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Effects of level of retrieval success on recall-related frontal and medial temporal lobe activations.

Brain dedicated single photon emission computed tomography (SPECT) was used to compare the neuroactivation produced by the cued recall of response words in a set of studied word pairs with that produced by the cued retrieval of words semantically related to unstudied stimulus words. Six of the 12 subjects scanned were extensively trained so as to have good memory of the studied pairs and the remaining six were minimally trained so as to have poor memory. When comparing episodic with semantic retrieval, the well-trained subjects showed significant left medial temporal lobe activation, which was also significantly greater than that shown by the poorly trained subjects, who failed to show significant medial temporal lobe activation. In contrast, the poorly trained subjects showed significant bilateral frontal lobe activation, which was significantly greater than that shown by the well-trained subjects who failed to show significant frontal lobe activation. The frontal activations occurred mainly in the dorsolateral region, but extended into the ventrolateral and, to a lesser extent, the frontal polar regions. It is argued that whereas the medial temporal lobe activation increased as the proportion of response words successfully recalled increased, the bilateral frontal lobe activation increased in proportion to retrieval effort, which was greater when learning had been less good.

Adult↗

Mapping the polarity profiles of general anesthetic target sites using n-alkane-(alpha, omega)-diols.

The effects of the homologous series of n-alkane-(alpha, omega)-diols have been studied on the inhibition of the purified firefly luciferase enzyme from Photinus pyralis, the inhibition of the purified bacterial luciferase enzyme from Vibrio harveyi, and the induction of general anesthesia in Xenopus laevis tadpoles. All but one of the diols tested were found to be reversible general anesthetics. The diols inhibited firefly luciferase by competing with its normal substrate firefly luciferin, and they inhibited bacterial luciferase by competing with the substrate n-decanal. For all but the smallest agent (1,4-butanediol), only a single diol molecule was found to be involved in the inhibition of the enzymes. Inhibition constants Ki were determined for the enzymes, and general anesthetic EC50 concentrations were determined for tadpoles. These data were then used in conjunction with previously determined n-alkane and n-alcohol data to calculate, as a function of chain length, the incremental standard Gibbs free energies delta (delta G0) for adding apolar -CH2- groups and for converting apolar terminal -CH3 groups to polar -CH2OH groups. The resulting plots of delta (delta G0) versus chain length gave a consistent mapping of the polarity profiles of the anesthetic-binding pockets. They clearly reveal the existence of two substantial and distinct polar regions in the anesthetic-binding pocket of firefly luciferase but only one such region for bacterial luciferase and for the unknown target sites underlying general anesthesia. The polarities and geometric properties of these different binding sites for straight-chain anesthetics are discussed in terms of simple models.

Anesthetics↗

Trophectoderm growth and bilateral symmetry of the blastocyst in the mouse.

BACKGROUND: The present study was undertaken to ascertain whether the polarized flow of cells from polar to mural trophectoderm is related to the axis of bilateral symmetry of the blastocyst in the mouse, and whether trophectoderm cells can initiate new cycles once they have left the polar region. METHODS AND RESULTS: Two different approaches were used to investigate the relationship of polar to mural flow of trophectoderm cells to the bilateral axis. One was to mark peripheral polar trophectoderm cells at one or both ends of the bilateral axis in early blastocysts and examine the distribution of their clonal descendants after further growth in culture. The other was to mark the two ends of the bilateral axis with small oil drops in the zona pellucida in blastocysts whose polar trophectoderm was then labelled globally with fluorescent latex microspheres before culture. In both cases, marking of additional blastocysts orthogonal to the bilateral axis was also done. The results show that the direction of polar to mural flow of cells is not random, and that the most distal mural trophectoderm cell could yield up to eight descendants during 45 h of culture. CONCLUSION: The findings are consistent with the polar to mural flow of trophectoderm cells being aligned with the bilateral axis. Moreover, trophectoderm cells can embark on new cycles even when remote from the inner cell mass.

Animals↗

Immunocytochemical localization of actin in Toxoplasma gondii.

The localization of actin in the trophozoites of Toxoplasma gondii was examined by means of immunogold staining for electron microscopy. The thin-sectioned specimens were incubated with the IgG fraction of polyclonal antibodies raised against Ascaris body-wall smooth-muscle actin following colloidal gold-conjugated protein A. Electron-dense gold particles were confined to the anterior polar region in the trophozoites: they were found in the conoid, preconoidal rings, possibly in the polar ring, and in the space between the anterior terminal of the inner membrane complex and conoid. The present experiments also suggest interactions of actin with subpellicular microtubules, leading to speculation that the association of actin with microtubules provides a link to myosin, a potential source of power for microtubule-dependent movements.

Actins↗

A model for astral stimulation of cytokinesis in animal cells.

A model is proposed in which stimulation of cortical cytoplasm occurs near the distal ends of astral rays. Levels of stimulation sufficient to cause furrowing occur only in equatorial zones between asters. The model can account for positioning of furrows in very large cells (fertilized eggs of amphibians, birds, and fish) and in cells with several mitotic apparatuses (insects). Finally, the model correctly predicts the positioning and occurrence of furrowing in two experiments in which cellular shape was manipulated into either an hourglass or a cylindrical form before division. These results are consistent with equatorial stimulation theories in which mitotic asters differentially stimulate the future furrow region (equatorial cortex). The results are not consistent with models requiring differential stimulation of nonfurrowing, polar regions of the cell.

Animals↗

Fine structural changes of Acetobacter suboxydans during growth in a defined medium.

Cytological differences were observed between stationary- and exponentialphase cells of Acetobacter suboxydans grown in a defined medium. Unstained cells observed with the light microscope just after entering the stationary phase differed from exponentially growing cells in that the former exhibited localized increases in density, particularly in the polar regions. Electron microscopy of thin sections revealed that early stationary-phase cells possessed predominantly polar complexes of intracytoplasmic membranes accompanied by polar increases in ribosomal material. When cultures were allowed to continue far into the stationary phase, cells contained extensive aggregations of membrane-like material as the predominant fine-structural feature. In contrast, thin sections of exponentially growing cells exhibited only occasional indications of intracytoplasmic membranes. Intracytoplasmic membranes heretofore have been observed only rarely in the heterotrophic Pseudomonadales.

Acetobacter↗

A reexamination of multiaxonal nerve endings innervating intrafusal muscle fibers of the Chinese hamster.

The multiaxonal nerve endings, consisting of a bundle of axons, were reexamined in the muscle spindles of the adult Chinese hamster. In addition to the frequent occurrence of the multiaxonal endings innervating the nuclear bag fibers in the equatorial region (Desaki and Uehara, 1988), these endings were also occasionally distributed not only in the equatorial region of the nuclear chain fibers, adjacent to annulospiral sensory endings, but also in the polar region of the same muscle fiber. They were derived from non-myelinated stem nerves becoming varicose along their course. Terminal axons composing the endings were almost naked, except at the initial muscle-contact portion, where they were surrounded by Schwann cell processes. They contained a significant number of clear synaptic vesicles and large granulated vesicles, and were closely apposed to each other and to the muscle cell surface without an interposing basal lamina. Some terminal axons formed synaptic contacts with each other and with the muscle cell surface. These structural features, including their wide distribution, suggest that the multiaxonal endings may belong to the autonomic nerves rather than the fusimotor nerves.

Animals↗

Permeability studies in neonatal rat lens epithelium.

Lanthanum nitrate (LN) and horseradish peroxidase (HRP) were used as tracers to study intercellular permeability as well as the existence and location of tight junctions and changes in them, if any, in the lens epithelium of Sprague Dawley rats. Thin sections of lenses taken from animals at birth and at three day intervals until the neonates were weaned at approximately 22 days were studied at the electron microscope level. At every age both tracers permeated the intercellular spaces of the anterior region epithelium and between the fiber cells of the equatorial region. The LN precipitates terminated near the apical part of the epithelium in the anterior polar region and were not seen at the epithelial-fiber interface or between fibers in this region. At the site of the tracer termination the intercellular space was found to be considerably constricted or completely sealed. The HRP precipitates, however, were present in these intercellular spaces and by using the HRP "washout" procedure could be washed out of the basolateral site of the intercellular spaces of the epithelium up to the point of constriction if a tight junction were present. The lens area occupied by epithelium with tight junctions was age related being very small in lenses from animals less than 24 hours old and becoming increasingly enlarged as the newborn aged. These findings further confirm the existence of a barrier in the form of tight junctions between epithelial cells in the central anterior region of the maturing rat lens. Moreover, our observations also provide information regarding the pattern of tight junction development and distribution during lens maturation.

Animals↗

The talpid2 chick limb has weak polarizing activity and can respond to retinoic acid and polarizing zone signal.

The talpid2 (ta2) chick mutant has wide, polydactylous wings and legs. Talpid2 limb cartilages have abnormal morphology and a very subtle anteroposterior polarity. Specifically, posterior ta2 limb structures are identifiable, while more anterior cartilages are less distinctive. Here, we investigate the development of anteroposterior limb pattern in the ta2 embryo. We show that ta2 posterior limb bud mesoderm is capable of respecifying the anteroposterior axis of a normal wing. However, the average duplication obtained after grafting a ta2 polarizing region was significantly less than the average duplication formed after a graft of normal wing bud polarizing zone. Thus, polarizing activity appears to be weak in ta2. Grafts of normal polarizing zone to the posterior edge of ta2 wing buds had no effect on the ta2 phenotype. This result suggests that a weakly functioning polarizing signal does not account for the altered anteroposterior polarity in ta2 limbs, and that normal polarizing zone activity is not sufficient for formation of normal limb bud cartilages. We demonstrated that transmission of a polarizing signal through the ta2 limb mesoderm was normal. In addition, ta2 anterior border mesoderm had no polarizing activity. We also assessed the ability of ta2 limb bud mesoderm to respond to a polarizing signal. Either normal polarizing zone tissue or a bead containing retinoic acid was placed at the anterior edge of ta2 wing buds at stages 18-23. Both polarizing zone and retinoic acid caused respecification of the ta2 wing anteroposterior axis. The result was that a ta2 ulna replaced the radius, and the most posterior digit was duplicated anteriorly. Limb cartilages with normal morphology never formed. When a bead containing retinoic acid was placed at the posterior edge of ta2 wing buds, there was no effect on anteroposterior pattern. However, beads with retinoic acid always caused a reduction in the number of ta2 wing digits which formed, whether the beads were placed at the anterior or posterior edge of the developing ta2 limb.

Animals↗

An ultrastructure investigation of intrafusal muscle fibers in myotonic dystrophy.

Ultrastructure features of muscle spindles from two patients with myotonic dystrophy are described. Intrafusal muscle fibers exhibit extensive splitting with nuclear bag fibers affected more so than nuclear chain fibers. No sensory endings are present on nuclear chain fibers nor on one nuclear bag fiber throughout the equatorial and myotube regions. Small motor end plates are evident on various segments of split intrafusal fibers in the polar region and some of these extend into the myotube region. Satellite cells are numerous on both nuclear bag and nuclear chain fibers. These frequently occupy the cleft space between segments of split intrafusal fibers. The myotonic dystrophy muscle spindle ultrastructure features seem to closely resemble the appearance of developing mammalian muscle spindles as illustrated with opossum fetal tissue.

Adult↗

Fluorescence polarization study on the dynamics and location of peroxidized fluorescent phospholipids in liposomes.

Motional properties of fluorescent substances produced by lipid peroxidation by a time-resolved fluorescence polarization technique were studied. When liposomes containing phosphatidylethanolamine (PE) and linoleic hydrocarbon chain were incubated at 37 degrees C, fluorophores absorbing maximally at 360 nm and emitting near 430 nm were produced. Their fluorescence anisotropy decay measured at 23 degrees C was fitted well with a sum of a fast relaxation and a time-independent residual term. With the increase of oxidation degree, the time constant of the relaxation term increased. This may be explained by alteration in the membrane structure or by modification of the fluorescent products themselves. Information on the location of the fluorescent products was obtained when their motional property was compared with those of various extrinsic probes that were incorporated at different positions of the lipid bilayer. It was found that the motional property of the fluorescent oxidation products is similar to that of 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene, a rod-shaped hydrophobic probe with a charged terminal. Other probes sensing the polar region or the hydrophobic region of the membrane were characterized by a lower order parameter. It is suggested that the fluorescent oxidation products have a polar moiety located at the membrane surface and attached to the amino group of PE while the tail part being buried in the hydrophobic region of the membrane. This picture is supported by fluorescence quenching experiments with the aqueous quencher Co2+. On the other hand, fluorophores produced by the reaction of malondialdehyde and PE suggested to have a chemical structure in which the angle between the absorption and emission dipole moments is very large. On the basis of these observations, the production pathway of fluorophores in oxidized membranes is discussed.

Fluorescence Polarization↗

Intracytoplasmic membrane formation and increased oxidation of glycerol growth of Gluconobacter oxydans.

Gluconobacter oxydans is well known for the limited oxidation of compounds and rapid excretion of industrially important oxidation products. The dehydrogenases responsible for these oxidations are reportedly bound to the cell's plasma membrane. This report demonstrates that fully viable G. oxydans differentiates at the end of exponential growth by forming dense regions at the end of each cell observed with the light microscope. When these cells were thin sectioned, their polar regions contained accumulations of intracytoplasmic membranes and ribosomes not found in undifferentiated exponentially growing cells. Both freeze-fracture-etched whole cells and thin sections through broken-cell envelopes of differentiated cells demonstrate that intracytoplasmic membranes occur as a polar accumulation of vesicles that are attached to the plasma membrane. When cells were tested for the activity of the plasma membrane-associated glycerol dehydrogenase, those containing intracytoplasmic membranes were 100% more active than cells lacking these membranes. These results suggest that intracytoplasmic membranes are formed by continued plasma membrane synthesis at the end of active cell division.

Cell Membrane↗

Partitioning and localization of spin-labeled amantadine in lipid bilayers: an EPR study.

EPR was used to study the distribution of the spin-labeled amantadine (AA-SL) between the bulk hydrophobic-hydrocarbon solvent, light paraffin oil, and water and between hydrophobic-hydrocarbon chain region of lipid membranes and water. The AA-SL molecules were soluble in both hydrophobic and polar regions of investigated systems. It was shown that the partition coefficient of AA-SL between the hydrocarbon region of the L-alpha-dimyristoylphosphatidylcholine fluid-phase membrane and water is much higher than between bulk hydrocarbon solvent and water. Furthermore, the partitioning of AA-SL into membranes of multilamellar liposomes made of L-alpha-dimyristoylphosphatidylcholine, L-alpha-dipalmitoylphosphatidylcholine, and L-alpha-distearoylphosphatidylcholine was studied as a function of temperature, indicating no abrupt change at the main phase transition of these membranes. It is also clear from our data that AA-SL can penetrate into the gel-phase membrane practically with the same partitioning as into the fluid-phase membrane. Furthermore, it was shown that at least part of the AA-SL molecule is deeply buried in the hydrocarbon chain region of the membrane.

Amantadine↗

X chromosome retains the memory of its parental origin in murine embryonic stem cells.

A cytogenetic and biochemical study of balloon-like cystic embryoid bodies, formed by newly established embryonic stem (ES) cell lines having a cytogenetically or genetically marked X chromosome, revealed that the paternally derived X chromosome was inactivated in the majority of cells in the yolk sac-like mural region consisting of the visceral endoderm and mesoderm. The nonrandomness was less evident in the more solid polar region containing the ectodermal vesicle, mesoderm and visceral endoderm. Since the same was true in embryoid bodies derived from ES cells at the 30th subculture generation, it was concluded that the imprinting responsible for the preferential inactivation of the paternal X chromosome that was limited to non-epiblast cells of the female mouse embryos, was stably maintained in undifferentiated ES cells. Differentiating epiblast cells should be able to erase or avoid responding to the imprint.

Animals↗

[Distribution of small hydrophobic molecules in membranes. I. Artificial lipid membranes].

A hydrophobic uncharged fluorescent probe of 4-dimethylaminochalcone (DMC) interacted with synthetic phospholipid membranes. Comparison of absorption spectra and fluorescence of DMC in the membranes and organic solvents shows that in the membranes the DMC molecules are located not in the hydrocarbon layer but in the polar regions near the surface. The probe is distributed regularly along the surface forming no dimers and clusters. Polar groups which surround the probe in the membrane are less mobile than the molecules of organic solvents at the same temperature. The evaluation shows that the relaxation time of polar groups in the probe environment is longer than 0.15-10(-9) sec. The DMC molecules may be located in different sites of the membrane surface, which seem to differ from one another in the mobility of polar groups.

Chalcone↗

The effect of interfacial parameters on cup-bone relative micromotions. A finite element investigation.

Achieving stability is a prerequisite for allowing bone to grow into the porous surface of non-cemented acetabular cups. The purpose of this study is to estimate the effects of interfacial characteristics on relative cyclical micromotion between cup and bone during gait in the immediate postoperative phase. The technique used is finite element analysis. Six models with different interfacial characteristics are created in order to study the effects of fixation technique. These include representation of a 1 mm press-fit, 2 mm press-fits (with and without an initial polar gap) and exact-fit conditions (with and without additional screw fixation). Although direct validation of the model has not been performed, the calculated micromotions under a static load of 1112 N are compared with appropriate experimental data. Generally, the model tends to underestimate micromotion and this underestimate is significant in the case of relative surface-normal micromotion in polar regions for models with low- and no-interference. The most likely cause of this significant underestimate is a failure of the model to accurately represent penetration of rough contacting surfaces under compression. Other types of micromotion, although low, are within standard deviations reported by Kwong et al. (1994 Journal of Arthroplasty 9, 163-170). Quasi-static joint contact and muscle forces, representative of the stance phase of gait are then applied and maximum micromotions are found to occur consistently prior to toe off: this being the point of maximum force. With regard to the press-fit simulations, good cup-bone contact in the superior region of the interface is required for stability and the greatest micromotions occur in the models with the larger interference and larger polar gaps. In contrast to the press-fit models, muscle activity in exact-fit models influences the calculations. Specifically, the early activity of m.semimembranosus modelled causes opening of the peripheral seal. Taken together it is found that polar gaps reduce the stability of the model and lack of pre-compresssion in the periphery allows this region of the interface to be opened up.

Acetabulum↗

[Histochemical study of the sensory endings of muscle spindles in rat longissimus muscles].

Most studies concerning the structure and function of muscle spindles have utilized the hind limbs of experimental animals. However, little is known about muscle spindles of the back muscles. The purpose of this study was to investigate the sensory innervation of muscle spindles of the paravertebral muscle in the rat. The subjects were 10 normal male rats. The longissimus muscles were isolated and frozen in cooled isopentane (-160 degrees C), and serial transverse sections were made with a cryostat. Histochemical preparations were then made using nicotinamide adenine dinucleotide tetrazolium reductase (NADH-TR) stain and modified Gomori-trichrome stain. The muscle spindles in each segment were identified microscopically by observing the equatorial and polar regions. NADH-TR staining was employed to distinguish nuclear bag1, nuclear bag2, and nuclear chain intrafusal muscle fibers. A total of 20 spindle poles were surveyed. The mean polar length of intrafusal fibers as well as that of each region (A, B, and C) were measured. NADH-TR staining also demonstrated the terminal sites of sensory fibers along intrafusal fibers. All spindle poles surveyed were innervated by secondary sensory fibers in addition to primary sensory fibers. Eight spindle poles were intermediate type muscle spindles that were innervated by one primary sensory fiber and one secondary sensory fiber. Twelve spindle poles were complex type muscle spindles that were innervated by one primary sensory fiber and multiple secondary sensory fibers. The mean length of the A region was 223.1+/-37.9 microm (n=8) for intermediate type spindles and 493.8+/-157.0 microm (n=12) for complex type spindles. The length of the A region was significantly longer in the complex type spindles than in the intermediate type spindles (p<0.001). The results suggest that the innervations of secondary sensory fibers were well developed in the longissimus muscle spindles in the rat. The morphological features of muscle spindles of the longissimus muscle may represent the structural basis for qualitatively different afferent discharges that relate to the characteristic types of locomotion served by paravertebral muscles.

Animals↗

S-100 protein-induced changes in the physical state of synaptosomal particulate fractions as monitored by spin labels.

This report documents changes in the physical state of synaptosomal particulate fractions (SYN) upon binding of S-100 protein, as monitored by spin labels. Studies were conducted on SYN labeled with either 5-doxylstearic acid or 16-doxylstearic acid, which probe the polar region and the hydrophobic core of the lipid bilayer, respectively. S-100 perturbs to some extent both the polar surface and the hydrophobic core of SYN in a time- and temperature-dependent manner. Ca2+ is essential for S-100 to perturb the membranes. K+ almost completely inhibits the S-100 perturbing effect if present in the incubation medium, but fails to reverse the S-100-induced changes if added after S-100 has interacted with SYN. At room temperature and below, the overall S-100 effect registered after about 30 min of association of the protein with SYN is an increase in the fluidity of both the surface and the interior of the membranes. Spectra registered at intervals at room temperature indicate that the S-100 perturbing effect on the membrane surface is practically monophasic, consisting of an increase in fluidity, while that on the membrane interior is multiphasic, consisting of a decrease in fluidity during the first 10 min of association, followed by an increase in fluidity during the subsequent 20 min and a return to starting values during the second 30 min of association. Around 37 degrees C, on the contrary, a decrease in fluidity is registered in both regions. The data suggest that S-100 induces a spatial rearrangement of membrane components (proteins) involved in the specific binding and/or partially penetrates into the lipid bilayer.

Animals↗