Cellular automaton simulation of tumour growth -- equivocal relationships between simulation parameters and morphologic pattern features.

OBJECTIVE: To develop an interpretation procedure which estimates simulation parameters (tumour cell motility, tumour cell adhesion, autocrine and paracrine growth control, stroma destruction) of simulated patterns solely based on morphometric features of the morphologic pattern. METHODS: A cellular automaton computer simulation program was developed which produces morphologic patterns by growth of a seed of tumour cells. At the beginning of each simulation run certain simulation parameters are assigned to the tumour cells. After the run has been completed, the resulting pattern is evaluated by a set of morphometric features. Simulation parameters and resulting morphometric features of 27,800 simulations were stored in a database and were used for the evaluation of potential relationships. RESULTS: Correlation analysis showed highly significant correlations between morphometric features on the one hand and the preset simulation parameters (tumour cell motility, tumour cell adhesion, autocrine and paracrine growth control, stroma destruction) on the other. Correlation coefficients, however, varied from 0.72 to 0.99. When only one simulation parameter varied while all others were kept constant, morphometric features yielded a highly reliable estimate of the particular simulation parameter. When variability was extended to 4 simulation parameters, morphometric features were less effective in estimating the setting of the parameters. Though in all patterns tested several possible simulation parameter constellations could be ruled out, morphometric features were usually compatible with more than one set of simulation parameters thus preventing a straightforward interpretation. CONCLUSIONS: Though simulation parameters significantly and reproducibly influence the resulting morphologic pattern as characterized by morphometric features, estimates of the simulation parameters based on morphometric features yield equivocal results.     

VARIABILITY OF NEOPLASTIC PARAMETERS IN COLON TUMOURS, AND ITS SIGNIFICANCE IN DIAGNOSTIC PRACTICE

We have reviewed the value of individual variability in the reaction of tissues to treatment with carcinogens, and the manifestation of this variability in different morphological (histological, morphometric, and ultrastructural), histochemical and immunohistochemical parameters generated in tumorous tissues. Moreover, we also reviewed data in the literature on individual variability in the manifestation of some biochemical and immunochemical markers which are accumulated in the serum of tumour-bearing patients and which are characteristic for the different phases of tumourigenesis. The high variability of different tumorous parameters suggests that none can be utilized alone as a conclusive marker of neoplasia and that only their combined use can give objective information. We also reviewed the impact of this variability in the evaluation of various pathological methods to detect different stages of neoplastic transformation in the colon. It has been concluded that none of the present approaches can be absolutely conclusive and without false results, and that objective information regarding early cancerous changes may be obtained only by use of combined analyses utilizing several laboratory methods. The diagnostic procedures should be based on the complex utilization of all appropriate methods using the quantitative interpretation of the obtained data. Multivariate analysis of many parameters should be very effective for the prediction of therapeutic results.     

Analysis of morphometric indices in mice for registration of induced mutations in germ cells

The new-born mice of the first generation have been examined for changes in morphometric indices after the influence of fopurine, cyclophosphamide and cadmium chloride on their fathers. It is shown that fopurine and cyclophosphamide, the mutagens for mammals, increase the variability of morphometric indices. The method to calculate the morphometric indices of mice may be used for evaluating genetic risk of the compounds for man.     

Stereologic correlates of steroid receptor concentration in invasive ductal breast cancer

The hypothesis was tested that morphometric parameters of tumor cell nuclei correlate with the steroid receptor concentration in mammary carcinoma. In 50 consecutive mastectomy specimens with a diagnosis of invasive ductal cancer in which estrogen receptor (ER) and progesterone receptor (PR) concentrations had been assayed quantitatively, morphometric measurements were performed on four visual fields of two sections per case. The fields were sampled from the most cellular regions of the tumor. The number of tumor cell nuclear profiles per tissue area, the nuclear profile area and the long and short nuclear profile axes and their ratios were measured with a semiautomatic image analysis system. Estimates of the number of tumor cell nuclei per tissue volume (Nv) and of the mean tumor cell nuclear volume (V) were obtained by standard stereologic techniques. Association between the morphometric and biochemical parameters was tested by Spearman's rank correlation coefficient. Nv correlated positively with the steroid receptor concentration whereas V correlated negatively with both ER and PR concentrations. A correlation of the receptor concentrations to the standard deviation of the nuclear area or the mean ratio of the nuclear axes could not be demonstrated. These results suggest that receptor-rich tumors have a large number of small tumor cell nuclei whereas receptor-poor tumors have a small number of large tumor cell nuclei per tissue volume in the actively proliferating, highly cellular regions. These differences are not accompanied by significant changes in nuclear size variability or nuclear shape.     

Morphometry of the Stone Loach, Barbatula Barbatula: Do Mensural Characters Reflect the Species' Life History Thresholds?

Growth variability in 23 mensural characters was examined in 387 specimens of stone loach, Barbatula barbatula, from England. The standard length (SL) of the specimens ranged from 15.3 to 115.4 mm. We tested the hypothesis that body proportions change abruptly, rather than gradually, at certain intervals of ontogeny by fitting linear, quadratic and split linear curves to plots of each variable against SL. Based on patterns of allometric growth, two groups and two subgroups of mensural characters have been found. Three characters were best explained by a linear regression, indicating isometric growth. Eight characters were best explained by a quadratic curve, indicating gradual allometry. The remaining 12 characters were best explained by a split regression, indicating mainly isometric growth with abrupt allometry occurring at a specific SL (breakpoint). The first shift in morphometric values (a transformation of the head; breakpoints in three characters) occurred at 26-35 mm SL, the second (a change in fin shape and size as well as body form; breakpoints in six characters) at 36-47 mm SL. The coincidence of shifts in body morphology with those in microhabitat use (between the respective size classes) suggests that thresholds (though not as sudden as those between embryo and early larva steps) do occur during this interval of stone loach life history. We suggest that the larva period ends with the completion of the first shift in relative growth (i.e. not later than at 35 mm SL, depending on individual variability), and that the second shift in morphometric values reflects a threshold between the first and the second step of juvenile period. The importance of changes in external morphology decreased as the fish grew bigger and older.     

Morphometric analysis of populations of Centris aenea Lepeletier (Hymenoptera: Apidae) from Northeastern Brazil

Centris aenea Lepeletier is a solitary bee that has raised interest in management to pollinate crops, such as acerola, Malpighia emarginata. This study investigated the level of morphometric variability among populations of C. aenea from Northeastern Brazil. Traditional and geometric morphometric analyses were used. Head length, leg length, wing length, and wing shape were measured in samples (5-10 females) from eight localities. We did not find statistically significant differences among the populations (P > 0.01). The partial wing warps were similar in the populations and indicated that the bees were not morphometrically different. Our results suggest that C. aenea shows low population morphometric variability and highlight the need for further investigations on population variation in this species, preferably including populations sampled at the extremes of their geographic distribution. Significant insight into the population variation of C. aenea will probably require the use of molecular markers to allow a comparative approach between morphometric variability and genetic variability.     

Effects of food variability on growth rates, fledging sizes and reproductive success in the Yellow-eyed Penguin Megadyptes antipodes

Effects of a change of diet on growth rates and fledging sizes of Yellow-eyed Penguins Megadyptes antipodes were examined at two breeding areas on South Island, New Zealand, during two breeding seasons. An adverse change in diet was observed in the second season. Evidence for this included depressed growth rates of weight, differential growth of weight and most morphometric parameters between one- and two-chick nests in the second season, lower fledging weights, lower adult body weights, delayed moult, higher chick mortality and higher adult mortality during moult. The change in diet is suggested as being from one including oil-rich prey species, to one of oil-poor species.
Growth rates of first- and second-hatched chicks, and of survivors and non-survivors within a brood were not significantly different in either season, and growth rates of two-chick broods were only slightly slower than one-chick broods for some parameters in the second season. This, and synchronous hatching of chicks, equal egg-size and lack of sibling competition during feeding sessions, suggests that brood reduction is not an option available to Yellow-eyed Penguins, and that food supply may not be a limiting factor in the majority of breeding seasons.
Few changes in growth rates of morphometric parameters at either breeding area, and similar absolute sizes at fledging, indicate that slowing of growth rates of morphometric parameters only occurs when feeding conditions are so bad as to result in mortality and that, although fledging periods may be longer, patterns of development remain essentially unchanged.     

Morphometric and hormonal changes during the chimpanzee menstrual cycle

BACKGROUND: Sex steroids affect many peripheral tissue sites in female mammals. Receptors for these hormones have been found in skin, fat, and bone. In women, these tissues can show morphological changes during the menstrual cycle that may be directly related to steroid secretion. METHODS: The present study was done on chimpanzees to document morphometric markers associated with these tissues (anogenital swelling volume, skin fold thickness as indicator of subcutaneous fat, bony diameters of mandible, wrist, and elbow) and to compare them with cyclic patterns of estradiol, progesterone, testosterone, gonadotropins, and prolactin. RESULTS: Swelling volume changed significantly over the menstrual cycle. All other morphometric parameters showed variation without statistical significance. Skin folds were thickest during the luteal phase. Bony diameters displayed similar but less distinctive changes. Testosterone correlated positively with diameter sites, inversely with subcutaneous fat. No relationships with either estradiol or progesterone were found. We assume that subcutaneous fat and morphometric bone parameters exhibit cycle-dependent changes that may be caused by changes in steroid secretion.     

Effects of an induced magnetic storm on the early ontogenesis of roach <Emphasis Type="Italic">Rutilus rutilus</Emphasis> (L.)

The influence that an induced magnetic storm (IMS) modeled in a confined volume has on the early ontogenesis of Rutilus rutilus was studied. The significant effects of the IMS were observed for the hatching rate of roach embryos, the size-weight and morphometric parameters of the fry, the variability of the vertebral column, and ontogeny stability in the roach offspring.     

Endogenous cytokinin levels and metabolism of zeatin riboside in genetic tumour tissues and non-tumourous tissues of tobacco

The cytokinin content of stem tissues, primary genetic tumours (excised from 2-month-old plants) and 3-week-old in vitro cultured genetic tumour tissues derived from Nicotiana glauca (Grah.) × langsdorffii (Weinm.) and N. suaveolens (Lehm.) × langsdorffii (Weinm.) hybrids and stem tissues derived from 2-month-old N. suaveolens and N. langsdorffii plants has been analysed by radioimmunoassay. Stem tissues of tumour-prone hybrids contain high cytokinin levels (3–3.7 nmol g⁻¹). This increase is caused mainly by increased levels of cytokinin nucleotides, particularly those of zeatin nucleotide (0.5 nmol g⁻¹) in stem tissues of parent plants and 2.4 nmol g⁻¹ in stem tissues of hybrids). All other tissues contain lower cytokinin levels (0.7–1.7 nmol g⁻¹). Cytokinin bases and ribosides are major compounds in cultured tumour tissues while the nucleotides are dominant cytokinins in all freshly excised tissues from parent plants and their hybrids. In a separate study, the metabolic fate of supplied [³Hj-zeatin riboside. which is inactivated mainly by sidechain cleavage, has been studied. The results collectively suggest that cytokinins may be involved in tumourigenesis.     

Sperm motility parameters and spermatozoa morphometric characterization in marine species: A study of swimmer and sessile species

The biodiversity of marine ecosystems is diverse and a high number of species coexist side by side. However, despite the fact that most of these species share a common fertilization strategy, a high variability in terms of the size, shape, and motion of spermatozoa can be found. In this study, we have analyzed both the sperm motion parameters and the spermatozoa morphometric features of two swimmer (pufferfish and European eel) and two sessile (sea urchin and ascidian) marine species. The most important differences in the sperm motion parameters were registered in the swimming period. Sessile species sperm displayed notably higher values than swimmer species sperm. In addition, the sperm motilities and velocities of the swimmer species decreased sharply once the sperm was activated, whereas the sessile species were able to maintain their initial values for a long time. These results are linked directly to the species-specific lifestyles. Although sessile organisms, which show limited or no movement, need sperm with a capacity to swim for long distances to find the oocytes, swimmer organisms can move toward the female and release gametes near it, and therefore the spermatozoa does not need to swim for such a long time. At the same time, sperm morphology is related to sperm motion parameters, and in this study an in-depth morphometric analysis of ascidian, sea urchin, and pufferfish spermatozoa, using computer-assisted sperm analysis software, has been carried out for the first time. A huge variability in shapes, sizes, and structures of the studied species was found using electron microscopy.     

Characterization of ram (Ovis aries) sperm head morphometry using the Sperm-Class Analyzer

Sperm morphology has been identified as a characteristic that can be useful in the prediction of fertilizing capacity. The aim of the current study was to characterize ram sperm heads morphometrically as a basis for future studies on the relationship between sperm quality and male fertility. For this purpose, ejaculates from 241 mature rams (Ovis aries) belonging to 36 different dairy herds were used to evaluate sperm head morphometry by means of the Sperm-Class Analyzer. Sperm samples, collected by artificial vagina, were diluted in phosphate buffered saline (PBS) for the analysis. A microscope slide was prepared from single-diluted fresh sperm samples. Slides were air-dried and stained with Hemacolor. A minimum of 115 sperm heads were analyzed from each male. Each sperm head was measured for four primary parameters (area, perimeter, length, width), and four derived parameters of head shape were obtained. Significant differences in sperm head morphometry were found between rams (CV for morphometric parameters ranging from 0.9 to 10.1), and there were marked differences in the sperm morphometric composition of the ejaculates. For all parameters, within-animal CVs were greater than between-animal CVs. Within-animal CVs ranged from 4.2 to 10.6, showing the high degree of sperm polymorphism present in the sheep ejaculate. Significant differences in sperm head morphometry were found between rams belonging to the different herds (i.e., origin). An important part of the variability observed on morphometric parameters was due to the male itself, with an explained variance ranging from 3.6% for regularity to 34.0% for p2a (perimeter²/[4 × π × area]). The explained variance by the herd of origin of the males ranged from 0.6% for regularity to 10.8% for area. Our results suggest that a genetic component might be responsible for the observed sperm head morphometry differences between herds.     

Altricial and precocial developmental features in modern meat-type chicks

Variability of egg weight, egg yolk content, neonatal growth rate and relationships of these parameters were studied in meat-type chicks. As it had been established the level of variability in neonatal growth traits was greater than variability of the egg morphology parameters. Egg weight had stronger influence on the chicks' neonatal growth rate than egg yolk content did. Low egg size was associated with limited neonatal growth rate variability, declined chick weight at hatching and increased relative growth rate throughout four days post hatch. Comparison of egg morphological parameters in two species having the same female definitive body weight--meat-type domestic fowl (precocial type) and brown pelican (altricial type) has shown, that, in contrary to predicted on the basis of avian developmental typology, egg weight to female body ratio was greater in brown pelican, egg yolk content was equal in both species.     

Morphometric and allozyme variation in central and southern Greek populations ofMicrotus (Terricola) thomasi

The endemic Balkan vole taxonMicrotus (Terricola) thomasi (Barrett-Hamilton, 1903) exhibits great karyological variability in Greece. In this study, populations belonging to two different karyotypic forms (‘atticus’ and ‘thomasi’) are examined both morphometrically and electrophoretically. A total of 140 individuals ofM. (T.) thomasi were collected from 6 localities of south and central Greece. For the morphometric analysis, 27 variables (external body and cranial characters) were examined and evaluated according to multivariate analyses (PCA, MANOVA, CVA and CLUS). For the electrophoretic analysis, 18 putative genetic loci were examined and the allozymic data were treated by the biostatistical package BIOSYS-1. According to the results obtained, all the populations studied show little overall morphometric variability, whereas they are characterized by high electrophoretic variability. The populations studied are not grouped according to the karyotypic form. In almost all the cases, in the two UPGMA-dendrograms plotted according to morphometric and electrophoretic distances (Mahalanobis’ and Nei’s distances, respectively), the populations branching together belong to different karyotypic forms. Conclusively, the morphometric and electrophoretic results of this study revealed that the two karyotypic forms should not be considered separate species or subspecies, as it has been proposed by some authors in the past, and the populations studied can be considered only as different local populations of the rather variable vole speciesM. (T.) thomasi.     

Factors influencing macrophyte metrics in Estonian coastal lakes in the light of ecological status assessment

Macrophytes and factors affecting their distribution were studied in 19 coastal lakes of Estonia. The aim of the study was to determine the factors influencing the distribution of macrophytes in coastal lakes and to assess the suitability of valid macrophyte metrics. Our hypothesis was that in coastal lakes most of the macrophyte distribution patterns are caused by lake-specific variables. Morphological, physico-chemical and catchment area characteristics of the lakes varied greatly. Lakes were in different development stages—lakes nearest to the Baltic Sea were younger and more influenced by brackish water and the furthest lakes were older with more freshwater. All that variability was reflected in macrophyte parameters. Factor analysis of environmental indices divided them into three groups—catchment area, morphometric and water chemistry factors. The first factor may be considered as a pressure and the other two as lake-type-specific factors. Lake catchment area parameters had an influence on Bolboschoenus maritimus, Chara tomentosa and Typha latifolia abundance. Morphometric parameters had an influence on the depth distribution of macrophytes and water chemistry factors on the abundance of helophytes. Current indicator species showed more variability associated with lake-specific factors than with changes in status or the influence of pressures.     

Hundred Years of Environmental Change and Phytoplankton Ecophysiological Variability Archived in Coastal Sediments

Marine protist species have been used for several decades as environmental indicators under the assumption that their ecological requirements have remained more or less stable through time. However, a growing body of evidence suggests that marine protists, including several phytoplankton species, are in fact highly diverse and may quickly respond to changes in the environment. Predicting how future climate will impact phytoplankton populations is important, but this task has been challenged by a lack of time-series of ecophysiological parameters at time-scales relevant for climate studies (i.e. at least decadal). Here, we report on ecophysiological variability in a marine dinoflagellate over a 100-year period of well-documented environmental change, by using the sedimentary archive of living cysts from a Scandinavian fjord (Koljö Fjord, Sweden). During the past century, Koljö Fjord has experienced important changes in salinity linked to the North Atlantic Oscillation (NAO). We revived resting cysts of Pentapharsodinium dalei preserved in the fjord sediments and determined growth rates for 18 strains obtained from 3 sediment core layers at salinity 15 and 30, which represent extreme sea-surface conditions during periods of predominantly negative and positive NAO phases, respectively. Upper pH tolerance limits for growth were also tested. In general, P. dalei grew at a higher rate in salinity 30 than 15 for all layers, but there were significant differences among strains. When accounting for inter-strain variability, cyst age had no effect on growth performance or upper pH tolerance limits for this species, indicating a stable growth response over the 100-year period in spite of environmental fluctuations. Our findings give some support for the use of morphospecies in environmental studies, particularly at decadal to century scales. Furthermore, the high intra-specific variability found down to sediment layers dated as ca. 50 years-old indicates that cyst-beds of P. dalei are repositories of ecophysiological diversity.     

Directionality theory and the evolution of body size

Directionality theory, a dynamic theory of evolution that integrates population genetics with demography, is based on the concept of evolutionary entropy, a measure of the variability in the age of reproducing individuals in a population. The main tenets of the theory are three principles relating the response to the ecological constraints a population experiences, with trends in entropy as the population evolves under mutation and natural selection. (i) Stationary size or fluctuations around a stationary size (bounded growth): a unidirectional increase in entropy; (ii) prolonged episodes of exponential growth (unbounded growth), large population size: a unidirectional decrease in entropy; and (iii) prolonged episodes of exponential growth (unbounded growth), small population size: random, non-directional change in entropy. We invoke these principles, together with an allometric relationship between entropy, and the morphometric variable body size, to provide evolutionary explanations of three empirical patterns pertaining to trends in body size, namely (i) Cope's rule, the tendency towards size increase within phyletic lineages; (ii) the island rule, which pertains to changes in body size that occur as species migrate from mainland populations to colonize island habitats; and (iii) Bergmann's rule, the tendency towards size increase with increasing latitude. The observation that these ecotypic patterns can be explained in terms of the directionality principles for entropy underscores the significance of evolutionary entropy as a unifying concept in forging a link between micro-evolution, the dynamics of gene frequency change, and macro-evolution, dynamic changes in morphometric variables.