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PubMed1974

Identification and quantitation of asbestos in talc.

The currently used analytical methods for identification, characterization and quantitation of asbestos fiber in consumer talcum products include polarized light microscopy, x-ray diffraction analysis, transmission electron microscopy with selected area electron diffraction and electron microprobe techniques. Light microscope methods have severe limitations imposed by the ultimate size resolution of the light-optical system. Small particles go unresolved; those marginally resolved may possess optical properties different from those properties cited in the literature; most optical properties, e.g., indices of refraction, are difficult to measure on small particles. In addition to these difficulties, talc fibers often possess optical properties different from those of talc plates, which further confound analysis. Light microscopy is recommended for use only as a preliminary tool on limited, large-sized, samples. Transmission electron microscopy is a good standard technique for visualization of contaminant asbestos fibers. Together with selected area electron diffraction, talc fibers may be easily differentiated from amphibole asbestos fibers on the basis of both morphological and structural characterization. Chrysotile fibers are easily distinguished on this basis as well. The amphibole asbestos minerals require chemical characterization to differentiate among the different fiber types. Probe analysis is mandatory for such fibers. The major drawbacks to electron beam instrumentation for the mineralogical characterization of talcum products are the time and effort required for data acquisition. These techniques do not lend themselves to routine study.X-ray diffraction analysis, utilizing the step-scan method, offers a relatively rapid, quantitative technique for gross fiber analysis. Based on comparison with standard specimens the fiber content of talcs may be quantitatively determined. It is essential to employ a specimen preparation technique which yields homogeneously dispersed particles. Tremolite may be determined at levels as low as 0.10% by weight, chrysotile 0.25%, and anthophyllite at 2.0% by weight occurrence in talc. The variance of these values depends upon many factors, including the mass absorption coefficient of the fiber types as compared to talc and selected diagnostic reflections and their relative intensities. Each of the above techniques is described in detail. A method for routine analysis of consumer talcum products is suggested.

Asbestos↗

PubMedDate not supplied

Effects of hepatocyte growth factor and platelet-derived growth factor on the repair of meniscal defects in vitro.

Injuries to the avascular region of the meniscus occur frequently and may be difficult to repair. This study was designed to determine whether growth factors could diffuse from a collagen sponge or a collagen gel into meniscal tissue and stimulate healing of defects using an in vitro model. The diffusion of platelet-derived growth factor (PDGF) from the collagen carriers into the medium was rapid with approximately 50% being released from the collagen sponge within the first hour. After 5 d of incubation, 8% of the PDGF was present in the meniscus, 11% in the collagen sponge, and 62% had been released into the medium. Similar results were obtained when a collagen gel was used as a carrier. Histological evaluation of the meniscal explants after 2 wk in culture revealed extensive proteoglycan staining in the areas surrounding defects treated with either hepatocyte growth factor (HGF) or PDGF compared with controls without growth factor. The HGF-PDGF treatment resulted in alignment and migration of meniscal cells toward the defect, which was not observed in untreated controls. At 3-7 d, increased number of cells were observed in defects treated with collagen gels (but not the sponge) with PDGF-HGF. At 4 wk, combined HGF-PDGF treatment resulted in the formation of tissue with birefringence by polarized microscopy, suggestive of organized collagen. The data suggest that use of specific PDGF-HGF may enhance the repair of meniscal injuries.

Animals↗

PubMed2004

Pneumoconiosis in rush mat workers exposed to clay dye "sendo" dust: clinical, radiologic, and histopathologic features in seven patients.

STUDY OBJECTIVES: The purpose of this study was to describe the clinical, chest radiographic, high-resolution CT, and histopathologic features of clay dye "sendo" dust pneumoconiosis in seven rush mat ("tatami") workers. PATIENTS: Seven patients with a history of occupational exposure to sendo dust and radiographic changes suggestive of pneumoconiosis were retrospectively reviewed. RESULTS: The duration of exposure ranged from 15 to 45 years (median, 30 years). Three patients had cough, and four patients had abnormal pulmonary function test results. Chest radiographs showed nodular opacities < 3 mm in diameter (types p and q) in all patients. The standard International Labor Office profusion score ranged from 0/1 to 1/1 (median, 1/0). High-resolution CT demonstrated small nodular opacities (types p and q) in all seven patients. In four patients, high-resolution CT demonstrated branching centrilobular structures, airway ectasia, airway wall thickening, and emphysematous changes. None of the patients had conglomerate nodules, large opacities, honeycombing, pleural effusion, or lymphadenopathy. Microscopic examination of the specimens obtained by open lung biopsy or transbronchial lung biopsy revealed nodular fibrosis with accumulation of dust-laden macrophages, but no silicotic nodules. Needle-like particles of 1 to 20 microm in length were evident among the dust deposits, and birefringent crystals were identified under polarizing microscopy. Four of seven patients showed intra-alveolar fibroblastic foci similar to Masson bodies, accompanied by dust deposition. CONCLUSION: Rush mat workers' sendo dust pneumoconiosis is caused by dust containing free silica. The radiographic and high-resolution CT findings consist of small nodular opacities < 3 mm in diameter and bronchial and bronchiolar abnormalities.

Aged↗

PubMed1978

Patterns of pulmonary involvement in systemic amyloidosis.

The clinical and histopathologic features of pulmonary amyloidosis were reviewed in 22 patients with systemic amyloidosis who came to autopsy. Eleven of 12 patients (92 percent) with primary amyloidosis had prominent interalveolar amyloid deposits. Symptoms attributable to these deposits were found in four cases (33 percent), while severe lung involvement was the apparent cause of death in one. Extensive deposition was noted in all three cases of amyloidosis associated with multiple myeloma or Waldenstrom's macroglobulinemia. Five of seven patients (71 percent) with secondary amyloidosis showed histologic lung involvement, which was perivascular or tracheobronchial in location, but not associated with symptoms. Histologic lung involvement is frequent in all forms of amyloidosis and lung tissue obtained from any patient with unexplained interstitial or reticular-alveolar pulmonary disease should be stained with Congo-red and viewed for green birefringence under polarizing microscopy for the presence of amyloid.

Adult↗

PubMed2005

DNA cholesteric pitch as a function of density and ionic strength.

The nature of chiral interactions among chiral biopolymers, such as DNA, protein alpha-helices, and rodlike virus particles, remains elusive. In particular, a satisfactory model connecting molecular chiral interactions and the pitch of the resulting chiral mesophases is lacking. We report the measurement of short-fragment (146-bp) DNA cholesteric spherulite pitch as a function of osmotic pressure, average DNA interaxial spacing, and salt concentration. We determined cholesteric pitch and interaxial spacing by polarizing optical microscopy and x-ray scattering, respectively, from which the twist-angle between DNA molecules can be calculated. Surprisingly, we found that decreasing ionic strength resulted in weaker chiral interactions between DNA chains, as evidenced by the decrease in the twist-angle, and consequent increase in the cholesteric pitch, for a fixed interaxial spacing. We propose that this behavior can be explained by increased smearing-out of the helical charge pattern along DNA as the Debye screening length is increased.

DNA↗

PubMed2004

Titanium pigment in tissues of drug addicts: report of 5 necropsied cases.

The aim of this report is to describe the anatomic-pathologic findings from necropsies of 5 drug addicts with titanium pigment in several organs after chronic intravenous injection of crushed propoxyphene hydrochloride tablets. Samples from liver, spleen, lungs, lymph nodes, and bone marrow were obtained, and after being grossly studied, they were submitted to evaluation using common light and polarized microscopy. In all 5 cases, a pigment with characteristics of titanium dioxide was found within tissue samples of the organs studied. Our findings suggest that research concerning titanium pigment within body tissues should be enhanced, considering the potential contribution of this morphologic data to forensic pathology.

Adult↗

PubMedDate not supplied

Calomys callosus: an alternative model to study fibrosis in schistosomiasis mansoni. The pathology of the acute phase.

Twenty Calomys callosus, Rengger, 1830 (Rodentia-Cricetidae) were studied in the early stage of the acute schistosomal mansoni infection (42nd day). The same number of Swiss Webster mice were used as a comparative standard. Liver and intestinal sections, fixed in formalin-Millonig and embedded in paraffin, were stained with hematoxilin and eosin, PAS-Alcian Blue, pH = 1.0 and 2.5, Lennert's Giemsa, Picrosirius plus polarization microscopy, Periodic acid methanamine silver, Gomori's silver reticulin and resorcin-fuchsin. Immunohistological study (indirect immunofluorescence and peroxidase labeled extravidin-biotin methods) was done with antibodies specific to pro-collagen III, fibronectin, elastin, condroitin-sulfate, tenascin, alpha smooth muscle actin, vimentin and desmin. The hepatic granulomas were small, reaching only 27% of the volume of the hepatic Swiss Webster granuloma. They were composed mainly by large immature macrophages, often filled by schistosomal pigment, characterizing an exsudative-macrophage granuloma type. The granulomas were situated in the parenchyma and in the portal space. They were often intravascular, poor of extracellular matrix components, except fibronectin and presented, sometimes alpha smooth muscle actin and vimentin positive cells. The C. callosus intestinal granulomas were similar to Swiss Webster, showing predominance of macrophages. Therefore, the C. callosus acquire very well the Schistosoma mansoni infection, without developing strong hepatic acute granulomatous reaction, suggesting lack of histopathological signs of hypersensitivity.

Acute Disease↗

PubMed2002

Intestinal fibrovascular nodules caused by Schistosoma mansoni infection in Calomys callosus Rengger, 1830 (Rodentia: Cricetidae): a model of concomitant fibrosis and angiogenesis.

Human schistosomiasis develops extensive and dense fibrosis in portal space, together with congested new blood vessels. This study demonstrates that Calomys callosus infected with Schistosoma mansoni also develops fibrovascular lesions, which are found in intestinal subserosa. Animals were percutaneously infected with 70 cercariae and necropsied at 42, 45, 55, 80, 90 and 160 days after infection. Intestinal sections were stained for brightfield, polarization microscopy, confocal laser scanning, transmission and scanning electron microscopies. Immunohistological analysis was also performed and some nodules were aseptically collected for cell culture. Numerous intestinal nodules, appearing from 55 up to 160 days after infection, were localized at the interface between external muscular layer and intestinal serosa, consisting of fibrovascular tissue forming a shell about central granuloma(s). Intranodular new vessels were derived from the vasculature of the external vascular layer and were positive for laminin, chondroitin-sulfate, smooth muscle alpha-actin and FVIII-RA. Fibroblastic cells and extracellular matrix components (collagens I, III and VI, fibronectin and tenascin) comprised the stroma. Intermixed with the fibroblasts and vessels there were variable number of eosinophils, macrophages and haemorrhagic foci. In conclusion, the nodules constitute an excellent and accessible model to study fibrogenesis and angiogenesis, dependent on S. mansoni eggs. The fibrogenic activity is fibroblastic and not myofibroblastic-dependent. The angiogenesis is so prominent that causes haemorrhagic ascites.

Animals↗

PubMed2000

Mechanisms involved in calcium oxalate endocytosis by Madin-Darby canine kidney cells.

Calcium oxalate (CaOx) crystals adhere to and are internalized by tubular renal cells and it seems that this interaction is related (positively or negatively) to the appearance of urinary calculi. The present study analyzes a series of mechanisms possibly involved in CaOx uptake by Madin-Darby canine kidney (MDCK) cells. CaOx crystals were added to MDCK cell cultures and endocytosis was evaluated by polarized light microscopy. This process was inhibited by an increase in intracellular calcium by means of ionomycin (100 nM; N = 6; 43.9% inhibition; P<0.001) or thapsigargin (1 microM; N = 6; 33. 3% inhibition; P<0.005) administration, and via blockade of cytoskeleton assembly by the addition of colchicine (10 microM; N = 8; 46.1% inhibition; P<0.001) or cytochalasin B (10 microM; N = 8; 34.2% inhibition; P<0.001). Furthermore, CaOx uptake was reduced when the activity of protein kinase C was inhibited by staurosporine (10 nM; N = 6; 44% inhibition; P<0.01), or that of cyclo-oxygenase by indomethacin (3 microM; N = 12; 17.2% inhibition; P<0.05); however, the uptake was unaffected by modulation of potassium channel activity with glibenclamide (3 microM; N = 6), tetraethylammonium (1 mM; N = 6) or cromakalim (1 microM; N = 6). Taken together, these data indicate that the process of CaOx internalization by renal tubular cells is similar to the endocytosis reported for other systems. These findings may be relevant to cellular phenomena involved in early stages of the formation of renal stones.

Animals↗

PubMed1980

Three-dimensional architecture and development of lumber intervertebral discs.

The three-dimensional architecture of the collagen framework of human lumbar intervertebral discs was studied with scanning electron microscopy and polarized light microscopy concentrating on the fibrillar interconnection between the intervertebral disc and the vertebral bodies. The fibrillar framework of the annulus fibrosus and the cartilage end-plates encircled the nucleus pulposus as a closed-pack system in the adult. This closed-pack system developed in the seventh embryonic month and was completed by the tenth month. In general, fibrils composing the framework in the fetus were thinner than in the adult. There was no fibrillar interconnection between the cartilage end-plate and subchondral bone. In the inner one-third of the annulus, obliquely oriented fibrillar lamellae interconnected with the cartilage end-plate. In the outer two-thirds, the fibrillar bundles of the lamellae were firmly anchored into the vertebral bodies. This fibrillar anchoring system was already present in the full-term fetus although the vertebral rim was not ossified.

Adolescent↗

PubMed2004

In vitro inhibition of caries-like lesions with fluoride-releasing materials.

The aim of this study was to compare the in vitro caries inhibition of various resin-based materials. Class V cavities were prepared in twenty-five freshly extracted human premolar teeth which were then restored with glass-ionomer cement (Chemfill II), compomer (Compoglass F, Dyract AP) and composite resin (Tetric Ceram and Z 100). The teeth were submerged in an acid gel for 6 weeks. Each specimen was sectioned. These sections were left in water for 24 hours, and then examined using polarized light microscopy. The lesion consisted of two parts, the outer surface lesion and the cavity wall lesion. There was no significant difference in the body depth of the outer lesion and in the depth of the wall lesion among teeth restored with Compoglass F, Dyract AP and Chemfill II (P > 0.05). There was a significant difference between those restored with Z 100 and Tetric Ceram (P < 0.05). The length of the wall lesion for the teeth restored with Chemfill II was significantly smaller than that in the remaining groups (P < 0.05). The length of the wall lesion for teeth restored with Tetric Ceram and Z 100 was significantly higher than in the remaining groups (P < 0.05). These results suggest that composite materials and compomer provide less caries inhibition than glass-ionomer cements.

Cariostatic Agents↗

PubMedDate not supplied

Histopathologic evaluation of the ethmoid bone in chronic sinusitis.

Chronic rhinosinusitis (CRS) is defined as a condition lasting for a period greater than 12 weeks, and manifested by an inflammatory response involving the mucous membranes of the nasal cavity and paranasal sinuses, fluids within these cavities, and/or the underlying bone. The mucosal changes that occur in CRS have been well described, and include edema, decreased number of ciliated cells, and goblet cell hyperplasia. However, the changes that may occur in the underlying ethmoid bone have only recently been investigated. We evaluated decalcified ethmoid bone specimens from 20 patients undergoing endoscopic sinus surgery for CRS. Our analysis revealed histopathologic changes consistent with varying grades of bone remodeling. Polarized light microscopy demonstrated changes in the extracellular matrix, such as bone resorption and neoosteogenesis. Preoperative clinical data and CT staging were recorded on all patients and correlated with the histopathologic findings. These findings suggest that CRS may be associated with osteitis of the underlying ethmoid bone.

Bone Remodeling↗

PubMed2007

Treatment of severe male infertility by micromanipulation-assisted fertilization: an update.

In the past 5-10 years the evolution of micromanipulation-assisted fertilization for the treatment of severe male infertility was marked by the introduction of new technical support, refinement of diagnostic methods for the evaluation of sperm developmental potential, and development of new treatment regimens for the newly discovered abnormalities. The new technical support involves the use of non-contact laser technology to assist micromanipulation for fertilization, the evolution of polarized microscopy-based optical systems to non-invasively detect the position of the meiotic spindle in living human oocytes, and the development of high-magnification optical systems for a better morphological selection of spermatozoa to be used for fertilization. Diagnostic approaches were enriched by commercial availability of kits for the analysis of sperm DNA integrity, leading to the definition of sperm nuclear DNA damage as a distinct cause of male infertility, and by the development of tests, based on heterologous ICSI, for detection of sperm failure to activate oocytes. Several treatment options for these conditions have been proposed and are currently being tested in larger-scale trials. Some technical improvement was also achieved in the field of in vitro maturation of germ cells from men with in vivo maturation arrest, but only a modest clinical improvement resulted from their application. As to the risk for the offspring, recent data are rather reassuring. Except for the risk of transmission of genetically based infertility, no straightforward evidence for a health risk derived from these techniques has been provided. Nevertheless, caution is necessary, particularly concerning the eventual increase in genomic-imprinting abnormalities.

DNA Damage↗

PubMed1993

The absence of correlations between a clinical classification and ultrastructural findings in amelogenesis imperfecta.

This study was performed to examine whether a clinical classification of different phenotypes of amelogenesis imperfecta could be discernible at the ultrastructural level. Seventeen primary teeth from 16 children with hypomineralization, hypomaturation, or hypoplastic variants of the disease were collected for histologic studies of the enamel by means of polarized light microscopy, scanning electron microscopy (SEM), and secondary ion mass spectrometry (SIMS). Polarization microscopy showed that the enamel was hypomineralized; in six teeth a wavy configuration of the enamel prisms also appeared. Three histomorphologic main types could be discerned. In 10 of the teeth extensive hypomineralization of the bulk of the enamel was found. One tooth had an unusually thick enamel with only a thin normally mineralized surface layer. SIMS images showed less pronounced signals from Ca2+ and Na+ but with stronger signals from Cl- and CN-, representing the organic component of enamel. The SEM images showed an irregular prism pattern with marked interprismatic areas. Irrespective of the clinical appearance or the hereditary pattern the main findings were hypomineralized enamel with or without wavy bands. Neither of the analytical methods used in this paper distinguishes between the clinical phenotypes of amelogenesis imperfecta.

Amelogenesis Imperfecta↗

PubMed1995

Dissolution of human cholesterol gallstones in bile salt/lecithin mixtures: effect of bile salt hydrophobicity and various pHs.

BACKGROUND: Unconjugated bile salts currently available for gallstone dissolution are poorly effective. We evaluated in vitro the litholytic potency of taurine-amidated bile salts against human cholesterol gallstones. METHODS: Seventy radiolucent gallstones with similar size and composition (cholesterol content, 70.1 +/- 0.9%) from a single patient were incubated in model biles composed of 100 mM of either taurochenodeoxycholate (TCDC), taurocholate (TC), taurohyodeoxycholate (THDC) or tauroursodeoxycholate (TUDC) and of 45 mM egg yolk lecithin in saline buffered with tris/HCl (at pHs 7 and 8) or phosphate (at pHs 4 and 6). Biles (total lipids, 10 g/dl; cholesterol saturation, 99%) were incubated at 37 degrees C for 40 days. Gallstones were periodically weighed and returned to the dissolution vials, and the biliary cholesterol concentration was monitored. RESULTS: Model biles remained optically clear during the initial 48 h of incubation. Then, biles containing THDC and TUDC, but not those with TC and TCDC, became progressively turbid until, after several days, a white precipitate surrounded the residual stone. Abundant liquid crytalline droplets were observed at polarizing microscopy in biles containing TUDC and THDC. Gallstone dissolution was closely related to cholesterol solubilization and decreased in the order TCDC > THDC > or = TC > TUDC, being highest at pH 8. At the physiologic pH of 7 THDC was more litholythic than TC. CONCLUSIONS: In vitro, the litholytic potency of bile salts on cholesterol gallstones primarily depends on their hydrophobicity. THDC is a new potential gallstone-dissolving agent, deserving in vivo studies.

Cholagogues and Choleretics↗

PubMedDate not supplied

Comparison of the pulmonary surfactant content in alveolar macrophages of newborn, young, and adult rats.

The phospholipid composition of the 150-g pellet containing macrophages from neonatal lung lavages resembles that of surfactant. To study whether this composition reflects the surfactant content of the macrophage, we isolated the alveolar phospholipids and macrophages from the lavage fluids of fetal, newborn, young, and adult Wistar rats. The alveolar surfactant phospholipids increased from fetal levels of 2.8 nmol/mg dry lung weight (DLW) to 39 nmol/mg DLW at day 1, decreased sharply within the first week, and stabilized at a level of 2-4 nmol/mg DLW after day 15. The number of alveolar macrophages increased significantly during the first postnatal day from approximately 750 to more than 5000 (per mg DLW), decreased during the next 4 days, and varied strongly at older ages. We estimated the surfactant content in the macrophages semiquantitatively by polarization microscopy. Birefringence augmented significantly during the first 1.5 days after birth and decreased after that concurrently with the amount of alveolar surfactant. However, only cells without birefringent inclusions sedimented at 150g, whereas the phospholipid composition of the pellets falsely suggested that large amounts of intracellular surfactant were present in its cells. At least two populations of macrophages (surfactant-rich and surfactant-poor) are present in the growing animal. We suggest that differences in function of these various types of macrophages also might depend on surfactant congestion.

Aging↗

PubMed1978

The multicomposite structure of tendon.

A revised morphological model for the crimp structure of tendon is presented. The 300-500 mu diameter tendons of the mature rat tail are comprised of from one to more than ten substructures, called fascicles, of 80-320 mu diameter. Fascicles each possess a "crimp structure" demonstrable in the polarizing microscope and neighboring fascicles within a tendon usually exhibit crimp registry. The fascicle itself is shown to be a cylindrical array of planar-zig-zag crimped 500-5000 A diameter collagen fibrils. The approximate cylindrical symmetry of the fascicle is domonstrated by SEM not equal to and polarizing optical microscopy. A method of replacing native water with other liquids of refractive index near to that of collagen is utilized to reduce or eliminate light diffusion and therby greatly improve OM observations. Small bunches of collagen fibrils removed from the tendon are shown to exhibit the simple planar zig-zag morphology described in previous literature. The planar crimping of collagen fibrils and their assemblage into cylindrically symmetric fascicles is verified by small angle X-ray diffraction.

Animals↗

PubMed1983

Birefringence of articular cartilage and the distribution on collagen fibril orientations.

Birefringence of the deep zone of pig tibial plateau cartilage has been measured by polarized light microscopy using monochromatic light and a Sénarmont compensator. Thin (20 micrometers) sections of the tissue were imbibed with mixtures of physiological saline and glycerol to separate the effects of form and intrinsic birefringence. Measured birefringence was plotted against liquid refractive index and the experimental points defined a parabola whose minimum corresponded to a refractive index for collagen of 1.48. When the tissue was imbibed with glycerol, the birefringence measurements provided a good estimate of intrinsic birefringence. The orientation distribution function, g(phi), of collagen fibrils in this zone was then determined by X-ray diffraction. Thus X-ray diffraction yielded the form of g(phi) which corresponded to the measured intrinsic birefringence. Using this result the intrinsic birefringence of a hypothetical cartilage site with perfectly oriented collagen fibrils was calculated to be (6 +/- 1) x 10(-4). Birefringence measurements on further sites could then be converted into standard deviations of g(phi), assuming a Gaussian distribution.

Animals↗