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PubMed1984

The use of semiautomatic morphometry in the study of normal rat gastrocnemius muscle fibers.

Based on a fixed number of EM photomicrographs, a semiautomatic image analysis is made of normal gastrocnemius muscle fibers to obtain reference values for comparison with pathologic data. Z disc, mitochondria, tubular system, and sarcoplasmic reticulum are measured. It is stated that no definite fiber type differentiation is possible based on these quantitative data. However, measurements of Z disc and mitochondria are suggestive of such a differentiation. On the other hand, all data reach a Gaussian distribution on a logarithmic scale after a 5% correction factor is introduced.

Animals↗
PubMed1989

Morphometry of terminal hepatic veins. 1. Comparative study in man and baboon.

In the present study, hepatic venous distribution per unit of liver surface area on normal wedge biopsies from man (n = 11) and baboon (n = 8) were analysed and compared. Terminal hepatic veins (THV - man: n = 100; baboon: n = 200) morphometric size variables were obtained with a Leitz ASM 68K morphometric equipment. THV, defined as hepatic veins up to 150 microns in internal diameter (ID), in the centrolobular position and with sinusoidal openings, represented 84% and 74% of hepatic veins of man and baboon, respectively. Four or more THV were generally found on 8 mm2 of liver surface. Transversely sectioned THV selected by the ratio IDminimum/IDmaximum greater than 0.67, was found to be only 25% of the total THV. In baboon, THV merge with other terminal veins and the interlobular veins present sinusoidal inlets. The baboon THV wall surface (WS) and wall thickness (WT) values were higher than in man. Positive correlations between the number of mesenchymal cells (Mc) in the vein wall and wall surface of terminal hepatic veins (man: r = 0.79; baboon: r = 0.83) and between wall surface and internal surface (IS) (man: r = 0.80; baboon: r = 0.72) were found. Two ratios were selected as the most reliable parameters: (1) for the THV wall rim, wall surface/internal surface (WS/IS - man: 0.43 +/- 0.16; baboon: 0.63 +/- 0.23), regarding transversely sectioned THV; and (2) for the evaluation of wall cell density (WS/Mc-man: 550 +/- 231; baboon: 558 +/- 183 micron 2/cell) as they did not depend on THV caliber.

Adult↗
PubMed1989

Morphometry of terminal hepatic veins. 2. Follow up in chronically alcohol-fed baboons.

Alcohol induced perivenular fibrosis of terminal hepatic veins (THV) is claimed to be a precursor lesion leading to fibrosis development in man and baboon. The nature and significance of the THV lesions found in four baboons chronically fed with alcohol were studied in liver biopsies obtained during a three year period. The surface of THV wall and the number of mesenchymal cells were assessed with a semi-automatic image analyser. Histologically, THV were characterized as (a) phlebitic, in the presence of an inflammatory cell infiltrate involving the venous wall; (b) oedematous, when the connective tissue of the THV wall was disrupted or dissociated and (c) fibrotic, when perivenular scarring appeared as an increased rim. These lesions were present simultaneously and their intensity and distribution were independent of the duration of alcohol intoxication. Morphometric data obtained from these THV confirmed the thickening of the THV wall (WS/IS in: oedematous 1.05 +/- 0.36; phlebitic 1.65 +/- 1.04; fibrotic 1.47 +/- 0.36); and revealed an increased number of mesenchymal cells in fibrotic (439 microns 2/cell; p less than 0.01) and in phlebitic THV (510 microns 2/cell; p less than 0.05). A constant relationship was found between phlebitis and the presence of inflammatory infiltrate within the hepatic acini. Fibrotic THV was a less frequent finding and the lesion disappeared in the sequential biopsies of one of the baboons. In conclusion, THV lesions were heterogeneous and the collagen deposition in the THV wall was potentially reversible during the three year alcohol intoxication period, regardless the inflammatory reaction and, thus, did not indicate early irreversible hapatic fibrosis.

Alcoholic Intoxication↗
PubMed1989

Different responses between the upper and the lower parathyroid gland in a state of secondary hyperfunction. A study on chronic renal failure by morphometry and nuclear DNA analysis.

The size of the parathyroid gland and the size, the numerical density and nuclear DNA-content of the parathyroid gland cells were evaluated in chronic renal failure (CRF) and revealed a difference between the upper and the lower glands in the manner of adaptation to a state of long-term hyperfunction, secondary to CRF. The parathyroid gland enlarged as a whole in CRF, an effect more marked in the lower gland, whereas individual parathyroid gland cell enlargement in CRF was mainly seen in the upper gland cells. The numerical density of the lower parathyroid gland cells was higher than that of the upper gland. Nuclear DNA-content of the parathyroid gland cells were increased in CRF and the lower gland tended to show hyperdiploid aneuploidy. These findings are probably related to the fact that parathyroid adenomas occur most often in the lower gland. The higher proliferative activity of the lower parathyroid gland in long-term hyperfunction may explain the higher risk for the lower gland in the occurrence of adenomas.

Autopsy↗
PubMed1987

Electron microscopical morphometry of GH producing pituitary adenomas in comparison with normal GH cells.

GH producing adenomas of patients with acromegaly (undifferentiated acidophil adenomas and well differentiated GH cell adenomas) were studied at the ultrastructural level and analysed morphometrically by the point counting method. They were compared with identically prepared GH cells of normal pituitaries from patients undergoing surgery for metastasizing cancer of the prostate. In the well differentiated GH cell adenomas significantly more points were counted on nucleoli, unorganized cytoplasm, rough endoplasmic reticulum, immature secretory granules, Golgi areas and on the plasma membranes, than in normal GH cells. Comparison of normal GH cells with tumour cells in undifferentiated acidophil adenomas demonstrated significantly larger volumes of nuclei, rough endoplasmic reticulum, Golgi fields, immature secretory granules and of the cell membranes, and also of nucleoli and of the mitochondria. Secretory granules and lysosomes were observed more frequently than in normal GH cells. In a comparison of both adenoma types, the well differentiated acidophil adenomas contained significantly larger volumes of the unorganized cytoplasm, secretory granules and of cell membranes, whereas more points were counted on the rough endoplasmic reticulum and on the mitochondria in undifferentiated acidophil adenomas. The differences between the normal GH cells and the GH cell in undifferentiated adenomas (mainly larger nucleoli, larger volumes of the rough endoplasmic reticulum and the lower volumes of secretory granules) indicate a higher secretory activity in the adenomas. The significant differences between the well differentiated and the undifferentiated adenomas (mainly the increased volumes of mitochondria and of the unorganized cytoplasm in the undifferentiated tumours) indicate a lower grade of differentiation and may be interpreted as signs of increased proliferation.

Adenoma↗
PubMed1986

Unmyelinated axons in a muscle nerve. Electron microscopic morphometry of the sternomastoid nerve in normal and sympathectomized rats.

In continuation of earlier studies on the innervation of the sternomastoid muscle of the rat, a detailed morphometric analysis was performed on the unmyelinated axons of the nerve, in normal rats and after extensive cervical sympathectomy. In 4 normal rats an average of 314 myelinated and 319 unmyelinated axons were present. 42 days after surgery, the 3 animals showed Horner's syndrome and a highly significant 40% loss of unmyelinated axons. We therefore suggest that 40% of the C-fibers in this nerve are postganglionic sympathetic efferents and that the remaining 60% are type IV fibers, i.e., unmyelinated afferents. Our counts also indicate that part of the Remak bundles of the Schwann cells contain only sympathetic axons, whereas others contain mixed groups of sympathetic and afferent axons. Myelinated nerve fibers were not lost due to sympathectomy. Unexpectedly, the 3 animals analyzed 7-13 days after surgery showed Horner's syndrome but only a 16% loss of unmyelinated axons, which was not even statistically significant. Morphological signs of degeneration and sprouting did not provide any clue, but a possible explanation would be that a transitory sprouting of the remaining afferent C-fibers or Schwann cells occurred.

Animals↗
PubMed1988

Differentiation of plasma cell infiltrates in the bone marrow. A clinicopathological study on 80 patients including immunohistochemistry and morphometry.

In 80 patients immunohistochemical, morphometrical and clinical studies were performed on routinely referred trephine biopsies of the bone marrow showing an abnormal increase in plasma cells. From the approximately determined density of plasma cell infiltrates two main groups were distinguished, the first with an involvement exceeding 20% and the second with less than 10% of the total marrow area involved. The first group (n = 30; 324 +/- 130 plasma cells per square millimeter bone marrow) consisted of patients with frank malignant myeloma (MM) by clinical and histomorphological diagnosis. The second group (n = 50; 132 +/- 54 plasma cells per square millimeter bone marrow) with plasmacytic differentiation of infiltrates, had to be further divided into one component with evidence for initial or residual MM following chemotherapy (n = 27), another with obviously monoclonal gammopathy of undetermined significance--benign monoclonal gammopathy (BMG, n = 6), and a final set of cases with a reactive plasmacytosis mostly associated with an inflammatory condition (n = 17). There was an excellent agreement between the intracellular immunoglobulin staining as defined by the immunoperoxidase technique and the serum or urinary M-component detected by immunoelectrophoresis. In MM significant correlations were found between osteoclastic activity (number of osteoclasts specifically stained by acid phosphatase) per trabecular bone area, presence of lytic bone defects and the density of plasma cell infiltrates in the marrow. This latter feature corresponded well with the titer of secreted serum M-components measured by quantitative immunoelectrophoresis. Using morphological data alone, BMG cases could not be discriminated with any certainty from initial or residual plasmacytic MM. They consequently need a prolonged clinical follow up to clarify the nature of the lesions.

Bone Marrow↗
PubMed1993

Glomerular morphometry in reflux nephropathy: functional and radiological correlations.

Using computerized digitometry, we investigated the relationships between renal size, glomerular filtration rate (GFR), proteinuria, incidence of segmental and global glomerulosclerosis, glomerular size, hilar arteriolar wall thickness and hyaline deposition in renal biopsies obtained from 24 children and adolescents with reflux nephropathy, of whom only 4 were hypertensive. Age-matched controls comprised minimal-change nephrotic syndrome (6) and recurrent haematuria with normal biopsy (13). The mean sectional area of patients' glomeruli was double that of controls. Glomerular size correlated with the amount of proteinuria (measured as protein/creatinine ratios in early morning urine) and inversely with renal size and GFR. Segmental sclerosis, invariably of hilar origin, was observed in 8 patients and the percentage of glomeruli affected correlated strongly with glomerular size and proteinuria. Global sclerosis was found equally in patients and controls, and showed no similar correlations. Compared with controls, patients' hilar arterioles showed increased wall thickness, more intramural hyaline deposits and decreased luminal diameter when related to glomerular size. The proteinuria and glomerular changes are consistent with hyperfiltration, while the previously undescribed hilar vascular changes, which both precede and accompany sclerosis, resemble abnormalities reported experimentally following renal ablation.

Adolescent↗
PubMed1991

The transverse tubular system of the hypertrophic myocardium: morphology and morphometry in spontaneous hypertensive rats (SHR).

We reported previously on a modified Golgi stain that, in conjunction with high voltage electron microscope stereoscopy, gives striking views of the elaborate network of the transverse tubular system (T system) in rat myocardium. In this report we used the same techniques to study three-dimensional arrangements of the T system in the left ventricular myocardium of spontaneous hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). High voltage electron microscope stereoscopy revealed distinctive morphological characteristics of the T system, such as undulating running, short dead-end branches, and labyrinth-like tubular aggregates in the hypertrophic myocardium of SHR. Quantitative analysis of the SHR T system indicated a surface area greater than that of WKY. These findings may support the hypothesis that making an additional T system membrane will compensate for the smaller surface-to-volume ratio. However, the normal regulatory mechanism required to maintain the surface-to-volume ratio does not function properly in SHR, resulting in morphological abnormalities and functional disturbances of the myocardium.

Animals↗
PubMed1984

Ultrastructural morphometry of collagen from lamina propria during experimental oral carcinogenesis and chronic inflammation.

Stereological point-counting methods were used to determine the volumetric alterations in collagen from the lamina propria immediately beneath the epithelial-connective tissue junction in hamster check-pouch mucosa treated with the chemical carcinogen DMBA. In addition, a non-neoplastic inflammatory control was evaluated in which a delayed hypersensitivity reaction was induced by the contact-sensitising agent DNCB. DMBA-treated tissues were assigned to histopathologically defined hyperplasia, dysplasia and carcinoma stages. The volume densities of collagen present in unit volume of extracellular lamina propria were found to decrease progressively and significantly in DMBA-treated tissues when compared with values obtained from normal untreated mucosa. Values from the inflammatory control were comparable with those from the dysplasia stage of carcinogenesis. The mechanisms responsible for these decreases in collagen volume density are unknown, but contributory factors might include collagen destruction by enzymes originating in either the epithelium or the cells of the inflammatory infiltrate, dilution of collagen produced by inflammatory oedema or alterations in the synthetic capabilities of fibroblasts.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1995

Nuclear morphometry and DNA densitometry of human gliomas by image analysis.

In 48 patients with gliomas in whom complete clinical follow-up was obtained, DNA ploidy was evaluated by using formalin-fixed paraffin-embedded tissues and by means of image analysis. The mean DNA indices, determined by averaging DNA indices of all tumor cells in a tumor, were mainly affected by mean DNA indices of the nuclei of SG2M phase tumor cell (including S phase and G2M phase cells) (SG2M DNA indices) and that mean DNA indices correlated with the SG2M phase fraction. The SG2M DNA indices and the percentage of tumor cells with S phase and G2M phase were higher in high grade gliomas including anaplastic glioma and glioblastoma multiforme than in low grade gliomas. Patients with G2M-hypertetraploid tumors demonstrated a shorter time to tumor progression than those with G2M-tetraploid in high grade glioma. Morphometrically, the nuclei of SG2M phase glioma cells were larger and more deformity than those of G0G1 phase (including G0 phase and G1 phase cells) cells. The G2M-hypertetraploid tumors were highly malignant and demonstrated large nuclei, greater nuclear deformity, and a higher proliferative potential. The G2M-tetraploid gliomas demonstrated a shorter time to tumor progression in cases whose the SG2M fraction was large. In contrast, G2M-hypotetraploid gliomas revealed an insignificant trend towards a longer time to tumor progression than those associated with tetraploid and hypertetraploid gliomas. We emphasize herein the prognostic importance of the SG2M phase cell, as well as other proliferation indices.

Adolescent↗
PubMed1977

Trabecular bone morphometry in beagles with chronic renal failure.

A morphometric study was performed on undecalcified sections of trabecular bone from the ribs of adult dogs that were controls (n = 8), or had various degrees of renal failure as a result of perinatal irradiation (n = 16). In a group identified as markedly uremic there was an increase in the proportion of surface with osteoid seams (P less than 0.01), osteoblasts (P less than 0.01) and osteoclastic resorption (P less than 0.01) as well as the % osteoid volume (P less than 0.01). Aside from a decrease in the % trabecular bone which was of unclear significance, the results agree well with those of similar studies in man. Within this group, 6 of the samples had a histologic pattern that was principally that of hyperarathyroidism and in the remaining 2 there was evidence of a mineralization defect and osteomalacic changes. These 2 dogs had the poorest renal function and one that had been biopsied 5 months earlier, when its renal function was less impaired, had had a hyperparathyroid pattern.

Animals↗
PubMed1992

Morphometry versus densitometry--a comparison by use of casts of human coronary arteries.

Geometric and densitometric methods for quantitative coronary arteriography have generally been compared by use of phantoms simulating arteries with circular lumina ('Hole phantoms'). We have used more adequate phantoms obtained by casting disease-free and atheromatous human coronary arteries. The phantoms, filled with contrast medium, were imaged digitally (1024 x 1024 x 10 matrix) under experimental conditions simulating routine coronary angiography. The angiographic 'diameters' and the densitometric cross-sectional areas of 59 marked lumina were determined in single plane and orthogonal biplane raw images. Geometric calibration was performed by help of a 7F coronary catheter. For the densitometric calibration, we used a 'hole phantom' attached to the image intensifier. The obtained luminal areas were compared to their true values determined previously by planimetry. The mean absolute error of single plane cross-sections obtained geometrically was 1.53 mm2. Biplane imaging reduced it by a factor 2.4 to 0.64 mm2. The corresponding mean absolute errors for densitometry were 0.56 mm2 and 0.51 mm2. Single plane 'diameter' measurements appear thus of very limited value for hemodynamic conclusions. In contrast, biplane geometric quantification was not markedly inferior to single plane and biplane densitometry.

Absorptiometry, Photon↗
PubMed1985

The axon tree of rat motor fibres: morphometry and fine structure.

A quantitative light and electron microscopic study of the axon tree of rat motor fibres was undertaken to supply data relevant to the understanding of the spread of excitation. Continuous proximodistal foreshortening of internode length and dwindling of fibre calibre were statistically verified. Internodes had progressively foreshortened geometric proportions (length/diameter quotient). The postbranching internodes were significantly shorter than would correspond to the general trend, but no such foreshortening was evident for the prebranching internodes. Relative sheath thickness (g-ratio, i.e. quotient axon diameter/fibre diameter) of filial fibres did not differ from the stem. Increase in axon area in filial fibres versus stem fibre averaged 1.3. The total proximodistal increase in axoplasmic area from the stem fibre to the distal branches was less than two since increases at branch points were compensated by the proximodistal dwindling of fibres. Nodal membrane area of branch points was greater than in nonbranching fibres, and there were larger than normal microvillar spaces. The majority of branches were approximately symmetric, but asymmetric branchings were also found. The thinner branches of these had wider nodal gaps than the thick branches. The observed changes were interpreted as adaptations to facilitate uniform spread of excitation along the axon tree; asymmetric branchings, however, may permit routing of impulses.

Animals↗
PubMed1979

Stereological approach to the study of synapse morphometry with particular regard to estimating number in a volume and on a surface.

The paper presents a system of stereological formulae which are suitable for the quantitative ultrastructural characterization of synapses. Attention is given first to the question of deciding which synaptic feature to adopt as the basis for measurement. Three counting units can be recognized on electron micrographs: the bouton, the total zone of apposition of pre- and postsynaptic membranes, and the zone of paramembranous density. These may provide different numerical results. Possible units of reference are considered also and whilst emphasis is given to a volume of nervous tissue and a membrane surface of postsynaptic element, interesting alternatives are presented for the sake of completeness. Stereological parameters available for quantifying synapses (for example boutons per tissue volume, paramembranous dense zones per soma surface) are described. In addition, some new and previously neglected possibilities are included and some errors found in the literature are corrected. Worked examples of the estimation of synaptic densities are provided.

Animals↗
PubMed1989

Studies of alveolar cell morphometry and mass clearance in the rat lung following inhalation of an enriched uranium dioxide aerosol.

Seventy-three rats were exposed to an aerosol of enriched uranium dioxide (UO2), giving initial lung burdens of 26 to 447 micrograms at 6 days post-inhalation (PI). At 7 days PI 35 of these rats were further exposed to thermalised neutrons at a fluence of 1 x 10(12) neutrons cm-2. There was no significant difference between the two groups in the clearance rate of the UO2 particles from the lung, up to 590 days PI. The particles cleared relatively slowly over this period with a retention half-time in the lung of 160 to 176 days. Transmission electron microscope (TEM) studies of tissue from the alveolar region at 8 days PI showed that inhalation of UO2 particles significantly increased the sizes of macrophage and type II cells, and the number of macrophage and type I cells. There was also a significant increase in the size of lysosomal granules within the macrophages after exposure to the UO2 particles. The exposure to UO2, neutrons and 235U fission fragments had no significant effect on any of the cells above that observed in the animals exposed to UO2 alone. Additional rats were exposed to the same neutron fluence without prior UO2 inhalation. The alveolar cells of neutron-only exposed rats were, in size and number, typically no different from those in the completely unexposed control rats.

Administration, Inhalation↗
PubMed1993

The prognostic value of basement membrane morphology, tumour histology and morphometry in superficial bladder cancer.

This analysis evaluated the prognostic importance of clinical, histological (WHO grade, papillary status, tumour-infiltrating lymphocytes) and morphometric factors (mitotic index, mean nuclear area, SD of nuclear area) in a cohort of 132 patients with a Ta-T1 transitional cell bladder cancer followed-up for over 8 years. Special attention was given on the state of the subepithelial basement membrane (BM) and subepithelial vascular density as demonstrated by type IV collagen immunostaining. Defects in the BM and dense vascular network were related to nodular growth pattern, high histological grade, rapid cancer cell proliferation and a dense inflammatory cell infiltrate (tumour-infiltrating lymphocytes, TIL) in tumour stroma. The most important predictor of progression was the mitotic index (P < 0.0001) followed by vascular density (P = 0.0054), the state of the BM (P = 0.0056), density of TIL (P = 0.0193), WHO grade (P = 0.0356) and papillary status (P = 0.0448). In univariate survival analysis the mitotic index (P < 0.0001), the state of the BM (P < 0.0001), subepithelial vascular density (P = 0.0013) and WHO grade (P = 0.01) predicted survival. In a multivariate analysis the mitotic index (P < 0.0001) and the state of the BM (P = 0.008) were independent predictors. The results show that the evaluation of the mitotic rate and investigation of the continuity of the BM should be done whenever the prognosis of Ta-T1 tumours of the bladder is assessed.

Aged↗