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PubMed1994

Morphometry of the kidney in rat pups from uninephrectomized mothers.

BACKGROUND: The present study was designed to explore whether maternal renal dysfunction affects fetal kidney development and if the effects are lasting during the postnatal development. METHODS: Kidneys of 1-day-old and 6-week-old pups from mothers which were uninephrectomized on day 5 of gestation were studied. RESULTS: One day after birth, both the number of immature glomeruli and average volume of mature glomeruli of the neonates from uninephrectomized mothers were significantly larger than those from sham-operated mothers. Six weeks after birth, no significant differences in parameters of the kidney were observed between the pups from uninephrectomized and those from sham-operated mothers. Furthermore, blood urea nitrogen (BUN) concentration in adult female rats was determined at various days after uninephrectomy. BUN concentration in uninephrectomized rats was significantly higher than that in sham-operated ones. CONCLUSIONS: These results suggest that the fetal kidney development is accelerated by the elevated BUN level following maternal uninephrectomy and that the renotropic activity does not last during the postnatal developmental period.

Animals↗
PubMed1995

Morphometry of rat germ cells during spermatogenesis.

BACKGROUND: There has never been a study of the components of germ cells as they progress through spermatogenesis. METHODS: The structural changes taking place in rat germ cells, from spermatogonia to late spermatids, were studied utilizing morphometric techniques conducted largely at the ultrastructural level. RESULTS: Volume and surface area parameters for virtually all cellular and subcellular features were obtained for nine periods during the spermatogenic cycle. Virtually all germ cell components show dynamic properties associated with specific phases of their development. CONCLUSIONS: The data provided can be used in an objective way to characterize structural changes taking place during spermatogenesis and to relate those structural changes to functional properties of germ cells.

Acrosome↗
PubMed1995

Cellular distribution patterns of lanthanum and morphometry of rat hearts exposed to different degrees of ischemic stress.

BACKGROUND: The element lanthanum (La) can be used as a tracer for verification of membrane permeability. The aim of this study was to establish whether 1) distribution of La in the myocardium of rat hearts depends on the degree of ischemic stress and 2) morphometrically determined cell and mitochondrial swelling correlates with the La distribution. MATERIALS AND METHODS: Isolated beating rat hearts were arrested by coronary perfusion with the cardioplegic solution Custodiol (controls) or by aortic cross clamping followed by exposure to different degrees of ischemic stress. The solutions for perfusion-and postfixation as well as for rinsing contained 1.1% La(NO3)3. Cellular and mitochondrial swelling were determined morphometrically and myocytes exhibiting intracellular La were quantified and stated as percentage of test fields. RESULTS: Immediately after cardiac arrest La was present as precipitates only in a few myocytes adjacent to the outer mitochondrial membrane as seen by cTEM and ESI. In such cells La was also detected by EELS in mitochondrial matrix and myofibrils. Advanced ischemic stress led to an increase of the percentage of myocytes containing detectable intracellular La. After 45 min ischemia at 30 degrees C, myocytes and mitochondria showed a remarkable edema and different intracellular distribution patterns of La. After 90 min of ischemia at 20 degrees C interruptions of sarcolemma could only be detected in a few of the swollen myocytes. Roundish La granules were seen in the myofibrils. The percentage of myocytes containing intracellular La and the extent of cellular and mitochondrial swelling showed a significant correlation. CONCLUSIONS: Patterns of intracellular La distribution depend on the degree of ischemic stress and correspond to the degree of cellular as well as mitochondrial edema. These results point at a direct relation between alterations of membrane permeability and development of edema.

Animals↗
PubMed2003

Ultrastructural morphometry of capillary basement membrane thickness in normal and transgenic diabetic mice.

Capillary basement membrane (CBM) thickening is an ultrastructural hallmark in diabetic patients and in animal models of diabetes. However, the wide variety of tissues sampled and diverse methods employed have made the interpretation of thickness data difficult. We showed previously that acellular glomerular BMs in OVE26 transgenic diabetic mice were thickened beyond normal age-related thickening, and in the current study we hypothesized that other microvascular BMs likewise would show increased widths relative to age-matched controls. Accordingly, a series of tissues, including skeletal and cardiac muscle, ocular retina and choriod, peripheral nerve, lung, pancreas, and renal glomerulus was collected from 300-350-day-old normal and transgenic mice. Transmission electron micrographs of cross sections through capillary walls were prepared, and CBM thickness (CBMT) was determined by the "orthogonal intercept" method. Morphometric analyses showed highly variable transgene-related BMT increases in the sampled tissues, with glomerular BM showing by far the greatest increase (+87%). Significant thickness increases were also seen in the retina, pulmonary alveolus, and thoracoabdominal diaphragm. BMT increases were not universal; however, most were modestly widened, and those that were thickest in controls generally showed the greatest increase. Although the pathogenesis of diabetes-related increases in CBM is poorly understood, data in the current study showed that in OVE26 transgenic mice increased BMT was a frequent concomitant of hyperglycemia. Accordingly, it seems likely that hyperglycemia-induced microvascular damage may be a contributing factor in diabetic BM disease, and that microvessel cellular and extracellular heterogeneity may limit the extent of CBM thickening in diverse tissues.

Animals↗
PubMed2004

Neurogenesis and stereological morphometry of calretinin-immunoreactive GABAergic interneurons of the neostriatum.

We determined the neurogenesis characteristics of a distinct subclass of rat striatum gamma-aminobutyric acidergic (GABAergic) interneurons expressing the calcium-binding protein calretinin (CR). Timed-pregnant rats were given an intraperitoneal injection of 5-bromo-2'-deoxyuridine (BrdU), a marker of cell proliferation, on designated days between embryonic day 12 (E12) and E21. CR-immunoreactive (-IR) neurons and BrdU-positive nuclei were labeled in the adult neostriatum by double immunohistochemistry, and the proportion of double-labeled cells was quantified. CR-IR interneurons of the neostriatum show maximum birth rates (>10% double labeling) between E14 and E17, with a peak at E15. CR-IR interneurons occupying the lateral half of the neostriatum become postmitotic prior to medial neurons. In the precomissural neostriatum, the earliest-born neurons occupy the lateral quadrants and the latest-born neurons occupy the dorsomedial sector. No significant rostrocaudal neurogenesis gradient is observed. CR-IR neurons make up 0.5% of the striatal population and are localized in both the patch and the matrix compartments. CR-IR neurons of the patch compartment are born early (E13-15), with later-born neurons (E16-18) populating mainly the matrix compartment. CR-IR cells of the neostriatum are a distinct subclass of interneurons that are born at an intermediate time during striatal development and share common neurogenesis characteristics with other interneurons and projection neurons produced in the ventral telencephalon.

Age Factors↗
PubMed2001

Quantitative analyses of anatomical and electrotonic structures of local spiking interneurons by three-dimensional morphometry in crayfish.

We quantitatively investigated the three-dimensional structure of the dendrites of local spiking interneurons using a confocal laser scanning microscope in the terminal abdominal ganglion of crayfish. We also studied their passive membrane properties electrophysiologically using the single-electrode current clamp techniques to analyze their electrotonic structure. All of the local spiking interneurons examined in this study lacked distinctive axonal structure and had a monopolar cell body that was connected with a fine primary process to a thick main segment. Numerous fine secondary processes projected from the main segment in the ganglionic neuropile. The average anatomical length of a secondary process from the main segment to its terminal was 261.9 +/- 15.2 microm. The average input resistance and membrane time constant of local spiking interneurons, obtained from their voltage responses to intracellular injection of step current pulses in the main segment, were 15.2 +/- 1.6 MOmega and 13.9 +/- 1.9 msec, respectively. Calculation of the electrotonic length of dendritic processes based on morphological and physiological data obtained in this study revealed that the average electrotonic length of secondary processes in local spiking interneurons was significantly longer than in local nonspiking interneurons, although both types of local interneurons showed apparently similar anaxonic structure. The steady-state voltage attenuation factors for the secondary processes of local spiking interneurons were significantly greater than those of local nonspiking interneurons in both centrifugal and centripetal directions. The larger electrotonic structure of local spiking interneurons compared to that of nonspiking interneurons appears to be compensated for by their excitable dendritic membrane.

Action Potentials↗
PubMed2004

Quantitative morphometry of hippocampal pyramidal cells: differences between anatomical classes and reconstructing laboratories.

The dendritic trees of hippocampal pyramidal cells play important roles in the establishment and regulation of network connectivity, synaptic plasticity, and firing dynamics. Several laboratories routinely reconstruct CA3 and CA1 dendrites to correlate their three-dimensional structure with biophysical, electrophysiological, and anatomical observables. To integrate and assess the consistency of the quantitative data available to the scientific community, we exhaustively analyzed 143 completely reconstructed neurons intracellularly filled and digitized in five different laboratories from 10 experimental conditions. Thirty morphometric parameters, including the most common neuroanatomical measurements, were extracted from all neurons. A consistent fraction of parameters (11 of 30) was significantly different between CA3 and CA1 cells. A considerably large number of parameters was also found that discriminated among neurons within the same morphological class, but reconstructed in different laboratories. These interlaboratory differences (8 of 30 parameters) far outweighed the differences between experimental conditions within a single lab, such as aging or preparation method (at most two significant parameters). The set of morphometrics separating anatomical regions and that separating reconstructing laboratories were almost entirely nonoverlapping. CA3 and CA1 neurons could be distinguished by global quantities such as branch order and Sholl distance. Differences among laboratories were largely due to local variables such as branch diameter and local bifurcation angles. Only one parameter (a ratio of branch diameters) separated both morphological classes and reconstructing laboratories. Compartmental simulations of electrophysiological activity showed that both differences between anatomical classes and reconstructing laboratories could dramatically affect the firing rate of these neurons under different experimental conditions.

Action Potentials↗
PubMed1984

Morphometry of intracellularly labeled neurons of the auditory nerve: correlations with functional properties.

Single auditory-nerve fibers were injected with horseradish peroxidase after their tuning properties, characteristic frequencies, and spontaneous discharge rates were measured. From these functional properties virtually all other aspects of auditory-nerve response can be predicted. Labeled fibers were reconstructed from the point of peripheral termination on cochlear hair cells to the point at which they enter the cochlear nucleus. Several morphological properties were measured at the light-microscopic level, including axonal diameter, axonal length, internodal distances, cell-body area, and cell-body shape. All of these parameters were correlated, though some weakly, with characteristic frequency. However, only axonal diameter was correlated with spontaneous discharge rate.

Animals↗
PubMed1990

Synaptic morphometry and synapse-to-neuron ratios in the superior colliculus of albino rats.

The superior colliculus of mammals is generally divided into seven layers on the basis of the distribution of myelinated fibers, which are densely packed in layers III, V, and VII but sparse in the other layers. The laminar distribution of afferents and efferents allows, in addition, for the distinction of a superficial visual zone (layers I-III) and a deeper multimodal and premotor zone (layers IV-VII). Collicular neurons, however, do not show a lamination pattern, but are rather homogeneously distributed with only gradual transitions (Albers et al.: J. Comp. Neurol. 274:357-370, '88). The present study analyses whether the distribution of collicular synapses is correlated with the laminar organization of collicular axons or rather with the more homogeneous distribution of collicular neurons. For this purpose, the size and density of synaptic terminals and contacts as well as synapse-to-neuron ratios were determined in all collicular layers of albino rats by means of quantitative analysis of electron microscopic pictures. The size of presynaptic terminals and contacts does not differ significantly between individual collicular layers. On average, presynaptic terminal diameter is 1,079 nm, and synaptic contact size 338 nm, while 23% of all contacts are of the symmetrical type with pleiomorphic vesicles. The average numerical synaptic density is 422 million per mm3. This value is significantly higher in layers I and II (on average 670 million per mm3) than in layers III-VII (on average 370 million per mm3). The synapse-to-neuron (S/N) ratios calculated show that collicular neurons have on average 6,120 synaptic contacts on their receptive surface. The S/N ratio is lowest in layer III (4,330), while this ratio is highest in layers I and VII (i.e., 8,970 and 8,560 respectively). Layer II has a significantly higher S/N ratio than layer III (i.e., 8,060 and 4,330, respectively). Our results show that the size of synaptic terminals and contacts is not correlated with the different connectivity patterns of the distinct collicular layers. However, the density of synapses as well as the synapse-to-neuron ratios show a certain degree of laminar differentiation. In particular the superficial visual zone appears to be inhomogeneous in this respect, since layers I and II have a significantly higher density of synapses and higher S/N ratios than layer III. The deeper collicular zone is more homogeneously organized with synaptic densities similar to that of layer III and gradually increasing synapse-to-neuron ratios from layer IV to layer VII.

Animals↗
PubMed1990

Single fiber studies of ascending input to the cuneate nucleus of cats: I. Morphometry of primary afferent fibers.

The morphology of afferent fibers ascending to the cuneate nucleus has been examined in this and the subsequent paper in order to quantify the pattern of arborization and bouton arrangement of selected classes of primary afferents and to compare these data with data from postsynaptic fibers ascending to the cuneate nucleus. Electrophysiologically identified G hair and Ia muscle afferent fibers in the cuneate fasciculus were intraaxonally injected with horseradish peroxidase. Cutaneous afferents terminated dorsal to proprioceptive afferents, especially at middle levels of the cuneate nucleus. The spacing of collaterals along G hair fibers was variable, but averaged 1.46 collaterals per mm; collateral density was higher at middle cuneate levels than in the rest of the nucleus. Collateral density of Ia fibers was lower than for G hair fibers and was lowest at caudal levels of the nucleus. Branches of G hair collaterals, though often initially diverging, usually converged to terminate in a single focus in the dorsal part of the nucleus. The probability of bifurcation of Ia collaterals decreased steadily at successive branch points. These collaterals branched less symmetrically than G hair collaterals, and terminated in the ventral cuneate with less dense arbors, stretched mediolaterally, but of comparable cross-sectional area. Individual G hair collaterals gave rise to more boutons than Ia collaterals; in both cases they were mostly of the en passant type. Boutons were restricted to distal branches of G hair collaterals, whereas boutons of Ia collaterals were also located on proximal branches. Bouton size was similar for the two classes of collaterals. The data reported here, in combination with the published literature, suggest that the collaterals of roughly 300 G hair fibers overlap at any given point at middle levels of the cuneate nucleus. This high degree of anatomical convergence is not predicted by the functional segregation described with electrophysiological mapping, implying the presence of intrinsic nuclear mechanisms enhancing response specificity.

Afferent Pathways↗
PubMed1984

Classifying breast carcinomas with DNA measurements and morphometry.

The relation between the DNA distribution pattern and the variables DNA content and nuclear area was investigated in 73 breast carcinomas. According to the DNA patterns, the tumors were subclassified into groups I, II, III, and IV (Auer's classification). The Auer's classes correlate with prognosis. In this investigation it was shown that Auer's classes strongly correlate with the standard deviation of DNA content and mean nuclear area. The carcinomas were subclassified using data of DNA content and nuclear area. For the establishment of this subclassification, multivariate analysis was applied. It was evident that Auer's group III was a hybrid group. Therefore a second classification rule was calculated without the data of Auer's group III. This second classification rule was even more successful in determining the Auer's group of each individual case. Our investigation shows that it is possible to subclassify breast carcinomas objectively using DNA content and nuclear area of the tumor cells.

Breast Neoplasms↗
PubMed1991

Prognostic value of morphometry and DNA flow-cytometry features of invasive breast cancers detected by population screening: comparison with control group of hospital patients.

Using the prognostic value of morphometric and flow-cytometric features, a group of patients with invasive breast cancers detected with population screening (PS, n = 70) has been evaluated and compared with a random control group in 2 hospitals (H group, n = 225) diagnosed in the same period. The results show that the PS patients had smaller tumors, less positive lymph nodes, better differentiated tumors with a lower mitotic activity index (MAI) and lower values of the morphometric prognostic index (MPI). Furthermore, the women more frequently had diploid tumors and tumors with small nuclei. The second purpose was to evaluate whether quantitative microscopical features, in comparison with other prognostic features such as size of primary tumor, nodal status and histologic grade, are as strong prognosticators in PS tumors as in H-detected breast cancers. In comparison with H tumors, morphometric and flow-cytometric features, as well as tumor size, had the same prognostic value for the PS tumors. In contrast, nodal status was not significant within the PS group, and the same phenomenon was found in a subgroup of H patients with similar sized tumors. Of all quantitative microscopical features (MPI, MAI, mean nuclear area (MNA) and DNA Index (DI], the MAI had the strongest prognostic value. DI showed additional prognostic value to the MAI for patients with small tumors and with small tumor-cell nuclei, because a diploid pattern in these cases (this combination occurred in 21 patients of the total group = 30%) was correlated with a 95% 10-year survival rate. Histologic grade, although significant within the large H group, was of no prognostic value within the PS group, and also not as in the H sub-group with small tumors. It is concluded from morphometric and DNA flow-cytometric criteria that these prognostic features in invasive breast cancers detected by PS were all more favorable than in randomly detected hospital breast cancers. This may account for the reported better survival rate of PS patients. Furthermore, the prognosis of patients with small invasive breast cancers detected by population screening can be more accurately deduced by quantitative microscopical features than by axillary-lymph-node status.

Aged↗
PubMed1992

DNA flow cytometry, nuclear morphometry, mitotic indices and steroid receptors as independent prognostic factors in female breast cancer.

Clinical features, 8 histological variables, 7 nuclear morphometric variables, 2 mitotic indices, oestrogen-receptor (ER) and progesterone-receptor content (PR), DNA ploidy and S-phase fraction (SPF) were entered in a Cox's model to assess their independent predictive value in 216 breast-cancer patients followed up for over 9 years. In the whole series, histological type (p = 0.007), volume-corrected mitotic index (M/V index) (p = 0.01), axillary-lymph-node (pN) status (p = 0.024) and the year of treatment (p = 0.045) predicted independently the recurrence-free survival (RFS). In a sub-analysis including SPF (n = 148), the year of treatment (p = 0.003), tumour diameter (p = 0.004), SPF (p = 0.022) and nuclear pleomorphism (p = 0.056) independently predicted the RFS. In a Cox's analysis of the whole series, tumour diameter (p less than 0.001), pN status (p = 0.001), PR status (p = 0.002) and the year of treatment (p = 0.021) were independent predictors of survival. In a separate analysis including also SPF (n = 148), tumour diameter (p less than 0.001), SPF (p = 0.003), pN status (p = 0.008) and the year of treatment (p = 0.015) proved to be independent prognostic factors. The results show that tumour diameter, pN status, M/V-index, histological type, SPF and PR status comprise a sufficient combination of prognostic factors in female breast cancer. In pN patients, age and SDPE may be of additional prognostic significance. The prognostic scores combining the independent prognostic variables reflecting both the proliferative rate and metastatic potential of the tumours are accurate predictors of the RFS and overall survival.

Adult↗
PubMed1995

Different risk groups in node-negative breast cancer: prognostic value of cytophotometrically assessed DNA, morphometry and texture.

Feulgen-stained imprints and fine-needle aspirates from 528 lymph-node-negative breast cancers were investigated by means of an image analysis system. Several DNA, morphometric and textural parameters were evaluated for each patient. The prognostic value of the parameters was investigated by multivariate Cox regression analysis. As prognostic criteria, a distant recurrence-free survival of 5 years and an overall survival of 8 years were considered. In multivariate analyses the anisokaryosis (standard deviation of nuclear radius, Rad-SD) was the strongest parameter in predicting the clinical course of node-negative patients. This was followed by a textural parameter (run-length, NR2 M) and the tumor size (pT). The DNA histogram type could also add prognostic information concerning distant recurrence-free survival, but not overall survival. In both approaches a multivariate prognostic factor was calculated for each of the node-negative patients by a linear combination of the selected variables. Using this factor, patients could be split into 5 subgroups with significantly different risks of distant metastases. Thus, a low-risk subgroup, with a 5-year distant recurrence rate of only 3%, and a subgroup with a considerably higher risk and a distant recurrence rate of 35%, could be distinguished. In survival analysis the low-risk group of node-negative patients showed an 8-year death rate of only 3%, whereas in the high-risk group 30% of the patients had died at 8 years. Thus DNA, morphometric and textural parameters can provide powerful prognostic information in node-negative breast carcinomas. The multivariate combination of the relevant variables may allow a better selection of those node-negative patients with a proven good prognosis, and of those who are at risk of distant recurrence and therefore may benefit from adjuvant treatment.

Adult↗
PubMed1981

Ultrastructure and morphometry of the alveolar lung of guinea pigs chronically exposed to diesel engine exhaust: six month's experience.

The impact of chronic inhalation of diesel exhaust (DE) on alveolar lung was studied in 24 Hartley guinea pigs. Groups were sacrificed sequentially at 2 weeks, 3 months and 6 months of exposure to either 750 micrograms or 1500 micrograms DE particles (DEP) per m3 along with age-matched concurrent controls. Although qualitative ultrastructural changes were noticed during this time interval and dosage schedule, there was no evidence of pathologic changes such as fibrosis or emphysema. The cellular uptake of DEP was striking. By 2 weeks three alveolar cell types (alveolar macrophages, epithelial type 1 cells and interstitial macrophages) plus one type of granulocytic leukocyte (eosinophils) confined DEP within phagosomes without evidence of cytotoxicity. A certain phagosomal DEP population had a bull's eye appearance and diameters of 0.0727 +/- 0.01 micron. Morphometric analysis applied to electron micrographs demonstrated that arithmetic mean tissue thickness of the air-blood barrier was generally increased (p less than 0.05) during DE exposure. For the 750 micrograms DE sets, the increase over control (1.56 micron) was 41% at 2 weeks, 46% at 3 months and 77% at 6 months while the 1500 micrograms DE set at 6 months exceeded control by 130%. Increases in absolute tissue volumes of interstitium and epithelial type 2 cells largely accounted for the increased tissue thickness. Harmonic mean tissue thickness for controls remained near 0.537 +/- 0.03 micron for the study interval, contrasting with values for 3 and 6 months 750 micrograms DE and 6 months 1500 micrograms DE sets which increased. However, the diffusion capacity of the lung determined morphometrically was not decreased in DE exposed sets. Although cellular uptake of DEP and increased prominence of secretory epithelium were dose/duration related, absence of linearity suggests adaptative responses.

Aerosols↗
PubMed1992

Flow cytometry, morphometry and histopathology as biomarkers of benzo[a]pyrene exposure in brown bullheads (Ameiurus nebulosus).

Brown bullheads were given a single intraperitoneal dose of 0, 5, 25 or 125 mg kg-1 benzo[a]pyrene (BaP), a carcinogenic polycyclic aromatic hydrocarbon, and evaluated over 18 months. Flow cytometric analyses of hepatocyte DNA content indicated an increase in DNA synthesis in BaP-exposed fish prior to day 14 post-exposure. Thereafter, all flow cytometric variables returned to initial levels. Histopathological evaluation of livers from fish sampled at 18 months revealed significant differences among treatments in the amount of hepatic macrophage ceroid pigmentation and basophilic staining intensity. No neoplasms or changes in blood cell DNA content were detected. Significant morphometric variations existed among fish, but differences between sexes overshadowed differences attributable to dose. Flow cytometry yielded no evidence of long-term DNA alterations from a single exposure to BaP; however, the differences detected by DNA analysis shortly after the toxic event suggest that flow cytometric cell cycle analysis may be useful for documenting continuing exposures.

Animals↗