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Comparative developmental genetics and the evolution of arthropod body plans.

The arthropods display a wide range of morphological diversity, varying tagmosis, as well as other aspects of the body plan, such as appendage and cuticular morphology. Here we review the roles of developmental regulatory genes in the evolution of arthropod morphology, with an emphasis on what is known from morphologically diverse species. Examination of tagmatic evolution reveals that these changes have been accompanied by changes in the expression patterns of Hox genes. In contrast, review of the modifications to wing morphology seen in insects shows that these body plan changes have generally favored alterations in downstream target genes. These and other examples are used to discuss the evolutionary implications of comparative developmental genetic data.

Animals↗

Low-temperature crystallization of calcium phosphate coatings synthesized by ion-beam-assisted deposition.

Crystallization temperature of the amorphous calcium phosphate coating synthesized by ion-beam-assisted deposition successfully was decreased to 400 degrees C for the making of orthopedic implants with better qualities. Transmission electron microscopy (TEM) and X-ray diffraction (XRD) were used to investigate the morphological and structural evolution of the crystals in the coating during post-heat treatment. The crystallization of calcium phosphate coating is a hydroxyl-diffusion-controlled process, which is thought to be the mechanism responsible for the decrease of the crystallization temperature. In addition, the detailed crystallization process of calcium phosphate coating is elaborated in the present paper. The results show that the crystallinity of the hydroxyapatite coating can be well controlled by adjusting the post-heat-treatment time.

Alloys↗

In vitro stability of plasma-sprayed hydroxyapatite coatings on Ti-6Al-4V implants under cyclic loading.

The success of hydroxyapatite (HA)-coated Ti-6Al-4V implants relies on the long-term stability of HA coatings. In this study, the mechanical stability of plasma-sprayed HA coatings on Ti-6Al-4V implants under four-point cyclic bending was systematically investigated in both air and simulated body fluid (SBF) environments at room temperature. To have a clear view of the microscale damage evolution, the surface morphology change of HA coatings during cyclic loading was carefully examined by scanning electron microscopy at the same locations on the coating surfaces after four-point bending for 4, 6.5, 8.5, and 10 million cycles. Also, possible changes of other characteristics such as thickness, weight, crystallinity, and residual stress of HA coatings were measured as a function of loading cycles. Up to 10 million cycles of bending in air and SBF, we found no significant microcracking or coating spalling on the surface of coatings, and no significant changes in thickness, weight, crystallinity, or residual stress of the plasma-sprayed HA coatings. The experiment results indicate that thickness and crystallinity had no effects on the stability of the HA coatings. HA coating resistance to the cyclic four-point bending might result from the stress shielding effects of preexisting microcracks in the coatings.

Biocompatible Materials↗

Genetic analysis of mutations that alter cell fates in maize leaves: dominant Liguleless mutations.

The three major components of the maize leaf are the blade, the sheath, and at their junction, ligular region. Each exhibits specific cell types and organization. Four dominant Liguleless (Lg) mutations (Lg3-O, Lg4-O, Lg*347, and Lg*9167) in at least three different genes cause a similar morphological phenotype in leaves, although each mutation affects a distinct domain of the blade. Mutant leaves display regions of altered cell fate in the blade, accompanied by elimination of ligule and auricle at their wild-type positions and development of ligule and auricle in the blade at the borders of the altered regions. The affected blade cells are transformed into sheath-like cells, as determined by morphological and genetic tests. Lg4-O expressivity is highly dependent on genetic background. For example, two different backgrounds nay specify converse patterns of phenotypic expression. Lg4-O expressivity is also affected by the heterochronic mutation Teopod2 (Tp2). Gene dosage experiments indicates Lg4-O is a neomorph. Interactions between recessive lg mutations (which eliminate ligular structures) and the dominant Lg mutations suggest that the lg+ genes act after the Lg mutations. Lg3-O and Lg4-O act semidominantly, and interact with each other and with other mutations in the Knotted1 (Kn1)-like family (a family in which dominant mutant alleles cause blade to sheath transformation phenotypes). These interactions suggest that the above Kn1-like mutations may function similarly in the leaf. We discuss the similarities between the Lg mutations and the other mutations of the Kn1-like family, which led us to postulate that Ig3 and Ig4 are members of a growing family of kn1-like (knox) homeobox genes that are identified by dominant mutant alleles causing leaf transformation phenotypes. We also propose that certain key characteristics of this family of dominant neomorphic mutations are important for generating meaningful morphological changes during evolution.

Alleles↗

A genetic attack on the defense complex.

An increasing number of "non-model" organisms are becoming accessible to genetic analysis in the field, as evolutionary biologists develop dense molecular genetic maps. Peichel et al.'s recent study(1) provides a microsatellite-based map for threespine stickleback fish (Gasterosteus aculeatus), and the first evidence for QTL affecting feeding morphology and defensive armor. This species has undergone rapid and parallel morphological and behavioral evolution, and there is now hope that some of the genes responsible for the divergence may soon be identified.

Animals↗

Similar morphologies of cichlid fish in Lakes Tanganyika and Malawi are due to convergence.

The species flocks of cichlid fishes in the lakes of East Africa are the most spectacular example of adaptive radiation among living vertebrates. Similar highly derived morphologies are found among species in different lakes. These similarities have been variously interpreted either as evidence for migration of ancestral species between the lakes, or of striking convergence of morphology. To distinguish among these competing hypotheses we sequenced a portion of the mitochondrial DNA control region from six pairs of morphologically similar taxa from Lakes Malawi and Tanganyika. Our results clearly indicate a separate origin of these morphologies in the two lakes. They also suggest that the Tanganyikan radiation is relatively old, and that the species flocks in lakes Malawi and Victoria are derived from one of the ancient lineages found in Lake Tanganyika. These findings have important implications for understanding the pattern of morphological and behavioral evolution in these fish.

Africa, Eastern↗

Evidence for three major clades within the snapping shrimp genus Alpheus inferred from nuclear and mitochondrial gene sequence data.

The snapping shrimp genus Alpheus is among the most diverse of caridean shrimps, and analyses of taxa separated by the Isthmus of Panama have been used to estimate rates of molecular evolution. Although seven morphological groups have been informally suggested, no formal phylogenetic analysis of the genus has been previously attempted. Here we infer the phylogenetic relationships within Alpheus using sequence data from two nuclear genes, glucose-6-phosphate isomerase and elongation factor-1alpha, and from the mitochondrial gene cytochrome oxidase I. Three major clades corresponding to previously noted morphological features were identified. Discrepancies between earlier informal morphological groupings and molecular analyses largely consisted of species whose morphologies were not entirely typical of the group to which they had been assigned. The traditional placements of shrimp with highly sessile lifestyles and consequently simplified morphologies were also not supported by molecular analyses. Phylogenies for Alpheus suggest that specialized ecological requirements (e.g., symbiotic associations and estuarine habitats) and modified claw morphologies have evolved independently several times. These new analyses also support the sister species status of transisthmian pairs analyzed previously, although very similar pairs were not always resolved with the more slowly evolving nuclear loci. In addition, six new cryptic species were identified in the course of these studies plus a seventh whose status remains to be determined.

Animals↗

Chronic myelogenous leukemia in blast crisis: retrospective analysis of prognostic factors in 90 patients.

Ninety patients with Philadelphia chromosome-positive chronic myelogenous leukemia in blast crisis were reviewed to identify significant prognostic associations. At diagnosis of blast crisis the main clinical, laboratory, and cytogenetic data were recorded and evaluated for prognostic significance. At the time of the analysis 89 patients had died, with a median survival of 11 weeks from diagnosis of blast crisis. Patient characteristics demonstrated in the univariate analysis to have significant association with shorter survival were: thrombocythemia, leukocyte count above 20 x 10(9), Karnofsky index < 50%, nonlymphoid blast cell morphology, cytogenetic clonal evolution, the presence of a double Philadelphia chromosome or trisomy 8, and no response to therapy. In 17 of 59 patients (29%) evaluable for response to therapy a complete or partial remission was achieved. These responders had a significantly longer median survival (25 weeks) as compared with nonresponders (9 weeks). Response to therapy was significantly better in lymphoid blast crisis and in patients without clonal evolution. In a multivariate analysis containing all significant variables of the univariate analysis two parameters retained their prognostic significance: response to therapy and trisomy 8. In spite of the short overall survival in blast crisis, the determination of prognostic factors may be a useful tool for the clinician planning therapy, especially new therapeutic approaches.

Adult↗

Assessment of ploidy stability of the somatic embryogenesis process in Quercus suber L. using flow cytometry.

Flow cytometry analyses were used to verify the ploidy stability of Quercus suber L. somatic embryogenesis process. Leaf explants of two adult cork oak trees (QsG0 and QsG5) of the North of Portugal were inoculated on MS medium with 2,4-D and zeatin. After 3 months, calluses with embryogenic structures were isolated and transferred to fresh MS medium without growth regulators and somatic embryo evolution was followed. Morphologically normal somatic embryos (with two cotyledons) and abnormal somatic embryos (with one or three cotyledons) were used in this assay. Flow cytometry combined with propidium iodide staining was employed to estimate DNA ploidy levels and nuclear DNA content of somatic embryos and leaves from mother plants. No significant differences (P< or =0.05) were detected among embryos, and between the embryos and the mother plants. Also, after conversion of these embryos, no significant morphological differences were observed among the somatic embryo-derived plants. These results and further studies using converted plantlet leaves and embryogenic callus tissue indicate that embryo cultures and converted plantlets were stable with regard to ploidy level. As no major somaclonal variation was detected our primary goal of "true-to-type" propagation of cork oak using somatic embryogenesis was assured at this level. The estimation of the 2C nuclear DNA content for this species is similar to the previously obtained value.

Flow Cytometry↗

Male and female effects on sperm precedence in the giant sperm species Drosophila bifurca.

Sperm competition is expected to be a driving force in sexual selection. In internally fertilized organisms, it occurs when ejaculates from more than one male are present simultaneously within the female's reproductive tract. It has been suggested that greater sperm size may improve the competitive ability of sperm, but studies provide contradictory results depending on the species. More recently, the role of females in the evolution of sperm morphology has been pointed out. We investigate here the male and female effects that influence sperm precedence in the giant sperm species, Drosophila bifurca Patterson & Wheeler. Females were mated with two successive males, and the paternity outcomes for both males were analyzed after determining sperm transfer and storage. We found very high values of last male sperm precedence, suggesting a strong interaction between rival sperm. However, the data also indicate high frequencies of removal of the sperm of the first male from the female reproductive tract prior to any interaction with the second male. This implies that successful paternity depends mainly on successful sperm storage. Knowing what happens to the sperm within females appears to be a prerequisite for disentangling post-copulatory sexual interactions between males and females.

Animals↗

Chromatin reorganization during senescence of proliferating cells.

It was previously proposed (Macieira-Coelho, 1979) that aging of proliferating cells is the result of genome reorganization taking place during the division cycle. This hypothesis was investigated and a reorganization could indeed be ascertained in the different hierarchical orders of DNA structure; a correlation was found between changes in chromatin organization and the impairment of cell cycle-related events. Indeed, like the latter, the reorganization of chromatin structure is characterized by a succession of subtle changes through the cell population life span, and a final short stage with abrupt events. The final events seem to concern mainly the organization of heterochromatin. The reorganization in the genome is accompanied by structural changes in the cellular scaffold and an evolution of cell morphology. The remodeling occurring in the cell through serial divisions seems to take place in such a way as to decrease the probability of further reorganizations, tending to a limit. The decline of the proliferative activity seems to be the result of the tendency to reach this limit.

Animals↗

Culture of neural cells on polymers coated surfaces for biosensor applications.

We focused our study on the olfactory cells growth on biocompatible polymer films electrodeposited on a silicon microsystem. Several substrates such as polyethyleneimine (PEI), polypropyleneimine (PPI), and polypyrrole (PPy), acting as potentially good candidates for cell culture, were tested in order to allow cells to adhere and proliferate. During their growth, the evolution of their morphology was monitored using both confocal microscope and immunohistochemistry, leading to the conclusion of a normal development. An estimation of the adhesion and proliferation rates of rat neuronal cell cultures indicated that PEI and PPI were the best substrates for cultivating olfactory cells.

Animals↗

Properties of after-discharges from cortical electrical stimulation in focal epilepsies.

OBJECTIVE: This study sought to determine whether certain aspects of after-discharges (ADs) obtained during cortical functional mapping provide better correlations between stimulus site and that of spontaneous seizures. Secondly, we wished to determine the percentage of stimulations evoking ADs and, of these, the percentage which clearly involves more than one electrode position, potentially inaccurately localizing cortical function. Thirdly, we wished to quantify the incidences of the several AD morphologies described by Jasper [in: Epilepsy and the functional anatomy of the human brain, 1954, p. 183; p. 692] and to assess whether certain morphologies had a greater tendency to evolve in frequency or morphology. METHODS: In these 29 patients requiring invasive recordings to determine principal epileptogenic areas, only subdural strips were placed in 19 patients, only grids in 2 patients, and both in 8 patients. A median of 21 electrodes per patient was stimulated of a median of 63 electrodes placed, with the following parameters: biphasic, monopolar, 50 Hz, 0.3 ms pulse duration, 1.5-18 mA. Coverage involved the frontal and parietal lobes (9 patients), frontal parietal temporal lobes (8), frontal temporal (3), temporal (2), occipital (2) and occipital temporal (2) with other combinations in 3 additional patients. Classification of AD morphologies was determined by a pilot study using IFSECN definitions [Electroenceph clin Neurophysiol 1974;37:538] and descriptions by Jasper [in: Epilepsy and the functional anatomy of the human brain, 1954, p. 183] and Gloor [in: Advances in neurology, vol. 8, 1975; p. 59]. RESULTS: Four hundred and two ADs (12%) were elicited by 3358 trains of electrical stimuli of which 260 (65%) clearly involved more than only the stimulated electrode position. Thus, 260 (8%) of 3358 stimulations evoked an AD that could mislocalize cortical function. The proportion of stimulating electrodes eliciting ADs ranged among patients from 4 to 83% (median 33%). Polyspike bursts and sequential spikes were the most common AD morphologies. Ten percent of ADs evolved in morphology, frequency or both. Evolution occurred more commonly (44%) with rhythmic waves than with other AD morphologies (7%). Neither evolving ADs, ADs producing clinical seizures, or ADs exceeding 10 s correlated topologically with spontaneous seizure origins. CONCLUSIONS: Although occurring in a minority of cortical electrical stimuli, ADs may involve more than the stimulus site and therefore may inaccurately localize cortical function. Our material failed to disclose any consistent relationship between the site of stimulus eliciting ADs and that of spontaneously appearing seizures, even when certain aspects of such ADs were analysed. SIGNIFICANCE: These data illustrate the need for scrutiny of the post-stimulus electrocorticogram for ADs and particularly those involving extra stimulus sites. Whether varying the stimulus parameters prospectively will disclose a better seizure localizing value for ADs remains to be determined.

Adolescent↗

Ventricular remodeling in Loeffler endocarditis: implications for therapeutic decision making.

BACKGROUND: Little is known about the morphological and functional evolution of ventricular abnormalities in Loeffler endocarditis. METHODS AND RESULTS: We describe 5 patients, including 3 with long-term echocardiographic follow-up, in whom apical obliteration due to fibro-thrombotic thickening of the endocardium showed favorable patterns of evolution. In one patient there was almost complete resolution of the obliterative process with consequently increased effective ventricular volume. In two patients formation of a flow-passage in the fibrocalcific apical 'floor' between the main medioventricular cavity and the apical chamber, leading to a 'double-chambered' left ventricle was observed. CONCLUSIONS: Medical therapy and appropriate anticoagulation, can induce favorable long-term ventricular remodeling in Loeffler endocarditis.

Adult↗

UVR-B induced cataract development in C57 mice.

The evolution of the morphological appearance and intensity of light scattering in C57 mice lenses after exposure to ultraviolet radiation type B (UVR-B) was investigated. A total of 80, 6-week-old female C57BL/6 mice were divided into four groups (n=20). One eye in each animal was exposed in vivo to UVR-B in the 300 nm wavelength region (UVR-B-300 nm) to a dose of 5 kJm(-2) for 15 min. The radiation output had lambda(max) at 302 nm with 5 nm [FWHM]. The animals were consecutively sacrificed at 1, 2, 4 and 8 days after the exposure. Macroscopic lens changes were documented using grid- and dark field illumination photography. Light scattering in the exposed and contralateral not exposed lens was measured quantitatively. Morphological lens changes were documented using grid- and dark field illumination photography. In vivo exposure to UVR-B-300 nm induced subcapsular cataract in all exposed lenses and occasionally cortical and nuclear cataract at all investigated time points. Exposed lenses scattered light significantly higher on all investigated days compared to contralateral non-exposed lenses. A transient increase of light scattering peaking at day 2 in exposed as well as in contralateral not exposed lenses was identified. Light scattering of the lenses varies with latency time after exposure. A dose of 5 kJm(-2) UVR-B-300 nm induces light scattering in C57 mice lenses. The increase has a transient peak at 2 days after exposure. The variation of light scattering among days 1, 2, 4, and 8 indicates a dynamic change of scattering characteristics in the mouse lens following unilateral in vivo exposure to 5 kJm(-2) UVR-B-300 nm.

Animals↗

Isolation of the LFY/FLO homologue in Orchis italica and evolutionary analysis in some European orchids.

It has been suggested that the evolutionary analysis of floral development genes could explain the divergences between the rates of morphological and molecular evolution. The LEAFY (LFY) gene of Arabidopsis thaliana is one of the central regulatory genes in the control of flower development. We have identified the homologue of this gene (OrcLFY) in Orchis italica using 5'/3'RACE and primer walking, and compared the coding sequences of several orchid species. We analyzed nonsynonymous and synonymous substitution rates between the OrcLFY coding regions of 14 species, and performed a McDonald-Kreitman test on Orchis morio and Orchis laxiflora populations, showing that purifying selection is acting on this gene in these orchids. We have performed a phylogenetic analysis showing that OrcLFY is a new useful marker to reconstruct molecular phylogenies at low taxonomic levels.

DNA, Plant↗

Hydrolysis of fluid supported membrane islands by phospholipase A(2): Time-lapse imaging and kinetic analysis.

The activity of phospholipase A(2) (PLA(2)) which catalyzes the hydrolysis of phospholipids into free fatty acids and lysolipids, depends on the structure and thermodynamic state of the membrane. To further understand how the substrate conformation correlates with enzyme activity, model systems that are based on time-resolved membrane microscopy are needed. We demonstrate a methodology for preparing and investigating the dynamics of fluid supported phospholipid membranes hydrolyzed by snake venom PLA(2). The method uses quantitative analysis of time-lapse fluorescence images recording the evolution of fluid bilayer islands during hydrolysis. In order to minimize interactions with the support surface, we use double bilayer islands situated on top of a complete primary supported membrane prepared by hydration of spincoated lipid films. Our minimal kinetic analysis describes adsorption of enzyme to the membrane in terms of the Langmuir isotherm as well as enzyme kinetics. We use two related models assuming hydrolysis to occur either at the perimeter or at the surface of the membrane island. We find that the adsorption constant is similar for the two cases, while the estimated turnover rate is markedly different. The PLA(2) concentration series is measured in the absence and presence of beta-cyclodextrin which forms water soluble complexes with the reaction products. The results demonstrate the versatility of double bilayer islands as a membrane model system and introduces a new method for quantifying the kinetics of lipase activity on membranes by directly monitoring the evolution in substrate morphology.

Adsorption↗

Contribution of glial cells to the development of amyloid plaques in Alzheimer's disease.

Amyloid plaques appear early during Alzheimer's disease (AD), and their development is intimately linked to activated astrocytes and microglia. Astrocytes are capable of accumulating substantial amounts of neuron-derived, amyloid beta(1-42) (Abeta42)-positive material and other neuron-specific proteins as a consequence of their debris-clearing role in response to local neurodegeneration. Immunohistochemical analyses have suggested that astrocytes overburdened with these internalized materials can eventually undergo lysis, and radial dispersal of their cytoplasmic contents, including Abeta42, can lead to the deposition of a persistent residue in the form of small, GFAP-rich, astrocytic amyloid plaques, first appearing in the molecular layer of the cerebral cortex. Microglia, most of which appear to be derived from blood monocytes and recruited from local blood vessels, rapidly migrate into and congregate within neuritic and dense-core plaques, but not diffuse plaques. Instead of internalizing and removing Abeta from plaques, microglia appear to contribute to their morphological and chemical evolution by facilitating the conversion of existing soluble and oligomeric Abeta within plaques to the fibrillar form. Abeta fibrillogenesis may occur largely within tiny, tube-like invaginations in the surface plasma membrane of microglia. These results highlight the therapeutic potential of blocking the initial intracellular accumulation of Abeta42 in neurons and astrocytes and inhibiting microglia-mediated assembly of fibrillar Abeta, which is particularly resistant to degradation in Alzheimer brain.

Alzheimer Disease↗