Increase in calcitonin gene related peptide (CGRP) and decrease in mast cells in dihydroepiandrosterone (DHEA)-induced polycystic rat ovaries.

UNLABELLED: The polycystic ovary is reported to correspond with a high density in intraovarian nerve fibers and their sympathetic hyperresponsiveness. Peptidergic nerves may also be involved in this process. An interaction between nerve fibers and mast cells is assumed because of nerve growth-factor production by mast cells. Here we investigated CGRP-positive nerve fibers and mast cells in polycystic ovaries induced in immature rats with dihydroepiandrosterone (DHEA). The DHEA treated ovaries contained less corpora lutea than controls (mean +/- SEM: 4.3 +/- 0.6 versus 11.3 +/- 0.9, P > 0.001) and less intact antral follicles (4.7 +/- 0.7 versus 8.1 +/- 1.1; P < 0.05) according to the histometric approach. By immunolabelling more CGRP-positive nerve fibers were found in the DHEA treated ovaries than in controls (mean +/- SEM per one section: 23.2 +/- 5.8 fibers versus 10.3 +/- 0.9 and 171 +/- 44.7 varicosities versus 84 +/- 9.5). This was confirmed by dot blot analysis, showing a significant higher CGRP signal intensity per microgram homogenized ovaries of the DHEA treated group compared to the untreated (P < 0.05). Toluidine-blue-stained mast cells populated the medulla in both groups, yet had strikingly decreased in the DHEA treated ovaries (23.5 +/- 3.9 versus 89 +/- 5.6, P < 0.005). CONCLUSION: The increase in CGRP-positive nerve fibers and the decrease of toluidine-blue-stained mast cells points to an altered neuroimmune function in DHEA-induced polycystic rat ovaries.

[Sex differences in morphologic aging processes]. / Geschlechtsunterschiede morphologischer Alternsprozesse.

According to Max Bürger (2) it is justified to speak of sexual divergence of biomorphosis. Morphometrical investigations have proved the existence of such sexual differences. The following results were obtained: The first example concerns the increase of fatty tissue in parathyroid glands which we measured on histological slides of 135 human samples by means of the point counting method. The fatty tissue of males was shown to increase continuously with statistic significance, whereas this process stops in females during their generative phase between the 2nd and 5th decade. Evidence for the sexual divergence was also obtained by investigating the fatty tissue between the bundles of muscle fibers of the tongue, however, without being able to statistically verify these results. We found the same sexual differences by measuring the muscle cell regression and connective tissue proliferation in the ciliary muscle of human eyes and tunica muscularis of the small intestine as well. All findings prove that the female is able to stay biologically younger during her generative activity compared to males. After menopause the aging processes speed up in female tissues without making up for the "age lead" of males. It is also worth mentioning that macroscopic anthropometric features do confirm the same fact, e.g., length and breadth of human auricles, ranging in age from 18 to 51 years.

[Genesis and significance of so-called asphyxial infiltrates in the placenta. I. Histologic results]. / Zur Genese und Bedeutung der sogenannten Asphyxieinfiltrate der Plazenta. I. Histologische Befunde.

The conventional conception of the cause and formal pathogenesis of so-called "asphyxial infiltrates" of the placenta is presented. Morphological examples are given. Our study included 561 newborn cases. Eighty two cases (14.6%) had such cell infiltrates, and there were 6 cases with phlegmoneous inflammation of the fetal membranes. These morphological findings correlated with the following clinical characteristics: term of birth, weight classes of newborns, Clifford-signs, clinical risks with respect to hypertension, the course of pregnancy and particularly the risk of inflammation. Additional correlates were the clinical statement of hypoxia and acidosis in newborns. We discovered relationships between these cell infiltrates and the Clifford-signs, and to some clinical risks (especially those for inflammation), but not to hypoxia and acidosis of the newborn. Our results show, that there are no relations between the conventional "Asphyxial infiltrates" of the placenta and the hypoxia and acidosis observed in newborns.

Anatomic basis of micturition and urinary continence. Muscle systems in urinary bladder neck during ageing.

Dorschner et al have described the unique function and form of several different muscle systems of the urinary bladder neck. If these systems have different functional responsibilities, then the muscles must undergo different ageing processes, as stated in the theory of function-dependent ageing. One characteristic of histologic ageing is the change over time in the proportion of muscle cells to connective tissue, a phenomenon we have demonstrated in both the ciliary muscle and in the two muscle systems of the small intestine. Using an SIS-Image Analysing System, we have now measured automatically the ratios of muscle cells to connective tissue in sections from several regions of the urinary bladder neck, taken from 50 male and 15 female cadavers. Our results confirm new functional explanations of the different muscle systems in the bladder neck. The relative volume of muscle cells in both the sphincter trigonalis m. and the dilator urethrae m. diminishes continuously with age. In the ejaculatorius m., however, the volume of muscle cells first increases until beginning at the end of the third decade, it decreases until senescence. As was presumed, the proportion of muscle cells in the detrusor vesicae m. does not decline during the later decades. The volume of muscle cells and fibers in both urethral sphincter muscles, however, decreases with age, beginning in early childhood.

[Age-related changes in systematic inhomogeneities of articular cartilage]. / Alternsveränderungen systematischer Inhomogenitäten des Gelenkknorpels.

We have considered the question of the quantification of differentiation processes. We investigated articular cartilage of the Caput femoris in 22 cases aged from 0 to 77 years. We measured cell size and volume density. During aging a shifting of the layer with the maximum cell size in relation to the joint surface as well as the formation of two layers with different volume densities take place. We believe that the changes in nutritional and load conditions during life play a role in these processes. We describe these with the term "diversification", characterizing the process of the changing of the degree of structural order.

[Quantitative analysis of a differentiation characteristic of hyaline cartilage in the aging process]. / Quantitative Analyse eines Differenzierungsmerkmals des hyalinen Knorpels im Alternsgang.

Changes in the distribution patterns of chondrocytes in the permanent hyaline cartilage were studied in order to analyze a process of differentiation quantitatively. Sections of 29 specimens of arytenoid cartilage and 42 specimens of Carina tracheae from newborn up to the age of 91 years were used. The chondrocytes were transferred via a mirror to paper, and the resulting patterns were analyzed. Various distribution functions were calculated. The most important one is the pair correlation function. The behavior of the differentiation is resembling in both organs. Especially in arytenoid cartilage there were found the following results: The clearest process of differentiation took place up to the 3rd decade of life. Initial hard core distribution is followed by soft core distribution up to the final of puberty. After the 2nd decade of life clustering was found. The grade of differentiation decreases in the 5th decade of life and later.

[Behavior of adiposity indices in relation to body weight and height between the ages of 3 and 17 years]. / Comportamiento de cuatro índices ponderales con relación al peso y a la altura desde los tres a los diecisiete años.

We studied the coefficients of correlation between weight and height and four adiposity indices (weight/height 0,5; weight/height; weight/height2; weight/height3), year to year, in 1676 schoolchildren of both sexes (663 boys and 1,013 girls), aged between 3 and 17 years. Weight-for-height index should be uncorrelated with height and highly correlated with weight. In our study, Quetelet index (w/h2) in females and Rohrer index (w/h3) in males, are independent from height. Between two indices, Quetelet index had better correlations with weight.

[Quantitative analysis of cell density and pattern of chondrocytes in transitory hyaline cartilage anlagen of human tarsal bones]. / Quantitative Analyse der Knorpelzelldichte und des Chondrozytenmusters im transitorischen hyalinen Knorpel der embryonalen Anlagen menschlicher Fusswurzelknochen.

Changes in the distribution patterns of chondrocytes in the embryonic transitory hyaline cartilage of the tarsus were studied using 56 sagittal section of tarsi from 28 human embryos, with a vertex-foot length from 7.0 to 44.0 cm. The os naviculare and os cuboideum were not sliced in all sections. A total of 93,931 cells were transferred via a mirror to paper, and the resulting patterns were analyzed. Determinations included the numerical surface and volume density of the chondrocytes and the cell distribution patterns using the dispersion index. In the case of the calcaneus, talus, os naviculare and os cuboideum, the number of sliced cells found over a constant area decreases at a continuous linear rate in the course of the embryonic period. In all tarsal primordia the distribution pattern developed from regular to random. Particular conditions prevail immediately below the perichondrium and near the ossification centers which can be found in the calcaneus from the 6th month onwards.

[Morphological studies of postnatal changes of the sound transport apparatus in the middle ear of humans and rabbits]. / Morphologische Untersuchungen über postnatale Veränderungen des Schallantransportapparates im Mittelohr bei Mensch und Kaninchen.

The dimensions of the tympanic membrane and middle ear ossicles, respectively, were measured, in order to investigate the postnatal development of sound transport function. In human we observed only a small increase of the tympanic membrane area between the 7th postconceptional month and the 32nd year of life. This growth has no functional effect on the sound transport. In contrast, in rabbits all measured parameters with the exception of the stapes plate area show an increase until the 12th day of life. Thus, the amplification factor doubles in this species during the first 2 weeks of life. The temporal parallelism of the middle ear development with the ontogenesis of the acoustic function seems to exist in humans, but does not in rabbits.

[Morphometric proof of function-related aging]. / Morphometrische Beweise für funktionsabhängiges Altern.

In a new book of W. Frolkis and co-workers there is emphasized the dependence of aging processes on the cell functions. Besides always demonstrable universal mechanisms each kind of cell shows its specific aging in dependence on its function. This is the case in different tissues and organs too. We show that in muscle tissues, in the comparison of the kidney with the submaxillary gland and in the appearance of collagen type I and III in the anterior and middle part of the human vocal cord.

[Random sample planning in morphometry]. / Stichprobenplanung in der Morphometrie.

This report tries to explain principles and presuppositions of planning sample sizes. The aim is estimation of the optimal, that means minimally necessary sample size, justifying an investigation ethically and economically. The problems are complicated because of the necessity of nested sampling or samples within samples. We describe the most important presuppositions using an example of mean value estimation. These are: establishing of demands for exactness, of error probability, and knowledge of variance of the parameters. It is explained in detail by means of binomially distributed variables as we find them in the point counting method.

[Quantitative analysis of chondrocyte distribution patterns in hyaline cartilage tissue--a comparison of different models]. / Quantitative Analyse von Chondrocytenverteilungsmustern im hyalinen Knorpelgewebe--Vergleich verschiedener Modelle.

The purpose of our investigations were aging changes in hyaline cartilage of the trachea and of the larynx. 42 samples of carina tracheae and 29 samples of arytenoid cartilage were used in both organs from the newborn age up to the age of 91 a. We have examined patterns of cell distribution with the help of the computer programme "Dichte" ("density") using the following methods of stochastic geometry: product density, L function, and others. Product density allows for a reliable destination between a regular pattern of cells (hard-core distribution), a random pattern (soft-core distribution), and a typical clustering of chondrocytes. 2 examples of arytenoid cartilage have been selected to demonstrate the possibilities of interpretation of product density supported by the L function. Soft-core and in few cases hard-core distribution have been found both with the carina tracheae and the cartilago arytaenoidea in the 1st decade of life. Beginning with the 2nd decade of life, the typical clustering of chondrocytes have been confirmed with both organs.

[Age changes of functional nuclear swelling and classes of nuclei in human hepatocytes]. / Zu Alternsveränderungen der funktionellen Kernschwellung und der Kernklassen menschlicher Hepatozyten.

The aim of the study was to describe aging changes in the functional nuclear edema and the classification of human hepatocyte nuclei, by determining three parameters--the size of the nucleus, the relative DNA quantity and the number of chromocenters. For this purpose, karyometry and DNA cytophotometry were performed on 10 human liver preparations. The data obtained was subjected to correlation, cluster and discriminance analysis. The results indicated a reduction in the capacity of liver cells for functional nuclear edema as aging progressed. Whereas at a young age there is only a loose correlation between nuclear size and DNA content, it becomes much stricter later on, partly caused by polyploidization. Cluster analysis, followed by discriminance analysis, is well suited for dividing the nuclei of human hepatocytes into two or three statistical populations provided the nuclear area, DNA quantity and number of chromocenters are all used as characteristics simultaneously. When allowance is made for functional edema, the biological interpretation of clusters from young liver preparations permits meaningful conclusions, but it appears problematic for old preparations. Here it might be more practical to analyze the mixed distributions resulting from a determination of the DNA quantity or nuclear size alone.

[Comparison of three histometric methods for the comprehension of stimulating effects on the rat thyroid gland]. / Vergleich von drei histometrischen Methoden zur Erfassung stimulierender Effekte auf die Rattenschilddrüse.

There exist some histometric methods for the morphological quantification of different strongly stimulating effects on the thyroid gland induced by drugs and/or other chemical substances in dependence upon dose and duration of application. But in respect of technical and temporal expense and also diagnostic statement, there are considerable differences between these recording procedures. Therefore we examined the 3 mostly used methods synchronously (i.e. determination of thyroid epithelial cell height, nuclear volume in thyrocytes, and estimation of relative volume parts in the thyroid gland by the point counting method) by investigating the thyroid glands of methylthiouracil-(MTU)-stimulated rats and corresponding controls in order to compare the diagnostic value and temporal expense. The largest temporal expense was required in the nuclear volume determination, the smallest in the point-counting method. On principle, all 3 procedures allow the determination of hypertrophic alterations but only by help of the point-counting method, also hyperplastic changes are recognizable. By nuclear volume determination, we found significant differences between central and peripheral parts of the thyroid gland. Therefore, to avoid the subjective error, it will be necessary to measure a large number of nuclei in many planes of the gland. Also the determination of epithelial cell high reinforces the subjective error because of the heterological structure especially in unstimulated thyroid gland. If the number of counting points is exactly determined and, full of sense, limited, the point-counting method allows a nearly complete measuring of the whole object to be tested within an acceptable investigation time. In this way, the heterological structure of thyroid gland will be regarded, and comparability and reproducibility are guaranteed on an high level.

[Quantitative detection of the differentiation processes of cartilage tissue]. / Zur quantitativen Erfassung eines Differenzierungsprozesses im Knorpelgewebe.

The onset of ageing during childhood and adolescence is accompanied by a variety of differentiation processes, whereas ageing later in life is characterized by dysdifferentiation. One of the subjects in the present discussion is the "Dysdifferentiation hypothesis of ageing". So far differentiation has been described only in qualitative terms. The paper uses the example of hyaline cartilage development (chondrogenesis) to show that a quantitative approach to differentiation is possible, which at the same time enables random-critical study. The method used for this is stochastic geometry.

[Biostatistical models in experimental gerontology]. / Biostatistische Modelle in der experimentellen Gerontologie.

Morphometric measuring values present themselves in a definite distribution which changes in the ageing process. This change of distribution can be clarified by distribution analyses. Another task within the analysis of frequency distributions is the decomposition of mixed distributions, as they may appear in various parameters in the course of life. The linear regression analysis is, for instance, to be used in the verification of sexual divergencies in the process of ageing. An adaptation of non-linear exponential functions to series of measuring values serves, among others, to make evidence concerning the ageing rapidity of the tissues. When in gerontological inquiries at the same time many different characteristics are determined also the degree of their ageing dependency is of interest. This is to be established by means of a special transformation within the factor analysis. Finally the study shows instances for the application of contingency tables in qualitative characteristics and the investigation of their frequency in the process of ageing as well as for the use of the cluster analysis.

[Discussion of errors and measuring strategies in morphometry using analysis of variance]. / Diskussion von Fehlern und Messstrategien in der Morphometrie mittels Varianzanalyse.

Statistical techniques known as the analysis of variance make it possible for the morphologist to plan work in such a way as to get quantitative data with the greatest possible economy of effort. This paper explains how to decide how many measurements to make per micrograph, how many micrographs per tissue block or organ, and how many organs or individuals are necessary for getting an exactness of sufficient quality of the results. The examples furnished have been taken from measuring volume densities of mitochondria in heart muscle cells and from cell counting in lymph nodes. Finally we show, how to determine sample sizes, if we are interested in demonstration of significant differences between mean values.