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Fine structure of the sensory receptors of Aglantha digitale (Hydromedusae: Trachylina).

Fine-structure studies of the sensory receptors of Aglantha digitale have been done by transmission- and scanning-electron microscopy. Aglantha has 8 club-shaped statocysts and three types of ciliated sensory receptors: ciliated comb pads, ciliated sensory cells with microvilli, and ciliated sensory cells without microvilli. The epithelium covering the statocyst has both ciliated sensory cells and epitheliomuscular cells. The ciliated comb pads are formed by a group of ciliated sensory cells, each having an apical kinocilium surrounded by a ring of microvilli. These microvilli show an increase in length towards the velum. The axons from the comb pad cells enter the inner nerve ring. The structure, function and the evolution of the statocysts and the possible role of the ciliated comb pads and other sensory receptors are discussed.

Animals↗

Neoplastic cell transformation by high-LET radiation: molecular mechanisms.

Experimental data on molecular mechanisms are essential for understanding the bioeffects of radiation and for developing biophysical models, which can help in determining the shape of dose-response curves at very low doses, e.g., doses less than 1 cGy. Although it has been shown that ionizing radiation can cause neoplastic cell transformation directly, that high-LET heavy ions in general can be more effective than photons in transforming cells, and that the radiogenic cell transformation is a multi-step process [correction of processes], we know very little about the molecular nature of lesions important for cell transformation, the relationship between lethal and transformational damages, and the evolution of initial damages into final chromosomal aberrations which alter the growth control of cells. Using cultured mouse embryo cells (C3H10T1/2) as a model system, we have collected quantitative data on dose-response curves for heavy ions with various charges and energies. An analysis of these quantitative data suggested that two DNA breaks formed within 80 angstroms may cause cell transformation and that two DNA breaks formed within 20 angstroms may be lethal. Through studies with restriction enzymes which produce DNA damages at specific sites, we have found that DNA double strand breaks, including both blunt- and cohesive-ended breaks, can cause cell transformation in vitro. These results indicate that DNA double strand breaks can be important primary lesions for radiogenic cell transformation and that blunt-ended double strand breaks can form lethal as well as transformational damages due to misrepair or incomplete repair in the cell. The RBE-LET relationship is similar for HGPRT gene mutation, chromosomal deletion, and cell transformation, suggesting common lesions may be involved in these radiation effects. The high RBE of high-LET radiation for cell killing and neoplastic cell transformation is most likely related to its effectiveness in producing DNA double strand breaks in mammalian cells. At present the role of oncogenes in radiation cell transformation is unclear.

Animals↗

Molecular-level investigation of the structure, transformation, and bioactivity of single living fission yeast cells by time- and space-resolved Raman spectroscopy.

The structure, transformation, and bioactivity of single living Schizosaccharomyces pombe cells at the molecular level have been studied in vivo by time- and space-resolved Raman spectroscopy. A time resolution of 100 s and a space resolution of 250 nm have been achieved with the use of a confocal Raman microspectrometer. The space-resolved Raman spectra of living S. pombe cells at different cell cycle stages were recorded in an effort to elucidate the molecular compositions of organelles, including nuclei, cytoplasm, mitochondria, and septa. The time- and space-resolved measurement of the central part of a dividing yeast cell showed continuous spectral evolution from that of the nucleus to those of the cytoplasm and mitochondria and finally to that of the septum, in accordance with the transformation during the cell cycle. A strong Raman band was observed at 1602 cm(-)(1) only when cells were under good nutrient conditions. The effect of a respiration inhibitor, KCN, on a living yeast cell was studied by measuring the Raman spectra of its mitochondria. A sudden disappearance of the 1602 cm(-)(1) band followed by the change in the shape and intensity of the phospholipid bands was observed, indicating a strong relationship between the cell activity and the intensity of this band. We therefore call this band "the Raman spectroscopic signature of life". The Raman mapping of a living yeast cell was also carried out. Not only the distributions of molecular species but also those of active mitochondria in the cell were successfully visualized in vivo.

Cell Cycle↗

Identification of myosin heavy chain in Saccharomyces cerevisiae.

Motility in biological systems is expressed in a variety of ways, such as cytoplasmic streaming, cell shaping, nuclear migration and muscle contraction. These functions are thought to be mediated by structural proteins, for example, myosin, actin and tubulin. The involvement of myosin in muscle contraction is well documented and this protein is implicated in generating the cleavage forces during cytokinesis in some non-muscle cells. Here, we report the isolation of a protein similar to myosin as judged by its biochemical and immunological properties, from the yeast Saccharomyces cerevisiae. Parts of the protein have been conserved through evolution at the protein and DNA sequence levels. The presence of this protein in the region bordering mother cell and bud, as revealed by immunofluorescence, suggests that it is involved in cell division.

Actomyosin↗

Cell structure of developing downfeathers in the zebrafinch with emphasis on barb ridge morphogenesis.

The present ultrastructural and immunocytochemical study on the embryonic feathers of the zebrafinch, an altricial passerine bird, describes cellular differentiation of developing downfeathers. Barb ridges are folds of the original epidermis of the embryonic feather germ in which the basal-apical polarity of epidermal cells is upset. The result is the loss of most germinal activity of basal cells of the barb ridges so that only the embryonic epidermal layers remain. The more external layer is the primary periderm, followed by 4-6 layers of inner-periderm cells that mature into feather sheath and barb vane ridge cells. The following layer, the subperiderm, produces a small type of beta-keratin typical of feathers. In barb ridges, the subperiderm layer is displaced to form barbule plates and barb cells. The formation of branching barbules occurs by the presence of barb vane ridge cells that function as spacers between barbule cells. The fourth layer is homologous to the germinal layer of the epidermis, but in barb ridges it rapidly loses the germinal capability and becomes the cyclindrical layer of marginal plates. The study indicates that a necrotic process determines the carving out of the final feather shape, although apoptosis may also play a role. In fact, after barb and barbule cells have formed a keratinized syncitium, retraction of the vascular bed determines anoxia with the resultant necrosis of all feather cells. Only those of the keratinized syncitium remain to form the feather while supportive cells disappear. The sheath covering the barb and barbule syncitium is lost by the formation of a sloughing layer following degeneration of external barb ridge vane cells and loss of the sheath. It is proposed that the evolution of the morphogenetic process of barb ridge formation was peculiar to tubular outgrowths of the integument of archosaurian reptiles that evolved into birds. Once established in the embryonic programmes of skin morphogenesis of ancient birds, variations in the process of barb ridge morphogenesis allowed the fusion of ridges into large or branched ridges that originated the rachis. This process produced pennaceous feathers, among which were those later used for flight. The present study stresses that the morphogenetic process of barb ridge formation determines the concomitant appearance of barbs and barbules. As a consequence, intermediate forms of evolving feathers with only barbs but not barbules are unlikely or are derived from alteration of the above basic morphogenetic mechanism.

Animals↗

The cytoskeleton, cellular motility and the reductionist agenda.

Eukaryotic cells depend on cytoskeletal polymers and molecular motors to establish their asymmetrical shapes, to transport intracellular constituents and to drive their motility. Cell biologists are using diverse experimental approaches to understand the molecular basis of cellular movements and to explain why defects in the component proteins cause disease. Much of the molecular machinery for motility evolved in early eukaryotes, so a limited set of general principles can explain the motility of most cells.

Actins↗

Shaping the genome--restriction-modification systems as mobile genetic elements.

A restriction enzyme gene is often linked to a modification methylase gene the role of which is to protect a recognition site on DNA from breakage by the former. Loss of some restriction-modification gene complexes leads to cell death through restriction breakage in the genome. Their behavior as genomic parasites/symbionts may explain the distribution of restriction sites and clarify certain aspects of bacterial recombination repair and mutagenesis. A comparison of bacterial genomes supports the hypothesis that restriction-modification gene complexes are mobile elements involved in various genome rearrangements and evolution.

Bacteria↗

The early life history of tissue oxygenation in crustaceans: the strategy of the myodocopid ostracod Cylindroleberis mariae.

We studied basic principles of respiratory physiology in Cylindroleberididae, Cylindroleberis mariae Baird 1850, which are millimetre-sized crustaceans (myodocop ostracod) having a fossil record dating back to about 425 millions years ago. Facing experimental changes of O2 partial pressures in the range 2-40 kPa (normoxia is 21 kPa), C. mariae lack any regulatory mechanism to adapt their ventilatory and circulatory activity. Thus, the oxygenation status of their internal milieu must follow, as a dependent variable, the ambient oxygenation. Freely behaving C. mariae exhibit a marked diurnal activity rhythm. They are actively swimming in the water column during night, where they inspire in normoxic-normocapnic water. They are resting in self-made nests during daytime, where they are rebreathing in a confined and hypoxic environment. By analogy to extensive previous literature data, we suggest that these changes of respiratory gas content, and the associated tissue gas status, participate to the shaping of their metabolic activity and behaviour. To conclude, as Cylindroleberididae are early crustaceans exhibiting a remarkable stasis since the Palaeozoic, present data illustrates how principles of tissue oxygenation strategy can cover an impressive time scale.

Analysis of Variance↗

Neuropathological and computerized tomographic findings in experimental brain abscess.

The neuropathological progression of brain abscess formation was studied experimentally at sequential stages in dogs, and the findings correlated with the appearance on computerized tomographic (CT) brain scans. The evolution of brain-abscess formation was divided into four stages based on histological criteria: early cerebritis (Days 1 to 3); late cerebritis (Days 4 to 9); early capsule (Days 10 to 13); and late capsule (Days 14 and later). The cerebritis stage was characterized by prominent perivascular cuffing by inflammatory cells in the area adjacent to the developing necrotic center. However, the early elements of capsule formation appeared with the presence of fibroblasts by Day 5. The CT scans showed ring-shaped contrast enhancement by Day 3. Delayed scans at 30 minutes revealed diffusion of the contrast material into the developing necrotic center, forming a solid lesion. In lesions that were well encapsulated (14 days and older), five distinct histological zones were apparent: 1) a well formed necrotic center; 2) a peripheral zone of inflammatory cells, macrophages, and fibroblasts; 3) the dense collagenous capsule; 4) a layer of neovascularity associated with continuing cerebritis; and 5) reactive astrocytes, gliosis, and cerebral edema external to the capsule. The CT appearance of well encapsulated abscesses showed a typical ring-shaped contrast-enhancing lesion. On the delayed scans, the "ring" did not fill in with contrast enhancement. The diameter of the ring correlated best with the presence of cerebritis (perivascular infiltrates in the adventitial sheaths of vessels surrounding the abscess). The discussion focuses on the relevance of this study to the current management of patients with brain abscess.

Animals↗

Genome and protein evolution in eukaryotes.

The past year has seen the completion of the genome sequence of the flowering plant Arabidopsis thaliana and the initial sequence reports of the human genome. The availability of completely sequenced eukaryotic genomes from disparate phylogenetic lineages has opened the door to comparative analyses and a better understanding of the evolutionary processes shaping genomes. Complex many-to-many relationships between genes from different species appear to be the norm, suggesting that transfer of detailed functional annotation will not be straightforward. In addition to expansion and contraction of gene families, new genes evolve from recombination of pre-existing domains, although some domain families do appear to have evolved recently and to be specific to restricted phylogenetic lineages. The overall picture is of a huge diversity of gene content within eukaryotic genomes, reflecting different functional demands in different species.

Animals↗

Relationship between the tissue-specificity of mouse gene expression and the evolutionary origin and function of the proteins.

BACKGROUND: The combination of complete genome sequence information with expression data enables us to characterize the relationship between a protein's evolutionary origin or functional category and its expression pattern. In this study, mouse proteins were assigned into functional and phyletic groups and the gene expression patterns of the different protein groupings were examined by microarray analysis in various mouse tissues. RESULTS: Our results suggest that the proteins that are universally distributed in all tissues are predominantly enzymes and transporters. In contrast, the tissue-specific set is dominated by regulatory proteins (signal transduction and transcription factors). An increased tendency to tissue-specificity is observed for metazoan-specific proteins. As the composition of the phyletic groups highly correlates with that of the functional groups, the data were tested in order to determine which of the two factors -- function or phyletic age -- is dominant in shaping the expression profile of a protein. The observed differences in expression patterns of genes between functional groups were found mainly to reflect their different phyletic origin. The connection between tissue specificity and phyletic age cannot be explained by the recent rate of evolution. Finally, although metazoan-specific proteins tend to be tissue-specific compared with phyletically conserved proteins present in all domains of life, many such 'universal' proteins are also tissue-specific. CONCLUSION: The minimal cellular transcriptome of the metazoan cell differs from that of the ancestral unicellular eukaryote: new functions were added (metazoan-specific proteins), whilst other functions became specialized and no longer took place in all cells (tissue-specific pre-metazoan proteins).

Amino Acid Substitution↗

Proceedings of the SMBE Tri-National Young Investigators' Workshop 2005. Lineage-specific expansions and contractions of the bitter taste receptor gene repertoire in vertebrates.

The sense of bitter taste plays a critical role in how organisms avoid generally bitter toxic and harmful substances. Previous studies revealed that there were 25 intact bitter taste receptor (T2R) genes in humans and 34 in mice. However, because the recent chicken genome project reported only three T2R genes, it appears that extensive gene expansions occurred in the lineage leading to mammals or extensive gene contractions occurred in the lineage leading to birds. Here, I examined the T2R gene repertoire in placental mammals (dogs, Canis familiaris; and cows, Bos taurus), marsupials (opossums, Monodelphis domestica), amphibians (frogs, Xenopus tropicalis), and fishes (zebrafishes, Danio rerio; and pufferfishes, Takifugu rubripes) to investigate the birth-and-death process of T2R genes throughout vertebrate evolution. I show that (1) the first extensive gene expansions occurred before the divergence of mammals from reptiles/birds but after the divergence of amniotes (reptiles/birds/mammals) from amphibians, (2) subsequent gene expansions continuously took place in the ancestral mammalian lineage and the lineage leading to amphibians, as evidenced by the presence of 15, 18, 26, and 49 intact T2R genes in the dog, cow, opossum, and frog genome, respectively, and (3) contractions of the gene repertoire happened in the lineage leading to chickens. Thus, continuous gene expansions have shaped the T2R repertoire in mammals, but the contractions subsequent to the first round of expansions have made the chicken T2R repertoire narrow. These dramatic changes in the repertoire size might reflect the daily intake of foods from an external environment as a driving force of evolution.

Animals↗

Molecular structure of 20S and 26S proteasomes.

Eukaryotic proteasomes are unusually large protein complexes with characteristic sets of subunits and have been classified into two isoforms with apparent sedimentation coefficients of 20S and 26S, respectively. The 20S proteasome (previously named the multicatalytic proteinase complex) is a cylindrical particle with a molecular weight (MW) of approximately 750 kD. It is a dimeric assembly of two symmetrical discs, each consisting of 7 alpha-type subunits and 7 beta-type subunits, having the molecular organization alpha n[1-7)beta n[1-7)beta n[1-7)alpha n[1-7), where 'n' indicates the number of heterogeneous 7 subunits with MWs of 21-32 kD. The alpha-type and beta-type subunits constitute a unique multi-gene family encoding previously unidentified, but homologous, polypeptides that have been conserved during evolution. Interestingly, some beta-type subunits with catalytic functions appear to be replaced by very homologous, but distinct, gene products that might be generated by gene duplication in response to extracellular signals, such as gamma-interferon, suggesting that the 20S proteasome exists in cells as a heterogeneous population with functional diversity. The 26S proteasome is a eukaryotic ATP-dependent protease, selectively degrading various cellular proteins with specific degradation signals such as a multi-ubiquitin chain. It is a cylindrical caterpillar-shaped complex with a MW of about 2,000 kD. The 26S proteasome is a symmetrical assembly of a central 20S proteasome and a large terminal polypeptide complex with an apparent sedimentation coefficient of 22S. The terminal 22S subset consists of multiple components with MWs of 30-110 kD, which possibly have regulatory functions, and contains multiple ATPases, a de-ubiquitinating enzyme and the recognition molecule(s) for the target proteins. Thus the 26S proteasome is a multi-molecular assembly, consisting of the 20S proteasome and the 22S regulatory subunit complex.

Adenosine Triphosphatases↗

Intelligence is universal in life.

Behaviorists assume that living things memorize random atoms of information (engrams), "reinforced" by success, just as in the neo-Darwinian mutation-selection process. On the contrary we have to recognize the existence of organized and systematic responses in the learning process (Krechevsky). The animals seek desperately to "understand the meaning" of the world around them, by widening its context. Intelligence is not an exclusive prerogative of human mind. The minds of insects operate in the same way as that of man. Even a cell has a sort of intelligence (Cuenot). Consciousness is a state of awareness associated with enhanced mental activity. It occurs also in other "higher" animals (Thorpe). However human themselves are non conscious of their basic underlying motivations. Unconscious or ineffable knowledge plays a great role in shaping our world-view and in determining our influence on the Gaian hierarchy.

Animals↗

[An electron microscopic study of the natural death of cyst cells in Sarcosporidia. II. Ultrastructural changes in the nuclei of cyst cells of Sarcocystis muris].

Nuclear changes in cyst cells, developing within 6 and 10 month old sarcocysts of Sarcocystis muris, were followed in terms of the programmed cell death phenomenon. This communication extends our previous studies in the cytoplasm of S. muris cyst cells (Radchenko et al., 1995) to include particular nuclear changes in different involve changes in nuclear configuration: the original spherical from is progressively substituted for irregular or lobulated shapes. This may suggest some corresponding changes in cytoskeleton, involved in formation and maintaining of some definite nuclear shape. In normal cyst cells, the nuclear chromatin appears as a filiform and reticulate structure with a few lumps made of granules and filaments. In the cyst stages subject to natural cell death, structural changes in nuclei involve disassembly of lumps into separate granules. Some spherical structures are seen outbudding from the nucleolus. These structures are presumably made of RNA-containing granules. The pattern of nucleolar segregation in S. muris cells resembles somewhat the changes in nucleoli reported for metazoan cells. However, the general picture of morphological evolution in the nuclei of S. muris cells, in the course of natural dying, differs from that in metazoan cell nuclei. No condensation of nuclear chromatin at the nuclear periphery, or blebbing of the nuclear and cytoplasmic membranes, so characteristic of the latter, was followed in the former. The peculiarities noticed in the sarcosporidia may reflect biological peculiarities of these specialized parasitic protozoa.

Animals↗

Evolution of cell adhesion systems: evidence for Arg-Gly-Asp-mediated adhesion in the protozoan Neoparamoeba aestuarina.

Developmental processes in multicellular organisms require structural elements, such as adhesion molecules, to stabilize cells at functional positions. In vertebrates, a series of extracellular matrix proteins, e.g. fibronectin and laminin, are involved in cell adhesion. These proteins contain Arg-Gly-Asp [RGD] at their binding sites. Here we show that at concentrations above 2 mM the peptide GRGDSPK, comprising the tripeptide RGD (Arg-Gly-Asp), prevents the adhesiveness of cells of the marine amoeba Neoparamoeba aestuarina. In addition, elevated levels of GRGDSPK cause cells to alter their shapes from those with digitiform subpseudopodia to rounded cells with small lobed pseudopodia. These cells detach from the substratum. These results are specific for the RGD sequence, because incubation in GRGESPK solution at the same concentrations had no effect on cell attachment or structure. From these data we suggest that the structural adhesion molecules identified in vertebrates show amino acid homologies with those found in unicellular protozoa.

Amino Acid Sequence↗

Developmental expression of the glial fibrillary acidic protein mRNA in the central nervous system and in cultured astrocytes.

The expression of glial fibrillary acidic protein (GFAP)-mRNA during mouse brain development and in astroglial primary cultures has been investigated by using two approaches: Northern-blot evaluation using a specific cDNA probe, and cell-free translation associated with immunoprecipitation. During brain maturation (4-56 days postnatal), the GFAP-mRNA underwent a biphasic evolution. An increase was observed between birth and day 15 (i.e., during the period of astroglial proliferation), which was followed by a decrease until day 56 (i.e., during astroglial cell differentiation). At older stages (300 days), an increase was observed, which might reflect gliosis. During astroglial in vitro development (7-32 days in culture), the GFAP-mRNA showed similar variations. An increase, observed during the period of astroglial proliferation (7-18 days), was followed by a decrease which occurred in parallel to marked changes in cell shape, cell process outgrowth, and the organization and accumulation of gliofilaments. During the same culture period (7-32 days), alpha-tubulin mRNA, which was used as an internal standard, did not vary significantly. These results show that the increase of the GFAP protein and of gliofilaments observed both in vivo and in vitro during astroglial differentiation cannot be ascribed to an accumulation of the GFAP-mRNA. It might be that more than one mechanism regulates the levels of free and polymerized GFAP and of its encoding mRNA.

Animals↗

Interleukin-1-beta activation of cultured glomerular epithelial cells.

In crescentic glomerulonephritis, crescent formation involves the active participation of glomerular epithelial cells (GEC) and macrophages recruited to the glomerulus during the evolution of the disease. Cytokines derived from macrophages may affect many functions of GEC. In this study, we found that interleukin-1 beta (IL-1 beta) inhibited GEC growth (DNA synthesis and cell number) in vitro in a dose- and time-dependent manner. This effect was not mediated by tumor growth factor beta (TGF beta) which is a potent inhibitor of GEC growth in vitro. Treatment of GEC with various concentrations of IL-1 beta induced morphologic changes consisting in the loss of their cobblestone shape and acquisition of a fibroblast-like appearance. Moreover, IL-1 beta was shown to stimulate the expression of interleukin-6 (IL-6) by GEC. The increase in IL-6 secretion by GEC treated with IL-1 beta was observed at both the protein and mRNA levels. IL-1 beta also affected the metabolism of laminin in cultured GEC, inducing a dose-dependent increase in laminin production in culture supernatants harvested from GEC. Finally, we investigated the expression of MHC class II antigens and intercellular adhesion molecule-1 (ICAM-1) in GEC, and found that unstimulated GEC are negative for MHC class II antigens, as detected by flow cytometry. In contrast to the induction of effector functions, expression of MHC class II antigens stringently required interferon-gamma. IL-1 beta did not induce MHC class I antigen expression. The regulation of ICAM-1 expression in that unstimulated GEC expressed ICAM-1, and this expression was upregulated by IL-1 beta. We conclude that IL-1 beta alters many functions of GEC, and these changes may be involved in the initiation and amplification of glomerular injury.

Cell Count↗