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Gallstones in cystic fibrosis: a critical reappraisal.

Radiolucent gallstones are common in young adults with cystic fibrosis. In the mid-1970s, it was suggested that gallstones are made of cholesterol, but this hypothesis has never been tested. Several recent studies have shown that the detection of cholesterol monohydrate crystals in bile has high sensitivity and specificity for the diagnosis of cholesterol gallstones. We therefore used this approach to study 17 young adults with cystic fibrosis, 10 of whom had radiolucent gallstones. The two groups of patients were comparable in age and gender (all patients but one were male). Duodenal bile was obtained after gallbladder contraction with intravenous cerulein; it was used for lipid and protein chemistry studies and for polarizing microscopy. The latter was performed both in whole bile and in the postultracentrifugation (100,000 g) sediment. Bile cholesterol saturation did not significantly differ between patients with (1.21 +/- 0.28) or without gallstones (0.99 +/- 0.54). Slight cholesterol supersaturation was found in 7 of 10 gallstone and three of seven nongallstone patients. At no time were cholesterol crystals detected in either the group, even after bile ultracentrifugation. Two more cystic fibrosis patients with gallstones died of severe bronchopneumopathy, and small pigment gallstones were obtained at autopsy. At stone analysis, cholesterol content was 44% and 28% of dry weight, respectively. Infrared spectroscopy of stone powder was compatible with the presence of calcium bilirubinate and proteins as major components. We conclude that radiolucent gallstones of cystic fibrosis are not of the conventional cholesterol type.

Adolescent↗

Physicochemical properties of calcific deposits isolated from porcine bioprosthetic heart valves removed from patients following 2-13 years function.

The purpose of this study was to characterize the physicochemical properties of calcific deposits that cause the failure of tissue-derived heart valve bioprostheses. This was done in an effort to understand the mechanism of pathologic biomineralization in the cardiovascular system and potentially prevent deterioration of bioprostheses. Calcific deposits taken from 10 failed bioprosthetic valves that had been implanted in patients for 2-13 years were characterized by chemical analysis, x-ray diffraction, FTIR spectroscopy, scanning electron microscopy, polarized light microscopy, and solubility measurements. The combined results identified the biomineral as an apatitic calcium phosphate salt with substantial incorporation of sodium, magnesium and carbonate. The average Ca/PO4 ratio for this "young" pathologic biomineral was approximately 1.3, considerably lower than approximately 1.7 found in mature atherosclerotic plaque biomineral and mature skeletal biomineral, both of which approximate hydroxyapatite in composition. Deproteinated calcific deposits from bioprostheses had thermodynamic solubilities comparable to those of both atherosclerotic plaque, typical pathologic biomineral and hydrolyzed octacalcium phosphate (OCP, Ca4H(PO4)3 x 2.5 H2O), a proposed precursor phase to biomineral apatite. This later finding, together with chemical composition and structural details of the bioprosthetic deposits themselves, supports a mechanism of cardiovascular calcification in which OCP plays a crucial role in the formation of the final apatitic phase. This suggests an approach toward prevention of bioprosthetic tissue calcification through control of the formation of the kinetically favored OCP precursor and/or its transformation into bioapatite.

Bioprosthesis↗

Effects of Er:YAG laser irradiation on human dentin: polarizing microscopic, light microscopic and microradiographic observations, and FT-IR analysis.

BACKGROUND AND OBJECTIVES: The effects of Er:YAG laser irradiation on dentin have not been sufficiently investigated. The purpose of this study was to investigate the effects of Er:YAG laser irradiation on dentin. STUDY DESIGN/MATERIALS AND METHODS: After cavities were prepared using Er:YAG laser irradiation or rotary cutting instruments, histological observations of cavity-floor dentin utilizing polarizing microscopy, microradiography and light microscopy, and analysis of composition of cavity-floor dentin using Fourier-transformed (FT-IR) spectrometry were conducted. RESULTS: In the laser-treated side, a deeply stained basophilic layer was observed. The number of odontoblastic processes present was obviously less in the laser-treated side than in the bur-treated side. FT-IR analysis revealed that compared to the bur-treated side, a broad background peak at around 1,600 cm(-1) was present. CONCLUSIONS: Er:YAG laser irradiation might have denatured the organic materials of dentin.

Basophils↗

Comparison of the thermal tissue effects produced by aged sapphire and silica hemispherical tips.

This study evaluated the performance of sapphire and fused silica hemispherical tips under the same exposure conditions. Lesions produced in the chicken breast and a blood field were sectioned for light and transmission polarizing microscopy. Lesion size and thermal damage area were recorded as a function of the tips accumulated exposure. The tips transmission was measured after every 1,000 J of exposure. Fused silica tips lasted for approximately 5,000 J and experienced significant surface and transmission deterioration. The sapphire hemispherical tips lasted for > 12,000 J with no surface and transmission deterioration. Lesions produced with the fused silica tips generally increased in depth with use, and depths of 6 mm were common. Lesions produced by the sapphire tips were subsurface spherical areas of coagulation with the tissue surface relatively intact. This difference in resulting lesions may be attributed to the higher thermal conductivity of sapphire.

Aluminum Oxide↗

Crystalluria in idiopathic recurrent calcium urolithiasis. Dependence on stone composition.

A retrospective study was done on the nature and degree of crystalluria in fasting and postprandial urine in patients with recurrent idiopathic calcium urolithiasis (RCU) for whom stone analysis was available. RCU was stratified into subgroups in accordance with stone analysis. The crystals were obtained and identified using a filter technique and polarization microscopy, respectively. Crystalluria score, relative saturation products (RSPs), and low-molecular-weight inhibitors were assessed. Calcium oxalate crystals were never observed in either male or female patients with stones composed exclusively of calcium oxalate, and only sporadically in patients with mixed stones (the additional component was calcium phosphate in most cases). Other crystalluria phases, such as amorphous calcium phosphate, a urate-containing phase, and a phase presenting as spherolytic particles, were slightly more frequent in patients with mixed stones. In contrast to crystalluria, RSPs and inhibitors differed in male and female patients, suggesting that crystalluria may not be under the exclusive control of these factors. The following conclusions were reached. (1) Calcium oxalate crystalluria is absent from RCU with pure calcium oxalate stones; hence, calcium oxalate crystalluria does not qualify as a diagnostic aid. (2) The co-existence of the isotropic phase and mixed stones may indicate that the formation of these concretions is characteristic for a major RCU subgroup. (3) On the basis of clinical chemistry and physicochemical data in urine and of crystalluria, it appears that the pathogenesis of RCU differs in male and female subjects.

Adult↗

Benign human mammary myoepithelioma.

A leiomyoma-like, multifocal tumour developed from intraductal papillomatosis in the breast of a 42 year old woman. The spindle-shaped tumour cells were examined by light, electron and polarizing microscopy, which revealed that they originated from immature "precursor" myoepithelial cells. The author suggests that the tumour be called "myoepithelioma". From the morphological characteristics of the tumour the myoepithelial cells appear to be capable of producing leiomyoma-like benign or malignant tumours. The role that has been attributed by some to the myoepithelial cell in the production of epithelial tumours is problematical, in the light of the present finding.

Adult↗

The characteristic three-banded birefringence of ligamentum nuchae elastic fibres indicates ordered micellar texture.

The ultrastructural organization of the elastic fibres of bovine ligamentum nuchae is still controversial (Kádár 1979; Sandberg 1976; Sandberg et al. 1981; Quintarelli et al. 1973; Gosline 1976). Polarization microscopy demonstrated convincingly, contrary to a recent negative report (Aaron and Gosline 1980), an ordered birefringent micellar texture (Schmidt 1939; Romhányi 1965; Fischer 1979). The three-banded anisotropic pattern of the fibres can be explained by the optical interference of two micellar textures of opposite orientation, one arranged circularly in the surface layer and the other oriented longitudinally in the axial core of the fibres (Romhányi 1965). Analysis of the underlying molecular mechanism for the anisotropic staining reaction of these fibres with Congo red led to the conclusion that this is due to a definite structural order of specific cross-linking segments within the random network of elastin. The anisotropic staining reaction of the elastic fibres with Congo red is described as the most simple method to demonstrate the three-banded birefringent pattern of ligamentum nuchae elastic fibres even in histologic sections.

Animals↗

Mechanism of chlorpromazine binding by gram-positive and gram-negative bacteria.

Chlorpromazine forms charge-transfer complexes with xanthene dyes in bacteria. These complexes permit the differentiation of Gram-positive and Gram-negative bacteria in both light and polarization microscopy. The birefringence induced by the charge-transfer complex might explain the molecular basis of bacterial staining. The charge-transfer complexes formed between chlorpromazine and xanthene dyes accumulate in the bacterial cell, mainly inside the bacterial cell wall. The complexes give the cells a color, which depends on the chemical composition of the staining structure, and in particular the polysaccharides of the cell wall in bacteria. Metachromatic granules were seen inside Gram-positive bacteria after chlorpromazine and rose bengal staining. Although the nature of these granules remains unclear, this type of binding may have a role in the inhibition of biochemical processes in the bacterial cells.

Binding Sites↗

Effects of pressure overload on the passive mechanics of the rat left ventricle.

Both myocyte growth and changes in the extracellular matrix may affect the passive mechanics of the left ventricle (LV). Pressure-volume (PV) relationships and midwall two-dimensional strains versus passive loading were measured in isolated rat hearts 2 and 6 weeks after ascending aortic banding. Collagen area fractions and perimysial fibril orientations were determined with picrosirius-polarization microscopy, and the equatorial region of the LV was modeled with finite element analysis of a transversely isotropic cylinder with the same material properties in hypertrophy and control. Compared with weight-matched shams, heart weight increased at 2 (19%) and 6 (22%) weeks, as did LV wall thickness (6% and 31%, respectively). The PV curve became less compliant with hypertrophy; only circumferential strain decreased after hypertrophy. Collagen area fractions were not different at either subendocardium or subepicardium (3.37 +/- 1.06 versus 3.96 +/- 0.76 at 2 weeks and 3.61 +/- 1.30 versus 4.22 +/- 1.50 at 6 weeks for banded and sham, respectively; subendocardium). Collagen and muscle fiber orientations also did not change with hypertrophy. The finite element model predicted trends in the strains similar to those found experimentally. Thus, in this model of pressure-overload hypertrophy, the decreases in compliance and circumferential strain of the passive LV are not due to changes in the percentage of extracellular matrix, but rather to global geometric changes.

Analysis of Variance↗

Enamel structure in some therapsids and mesozoic mammals.

The distribution of enamel tubules, the shapes and arrangements of prisms, and the orientation of crystals in ground sections from several therapsids and mesozoic mammals have been investigated by conventional and polarizing microscopy. Along each of three separate phylogenetic lines which evolved occluding teeth, there was a progressive increase in the numbers of enamel tubules. In the investigation, the arcade-shaped prisms typical of recent mammals were first seen in material from the Cretaceous period. All the enamels investigated from the Triassic contained columns of crystals, which were deduced as hexagonal. The inner ends of the crystals within each column deviated towards the center of the column. It is concluded that the existence of an interprismatic region provides the most important distinction between prismatic enamels and the hexagonal columns of crystals in the Triassic material.

Animals↗

Persistence of bone collagen cross-links in skeletons of the Nuraghi population living in Sardinia 1500-1200 B.C.

Bone collagen has a specific molecular ultrastructure which can be proved by birefringence. This protein, forming the main organic component of bone tissue, is known to survive millennia in paleontological bones and teeth. Birefringence of bone collagen obtained from the skeletons of the Nuraghi population living in Sardinia c-ca 1500 years B.C. was found previously by the use of polarizing microscopy [1]. In this paper, using high pressure liquid chromatography (HPLC) techniques, we show the existence of bone collagen cross-links preserved in Nuraghi skeletons after more than 3000 years.

Amino Acids↗

Stone analysis.

The definition and the frequencies of the components of urinary stones are presented on the basis of more than 110,000 analyses. The stone morphologies are described. The methods of stone analysis X-ray diffraction, infrared spectroscopy and polarization microscopy and the most recent developments are presented. The benefits and disadvantages of different methods are compared. The results of ring trials show that the X-ray diffraction method is the best method with regard to correctness.

Kidney Calculi↗

[Changes in the collagen structures of human heart valves in relation to age and hypertension].

Using polarization microscopy and morphometrical methods, the influence of hypertension on the collagenous structures in the Lamina fibrosa of bicuspid valves of 14 middle-aged persons (30 to 50 years) were examined. The measurements were performed on histological sections. A group of 14 middle-aged subjects free from heart disease served as the control group. Furthermore, the results were compared to earlier findings on the histological biomorphosis of the atrioventricular valves. The following changes were observed and quantitatively determined: a) an enhancement of the percentage of the mechanically more resistant collagenous fibers (unsilvered, type I collagen), b) a stronger lateral aggregation of the collagenous fibrils of both types I and III collagen, c) a significant decrease of the total numbers of collagenous fibers per measuring area, and d) a significant decrease of the portion of silvered fibers (type III collagen) per measuring area for both sexes. The fiber density distributions confirm the observed changes: in the hypertension group the collagenous fibers are less densely distributed than in the control group. The observed regression of the content of collagenous fibers in the hypertension group is probably due to the anti-hypertensive treatment. Following the findings the turn-over of type III collagen is stronger influenced than that of type I collagen. The initial results determined in human heart valves confirm findings in animal models and characterize them with regard to the collagen types mentioned.

Adult↗

Physiochemical principles of cardiovascular calcification.

OBJECTIVES: The objectives of this review article are to provide detailed physicochemical information on the nature of pathological cardiovascular deposits, PCD, isolated from different sites of the human cardiovascular system. In order to help to understand the complex mechanism of the formation of PCD, special emphasis is focused on the attempt to recognize early precursor(s) of PCD by utilizing combined in vivo and in vitro studies. BACKGROUND: The common idea is that PCD is hydroxyapatite (HAP), Ca5(PO4)3OH; however, this is questionable and deserves critical evaluation. METHODS: After isolation of PCD, deproteination followed, producing pure inorganic fraction of deposits that were subject to chemical analyses, x-ray diffraction (XRD), FT-IR spectroscopy, optical, scanning electron microscopy (SEM), polarizing microscopy, energy dispersive x-ray micro-analyses (EDS) and thermodynamic solubility measurements. The same methods were used to characterize the in vitro and in vivo formed calcific deposits. RESULTS: The results of chemical analyses provided essential information that PCD contained significant organic fraction; inorganic fraction appears to be bioapatite, corresponding to defect HAP with substantial macro incorporation of sodium, magnesium, carbonate and fluoride. Structural XRD data show apatitic pattern; however, variable crystallinity of PCD suggests that crystallization is a time and flow dynamics-dependent process. Critical thermodynamic solubility measurements unequivocally prove that PCD are significantly more soluble than HAP. CONCLUSIONS: The PCDs are morphologically and chemically heterogeneous products which may be a consequence of time-dependent hydrolytic transformation of precursors that may include amorphous calcium phosphate and octacalcium phosphate (OCP) Ca4H(PO4)3.

Animals↗

In vivo light-microscopic documentation for primary calcification processes in the hermatypic coral Stylophora pistillata.

Skeletogenesis in the hermatypic coral Stylophora pistillata was studied by using the lateral skeleton preparative (LSP) assay, viz., a coral nubbin attached to a glass coverslip glued to the bottom of a Petri dish. Observations on tissue and skeletal growth were made by polarized microscopy and by using vital staining. The horizontal distal tissue edges developed thin transparent extensions of ectodermal and calicoblastic layers only. Four stages (I-IV) of skeletogenesis were observed at these edges, underneath the newly developed tissue. In stage I, a thin clear layer of coral tissue advanced 3-40 microm beyond the existing LSP peripheral zone, revealing no sign of spiculae deposition. At stage II, primary fusiform crystals (1 microm each) were deposited, forming a primary discontinuous skeletal front 5-30 microm away from the previously deposited skeleton. During stage III, needle-like crystals appeared, covering the primary fusiform crystals. Stage IV involved further lengthening of the needle-like crystals, a process that resulted in occlusion of the spaces between adjacent crystals. Calcification stages I-III developed within hours, whereas stage IV was completed in several days to weeks. Two basic skeletal structures, "scattered" and "laminar" skeletons, were formed, integrating the growth patterns of the needle-like crystals. High variation was recorded in the expression of the four calcification stages, either between different locations along a single LSP or between different preparations observed at the same diurnal time. All four skeletogenesis stages took place during both day and night periods, indicating that an intrinsic process controls S. pistillata calcification.

Animals↗

Oxalate mediated nephronal impairment and its inhibition by c-phycocyanin: a study on urolithic rats.

The assumption of oxidative stress as a mechanism in oxalate induced renal damage suggests that antioxidants might play a beneficial role against oxalate toxicity. An in vivo model was used to investigate the effect of C-phycocyanin (from aquatic micro algae; Spirulina spp.), a known antioxidant, against calcium oxalate urolithiasis. Hyperoxaluria was induced in two of the 4 groups of Wistar albino rats (n = 6 in each) by intraperitoneally injecting sodium oxalate (70 mg/kg body weight). A pretreatment of phycocyanin (100 mg/kg body weight) as a single oral dosage was given, one hour prior to oxalate challenge. An untreated control and drug control (phycocyanin alone) were employed. Phycocyanin administration resulted in a significant improvement (p < 0.001) in the thiol content of renal tissue and RBC lysate via increasing glutathione and reducing malondialdehyde levels in the plasma of oxalate induced rats (p < 0.001), indicating phycocyanin's antioxidant effect on oxalate mediated oxidative stress. Administering phycocyanin after oxalate treatment significantly increased catalase and glucose-6-phosphate dehydrogenase activity (p < 0.001) in RBC lysate suggesting phycocyanin as a free radical quencher. Assessing calcium oxalate crystal retention in renal tissue using polarization microscopy and renal ultrastructure by electron microscopy reveals normal features in phycocyanin-- pretreated groups. Thus the study presents positive pharmacological implications of phycocyanin against oxalate mediated nephronal impairment and warrants further work to tap this potential aquatic resource for its medicinal application.

Animals↗

Thermal and structural behavior of natural cerebroside 3-sulfate in bilayer membranes.

Differential scanning calorimetry (DSC), polarizing microscopy and X-ray diffraction studies have been performed on dry and hydrated natural bovine brain sulfatides. Dry sulfatide fractions exhibit a high temperature transition (delta H = 6.6 kcal/mol sulfatide) at 87.3 degrees C. X-ray diffraction shows this transition to be associated with a hydrocarbon chain order-disorder transformation between two lamellar phases. Hydrated sulfatide dispersions undergo a complex chain order-disorder transition (delta H = 7.5 kcal/mol sulfatide) at 32 degrees C with two peak temperatures at 35 degrees C and 47 degrees C. Structural studies performed on hydrated liquid-crystal sulfatide dispersions at 75 degrees C verify the existence of a bilayer structure over the 16 wt.% to 50 wt.% phosphate buffer (pH = 7.4) range. The interbilayer separation between galactosyl-3-sulfate groups averages 48 A as the multilamellar bilayers swell with the addition of phosphate buffer. The formation of micellar phases is not observed at high water contents. The comparison of the structural characteristics of dry and hydrated sulfatides with structural data for dry and hydrated bovine brain non-sulfated glycolipid (cerebroside) is discussed in molecular terms.

Animals↗

Phase behaviour and molecular species composition of oat root plasma membrane lipids. Influence of induced dehydration tolerance.

Tolerance to dehydration induced by repeated water-deficit stress is well correlated to changes in the lipid composition of oat root cell plasma membranes. The molecular species of the two dominant phospholipids phosphatidylcholine and phosphatidylethanolamine were determined. The four major species were 16:0/18:2, 16:0/18:3, 18:2/18:2 and 18:2/18:3. In contrast to the large changes in plasma membrane lipid composition in other respects, induced tolerance resulted in very weak alterations concerning the phospholipid molecular species pattern. Only minor alterations, appearing as a decrease in the 18:3-containing lipids, occurred. Total lipids of microsomes and isolated plasma membranes of root cells were analysed by X-ray crystallography at different degrees of hydration. The lipid phase behaviour at different degrees of hydration was further confirmed by polarization microscopy. In the presence of excess water all membrane lipids adopted a reversed micellar configuration. The plasma membrane lipids from root cells with induced dehydration tolerance formed upon dehydration two coexisting lamellar structures. The importance of the phase behaviour at different degrees of hydration for the membrane properties and the relation to membrane composition is discussed.

Edible Grain↗