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PubMed2002

Fine reconstruction of the pancreatic ductular system at the onset of pancreatitis.

The three-dimensional structure of the pancreatic ductular system (from the intercalated duct to the intercellular secretory canaliculus) is controversial and unclear. The aim of this study is to reveal the three-dimensional structure of the pancreatic ductular sysytem at the onset of pancreatitis. One day following rat pancreatic duct ligation, dilated lumina from the pancreatic ductular system were reconstructed by light microscopic and scanning electron microscopic examination of pancreatic tissue serial sections. The existence of the intra-acinar duct, which is formed only by centroacinar cells and interconnects the adjacent central lumina in an acinus, was demonstrated. The intercellular secretory canaliculi, which are the terminal parts of the pancreatic ductular system, anastomose and end blindly in the intercellular space located between adjacent lateral surfaces of the acinar cells. The intercalated ducts, the intra-acinar ducts, the central lumina, and the intercellular secretory canaliculi are arranged together in a complex connecting and branching system. However, there were no anastomoses found among the central lumina or acini.

Animals↗
PubMed2005

A 16-year study of the microgap between 272 human titanium implants and their abutments.

A microgap has been described at the level of the implant-abutment connection. This microgap can be colonized by bacteria, and this fact could have relevance on the remodeling of the peri-implant crestal bone and on the long-term health of the peri-implant tissues. The authors report on 272 implants with screw- or cement-retained abutments retrieved from humans for different causes during a 16-year period. In the implants with screw-retained abutments, a 60-microm microgap was present at the level of implant-abutment connection. In some areas the titanium had sheared off from the surface and from the internal threads. The contact between the threads of the implant and those of the abutment was limited to a few areas. Bacteria were often present in the microgaps between implant and abutment and in the internal portion of the implants. In implants with cement-retained abutments, a 40-microm microgap was found at the level of the implant-abutment connection. No mechanical damage was observed at the level of the implant or of the abutment. All the internal voids were always completely filled by the cement. No bacteria were observed in the internal portion of the implants or at the level of the microgap. The differences in the size of the microgap between the two groups were statistically significant (P < .05). In conclusion, in screw-retained abutments the microgap can be a critical factor for colonization of bacteria, whereas in cement-retained abutments all the internal spaces were filled by cement. In these retrieved implants, the size of the microgap was markedly variable and much larger than that observed in vitro.

Bacteria↗
PubMed2006

Influence of cervical preflaring on determination of apical file size in maxillary molars: SEM analysis.

The purpose of this study was to investigate the influence of cervical preflaring on determination of the initial apical file in mesiobuccal roots of maxillary molars. Fifty first molars with degree of curvature of the mesiobuccal root between 10 degrees and 15 degrees were utilized. After standard access opening and removal of pulp tissue, the working length (WL) was determined at 1 mm from the root apex. Five groups (n=10) were formed at random, according to the type of cervical preflaring performed. In group 1, the size of the initial apical file was determined without cervical preflaring. In groups 2 to 5, the cervical and middle thirds of the canals were preflared with Gates-Glidden drills, K3 Orifice Opener instruments, ProTaper instruments and LA Axxess burs, respectively. Canals were sized manually with K-files, starting with size 08 K-files, inserted passively up to the WL. File sizes were increased until a binding sensation was felt at the WL and the size of the file was recorded. The binding instruments were fixed into the canals at the WL with methylcyanacrylate. Cross-sections of the WL region were examined under scanning electron microscopy and the discrepancies between the canal diameter and the diameter of first file to bind at the WL were calculated using UTHSCSA ImageTool software. Data were analyzed statistically by ANOVA and multiple comparisons were done by Tukey's HSD post-hoc test. Significant differences (p<0.001) were found among the groups. The major discrepancy was observed for the group without preflaring (mean 0.1543 mm +/- 0.0216). Cervical preflaring with LA Axxess burs produced the least discrepancies between the canal size and the diameter of the initial apical instrument (mean 0.026 mm +/- 0.0037), followed by ProTaper files (mean 0.0567 mm +/- 0.0354). Canals preflared with Gates-Glidden drills and K3 Orifice Opener instruments showed statistically similar discrepancy results to each other (p>0.05) (means 0.1167 mm +/- 0.0231 and 0.1313 mm +/- 0.0344, respectively). In conclusion, preflaring of the cervical and middle thirds improved the determination of the initial apical instrument. Canals preflared with LA Axxess burs showed a more accurate binding of the files to the anatomical diameter at the WL in the mesiobuccal roots of maxillary first molars.

Cyanoacrylates↗
PubMed1996

Lysosomal cysteine and aspartic proteinases and ubiquitin in rat and human urinary bladder epithelium.

To examine localization of cysteine and aspartic proteinases, and ubiquitin in rat and human urinary bladders, immunocytochemistry was applied to the tissues. In semi-thin sections, immunoreactivity for cathepsins B and D was densely localized throughout epithelial layers of rats and humans, while that for cathepsins H and L was mainly localized in rat superficial and human intermediate cells. Immunoreactivity for cathepsin C was relatively high in rat and human epithelia, especially in humans. Immunoreactivity for ubiquitin was detected in rat and human epithelial cells. By electron microscopy, vesicular or heterogeneously dense lysosomes labeled with immunogold particles indicating cathepsin B were seen in rat and human epithelial cells; particularly, they often appeared near fusiform vesicles in rat superficial cells and in human intermediate and superficial cells. By double immunostaining, lysosomes with or without vesicular structures were co-labeled with immunogold particles showing both cathepsin B and ubiquitin. The results suggest that cathepsins B, C, H, and L, and cathepsin D are involved in the lysosomal system of rat and human bladder epithelia. Moreover, considering that ubiquitin is a cofactor in the soluble ATP-dependent proteolysis, the results may also indicate that epithelial cells actively form autophagolysosomes.

Animals↗
PubMed2001

Three-dimensional analysis of nephrogenesis in the neonatal rat kidney: light and scanning electron microscopic studies.

In order to clarify the process of renal development more precisely than previously, the present study observed the rat neonatal kidney by scanning electron microscopy (SEM) of KOH digested tissue as well as by light microscopy of plastic sections. In the subcapsular region, aggregation of the mesenchymal cells was closely associated with the upper side of the ureteric duct ampulla. These mesenchymal cells projected a number of fine irregular processes at the basal portion facing the ureteric duct. A spherical cluster transformed from the mesenchymal cell aggregation was found on the lower side of the terminal ampulla, and was differentiated into the renal vesicle. Some cells at the top of the renal vesicle formed a cone-shaped projection and invaded the ureteric duct ampulla, forming a connection with it. In the advanced stage, a shallow transverse cleft appeared on the outer lateral side of the renal vesicle, and a second cleft was formed on the opposite side close to the junction between the renal vesicle and the ampulla. As the two clefts deepened, the vesicle assumed the well-known S-shaped body. In the advanced S-shaped body, the lower limb became cup-shaped, while the segment between the middle and lower limbs of the "S" elongated to form a tubular structure (i.e., the prospective proximal tubule and Henle's loop). The upper limb of the "S" also increased its length to form a distal tubule. The middle limb of the "S", however, was attached firmly to the cup-shaped lower limb (i.e., the prospective renal corpuscle) and was considered to become the macula densa of the mature nephron. In the maturing renal corpuscle, irregularly shaped cells were observed as a sheet-like aggregation at its vascular pole and were continuous with the vascular smooth muscle cells. These findings will help toward a better understanding of the morphological complexities of nephrogenesis.

Animals↗
PubMed1990

Atrial structure and plasma ANF levels in rats with chronic diabetes mellitus.

The effect of streptozocin-induced diabetes (STZ-D) on right atrial structure was investigated in male Wistar rats. STZ (55 mg/kg) or saline (1 ml/kg) was administered by intravenous injection 12 wk before the experimental studies. Tissue was sampled from four regions of the atrium, processed, and embedded in plastic. Quantitative stereological analysis indicated that in STZ-D rats, there was a significant diminution in size of the musculi pectinati (muscular ridges), which form a network making up the wall of the atrium. In addition, within the muscular ridges, there was a significant reduction in the relative proportion of cardiocytes within the cardiac tissue. The rest of the cardiac tissue consisted of interstitial regions, connective tissue, and blood vessels, which correspondingly increased. This suggests there was some form of cardiomyopathy. When atrial granularity was determined relative to cardiocyte volume density, a significant decrease (54%) was found in tissue from STZ-D rats. The blood pressure of conscious STZ-D rats was significantly lower than control rats, whereas right atrial pressure was not different. The level of resting plasma immunoreactive atrial natriuretic factor (ANF) in conscious STZ-D rats (98 +/- 5 pg/ml) was significantly higher than in control rats (52 +/- 7 pg/ml). The decreased atrial granularity could be related to the higher resting plasma ANF levels, suggesting a more rapid turnover or increased synthesis bypassing storage in the granular form.

Animals↗
PubMed2004

Correlation of computed tomographic images with anatomic features of the abdomen of ringed seals (Phoca hispida).

OBJECTIVE: To correlate anatomic features of the abdomen of the ringed seal (Phoca hispida) identified in plastinated cross-sections with images obtained via computed tomography (CT) and thereby establish reference standards for normal abdominal organ size and position in this species. SAMPLE POPULATION: 2 adult male ringed seal cadavers. PROCEDURE: With the seal in sternal recumbency, CT images of the abdomen were acquired by use of a 4th-generation CT scanner. Image slice thickness was 1 cm, with no interslice gap. After imaging, the abdominal region was sectioned transversely into 4-cm slices, which were plastinated and photographed. Plastinated slices were matched to their corresponding CT images in preparation for anatomic descriptions. RESULTS: Relevant anatomic features were identified and labeled on both the plastinated tissue slice and corresponding CT image. Normal abdominal organ size and position were assessed, and topographic relationships among organs were ascertained. CONCLUSIONS AND CLINICAL RELEVANCE: The data obtained provide some reference standards for normal abdominal organ size and position in ringed seals. This information may aid researchers of future physiologic and clinical studies in this species.

Abdomen↗
PubMed1986

Intra- and inter-observer variability and accuracy in the determination of linear and angular measurements in computed tomography. An in vitro and in situ study of human mandibles.

The observer variability and accuracy of linear and angular computed tomography (CT) software measurements in the transaxial plane were investigated for the temporomandibular joint with the General Electric 8800 CT/N Scanner. A dried and measured human mandible was embedded in plastic and scanned in vitro. Sixteen observers participated in the study. The following measurements were tested: inter- and extra-condylar distances, transverse condylar dimension, condylar angulation, and the plastic base of the specimen. Three frozen cadaveric heads were similarly scanned and measured in situ. Intra- and inter-observer variabilities were lowest for the specimen base and highest for condylar angulation. Neuroradiologists had the lowest variability as a group, and radiology residents and paramedical personnel had the highest, but the differences were small. No significant difference was found between CT and macroscopic measurement of the mandible. In situ measurement by CT of condyles with structural changes in the transaxial plane was, however, subject to substantial error. It was concluded that transaxial linear measurements of the condylar processes free of significant structural changes had an error and an accuracy well within acceptable limits. The error for angular measurements was significantly greater than the error for linear measurements.

Child↗
PubMedDate not supplied

Patterns of hair cell loss in chick basilar papilla after intense auditory stimulation. Exposure duration and survival time.

Ten-day-old chicks were exposed to a pure tone (1.5 kHz) or white noise at 125 dB SPL avg. RMS for 4 to 24 hours, and were sacrificed either 10, 30 or 60 days after exposure. The basilar papillae were embedded in plastic, sectioned, and hair cells were counted at 100-micrometer intervals throughout the length of the papilla. As sound exposure duration increased, both the maximum number of hair cells lost, and the extent of the damaged area along the basilar membrane increased. Short hair cells situated on the free area of the basilar papilla were more susceptible to damage than were tall hair cells. The location of hair cell loss varied as a function of frequency band of exposure; the pure tone produced a well localized basal lesion, while wide-band noise produced a more general lesion which extending toward the apex. Degeneration continued with increased survival time up to 30 days. It is concluded that avians respond to acoustic overstimulation in a manner very similar to mammals. The convenience of this preparation along with the relative simplicity of its cochlea may render it useful for future investigations of the mechanisms of acoustic trauma.

Animals↗
PubMed1982

Patterns of hair cell loss in chick basilar papilla after intense auditory stimulation. Frequency organization.

Ten-day-old chicks were continuously exposed to pure tones (500, 1 500, or 3 000 Hz) at 125 dB SPL for 12 hours and sacrificed 10 days after exposure. The basilar papillae were embedded in plastic, sectioned and hair cells were counted at 100-mum intervals throughout the length of the papilla. The position of hair cell loss along the basilar membrane varied systematically as a log-linear function with the frequency of stimulation. This systematic relationship was used to predict the frequency organization of the chick basilar papilla. It is concluded that although the avian basilar papilla differs greatly in morphological detail from the mammalian cochlea, its response to intense acoustic stimulation is quite similar.

Acoustic Stimulation↗
PubMed1987

Observer variability in microcomputer-assisted morphometric study of nuclear parameters.

To evaluate observer variability during microcomputer-assisted planimetry, nuclear features (area, perimeter, and form factor) were studied in a case of malignant lymphoma. Tissue fixed in glutaraldehyde, embedded in plastic and sectioned at 1 micron was studied using a Zeiss Kontron IBAS 2000 image analysis system. Manual tracing of nuclear outline was performed to obtain the parameters. Both interindividual and intraindividual reproducibilities were evaluated, the former among 8 observers and the latter by the observers repeating the measurements. With the present technique, intraindividual reproducibility was high and correlation was excellent for area, perimeter and form factor measurements. A high degree of interindividual inconsistency was demonstrated for all parameters. Interindividual correlation was poor for form factor. It is concluded that poor interindividual reproducibility even when the objective method of morphometry is used may be a significant problem in tissue investigations, especially when results from different laboratories are being compared.

Cell Nucleus↗
PubMed1988

Response of the tracheobronchial mucociliary clearance system to repeated irritant exposure: effect of sulfuric acid mist on function and structure.

This study was designed to determine quantitative and temporal alterations in tracheobronchial mucociliary clearance function and structure due to repeated inhalation exposures to a common irritant, sulfuric acid mist. Rabbits were exposed to 250 micrograms/m3 sulfuric acid (0.3 micron) for 1 h/day, 5 days/week, for up to 1 year, with some animals allowed a 3-month recovery period following the end of the acid exposures. Control animals received temperature- and humidity-conditioned water vapor. At intervals of 2 to 4 weeks, animals inhaled a radioactively tagged tracer aerosol (ferric oxide microspheres, 4.5 micron), and its clearance via mucociliary transport from the thorax was monitored by external serial counting. Clearance became slower during the first month of acid exposure, and this slowing became progressive with time through the end of the 12-month exposure period. After cessation of acid exposure, clearance became extremely slow and did not return to normal by the end of the follow-up period. To assess specific histological changes in the tracheobronchial tree, groups of rabbits were killed after 4, 8, or 12 months of exposure and after the follow-up period. Tissue samples from each lung were embedded in plastic, sectioned at 3 micron, and stained with hematoxylin and eosin or alcian blue/periodic acid-Schiff (AB/PAS). Acid exposure changed the airway diameter distribution compared to the control; except for the follow-up group, all acid-exposed animals had a shift to smaller airways. Acid inhalation also caused an increase in epithelial secretory cell density and a shift from PAS to AB staining of glycoprotein within secretory cells, both of which were unresolved by 3 months after the exposures ceased. No evidence of inflammation was found in any of the animals. Thus, repeated exposures to H2SO4 resulted in a slowing of mucociliary clearance that was associated with alterations in airway morphometry and morphology. Such changes may be involved in the early pathogenesis of chronic airway disease.

Aerosols↗
PubMedDate not supplied

Ultrastructural confirmation of malignant melanoma.

We have recently had the opportunity to examine two cases, where malignant melanoma was suspected on light microscopy and Fontana-Masson stain was focally and faintly positive. Electron microscopy of four to five different blocks failed, however, to demonstrate the presence of premelanosomes and thereby to confirm the diagnosis of malignant melanoma. The areas in the paraffin blocks that on Fontana-Masson stain had showed a positive reaction were deparaffinized, embedded in plastic, and examined by electron microscopy. Now the characteristic premelanosomes were found and the diagnosis of malignant melanoma was confirmed.

Adult↗
PubMed1987

Electron spectroscopic imaging.

Electron spectroscopic imaging (ESI) is a short term for electron energy loss spectroscopic imaging. ESI is to date the most sensitive electron microscopic analytical technique and has a spatial resolution of 0.5 nm and a mass resolution of less than 50 atoms. ESI takes advantage of filtered, inelastically scattered, transmitted electrons to form spectroscopic images. This analytical mode was developed by F. P. Ottensmeyer of the Ontario Cancer Institute in Toronto, Canada. Specimens to be analyzed by ESI should not exceed 30 nm in thickness so that multiple scattering of electrons is avoided. Isolated macromolecules constitute ideal specimens. For the analysis of diffusable elements, elaborate techniques involving quick-freezing, drying and embedded in plastic of the specimens have to be used. An example is given in which calcium was analyzed in the mitochondria of the convoluted proximal tubules in the rat kidney in normal and acute ischemic conditions. The example shows that calcium was not extracted during the preparation of the tissue. The high resolution of ESI allows the localization of calcium inside mitochondria. Experiments in progress using homogenous standards will soon render ESI entirely quantitative. It is concluded that ESI will become more and more important as a tool for the study of the pathogenesis of conditions in which modification of the distribution and concentration of elements are involved.

Animals↗
PubMedDate not supplied

Cryofixation, cryosubstitution, and immunoelectron microscopy: potential role in diagnostic pathology.

In immunoelectron microscopic investigations, retention of antigenic sites is crucial. Methods for preparing samples for conventional electron microscopy involve chemical fixatives followed by dehydration in organic solvents and embedding in plastic resins, all procedures potentially detrimental to antigenicity. Cryomethods provide a physical fixation alternative for the preparation of biological samples for ultrastructural, immunocytochemical, and microanalysis studies without the use of any chemicals. This can be particularly useful in diagnostic pathology, providing an alternative to conventional fixation methods which sometimes destroy the antigen in question. The recent development of a portable cryofixation device, the PS1000 Portable Metal Mirror Ultra-Rapid Cryofixation Unit (Delaware Diamond Knives, Inc., Wilmington, DE, USA), provides an opportunity to freeze tissue immediately after procurement for use in diagnostic immunocytochemistry studies. This feasibility study examined the quality of tissue preservation with this device, in terms of both preservation of cellular ultrastructure and immunolabeling. Human tonsil and thymus tissue was slam frozen and, after cryosubstitution in Lowicryl K11M, was examined by immunoelectron microscopy. Good ultrastructural preservation was obtained and reasonable immunolabeling with antibodies to AE1/AE3 keratin filaments was also observed.

Cryopreservation↗
PubMed1982

A simple fluorescence technique for light microscopy of the crystalline lens.

Lenses fixed in formaldehyde or glutaraldehyde can be embedded in plastic, and sections as large as 3-4 mm across can be cut with glass knives on an ultramicrotome. When these sections are stained with 25% sodium fluorescein and allowed to bleach in a fluorescence microscope, high contrast images of essentially every cell in the section, including cells very near the lens center, can be seen. The method is effective with lenses from a number of species, and should be useful for a wide variety of morphological studies of normal and cataractous lenses.

Animals↗