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At least 631 records · Page 35Linked to original sources
PubMed2003

Native structural propensity in cellular retinoic acid-binding protein I 64-88: the role of locally encoded structure in the folding of a beta-barrel protein.

A central question in protein folding is the relative importance of locally encoded structure and cooperative interactions among residues distant in sequence. We have been exploring this question in a predominantly beta-sheet protein, since beta-structure formation clearly relies on both local and global sequence information. We present evidence that a 24-residue peptide corresponding to two linked hairpins of cellular retinoic acid-binding protein I (CRABP I) adopts significant native structure in aqueous solution. Prior work from our laboratory showed that the two turns contained in this fragment (turns III and IV) had the highest tendency of any of the eight turns in this anti-parallel beta-barrel to fold into native turns. In addition, the primary sequence of these two turns is well conserved throughout the structural family to which CRABP I belongs, and residues in the turns and their associated hairpins participate in a network of conserved long-range interactions. We propose that the strong local-sequence biases within the chain segment comprising turns III and IV favor longer-range interactions that are crucial to the folding and native-state stability of CRABP I, and may play a similar role in related intracellular lipid-binding proteins (iLBPs).

Amino Acid Sequence↗
PubMed1981

[Acid and alkaline phosphatases in the cerebellar structures in autosomal trisomy syndromes].

The activity of nonspecific phosphomonoesterases in cerebellar structure of newborns and infants with trisomies in the 13th (5 cases) 18th (4) and 21st (14) chromosomes was investigated. In all trisomies, changes of phosphatase activity in the cerebellar structures were demonstrated which were particularly marked with alkaline phosphatase. The activity of this enzyme increased not only in the vascular endothelium but in a number of cellular structures as well: in the external embryonal and internal granular layers of the cortex, in the nuclei of Purkinje cells and neurons of the nucleus dentatus, in the foci of heterotopia of cortical cells into the white matter and nucleus dentatus. In Edwards' syndrome the activity of alkaline phosphatase in cellular elements of the cerebellum was slightly higher than in the control group. Changes in the activity was observed in the cells of the external embryonal layer, neurons of the nucleus dentatus and in the cells of the heterotopic foci. In autosomal trisomies dystopic cells have dissimilar levels of phosphatase activity and stain mosaically which seems to confirm the assumption of their dissimilar differentiation.

Acid Phosphatase↗
PubMed1996

Microwave absorption by magnetite: a possible mechanism for coupling nonthermal levels of radiation to biological systems.

The presence of trace amounts of biogenic magnetite (Fe3O4) in animal and human tissues and the observation that ferromagnetic particles are ubiquitous in laboratory materials (including tissue culture media) provide a physical mechanism through which microwave radiation might produce or appear to produce biological effects. Magnetite is an excellent absorber of microwave radiation at frequencies between 0.5 and 10.0 GHz through the process of ferromagnetic resonance, where the magnetic vector of the incident field causes precession of Bohr magnetons around the internal demagnetizing field of the crystal. Energy absorbed by this process is first transduced into acoustic vibrations at the microwave carrier frequency within the crystal lattice via the magnetoacoustic effect; then, the energy should be dissipated in cellular structures in close proximity to the magnetite crystals. Several possible methods for testing this hypothesis experimentally are discussed. Studies of microwave dosimetry at the cellular level should consider effects of biogenic magnetite.

Acoustics↗
PubMed1995

Freeze-fracture, deep-etch, and freeze-substitution studies of olfactory epithelia, with special emphasis on immunocytochemical variables.

Freeze-fracturing and deep-etching are a well-suited set of methods to study membrane and cytoplasmic features. Various approaches are available. Possible variables include tissue preparation, fracturing only or fracturing followed by etching, modes and materials of replication, and various ways of combining freeze-fracturing and/or deep-etching with (immuno)cytochemistry. Freeze-substitution, in particular combined with embedding in methacrylate resins such as the Lowicryls, is becoming rather widely accepted for purposes of ultrastructural (immuno)cytochemistry. Most investigators active in this field agree that this combination yields superior results compared to (immuno)cytochemistry combined with more conventional means of thin section transmission electron microscopy. Yet relatively little information is available on the variations that can occur with different approaches of freeze-substitution immunocytochemistry. This review deals with some of the variations in freeze-fracturing, freeze-etching, and freeze-substitution as applied to olfactory epithelial structures and with the effectiveness of observations obtained by application of the above sets of methods in relating the special morphology of olfactory epithelial cellular structures with those obtained by other approaches. Indeed, the data obtained continue to provide an integral image in which that morphology can be related to the special biochemistry, cell and molecular biology, and electrophysiology of olfactory epithelial structures.

Animals↗
PubMed2002

DNA-membrane complexes, mitochondria and aging.

The results of extensive in vitro studies of DNA-lipid complexes allowed us to propose a model for the structure of such complexes and their involvement in the formation of DNA-membrane complexes (DMC). DMC seem to form the basis for such cellular structures as Bayer's junctions and nucleoid of bacteria, the nuclear pores, annulate lamellae and nucleoid of eucaryotes. The role of DMC in gene expression is discussed.Numerical density of mitochondria during cell aging correlates with the density of bacteria in batch culture. It is concluded that aging is caused by the unlimited growth of mitochondria and their subsequent degradation. The role of DMC in mitochondrial DNA damage at aging is discussed. The way of increasing the life span by controlling the density of mitochondria in a cell volume is likewise discussed. DMC formed between any two intracellular membranes can serve the basis for the membrane continuum in a cell.

Aging↗
PubMed2006

Probing the role of multicellular organization in three-dimensional microenvironments.

Successful application of living cells in regenerative medicine requires an understanding of how tissue structure relates to organ function. There is growing evidence that presentation of extracellular cues in a three-dimensional (3D) context can fundamentally alter cellular responses. Thus, microenvironment studies that previously were limited to adherent two-dimensional (2D) cultures may not be appropriate for many cell types. Here we present a method for the rapid formation of reproducible, high-resolution 3D cellular structures within a photopolymerizable hydrogel using dielectrophoretic forces. We demonstrate the parallel formation of >20,000 cell clusters of precise size and shape within a thin 2-cm(2) hydrogel and the maintenance of high cell viability and differentiated cell markers over 2 weeks. By modulating cell-cell interactions in 3D clusters, we present the first evidence that microscale tissue organization regulates bovine articular chondrocyte biosynthesis. This platform permits investigation of tissue architecture in other multicellular processes, from embryogenesis to regeneration to tumorigenesis.

Animals↗
PubMed1990

[Morphological skin changes in the action of complex mineral fertilizers].

Experimental study of the skin structure in exposure to mineral fertilizers (ammophos, nitroammophos) was carried out. Saturated solutions of fertilizers were applied to intact and scarified free of hair skin of the back of guinea pigs. Skin structure changes were more significant when the fertilizers were applied to scarified skin. The findings evidence the injurious toxic effects of mineral fertilizers on cellular structural components.

Animals↗
PubMed1989

[A quantitative analysis of regional features in the structure of aortal endothelial cells in short-term hypertension].

The results of quantitative analysis of regional differences in the structure of thoracic aorta endotheliocytes in normal and hypertensive rats are presented. Scanning electron microscopy and semiautomatic image analyser were employed in studying the cellular structure in three regions of aorta endothelial layer: dorsal and ventral surfaces and region of the intercostal arterial ostia. The data obtained show that in cases of a short-duration hypertension the most pronounced are the alterations in endotheliocytes of the dorsal surface. Thus, the results confirm the heterogeneity of the aortic endothelium under normal conditions and demonstrate that differences in overlapping of neighbouring endotheliocytes is the cause of the observed alterations in cases of hypertension.

Animals↗
PubMed1996

Formation of the light-harvesting complex I (B870) of anoxygenic phototrophic purple bacteria.

The light-harvesting (LH) complex I (B870) of anoxygenic photosynthetic purple bacteria is the oligomeric form of its subunit B820 consisting of the low-molecular-weight polypeptides alpha, beta, bacteriochlorophyll (BChl), and carotenoids in the stoichiometric ratio [alpha1 beta1 (BChl2) Crt1-2]n. LHI surrounds the photochemical reaction center (RC). The major absorption band of the LHI complex is species-specific and is found at 870-890 nm; those of the subunit and the monomeric BChl a (dissolved in methanol) absorb at 820 and 770 nm, respectively. The isolated LHI complex can be reversibly dissociated to the B820 subunit or to the polypeptides and pigments by addition of detergents. Reconstitution of the B820 or the functional B870 complex is still possible after partial truncation of the N- or C-terminal regions of the alpha- or beta-polypeptide or of the beta-polypeptide only. The minimal structural requirements for reconstitution of a spectrally wild-type form after truncation of the polypeptides and/or modifications of the BChl molecule are described. The insertion of the LHIalpha- and LHIbeta-polypeptides into the membrane and the in vivo assembly of LHI, studied in a cell-free system and in whole cells of Rhodobacter capsulatus, depend on the primary structures of both polypeptides, BChl, the chaperones DnaK and GroEL, membrane-bound proteins, and energized membranes. Exchanges, deletions, or insertions of amino acyl residues, especially in the conserved region of the N-terminus of the LHIalpha-polypeptide, prevent or reduce the efficiency and stability of the LHI assembly. Therefore, reconstitution of LHI in a detergent micelle does not exactly reproduce the formation of the LHI complex in the photosynthetic membrane in vivo. The N-terminal domains play a crucial role in the formation of the oligomeric protein scaffold and of the pigment array. Facultatively phototrophic bacteria such as Rhodospirillum (Rsp.) rubrum or Rhodobacter (Rba.) capsulatus can adjust to changes in oxygen tension, light intensity, temperature, and substrates to grow under chemotrophic or phototrophic conditions. The photosynthetic apparatus (PSA), localized mainly on intracytoplasmic membranes (ICM), is usually synthesized only under low oxygen partial pressure. The cellular amount and composition of the PSA are modified upon changing light intensity in relation to cell growth (Drews and Golecki 1995). The morphogenesis of cellular structures like ICM is quite different from self-assembly. Self-assembly is a reversible process of aggregation of the constituents of a complex structure without protein synthesis and is driven by weak or strong forces in the interactions of the constituents. Morphogenesis results from the interplay of numerous gene products and the cellular organization and is always dependent upon pre-existent structures (Harold 1995). The morphogenesis of the photosynthetic membrane in purple bacteria has been studied in its different steps. The regulation at the transcriptional and post-transcriptional levels in purple bacteria, and the structure and morphogenesis of the ICM have been described recently (Armstrong 1995; Bauer 1995; Biel 1995; Drews and Golecki 1995; Klug 1995). In this mini-review, I will focus on the minimal requirements for the in vitro assembly of light-harvesting (LH) complex I (B870) from its constituents in detergent micelles and compare the results with observations on the complex process of targeting and import of LHI polypeptides into the membrane and assembly of B870.

Amino Acid Sequence↗
PubMed1996

Non-isotopic electron microscope in situ hybridization for studying the functional sub-compartmentalization of the cell nucleus.

Post-embedding electron microscope in situ hybridization using gold particles as label permits the clear identification of the cellular structures which contain the nucleic acid molecules under study. It has yielded information on the distribution of defined nucleic acid sequences of different origins-cellular or viral, DNA or RNA, single- or double-stranded molecules-which has revolutionized the study of the nucleus. Application of this powerful technique in combination with other refined techniques to studies on transcription and replication of cellular and viral genes has augmented our knowledge of the functional organization of the cell nucleus. One can now ask mechanistically meaningful questions concerning the successive steps of gene replication and expression not only under normal conditions of cell growth, but also when the cellular metabolism is altered by a drug treatment or a viral infection. This chapter aims (a) to present the established methods of post-embedding electron microscope in situ hybridization for localizing, precisely and specifically, a nucleic acid target in its normal environment and (b) to present some contributions of this technique to investigations of the functional compartmentalization of the cell nucleus and to elucidate the cell-virus relationships in infected cells.

Animals↗
PubMed1979

Virus-simulating structures in the optic nerve head in Creutzfeldt-Jakob disease.

A 68-year-old man was treated for and died of Creutzfeldt-Jakob disease. At autopsy we found multiple virus-like particles in the optic nerve head, but saw no similar structures in the cornea. Although these particles were morphologically similar to those previously reported in brain, we believe that they are not virions but unrelated cellular structures. We speculate that the causative agents may be naked membrane bound nucleic acids rather than true viruses. We found no optic atrophy or other specific pathologic changes in the eyes; severe occipital cortical degeneration was responsible for the patient's visual loss.

Aged↗
PubMed1985

The mechanical behaviour of cancellous bone.

Cancellous bone has a cellular structure: it is made up of a connected network of rods and plates. Because of this, its mechanical behaviour is similar to that of other cellular materials such as polymeric foams. A recent study on the mechanisms of deformation in such materials has led to an understanding of how their mechanical properties depend on their relative density, cell wall properties and cell geometry. In this paper, the results of this previous study are applied to cancellous bone in an attempt to further understand its mechanical behaviour. The results of the analysis agree reasonably well with experimental data available in the literature.

Animals↗
PubMed1986

The Antarctic cold desert and the search for traces of life on Mars.

The cryptoendolithic microorganisms that live inside rocks in the frigid Ross Desert of Antarctica can serve as a terrestrial model for what may have happened to life forms on Mars when the planet became dry and cold. Trace fossils of microbial rock colonization exist in Antarctica, and similar structures could have formed on Mars. In some respects, such trace fossils could be an easier target for life-detection systems than fossils of cellular structures.

Adaptation, Biological↗
PubMed1998

Central olfactory structures in Pax-6 mutant mice.

During the development of the olfactory system, cells located in the olfactory placode/olfactory pit send their axons toward the rostral part of the telencephalic vesicles (TVs). Some of these enter the TV inducing the formation of the olfactory bulbs (OBs), whereas, mitral and tufted cell axons form the lateral olfactory tract (LOT). Our recent studies have shown that the beginning of the central olfactory projections is independent of the arrival of olfactory receptor neuron (ORN) axons to the TV. Here we have used the mouse carrying a mutation in the Pax-6 gene to study whether the nasal olfactory structures intervene in the formation of central olfactory structures. This mutant as well as lacking a nose and eyes, is reported to lack olfactory epithelium and OB. However, we have found an ovoid cellular structure localized in the rostral part of the brain, and some cells in this structure project axons toward the piriform cortex forming a presumptive LOT. We conclude that the referred structure is an OB, which fails to develop because the mutation in the Pax-6 gene affects the formation of nasal structures. As such, fibers of the ORNs are necessary for the protrusion and layered formation of the OB, but these inputs are not necessary for the establishment of the central olfactory projections.

Animals↗
PubMedDate not supplied

Multidrug resistance in heart transplant patients: a preliminary communication on a possible mechanism of therapy-resistant rejection.

Multidrug resistance refers to a complex cellular phenotype, the hallmark of which is cross-resistance to multiple drugs, for example, chemotherapeutic agents, that are unrelated to the selecting agent in structure, cellular target, and mode of action. The expression of this multidrug resistance is connected with the overexpression of P-glycoprotein. By applying the method of immunocytochemical assay, we have demonstrated the appearance of the multidrug-resistant phenotype (P-glycoprotein+ cells, multidrug-resistant cells) in mononuclear cells of the peripheral blood from 32/49 patients receiving triple-drug (azathioprine, steroids, cyclosporine) immunosuppressive therapy after heart transplantation. In the group of patients showing not only the presence of cells with multidrug-resistant phenotype in the peripheral blood, but also a significant increase in the number of these cells during the interval of observation (0 to 767 days)-16/32/49 cases--a significantly increased incidence of acute rejection episodes could be demonstrated. This supports the hypothesis of a possible existence of a therapy-resistant form of acute rejection, with an involvement of mechanisms of multidrug-resistance playing a role in its causal development.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1985

Structural and histochemical study of the effect of 1,25-dihydroxyvitamin D3 on long-bone growth center in suckling mice.

The in vivo effects of 1 alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3] on cellular structure and response, matrix metachromasia and mineralization have been studied in the epiphysis and growth plate of humeri in normal neonatal mice. A relatively low dose of the metabolite, 50 ng/kg body weight, significantly increased the overall size of humeral growth plate, the zone of proliferating cells and that of hypertrophic chondrocytes. The response of the tissue to the metabolite changed with the increase in dose administered so that it was only the zone of proliferation that still showed increases in size in comparison to untreated controls. 1,25 (OH)2D3 led to an increase in the metachromatic reaction of the cartilage matrix in the chondroblastic zone, and to marked increase in matrix mineralization in the hypertrophic zone. Qualitative changes were also noted in the structure of chondroblasts and hypertrophic chondrocytes. 1,25(OH)2D3 affected the osteoblastic and osteocytic populations of cells along the metaphysis and diaphysis of the humerus. High doses of 1,25(OH)2D3 brought about distinct atrophic changes in the above cells. Chondrocytes in the epiphysis were found to synthesize type I collagen and fibronectin. These findings indicate that excessive doses of 1,25(OH)2D3 in an intact growing animal affect the normal differentiative pathway of prechondroblasts and thereby affect long bone development.

Animals↗
PubMed2006

Change in shear stress (Deltatau)/hydraulic conductivity (Lp) relationship after pronase treatment of individual capillaries in situ.

A complex glycoprotein meshwork covers the inner wall of blood vessels and is implicated in mechanotransduction of fluid shear stress (tau). A relationship between Deltatau and capillary Lp has been established. The purpose of this study was to evaluate Lp in response to Deltatau after exposing the capillary lumen to a mild, non-specific protease selected to disrupt its inner matrix. We hypothesized that Lp would not correlate with Deltatau after enzyme treatment. Frogs (Rana pipiens, n=69) were pithed and the mesentery was exteriorized. Lp was assessed at 30 cm H2O using the modified Landis technique after an abrupt, square wave Deltatau produced by a physiologically relevant increase in pressure. Perfusate solutions were 10 mg ml-1 BSA/frog Ringer's (Control) or 0.1 mg ml-1 pronase in BSA/Ringer's (1 min) then BSA/Ringer's alone (Test). Mean (+/-SE) control Lp following Deltatau was 2.2+/-0.2 x 10(-7) cm s-1 cm H2O-1 and individual values correlated positively with Deltatau (r=0.85, P<0.0001, n=41). After pronase, mean Test Lp (17.6+/-2.5 x 10(-7) cm s-1 cm H2O-1) was higher compared to control and Deltatau/Lp plots revealed two subsets of capillaries. Lp correlated strongly with Deltatau in capillaries with diameters 15 microm (r=0.96, P=0.0001, n=8). Slopes were 3.9- and 8.7-fold higher, respectively, compared to control. These data suggest a protective role for luminal constituents of intact capillaries. Mechanisms involved in capillary responses to flow-induced, mechanical stimuli may be located in the cellular structures that form capillaries.

Algorithms↗
PubMed2002

Role of subcellular shear-stress distributions in endothelial cell mechanotransduction.

The endothelium of blood vessels presents a wavy surface to the flowing blood. The subcellular distribution of shear stress depends on the shape and orientation of the cells and on their spatial arrangement within the monolayer. By studying details of the distribution of stress at this scale and the morphological responses that serve to modify the distribution, we can gain insight into the physical mechanisms by which the cell senses its fluid mechanical environment. The rapidly growing body of evidence indicates that endothelial cells discriminate between subtle variations in the exact loading conditions including differences in temporal and spatial gradients of shear stress, steady and pulsatile laminar flow, and laminar and turbulent flows. While in a few studies the effects of these individual flow characteristics have been carefully isolated, it is difficult to assess the relative importance of any one parameter. To interpret the relationships between isolated flow characteristics or the integrated effects of combined loading conditions and the biochemical signaling events that mediate the cell response, a full stress analysis of the cell is needed. The microscopic distribution of shear stress acting upon the cell surface provides the boundary condition for such an analysis. Experimental and analytical tools are being developed to assess the stress distribution throughout the cellular structures that might be involved in mechanotransduction.

Arteries↗