth profiles were acquired using a 3.5kV Ar(+) ion beam that was calibrated using a SiO2 standard. Via electron spectroscopy for chemical analysis, the brown stains contained Fe and Cr decomposition products in which three carbon species were present. Proteinaceous N was found as amines or amides. No Ni was present because it had solubilized. The Cr:Fe ratio indicated severe Cr depletion in the stained regions (0.2) versus the control regions (1.3). The stained regions appeared mottled, having both dark and light areas. Via AES, the dark versus light areas of the stained regions indicated that there was an absence versus a presence of both Cr and Ni. In the dark areas corrosion penetrated 700 nm; in the light areas the depth equaled 30 nm. By comparison, the passivated layer of the control retainer was 10-nm thick. After sputtering away the affected areas, all specimens had similar spectra as the control regions. The bacterial environment created the mottled appearance and induced electrochemical potential differences so that, upon reducing the passivated layer, an otherwise corrosion-resistant alloy became susceptible to rampant corrosion. An integrated biological-biomaterial model is presented for the classic case of an orthodontic acrylic-based stainless steel retainer subject to crevice corrosion.


Subject(s)
Orthodontic Retainers , Stainless Steel/chemistry , Acrylates , Corrosion , Equipment Failure Analysis/methods , Humans , Hydrocarbons/analysis , Metals/analysis , Microscopy, Electron, Scanning , Organic Chemicals/analysis , Oxidation-Reduction , Spectrum Analysis , Stainless Steel/analysis , X-Rays
2.
Oral Microbiol Immunol ; 16(3): 163-9, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11358538

ABSTRACT

Coaggregation between Porphyromonas gingivalis and Fusobacterium nucleatum strains was previously studied using either a semi-quantitative macroscopic assay or radioactive tracer assays. A new automated microtiter plate assay is introduced, in which the plate reader (Vmax) was adapted to allow quantitative evaluation of the kinetics of coaggregation. F nucleatum PK 1594 coaggregated with P. gingivalis HG 405 with a maximal coaggregation rate of 1.05 mOD/min, which occurred at a P. gingivalis to F. nucleatum cell ratio of 1 to 2. F. nucleatum PK 1594 failed to do so with P. gingivalis strains A 7436 or ATCC 33277. Galactose inhibition of this coaggregation could be quantitatively measured over a wide range of concentrations to demonstrate its dose-dependent manner. P. gingivalis HG 405 failed to coaggregate with F. nucleatum strains ATCC 25586 and ATCC 49256. The assay used in the present study is a sensitive and efficient quantitative automated tool to study coaggregation and may replace tedious radioactive tracer assays.


Subject(s)
Bacteriological Techniques , Fusobacterium nucleatum/cytology , Porphyromonas gingivalis/cytology , Automation , Bacterial Adhesion/drug effects , Dose-Response Relationship, Drug , Fusobacterium nucleatum/classification , Fusobacterium nucleatum/drug effects , Galactose/administration & dosage , Galactose/pharmacology , Humans , Kinetics , Porphyromonas gingivalis/classification , Porphyromonas gingivalis/drug effects , Sensitivity and Specificity , Statistics as Topic , Titrimetry
3.
Dent Mater ; 15(4): 282-95, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10551097

ABSTRACT

OBJECTIVES: Recently, a new generation of simplified one-bottle dentin bonding systems, sensitive to variations in the degree of substrate moisture, was introduced. This in vitro project compared the dentin bond strengths and interfacial ultra-morphology formed by three one-bottle bonding systems [OptiBond SOLO (ethanol-based), Prime&Bond 2.1 (acetone-based), and Single Bond (ethanol- and water-based)]. The null hypothesis tested was that re-wetting a dried dentin surface with a HEMA aqueous solution would not result in bond strengths, and resin impregnation into demineralized dentin, comparable to those obtained for moist dentin. METHODS: Dentin specimens were assigned to the following three etched surface conditions: moist dentin-control group; dentin dried for 5 s; and dentin dried for 5 s and re-moistened with a commercial 35% HEMA aqueous solution. Mean shear bond strengths were calculated and analyzed with one- and two-way ANOVA. Dentin discs treated with the same combination of surface condition/adhesive were processed and observed under both transmission and scanning electron microscopes. RESULTS: For moist dentin, the morphology of the resin-dentin interfaces showed penetration of the dentin adhesives to the depth of the transition between demineralized and unaffected dentin. Drying dentin for 5 s resulted in a significant decrease in mean bond strengths and an incompletely infiltrated collagen structure with areas of unveiled collagen fibers, regardless of the solvent. Re-wetting dentin with the aqueous HEMA solution re-established the level of bond strengths obtained to moist dentin and resulted in a raise of the fiber network with simultaneous increase in interfibrillar space dimensions. SIGNIFICANCE: The results suggest that the use of an aqueous HEMA solution might compensate for the dryness induced on dentin surfaces by using air blasts from an air syringe, after rinsing off the etchant. As the behavior of the material that contained water was also affected by surface dryness, the percentage of water included in the composition of current ethanol- and water-based adhesives, such as Single Bond, may not be enough to compensate for the collapse of the collagen filigree upon drying.


Subject(s)
Dental Bonding , Dentin-Bonding Agents/chemistry , Wetting Agents , Analysis of Variance , Animals , Bisphenol A-Glycidyl Methacrylate/chemistry , Cattle , Composite Resins/chemistry , Dentin/drug effects , Dentin/ultrastructure , Methacrylates/chemistry , Microscopy, Electron , Microscopy, Electron, Scanning Transmission , Polymethacrylic Acids/chemistry , Resin Cements/chemistry
4.
Am J Orthod Dentofacial Orthop ; 114(5): 558-72, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9810052

ABSTRACT

The static and kinetic frictional coefficients of commercially pure titanium brackets were evaluated in the passive configuration in the dry and wet states against stainless steel, nickel-titanium, and beta-titanium arch wires. For comparison, stainless steel brackets were evaluated under identical conditions. Titanium brackets were grayer in color and rougher in texture than the stainless steel brackets. Bracket slots were up to 0.002 inch greater than the nominally stated values. Remarkably, the static and kinetic frictional coefficients of the couples formed by titanium and stainless steel brackets were comparable. When evaluated against stainless steel and nickel-titanium arch wires in the dry state at 34 degrees C, the static coefficient averaged.12 and.20, respectively, independent of bracket alloy. When evaluated against stainless steel and nickel-titanium wires in the wet state at 34 degrees C using human saliva, the static coefficient averaged.15 and.20, respectively, independent of bracket alloy. Only the beta-titanium arch wires increased by about 15%, when tested in either the dry or the wet state against titanium versus stainless steel brackets. Noteworthy, too, was the decrease of both coefficients in the beta-titanium wire couples from their previously reported values. Analyses of electron spectroscopy for chemical analysis spectra and depth profiles show that these new brackets are titanium only in the bulk. Indeed the immediate surfaces are composed of, at least, 80 atomic percent (at.%) carbon and oxygen; whereas, the titanium that is present (>11 at.%) is mostly in the form of titanium dioxide. The presence of this quite thin passivating layer, which resides on top of an oxygen-hardened titanium substrate, reduces the galling and fretting that would normally be expected in such materials. Pending the outcome of future angulation tests, these frictional measurements show that titanium brackets are not only comparable to stainless steel brackets but also are more biocompatible with nickel having been eliminated from their constitution.


Subject(s)
Dental Alloys/chemistry , Orthodontic Brackets , Titanium/chemistry , Education, Dental, Continuing , Electron Probe Microanalysis , Friction , Linear Models , Microscopy, Electron, Scanning , Nickel/chemistry , Orthodontic Brackets/statistics & numerical data , Orthodontic Wires/statistics & numerical data , Orthodontics/education , Stainless Steel/chemistry , Surface Properties
5.
Proc Soc Exp Biol Med ; 217(3): 345-50, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9492346

ABSTRACT

The ovariectomized (OVX), lactating rat model has been used to investigate the skeletal effects of the plant estrogen, genistein, over a 14-day period. The OVX, lactating rat on a low-calcium diet loses slightly more than 50% of its bone mineral mass during the first 2 weeks of lactation, and we have demonstrated that estrogen treatment can significantly reduce the loss of femoral mass (ash weight). Following OVX, the rats were assigned to treatment or control groups (both placebo and positive control with estrogen replacement). The treatment groups received one of three doses of a genistein-rich preparation each day via the feed for 2 weeks, after which time the pups began to have an interest in solid feed. A positive control group received conjugated estrogen in the feed. The genistein doses were: low (0.5 mg/d); intermediate (1.6 mg/d); and high (5.0 mg/d). Measurements included ash weights of the femur, scanning electron microscopy (SEM) of the proximal tibia, and uterine weights. SEM results were as follows: (1) at the low dose genistein was approximately equally effective to estrogen in the retention of cancellous bone tissue, as reflected in the number and density of trabeculae in hemisections of the tibial subepiphyseal region, but at high doses genistein was less effective; and (2) rats treated with low-dose genistein, like estradiol, had rougher endosteal surfaces and smaller pores on these surfaces than untreated control rats. Mean ash weights of the entire femur were highest in the rats treated with the low dose compared to control rats (P < 0.05), and they were higher than ash weights of rats administered the intermediate or high doses of genistein. The mean ash weights of the femurs were consistent with the genistein effects on the tibias observed by SEM. In summary, a biphasic response to the genistein preparation was found in this OVX rat model. Interpretation of the results suggests that, at the low dose, genistein appears to be an agonist at the estrogen receptor locus, whereas at higher doses the genistein is less effective and may even have adverse effects on bone cells. These findings of a biphasic effect of genistein (i.e., an inverted U effect) are consistent with those of other recent reports in the literature on isolated bone cells and on reproductive tissues. In summary, lower doses of genistein from soy foods would be expected to act similarly to estrogens with a beneficial effect on bone tissue, but at high doses that are unlikely to be consumed in human diets, this soy derivative may have potentially adverse effects on bone cell functions and thereby on bone tissue.


Subject(s)
Bone and Bones/drug effects , Genistein/pharmacology , Animals , Bone Density/drug effects , Dose-Response Relationship, Drug , Female , Lactation , Ovariectomy , Rats , Rats, Sprague-Dawley , Uterus/drug effects
6.
Am J Dent ; 11(6): 291-301, 1998 Dec.
Article in English | MEDLINE | ID: mdl-10477981

ABSTRACT

PURPOSE: To evaluate the effects of a carbamide peroxide bleaching agent on interfaces formed by two one-bottle dental adhesives to etched enamel. The null hypotheses tested in this study were that vital bleaching with a commercial 10% carbamide peroxide gel would not (1) increase the concentration of oxygen in the superficial layer of enamel or (2) induce ultra-morphological changes in resin-enamel interfaces. MATERIALS AND METHODS: Five extracted human incisors were treated with 10% carbamide peroxide (Opalescence) for 4 h/day for 1 week and were compared with non-bleached teeth for oxygen, calcium, and phosphorus relative concentration using EDS. Mean elemental concentrations were analyzed using a t-test (bleached vs. unbleached enamel), one-way ANOVA (for surface location and also for depth) and three-way ANOVA (with bleaching treatment, surface location, and depth as the main factors). For TEM, fifteen extracted human molars were sectioned to obtain two crown halves. After roughening the occlusal surface, one half was bleached with Opalescence while the other half was stored in artificial saliva for 1 week. Enamel was etched for 15 s with a 35% phosphoric acid and bonded with one of three adhesives (Prime & Bond 2.1, Syntac Single-Component, or Scotchbond Multi-Purpose Adhesive-control) and a composite resin (Protect Liner F). Small enamel/resin sticks with a cross-section of 1.0 mm x 1.0 mm were removed and the specimens were processed for TEM observation. RESULTS: Vital bleaching with 10% carbamide peroxide caused no significant changes in relative oxygen concentration of enamel. For calcium and phosphorus, bleaching resulted in significant decreased relative concentrations. Bleaching also resulted in morphological alterations in the most superficial enamel crystallites. Some altered crystallites exhibited electron-lucent cores and reduced thickness of material around the core.


Subject(s)
Dental Enamel/drug effects , Dentin-Bonding Agents/chemistry , Peroxides/chemistry , Tooth Bleaching , Urea/analogs & derivatives , Acid Etching, Dental , Analysis of Variance , Calcium/analysis , Carbamide Peroxide , Composite Resins , Dental Enamel/chemistry , Dental Enamel/ultrastructure , Drug Combinations , Humans , Incisor , Materials Testing , Microscopy, Electron, Scanning Transmission , Molar , Oxygen/analysis , Phosphorus/analysis , Saliva, Artificial , Surface Properties , Tensile Strength , Tooth Bleaching/adverse effects , Urea/chemistry
7.
Int J Oral Maxillofac Implants ; 11(4): 456-65, 1996.
Article in English | MEDLINE | ID: mdl-8803341

ABSTRACT

Bone formation at implant surfaces may be directly influenced by effects of the implant material on osteoblast behavior. Cell culture models of osteoblast physiology may be used to investigate the interaction of osteoblastic cells with various surfaces. In this study, primary cultured fetal bovine mandibular osteoblastic cells were cultured on titanium, ceramic hydroxyapatite, and glass coverslip surfaces to allow for the comparison of the mineralizing matrix elaborated by osteoblasts grown on different implant material surfaces. Morphologic and immunohistochemical analysis revealed the similar formation of multilayered, mineralizing cultures on these three surfaces. The qualitative similarity of the matrix formed on these culture surfaces may reflect similar qualitative in vivo responses of bone to titanium and hydroxyapatite implants.


Subject(s)
Calcification, Physiologic , Durapatite/chemistry , Glass/chemistry , Mandible/physiology , Osteoblasts/physiology , Titanium/chemistry , Animals , Bone Matrix/cytology , Bone Matrix/physiology , Cattle , Cell Adhesion , Cells, Cultured , Ceramics/chemistry , Culture Media , Dental Implants , Immunohistochemistry , Mandible/cytology , Microscopy, Electron , Microscopy, Electron, Scanning , Osteogenesis , Surface Properties
8.
Cutis ; 57(2 Suppl): 13-8, 1996 Feb.
Article in English | MEDLINE | ID: mdl-8646864

ABSTRACT

The cosmetic and physicochemical properties of six topical corticosteroid creams were evaluated and compared. The following creams were provided in blinded tubes: Elocon, Westcort, Lidex, Kenalog, Valisone, and Cutivate. The following properties were evaluated in vitro: stiffness (hardness), grittiness, color, odor, homogeneity (phase separation), pH, weight loss, and tackiness (stickiness). Samples of the creams were evaluated by light microscopy and scanning electron microscopy to identify particle and droplet distribution, particulate contamination, and microscopic homogeneity of the products. Cutivate ranked number 1 in each category and received the best overall score for each of the cosmetic and physicochemical properties evaluated. The cosmetic and physicochemical properties of Elocon, Westcort, Lidex, and Kenalog were found to be similar to one another with regard to overall score but inferior to Cutivate. Valisone was also good with regard to overall score but was ranked less acceptable due to a strong odor.


Subject(s)
Anti-Inflammatory Agents/administration & dosage , Administration, Topical , Androstadienes/administration & dosage , Chemical Phenomena , Chemistry, Physical , Fluocinonide/administration & dosage , Fluticasone , Glucocorticoids , Mometasone Furoate , Ointments , Particle Size , Pregnadienediols/administration & dosage , Triamcinolone Acetonide/administration & dosage
9.
Int J Oral Maxillofac Implants ; 10(6): 655-65, 1995.
Article in English | MEDLINE | ID: mdl-8530168

ABSTRACT

In this study, the primary culture of bovine mandibular osteoblast cells in a microculture assay has been used to further investigate the interaction of mineralizing osteoblast cultures with implant surfaces by using correlative microscopic techniques. Rapid differentiation and mineralization of osteoblast cultures grown on titanium alloy surfaces was observed. The successful short-term culture of mineralizing mandibular osteoblasts on titanium alloy surfaces occurred without the formation of a tenacious adhesive interface between the alloplastic material and the multilayered cell culture.


Subject(s)
Osteoblasts/cytology , Animals , Cattle , Cell Adhesion , Cells, Cultured , Microscopy, Electron, Scanning , Osteoblasts/physiology , Osteoblasts/ultrastructure , Titanium
10.
Appl Environ Microbiol ; 57(11): 3086-92, 1991 Nov.
Article in English | MEDLINE | ID: mdl-16348576

ABSTRACT

The nonheterocystous filamentous cyanobacterial genus Lyngbya is a widespread and frequently dominant component of marine microbial mats. It is suspected of contributing to relatively high rates of N(2) fixation associated with mats. The ability to contemporaneously conduct O(2)-sensitive N(2) fixation and oxygenic photosynthesis was investigated in Lyngbya aestuarii isolates from a North Carolina intertidal mat. Short-term (<4-h) additions of the photosystem II (O(2) evolution) inhibitor 3(3,4-dichlorophenyl)-1,1-dimethylurea stimulated light-mediated N(2) fixation (nitrogenase activity), indicating potential inhibition of N(2) fixation by O(2) production. However, some degree of light-mediated N(2) fixation in the absence of 3(3,4-dichlorophenyl)-1,1-dimethylurea was observed. Electron microscopic immunocytochemical localization of nitrogenase, coupled to microautoradiographic studies of CO(2) fixation and cellular deposition of the tetrazolium salt 2,4,5-triphenyltetrazolium chloride, revealed that (i) nitrogenase was widely distributed throughout individual filaments during illuminated and dark periods, (ii) CO(2) fixation was most active in intercalary regions, and (iii) daylight 2,4,5-triphenyltetrazolium chloride reduction (formazan deposition) was most intense in terminal regions. Results suggest lateral partitioning of photosynthesis and N(2) fixation during illumination, with N(2) fixation being confined to terminal regions. During darkness, a larger share of the filament appears capable of N(2) fixation.

11.
Brain Res ; 536(1-2): 335-8, 1990 Dec 17.
Article in English | MEDLINE | ID: mdl-1964833

ABSTRACT

Cytochrome oxidase activity was investigated histochemically in the choroid plexus epithelium. Intense staining for the enzyme was exclusively limited to the mitochondria. Rats treated with octanoic acid displayed extensive ultrastructural disruptions in the epithelial cells of the choroid plexus. Mitochondria were fewer in number and more disrupted compared to the control. The enzyme activity was greatly reduced. However, pretreatment with an equimolar dose of L-carnitine followed by octanoic acid injection produced little alteration of either ultrastructure or enzyme staining. This study suggests that L-carnitine supplementation may restore mitochondrial function of the choroid plexus subjected to toxic organic anions in metabolic disorders, and may be useful in the prevention of metabolic encephalopathy.


Subject(s)
Caprylates/antagonists & inhibitors , Carnitine/therapeutic use , Choroid Plexus/drug effects , Mitochondria/drug effects , Animals , Choroid Plexus/enzymology , Electron Transport Complex IV/drug effects , Male , Mitochondria/enzymology , Rats , Rats, Inbred Strains
12.
J Bone Miner Res ; 5(2): 123-32, 1990 Feb.
Article in English | MEDLINE | ID: mdl-2316400

ABSTRACT

Measurements by scanning electron microscopy (SEM) of femoral hemisections confirmed and amplified results by single-photon absorptiometry that had shown a marked increase in lactation osteopenia in rats fed a low-calcium diet (LCD, 0.04% Ca) as compared with a medium-(adequate) calcium diet (ACD, 0.4% Ca). SEM of bones from rats at the end of lactation on either diet showed a large loss of trabecular bone, increased porosity of endosteal surfaces, and cortical thinning. These changes were much more striking in LCD rats than in ACD rats. Backscattered electron imaging of cross sections of the femora revealed marked cortical thinning at midshaft after lactation, especially in rats on the LCD; this method also showed a marked increase in newly formed, less dense diaphyseal bone on the endosteal surface when dietary calcium had been made available to the LCD rats after lactation ceased. Unlike the rats fed the ACD after lactation, the rats continued on the LCD for the first 3 weeks postlactation failed to recover bone mineral, even though there was a marked decrease in resorbing surfaces of the femora as revealed by morphologic examination. When the diet was changed from the LCD to the ACD for the second 3 weeks postlactation (week 4-6), the bone mineral increased substantially. Overall, these results demonstrate the marked loss of bone during lactation, especially severe in rats fed a low-calcium diet, and the rapid postlactational recovery of bone when adequate dietary calcium was made available, even if the recovery had been delayed for the first 3 weeks by feeding a diet very low in calcium.


Subject(s)
Bone Diseases, Metabolic/pathology , Lactation/physiology , Absorptiometry, Photon , Animals , Bone Density , Bone Diseases, Metabolic/etiology , Bone Diseases, Metabolic/metabolism , Calcium/blood , Female , Femur/ultrastructure , Microscopy, Electron, Scanning , Pregnancy , Rats , Rats, Inbred Strains
13.
J Bone Miner Res ; 4(3): 399-405, 1989 Jun.
Article in English | MEDLINE | ID: mdl-2763875

ABSTRACT

The intracellular localization of the tartrate-resistant purple acid phosphatase in osteoclasts of developing rat bone has been determined immunocytochemically using an antiserum to the purified bone-derived purple acid phosphatase. The localization of the immunoreactivity was compared with the results of enzyme histochemistry using p-nitrophenylphosphate as substrate and 10 mM tartrate. Both methods revealed the presence of the enzyme in numerous vesicles of various sizes up to 2-3 microns in diameter and in granules. There was no immunoreactivity in the Golgi apparatus, and tartrate completely inhibited the histochemical activity of this organelle. No consistent extracellular activity could be detected, nor was any reaction product observed at the ruffled border. The localization of the tartrate-resistant purple acid phosphatase in osteoclasts is consistent with an intracellular function for this enzyme.


Subject(s)
Acid Phosphatase/analysis , Osteoclasts/enzymology , Animals , Histocytochemistry , Immunohistochemistry , Microscopy, Electron , Osteoclasts/ultrastructure , Rats , Tartrates/pharmacology
14.
Toxicol Appl Pharmacol ; 90(3): 436-44, 1987 Sep 30.
Article in English | MEDLINE | ID: mdl-2444018

ABSTRACT

2,4,5-Trichlorophenoxyacetic acid (2,4,5-T) reduced the uptake of 5-hydroxy-3-indoleacetic acid (5-HIAA) by the choroid plexus in a dose-related manner, while treatment with quinolinic acid at comparable concentrations did not inhibit 5-HIAA uptake. The role of carrier-mediated transport in the clearance of 5-HIAA from cerebrospinal fluid (CSF) was also evaluated in vivo by ventriculocisternal perfusion. Steady-state clearance of 5-HIAA from CSF exceeded that of inulin and was reduced competitively in the presence of 2,4,5-T. However, the clearance was not affected by quinolinic acid. The effect of 2,4,5-T on transport enzyme systems was also studied by electron microscopic cytochemistry. Na+-K+-ATPase and cytochrome oxidase activities in the choroid plexus were reduced by 2,4,5-T. Since this transport system in the choroid plexus is normally responsible for the excretion of the serotonin metabolite from the brain to the plasma, accumulation of endogenously produced organic acids in the CSF and the brain, secondary to reduced clearance by the choroid plexus, could be a contributing factor in the development of neurotoxicity.


Subject(s)
2,4,5-Trichlorophenoxyacetic Acid/toxicity , Choroid Plexus/metabolism , Hydroxyindoleacetic Acid/metabolism , Pyridines/toxicity , Quinolinic Acids/toxicity , Animals , Biological Transport/drug effects , Brain/drug effects , Carnitine/pharmacology , Choroid Plexus/drug effects , Electron Transport Complex IV/analysis , Female , Histocytochemistry , In Vitro Techniques , Male , Metabolic Clearance Rate , Microscopy, Electron , Quinolinic Acid , Rabbits , Sodium-Potassium-Exchanging ATPase/analysis
15.
Scan Electron Microsc ; (Pt 1): 311-21, 1983.
Article in English | MEDLINE | ID: mdl-6415807

ABSTRACT

Medusa cells, amoeboid variants of the eosinophil with pseudopod-like processes, were examined by light microscopy (LM), transmission electron microscopy (TEM), the secondary electron imaging (SEI) and the backscattered electron imaging (BEI) modes of the scanning electron microscope. TEM was performed on rare medusa cells found in leukocyte concentrate preparations where the ion contents of the collection and fixation media were balanced so that divalent cations such as calcium and magnesium were not sequestered. LM, SEI and BEI studies were performed principally on cytochemically-stained films of leukocyte concentrate preparations on microscope slides or coverslips. These films of patients with eosinophilia contained many medusa cells and much higher ratios of medusa cells to eosinophils than critical point-dried specimens, if they were prepared as for routine hematologic examination, and precautions were taken to insure that calcium and magnesium ions in collection and fixation media were not sequestered. After brief glutaraldehyde fixation, the smears were stained with either osmium tetramethylethylenediamine (Os-TMEDA) for acid mucopolysaccharides, or 3,3'-diaminobenzidine (DAB)/hydrogen peroxide medium for peroxidases. The Os-TMEDA was sufficiently conductive for SEM. Chelation of the oxidatively-polymerized DAB dye with copper nitrate rendered it conductive. These conductive and electron-opaque stains permitted the correlation of SEI with BEI on individual cells, their unambiguous identification as eosinophils or medusa cells and their differentiation from other leukocytes by virtue of content and/or size of their granules and their degree of nuclear segmentation.


Subject(s)
Eosinophils/ultrastructure , Cell Separation , Eosinophils/cytology , Eosinophils/enzymology , Glutaral , Histocytochemistry , Humans , Microscopy, Electron/methods , Microscopy, Electron, Scanning/methods , Peroxidases/blood
16.
Acta Neuropathol ; 51(2): 145-53, 1980.
Article in English | MEDLINE | ID: mdl-6969018

ABSTRACT

C57BL/KsJ db/db inbred mice have an hereditary autosomal recessive disease resembling in some respects maturity onset human diabetes mellitus. At 8--11 months of age, they displayed intermittent symptoms suggestive of a mild sensory neuropathy. These symptoms consisted of adduction of their hind limbs and flexing hind paws when raised by the tail, and inability to maintain their position on the roto wheel. Peripheral nerves and sensory ganglia of the diabetic mice were compared with those of the unafflicted littermates and studied with respect to Schwann cell counts and myelinated nerve fiber diameter measurements. In addition, teased fibers of peripheral nerves were compared for obvious changes in internodal distance and demyelination. Chromatolytic neurons were moe abundant in lumbosacral spinal ganglia of diabetic mice than in corresponding ganglia of controls or in more anterior spinal ganglia and trigeminal ganglia of diabetics. Histologic studies showed an increase in Schwann cell counts in longitudinal sections of peripheral nerves. A similar but larger increase was observed in peripheral nerves of mice affected with an hereditary sensory neuropathy, dystonia musculorum. A small but general decrease in myelinated fiber diameter was observed in sensory and motor nerves.


Subject(s)
Diabetes Mellitus, Experimental/pathology , Diabetic Neuropathies/pathology , Animals , Cell Count , Dystonia/pathology , Ganglia, Spinal/pathology , Mice , Mice, Inbred C57BL/genetics , Mice, Mutant Strains , Nerve Fibers, Myelinated/pathology , Peripheral Nerves/pathology , Schwann Cells/pathology , Trigeminal Ganglion/pathology
17.
Histochemistry ; 68(2): 99-118, 1980.
Article in English | MEDLINE | ID: mdl-7419441

ABSTRACT

Circulating androgens are known to effect a sexual dimorphism of the submandibular gland and kidney of the mouse. Enzyme histocytochemical differences that correlate with these structural changes have been the subject of much study, especially in the kidney. In the present study, emphasis was placed on the hypogonadic effects of diabetes mellitus on the submandibular gland and kidney of C57Bl/KsJ db/db inbred mice with an autosomal recessive disease resembling maturity onset human diabetes mellitus. These glands of adult diabetic mice of both sexes were compared with those of unafflicted heterozygous littermates. The mitochondrial cytochrome oxidase and peroxisomal and cytoplasmic catalase were studied in their submandibular glands and kidneys. The parasympathetic innervation of the submandibular glands was studied by a histochemical method for acetylcholinesterase. The extensive differentiation of striated ducts of the submandibular gland into granular tubules in the postpubertal male mouse was readily evident with the cytochrome oxidase procedure. This differentiation resulted in ductal staining patterns characteristic of the sexes. Alteration of these patterns suggested that demasculinization or feminization was occuring in the male diabetic mice and that masculinization or virilization (defeminization) was occurring in the female diabetics. Similarly, in kidney, study of the parietal epithelium of Bowman's capsule revealed feminization in the male diabetics and masculinization in the female diabetics. With the catalase procedure, a dramatic sexual dimorphism was observed in the kidneys of the heterozygous unafflicted mice. Peroxisomal staining of epithelial cells of the proximal convoluted tubules was much more intense in the outer medulla of the male than of the female. In kidneys of the diabetics, the staining patterns again suggested that feminization of the male and masculinization of the female kidneys had occurred. On the other hand, neither a sexual dichotomy nor effects due to diabetes could be observed in the characteristic catalase staining observed in the luminal epithelial cells of submandibular gland distal ducts. The parasympathetic innervation of the submandibular gland, as revealed by the acetylcholinesterase method, was also markedly sexually dimorphic in the unafflicted mice. This was due to the more extensive innervation of the larger granular ducts characteristic of male than of the smaller striated ducts of the female. As a result of diabetes, the innervation and duct size decreased in the submandibular gland of the male, suggesting feminization, whereas they increased in the female suggesting masculinization. These changes were consistent with those observed in sumandibular gland with the cytochrome oxidase procedure. Attempts were made to interrelate all of the enzyme histochemical changes observed in submandibular gland and kidney with the weights of these glands, sex, gonadal weights, diabetic status and urinary protein excretion...


Subject(s)
Diabetes Mellitus/pathology , Hypogonadism/pathology , Kidney/pathology , Submandibular Gland/pathology , Acetylcholinesterase/metabolism , Animals , Diabetes Mellitus/genetics , Diabetes Mellitus/urine , Female , Histocytochemistry , Hypogonadism/urine , Kidney/enzymology , Male , Mice , Organ Size , Oxidoreductases/metabolism , Submandibular Gland/enzymology
18.
Cancer Res ; 39(5): 1635-9, 1979 May.
Article in English | MEDLINE | ID: mdl-85486

ABSTRACT

Hydroperoxidase-positive Phi bodies and rods are much more prominent and prevalent than rods visualized with a Romanovsky-type stain (Auer rods) in immature leukocytes of patients with active acute myelogenous leukemia (AML). They are readily observed with the light microscope in peripheral blood or marrow films of AML patients stained to show their peroxidatic activity. In many of these patients, Auer rods, which apparently constitute only a small subpopulation of the hydroperoxidase-positive Phi bodies and rods, were detected with difficulty, if at all. The hydroperoxidase-positive Phi bodies and rods were observed in 92% of 36 patients with active disease. They were never observed in leukocytes of patients with other hematopoietic disorders or of normal individuals. Thus, they facilitated the distinction of AML from acute lymphocytic leukemia and chronic granulocytic leukemia in blast crisis. They were absent in full clinical remission after chemotherapy and were greatly diminished in partial remission. They were present in disease relapse and reappeared in five patients who had been in full remission. These results suggest that these hydroperoxidase-positive enlarged particles are pathognomonic of AML and that monitoring them with the light microscope may aid in guiding its clinical management.


Subject(s)
Leukemia, Myeloid, Acute/diagnosis , Adolescent , Adult , Aged , Catalase , Cytoplasmic Granules/pathology , Histocytochemistry , Humans , Leukemia, Myeloid, Acute/blood , Middle Aged , Peroxidases , Staining and Labeling
19.
Histochem J ; 10(6): 711-20, 1978 Nov.
Article in English | MEDLINE | ID: mdl-83316

ABSTRACT

Acetylcholinesterase (AChE) activity of primary sensory neurons of the cat has been quantitated and correlated with cell size. Dorsal root ganglia of the fourth and fifth thoracic spinal levels were studied. Frozen longitudinal and cross-sections were collected serially and stained with Cresyl Violet for total cell counts of the ganglia on the left; the average count was 3375 cells. Ganglia from the right were stained for AChE after the method of Karnovsky & Roots (1964) as modified by El Badawi & Schenk (1967), and counterstained with Haematoxylin. Cells were counted in every fourth section and the diameter of each was recorded. AChE-positive cells were classified as brown (B1, B2, B3) and AChE-negative ones as blue (BL). An inverse correlation exists between cell size and AChE activity. High activity was demonstrated in 29% of the cells (B1), moderate activity in 52% (B2), minimal activity in 15% (B3) and 4% were classified as AChE-negative (BL). Small cells with high activity were centrally located in the ganglia whereas large AChE-negative cells were peripherally distributed. Chi-Square analysis revealed that the size of the cell was not independent of the enzyme colour category.


Subject(s)
Acetylcholinesterase/isolation & purification , Cats/anatomy & histology , Ganglia, Spinal/cytology , Neurons/enzymology , Animals , Cell Count , Female , Male , Staining and Labeling
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