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Carbonated apatite-induced arthropathy: a consideration in cases of polyarthritis.

BACKGROUND: A 79-year-old woman was referred for evaluation of her painful and swollen joints. She had a medical history of congestive heart failure, renal insufficiency and peptic ulcer disease. For the past 3 years she had experienced recurrent bouts of debilitating arthritis, lasting approximately 3-4 weeks at a time. The symptoms were most severe in the hands and knees, where the joints were warm, swollen and tender. During each flare-up, the patient was housebound and required therapeutic dosing of nonsteroidal anti-inflammatory drugs and codeine to control joint pain. INVESTIGATIONS: Physical examination, fine-detailed radiographs of the hands, standing radiographs of the knees, arthrocentesis including cell count and gram stain, compensated polarized light microscopy, alizarin-red staining, X-ray diffraction, scanning and transmission electron microscopy with energy dispersive spectrometry, electron microprobe analysis with energy dispersive spectrometry, Fourier transform infrared spectroscopy, and atomic force microscopy. DIAGNOSIS: Carbonated-substituted apatite arthropathy. MANAGEMENT: Both knees were aspirated and large volumes of a straw-colored synovial fluid was removed. The knees were injected with corticosteroid, resulting in excellent symptomatic response.

Aged↗

Configuration of the two kinesin motor domains during ATP hydrolysis.

To understand the mechanism of kinesin movement we have investigated the relative configuration of the two kinesin motor domains during ATP hydrolysis using fluorescence polarization microscopy of ensemble and single molecules. We found that: (i) in nucleotide states that induce strong microtubule binding, both motor domains are bound to the microtubule with similar orientations; (ii) this orientation is maintained during processive motion in the presence of ATP; (iii) the neck-linker region of the motor domain has distinct configurations for each nucleotide condition tested. Our results fit well with a hand-over-hand type movement mechanism and suggest how the ATPase cycle in the two motor domains is coordinated. We propose that the motor neck-linker domain configuration controls ADP release.

Adenosine Triphosphate↗

Structural and molecular hair abnormalities in trichothiodystrophy.

We examined hair from 15 patients with trichothiodystrophy (TTD), a rare inherited disorder with brittle, cystine-deficient hair. They had a wide variety of phenotypes, from brittle hair only to severe intellectual impairment and developmental delay. Polarizing light microscopic examination showed alternating light and dark (tiger tail) bands under polarizing microscopy. Confocal microscopy captured structural features of breaks in intact TTD hairs. The autofluorescent appearance was regular and smooth in normal donors and markedly irregular in sections of TTD hairs possibly reflecting abnormalities in melanin distribution. Scanning electron microscopy revealed numerous surface irregularities. All TTD hair samples had reduced sulfur content. We observed an inverse correlation (R(val)=0.9) between sulfur content and percent of hairs with shaft abnormalities (trichoschisis, trichorrhexis nodosa, or ribbon/twist). There was no association between clinical disease severity and percent of abnormal hairs. Raman spectra of hairs from TTD patients and normal donors revealed a larger contribution of energetically less favored disulfide conformers in TTD hairs. Our data indicate that the brittleness of the TTD hair is dependent upon abnormalities at several levels of organization. These changes make TTD hairs excessively prone to breakage and weathering.

Adolescent↗

Shear-banding and microstructure of colloids in shear flow.

We shall discuss the following phenomena found in various colloidal systems in shear flow. We recently observed shear-banding in suspensions of fd-virus in a cylindrical shear cell. Small angle light scattering experiments revealed that the shear-banding transition is preceded by a relatively fast process (minutes) of nematic-to-paranematic phase separation during which inhomogeneities on the micrometer length scale are formed. After the formation of these inhomogeneities, a slow (hours) appearence of shear-bands is observed, which have a height of a few mm. In the stationary state it is found, by means of polarization microscopy, that inhomogeneities exist within the bands. Small angle, time resolved light scattering experiments on near-critical microstructural order in a mixture of colloidal spheres and free polymer under stationary shear flow are discussed. The unexpected distortion of microstructure in directions perpendicular to the flow direction is quantitatively explained by extending an already existing theory, to include shear-induced short-ranged microstructural distortion. In colloidal systems consisting of "hairy colloids", where a spherical core is decorated with relatively long polymers, shear-induced polymer brush deformation might be important for its structural and rheological behavior. Preliminary neutron scattering and rheology experiments are performed to study polymer brush deformation in (semi-) dilute suspensions.

Journal Article↗

Mechanical manipulation of molecular lattice parameters in smectic elastomers.

Smectic liquid crystalline elastomers (SLCE) represent unique materials that combine a 1-D molecular lattice arrangement and orientational order with rubber-elasticity mediated by a polymer network. Such materials may exhibit large thermo-mechanical, opto-mechanical and electro-mechanical effects, due to the coupling of macroscopic sample geometry and microscopic structural features. It is shown that the molecular layer dimensions in the smectic phases can be influenced reversibly by macroscopic strain of the material. We present a microscopic model on the basis of experimental results obtained by mechanical dilatation measurements, optical interferometry, X-ray scattering, (13)C NMR, FTIR and polarizing microscopy data. The model gives an explanation of the controversial results obtained in different types of smectic elastomers.

Computer Simulation↗

Cholesterol gallstone pathogenesis: a study of potential nucleating agents for cholesterol crystal formation in bile.

Cholesterol monohydrate crystal formation was measured quantitatively in model bile solutions, which were supersaturated with cholesterol, by a radiochemical method and qualitatively in human gallbladder bile by polarizing microscopy. Various agents, which have been postulated to act as nucleating factors for cholesterol crystal and gallstone formation, were added to bile and their effect on the appearance of cholesterol crystals was determined. These agents included calcium salts found in gallstones (calcite, aragonite, apatite, bilirubinate), Escherichia coli bacteria, pigment residues from cholesterol gallstones, bilirubin and several mucin preparations. Human gallbladder bile, which was collected from patients with and without cholesterol gallstones, was also mixed with model bile to examine whether nucleating or anti-nucleating factors were present. None of the agents tested markedly and consistently promoted cholesterol monohydrate crystal formation in model or human bile, except seed crystals of cholesterol monohydrate which were used as a control. Human gallbladder bile from obese patients without gallstones delayed the appearance of cholesterol crystals in model bile solutions, whereas gallbladder bile from gallstone patients did not. These results do not provide experimental support for the hypothesis that calcium salts and pigment material found in gallstones, or gallbladder mucin at concentrations less than 10 mg/ml, act as nucleating agents for cholesterol crystal and stone formation. The difference between gallbladder biles from patients with and without gallstones in their propensity to form cholesterol crystals may be due to the presence of an anti-nucleating factor in normal bile.

Animals↗

Rigid endosseous implants for orthodontic and orthopedic anchorage.

Clinical rigidity ("osseointegration") was achieved by 94 percent of the titanium implants placed in dog mandibles. All loaded implants remained rigid. Inadvertent impingement on the periodontal ligament and roots of adjacent teeth was uneventful. Microradiographic imaging was more reliable than polarized light microscopy for assessing bone contact at the endosseous interface. Implants with less than 10 percent of the endosseous interface in direct contact with bone successfully resisted a continuous load of 3 N (greater than 300 grams) for 13 weeks.

Acid Etching, Dental↗

Histological features of artificial secondary caries adjacent to amalgam restorations.

Artificially induced carious lesions on either side of in vitro un/restored amalgam cavities were examined to establish the degree of randomness of caries development. Class I cavities were cut in 132 extracted premolars: twelve teeth were not restored, and 120 teeth were restored with one of 20 different restoration combinations of silver amalgam, base and varnish. After ageing for periods of 3 months and 1 year, the 12 unrestored teeth and 80 of the restored teeth were subjected to an in vitro bacterial challenge for 36 days, while the other 40 specimens were challenged in acidified broth (pH 4.0). Sections were then prepared for polarized light microscopy. Carious lesion configuration on either side of the cavity was noted, and outer, wall and dentine lesions measured. Data underwent Fisher's exact test, a chi-squared goodness of fit test and a Student's paired t-test with P<0.05. Except for dentine lesions, acid broth and unrestored specimens showed carious lesions having similar size and occurrence on either side of the cavity. Two unrestored cavities showed caries resistance. Restored, bacterially challenged specimens were significantly different regarding total and wall lesion distribution and wall lesion width and area on either side of the cut cavity. Acid broth challenge will promote regular caries development at the tooth-restoration interface. The random caries pattern which developed in restored bacterially challenged specimens indicates that the tooth-restoration interface forms a diverse environment providing sites of varying susceptibility to caries.

Bicuspid↗

Characterization of proteoglycans and glycosaminoglycans in splenic AA amyloid induced in mink.

Amyloidosis of the protein AA type is readily induced in mink using repeated injections of bacterial lipopolysaccharide (LPS). We have characterized splenic proteoglycans/glycosaminoglycans (PGs/GAGs) in mink during amyloidogenesis. Moderate to rich amounts of amyloid exhibiting green birefringence was demonstrated by polarization microscopy of the splenic section stained with Congo red in seven out of eight minks after 10 weeks of LPS-treatment, and a significant increase in the total amount of PGs and GAGs in AA amyloid spleens was observed (two to eight times that in unstimulated animals). Intact PGs as well as free GAGs were extracted, and heparan sulfate (HS) was the most abundant GAG in the amyloid as well as in the control spleens. The GAGs showing the most pronounced increase in the amyloid spleens was of the chondroitin sulfate/dermatan sulfate (CS/DS) type and these were extracted in the form of free GAG chains. We conclude that there is a selective enrichment of PGs/GAGs in extracted splenic amyloid in the mink, which confirms to previous observations in human amyloid as well as in other animal species, supporting their pathogenic significance in the formation of AA amyloid.

Amyloidosis↗

A new variant of trichothiodystrophy with recurrent infections, failure to thrive, and death.

Two brothers demonstrated a severe variant of trichothiodystrophy. Both had brittle hair, developmental delay with severe failure to thrive, recurrent infections, cataracts, and angioendotheliomas of the liver at autopsy. The elder died at 12 weeks, the younger at 6 months. The younger had the typical appearance of banded hair on polarizing microscopy and a low cystine content measured by ion exchange chromatography. The history, clinical findings, and basic defects of trichothiodystrophy are discussed.

Bacterial Infections↗

Gout and other crystal-associated arthropathies.

Intra-articular crystals (monosodium urate monohydrate, calcium pyrophosphate dihydrate, basic calcium phosphates) can cause acute and chronic inflammation and joint damage. Identification of the crystals by polarized microscopy is the key step in diagnosis but improved reliability of synovial examination is required. Treatment of disorders associated with gout or calcium pyrophosphate deposition may reduce non-joint morbidity and assist treatment of the arthritis. Various forms of anti-inflammatory therapy work for acute crystal-induced arthritis; prompt commencement is usually more important than which option is used. In gout, recurrent attacks are usual, but hypouricaemic therapy is almost never urgent, is life-long, and is too often negated by poor compliance. In most patients, allopurinol or any of the potent uricosuric drugs will allow maintenance of normouricaemia but renal failure, renal calculi, transplantation, and allopurinol allergy narrow the options and complicate management.

Calcium↗

Histochemical studies of the extracellular matrix of human articular cartilage--a review.

OBJECTIVE: This paper reviews the histochemistry of the extracellular matrix of human articular cartilage. No systematic review of histochemical knowledge and techniques in the study of articular cartilage has been published previously. METHODS AND RESULTS: Literature was searched in the Winspirs Medline database from 1960 to 2000. Only techniques applicable for bright field or polarization microscopy were considered. Unless otherwise noted, all applies to hyaline cartilage. The most widely used fixatives are adequate for routine staining of proteins, but proteoglycan fixation is problematic, and no one fixative can be recommended. Proteoglycan can be stained reliably but it is problematic that, at low substrate concentrations, these methods are not stoichiometric. Collagen can be stained efficiently, although attempts to differentiate collagen types have not been successful. CONCLUSIONS: Detailed studies of fixation and staining procedures should be carried out and standards for cartilage sampling, handling and evaluation agreed upon if results from different laboratories are to be compared.

Cartilage, Articular↗

Characteristics of topographical heterogeneity of articular cartilage over the joint surface of a humeral head.

OBJECTIVE: To investigate the topographical variations among the morphological and physical properties of articular cartilage over a canine humeral head. DESIGN: Nine side-by-side specimens from a canine humeral head were used in the combined polarized light microscopy (PLM) and microscopic magnetic resonance imaging (muMRI). Sixteen images were acquired from each specimen in muMRI. Subsequently, ten to fourteen histological slices were prepared at the location of the muMRI slice from each specimen. Four optical images were acquired from each histological slice. Using these images, the images of the T(2) relaxation, the optical retardation, and the collagen-fiber orientation were constructed in two dimensions. RESULTS: Along the medial/lateral direction of the humeral head, we have observed a number of topographical variations. These include the total thickness of the cartilage tissue, the thickness of the individual histological zones, the depth of the minimum optical retardation in the tissue, and the depth of the maximum T(2) relaxation in the tissue. We found that the depth of the minimum retardation from PLM occurs at the geometric middle point of the transitional zone in histology, and that the depth of the maximum T(2) relaxation from muMRI is closely correlated with the minimum retardation depth in PLM. In addition, although the thickness of the total tissue increases approximately by the same amount on both the lateral and medial sides of the joint, a slight asymmetry exists between the measurements from the medial and lateral halves of the humerus. CONCLUSIONS: Cartilage from different sites over a single joint could have different properties, possibly the consequence of the variation in load-bearing status. Because of these topographical variations, one must keep in mind that a different sampling site within a relatively small area of the same joint may significantly influence the results of the study.

Animals↗

Equation of state and structural changes in diaminodinitroethylene under compression.

Structural changes in 1,1-diamino-2,2-dinitroethylene (DADNE, FOX-7) compressed to high pressure in diamond anvil cells were investigated using angle-dispersive x-ray diffraction analysis, Raman spectroscopy, and optical polarizing microscopy. The x-ray results show several changes above 1 GPa. When the x-ray data are indexed according to the ambient-pressure structure, DADNE shows anisotropic compression, with higher compression along the b axis than along the a or c axis. An ambient-temperature isothermal equation of state of DADNE was generated from these data. In addition, the experimentally obtained Raman spectra were matched with vibrational normal modes calculated using quantum chemistry calculations. The shifts in vibrational modes indicate changes in H-wagging vibrations with pressure.

Journal Article↗

The effects of argon laser irradiation on enamel decalcification: An in vivo study.

Enamel decalcification is a significant problem in orthodontic patients. The argon laser has been shown to reduce decalcification during an acidic challenge in vitro. The purpose of this study was to investigate the in vivo effects of argon laser irradiation on enamel decalcification during orthodontic treatment. Nine volunteers whose treatment plans included 4 first premolar extractions were enrolled in the study. The 36 extracted premolars were assigned to 1 of the following 4 groups: group 1, control group with no treatment; group 2 (pumice-laser), teeth were pumiced for 3 seconds and treated with a 325 mW, 5-mm diameter laser beam for 60 seconds; group 3 (pumice-etch-laser), teeth were pumiced for 3 seconds, acid-etched with 30% phosphoric acid for 30 seconds, and treated for 60 seconds with laser; and group 4 (laser only), teeth were treated for 60 seconds with laser. A specially designed (oversized) orthodontic band was fitted on each of the premolars to create a pocket for decalcification. The bands were cemented in place for 5 weeks. After extraction, the teeth were sectioned and examined under polarized light microscopy. Images of lesions were digitally analyzed and measured. Average lesion depths were calculated from 3 depth measurements recorded 10 microm apart. Average lesion area was calculated with the aid of imaging analysis software. Data were analyzed with analysis of variance (P <.05) and Student t tests. Significant differences were found in lesion depth (P <.001) and lesion area (P <.01) among the 4 test groups. The average lesion depths were 15.93 +/- 9.31 microm (control), 6.45 +/- 8.70 microm (pumice-laser), 1.71 +/- 4.82 microm (pumice-etch-laser), and 1.34 +/- 3.80 microm (laser only). The average lesion areas were 1028.67 +/- 725.68 microm (2) (control), 555.49 +/- 948.20 microm (2) (pumice-laser), 79.91 +/- 226.03 microm (2) (pumice-etch-laser), and 55.71 +/- 157.59 microm (2) (laser only). The average lesion depth in the laser-only group was reduced by 94.1% and the average lesion area was reduced by 94.4% when compared with the control group. In the pumice-etch-laser group, the average lesion depth was reduced by 89.1% and the average lesion area was reduced by 92.2% when compared with the control group. There were no significant differences in lesion depth and lesion area between maxillary and mandibular teeth (P <.06 and P <.08, respectively) and between the teeth on the right and left sides (P <.68 and P <.55, respectively). These results show that argon laser irradiation is effective in reducing enamel decalcification during orthodontic treatment. Pumicing and etching do not appear to reduce the effect of laser on enamel solubility.

Acid Etching, Dental↗

In vitro evaluation of fluoride varnish on overdenture abutments.

STATEMENT OF PROBLEM: No clinical study has evaluated the efficacy of fluoride varnish as a topical fluoride agent for the protection of overdenture abutments from dental caries. PURPOSE: This study assessed the efficacy of a fluoride varnish in protecting overdenture abutments from demineralization in an in vitro environment. MATERIAL AND METHODS: Eighty caries-free, extracted teeth were sectioned and domed immediately below the cementoenamel junction, simulating overdenture abutment preparations. Acid resistant nail varnish was painted on all surfaces except for a window (1 x 4 mm) on the prepared occlusal dentin surfaces. Teeth were randomly divided into 4 groups of 20 specimens: control, washed with deionized/distilled water; Daily Gel, treated with Karigel-N (5,000 ppm) for 4 minutes daily; Weekly Gel, treated with Karigel-N for 4 minutes weekly; and Weekly Varnish, treated with Duraflor (22,600 ppm) weekly. The varnish was removed using a blade 24 hours after each application to simulate the clinical loss of the varnish. Teeth were then placed in a cycle of demineralization (2.2 mMol/L CaCl(2)2H(2)O, 2.2 mMol/L KH(2)PO(4), and 50 mMol/L acetic acid at pH 4.3) for 6 hours and remineralization (1.5 mMol/L CaCl(2)2H(2)O, 0.9 mMol/L KH(2)PO(4), and 150 mMol/L KCl at pH 7.0 for 17 hours) for 21 days. Half the teeth in each group were brushed with no dentifrice for 10 seconds, twice daily. Teeth were sectioned at 100 +/- 10 microm buccolingually and evaluated under polarized light microscopy and contact microradiography. The depth of each lesion and the width of the remineralization bands were measured (mm). Analysis of variance models and T-tests were used to assess the effects of different treatments (alpha=.05). Duncan multiple range tests were then chosen as post hoc tests to evaluate the statistical significance of all pairwise comparisons. RESULTS: The control group had the deepest lesions (mean depth 400 +/- 39 microm). The lesions from the varnish and daily gel groups were significantly shallower than the lesions in both the control group and the weekly gel group. The remineralization bands in the varnish group were 52% wider than the control group bands. However, the daily gel group had the largest increase in band width (117% increase) compared with the control group. There were significant differences between all groups for net depth (lesion depth minus the remineralization band) (P<.0001). Daily application of fluoride gel was significantly more effective than the other treatments tested. Brushing was not a significant factor. CONCLUSION: Within the limitations of this in vitro study, treatment of overdenture abutment with various topical fluorides significantly inhibited demineralization and enhanced remineralization of cut occlusal dentin surfaces. Daily use of Karigel-N was the most effective treatment, followed by the weekly application of Duraflor varnish.

Analysis of Variance↗

The role of fibrinolytic system proteins in cholesterol gallstone formation.

BACKGROUND: Accelerated nucleation, supersaturation of bile, and biliary stasis are known to be key factors in cholesterol gallstone formation. The mechanisms through which these factors interact to form stones are still incompletely understood. Among the proteins now known to be present in bile are several components of the fibrinolytic system: tissue plasminogen activator, urokinase-like plasminogen activator, and plasminogen activator inhibitors 1 and 2. The concentrations of plasminogen activator inhibitors 1 and 2 in gallbladder bile are increased in patients with gallstones. The aim of this study was to determine whether these fibrinolytic system proteins act as pro-nucleating agents for cholesterol gallstone formation. METHODS: Nucleation assays were done on gallbladder bile from eight cholesterol stone patients and eight control patients. The effects of tissue plasminogen activator, urokinase-like plasminogen activator, and plasminogen activator inhibitors I and 2 on cholesterol crystal appearance time (CCAT) were tested, by direct observation using polarizing microscopy, after measurement of biliary lipids and calculation of cholesterol saturation indices. RESULTS: There was no significant difference in cholesterol saturation indices between bile that nucleated and bile that did not (mean, 2.0 +/- 1.5 versus 1.8 +/- 0.5). When all samples in which nucleation occurred were compared, tissue plasminogen activator significantly shortened CCAT median from 4.75 days (range, 2-21) to 3.5 days (2.5-18) (P < 0.05). This was similar to the effect of fibronectin (3.75 days; range, 2-20), a known pro-nucleator used as a nucleation accelerating control (P < 0.05). None of the other fibrinolytic system proteins significantly accelerated CCAT. CONCLUSIONS: The results of this study suggest that tissue plasminogen activator may act as a pro-nucleating agent for cholesterol gallstone formation in gallbladder bile.

Adult↗

Bacterial or crystal-associated arthritis? Discriminating ability of serum inflammatory markers.

A retrospective study of patients with culture-verified septic arthritis (n = 54) and polarizing microscopy verified crystal-associated arthritis (n = 34) was conducted with the objective to identify discriminating laboratory parameters in serum. Serum CRP levels (p = 0.002) and ESR (p = 0.03) were significantly higher on admission in patients with septic arthritis than in those with crystal-associated arthritis. The peripheral WBC counts did not differ between the two groups, nor did the lactoferrin or procalcitonin (PCT) levels. Serum TNFalpha concentrations on admission were higher in patients with septic arthritis than in those with crystal-associated arthritis (p = 0.0008). Significant differences were also found for IL-8 (p = 0.01) and G-CSF (p = 0.002), but not for IL-6 (p = 0.5). However, extensive overlap between the groups was present, resulting in low sensitivity, specificity and predictive value for each test. Determining serum levels of acute phase reactants, including cytokines, does not replace careful synovial fluid examination, including direct microscopy and cultivation.

Acute-Phase Proteins↗