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Electrostatic Interaction between Two Ion-Penetrable Charged Spheroids

The electrostatic interaction between two ion-penetrable, charged spheroidal particles is examined theoretically. These particles can assume different sizes and an arbitrary spatial orientation. The electrical potential distribution is derived analytically under the Debye-Huckle condition. The results for two interaction spheres, one spheroidal particle and a planar surface, and rigid particles covered by an ion-penetrable membrane can be recovered as the special cases of the present general problem. We show that, for a fixed center-to-center distance between two particles, regardless of their relative sizes, the interaction free energy is the greatest if their major axes lie on the same line (head-to-head), and the smallest if their major axes are perpendicular to each other but not on the same plane (perpendicular).

Journal Article↗

The nuclear magnetic resonance solution structure of flavoridin, an antagonist of the platelet GP IIb-IIIa receptor.

The snake venom protein flavoridin, a polypeptide of 70 amino acid residues, is a potent inhibitor of blood platelet aggregation. It binds to cell-surface integrin receptors such as the fibrinogen receptor glycoprotein IIb/IIIa. The inhibitory properties of flavoridin have been attributed to the tripeptide segment Arg-Gly-Asp (residues 49 to 51). This paper describes the determination of the three-dimensional structure of flavoridin in aqueous solution based on two-dimensional nuclear magnetic resonance spectroscopy. A family of 18 conformers was selected to characterize the solution structure. The molecule comprises two structural domains, an N-terminal unit extending from residues 1 to 25, and a C-terminal unit from residues 26 to 70. Whereas the mutual spatial orientation of these regions is not well defined, each one is well organized within itself. The segment 26 to 70, which is homologous to the sequence of the snake toxins echistatin and eristostatin, shows an average value of 1.0 A for the root-mean-square deviations of the backbone atoms among the 18 conformers. The structure of flavoridin consists essentially of non-repetitive elements such as tight turns and loops, whose location and conformation are characterized in this paper. With the exception of two short regions of antiparallel beta-sheet, no classic element of protein secondary structure is present. The six disulphide bridges, which have been mapped by applying a novel computational strategy (see accompanying paper), are the dominant organizational feature of the polypeptide fold of flavoridin. Two of the bridges are located in the N-terminal domain, three in the C-terminal domain and one connects the two structural units. The mobile RGD recognition sequence for integrins is located peripheral to the core region of the C-terminal domain at the most exposed end of a nine residue loop structure, which is attached to a short beta-sheet. The C terminus is close to this loop structure.

Amino Acid Sequence↗

Collagen fibril structure in lamprey.

X-ray diffraction and electron microscopy are used to compare the molecular and higher order structure of collagen fibrils in three tissues of the lamprey: the dermis, perinotochord and notochord sheath. These lamprey tissues are known to contain five distinct genetic types of fibrillar collagen. The modes of axial and lateral packing of collagen molecules in fibrils of the lamprey tissues demonstrate the three major motifs seen in higher vertebrate D-periodic collagen fibrils. In particular, lamprey dermis was found to have a decreased D period resulting from a molecular tilt that is seen in skins of higher vertebrates. Our results suggest the molecular-packing motifs for collagen fibrils were in place at the dawn of vertebrate evolution and have been conserved since. In contrast, the diameters of fibrils and their spatial orientation in lamprey tissues do not in general, correspond to features found in mammalian tissues. Only for lamprey notochord is there a strong similarity, with fibril diameters and organization closely resembling those seen in type II tissues of higher vertebrates. This suggests that for all tissues except those with type II collagen, higher level organization of fibrils evolved along with the diversification of vertebrates.

Animals↗

Peptide carriers: A helicoid-type sequential oligopeptide carrier (SOC(n)) for multiple anchoring of antigenic/immunogenic peptides.

A new peptide carrier with three-dimensional predetermined structural motif has been constructed by the repetitive Lys-Aib-Gly moiety. The sequential oligopeptide carrier (SOC(n)), (Lys-Aib-Gly)(n), adopts a distorted 3(10)-helical conformation and the Lys-N(epsilon)H(2) anchoring groups exhibit defined spatial orientations. Conformational analysis of the SOC(n) conjugates showed that the coupled peptides retain their initial "active" structure, while prevalence of one conformer was also observed. It is concluded that the beneficial structural elements of SOC(n) induce a favorable arrangement of the conjugated peptides, so that potent antigens and immunogens are generated.

Amino Acid Sequence↗

Human vestibular cortex as identified with caloric stimulation in functional magnetic resonance imaging.

Anatomic and electrophysiological studies in monkeys have yielded a detailed map of cortex areas receiving vestibular afferents. In contrast, comparatively little is known about the cortical representation of the human vestibular system. In this study we applied caloric stimulation and fMRI to further characterize human cortical vestibular areas and to test for hemispheric dominance of vestibular information processing. For caloric vestibular stimulation we used cold nitrogen to avoid susceptibility artifacts induced by water calorics. Right and left side vestibular stimulation was repetitively performed inducing a nystagmus for at least 90 s after the end of the stimulation in all subjects. Only the first 60 s of this nystagmus period was included for statistical analysis and compared with the baseline condition. Activation maps revealed a cortical network with right hemispheric dominance, which in all subjects comprised the temporoparietal junction extending into the posterior insula and, furthermore, the anterior insula, pre- and postcentral gyrus, areas in the parietal lobe, the ventrolateral portion of the occipital lobe, and the inferior frontal gyrus extending into the inferior part of the precentral sulcus. In conclusion, caloric stimulation in fMRI reveals a widespread cortical network involved in vestibular signal processing corresponding to the findings from animal experiments and previous functional imaging studies in humans. Furthermore, this study demonstrates a strong right hemispheric dominance of vestibular cortex areas regardless of the stimulated side, consistent with the current view of a rightward asymmetrical cortical network for spatial orientation.

Adult↗

Dissociable neural pathways are involved in the recognition of emotion in static and dynamic facial expressions.

Facial expressions of emotion powerfully influence social behavior. The distributed network of brain regions thought to decode these social signals has been empirically defined using static, usually photographic, displays of such expressions. Facial emotional expressions are however highly dynamic signals that encode the emotion message in facial action patterns. This study sought to determine whether the encoding of facial expressions of emotion by static or dynamic displays is associated with different neural correlates for their decoding. We used positron emission tomography to compare patterns of brain activity in healthy men and women during the explicit judgment of emotion intensity in static and dynamic facial expressions of anger and happiness. Compared to judgments of spatial orientation for moving neutral facial expressions, the judgment of anger in dynamic expressions was associated with increased right-lateralized activity in the medial, superior, middle, and inferior frontal cortex and cerebellum, while judgments of happiness were associated with relative activation of the cuneus, temporal cortex, and the middle, medial, and superior frontal cortex. In contrast, the perception of anger or happiness in static facial expressions activated a motor, prefrontal, and parietal cortical network previously shown to be involved in motor imagery. The direct contrast of dynamic and static expressions indicated differential activation of visual area V5, superior temporal sulcus, periamygdaloid cortex, and cerebellum for dynamic angry expressions and differential activation of area V5, extrastriate cortex, brain stem, and middle temporal cortical activations for dynamic happy expressions. Thus, a distribution of neural activations is related to the analysis of emotion messages in the nearly constant biological motion of the face and differ for angry and happy expressions. Static displays of facial emotional expression may represent noncanonical stimuli that are processed for emotion content by mental strategies and neural events distinct from their more ecologically relevant dynamic counterparts.

Adult↗

A behavioral study of alpha-1b adrenergic receptor knockout mice: increased reaction to novelty and selectively reduced learning capacities.

Knockout mice lacking the alpha-1b adrenergic receptor were tested in behavioral experiments. Reaction to novelty was first assessed in a simple test in which the time taken by the knockout mice and their littermate controls to enter a second compartment was compared. Then the mice were tested in an open field to which unknown objects were subsequently added. Special novelty was introduced by moving one of the familiar objects to another location in the open field. Spatial behavior and memory were further studied in a homing board test, and in the water maze. The alpha-1b knockout mice showed an enhanced reactivity to new situations. They were faster to enter the new environment, covered longer paths in the open field, and spent more time exploring the new objects. They reacted like controls to modification inducing spatial novelty. In the homing board test, both the knockout mice and the control mice seemed to use a combination of distant visual and proximal olfactory cues, showing place preference only if the two types of cues were redundant. In the water maze the alpha-1b knockout mice were unable to learn the task, which was confirmed in a probe trial without platform. They were perfectly able, however, to escape in a visible platform procedure. These results confirm previous findings showing that the noradrenergic pathway is important for the modulation of behaviors such as reaction to novelty and exploration, and suggest that this is mediated, at least partly, through the alpha-1b adrenergic receptors. The lack of alpha-1b adrenergic receptors in spatial orientation does not seem important in cue-rich tasks but may interfere with orientation in situations providing distant cues only.

Animals↗

Characterization of a porcine enterocyte receptor for group A rotavirus.

We have identified, purified to apparent homogeneity and chemically characterized a biologically-relevant porcine enterocyte receptor for group A porcine rotavirus. Ceramide glycanase digestion followed by acid hydrolysis and monosaccharide compositional analyses indicated the receptor is a family of two GM, gangliosides, one containing N-glycolyl-neuraminic acid and the other N-acetylneuraminic acid. Both gangliosides displayed dose-dependent inhibition of rotavirus binding to, and infectivity of, host cells. Inhibition of infectivity in a focus-forming-unit-reduction assay was achieved with as little as 2 nmols of NeuGcGM3 (50% inhibition with 3.97 nmol) or NeuAcGM3 (50% inhibition with 9.84 nmol) per 10(4) FFU of virus. Preliminary data suggest specific porcine GM3 carbohydrate fine structure or spatial orientation of the sialyloligosaccharide epitopes of the holoGM3 gangliosides may be crucial to enterocyte receptor recognition by rotavirus. We have quantified both NeuGcGM3 and NeuAcGM3 in enterocytes of various-aged pigs from newborn through 16 weeks and have found with increasing age the amount of both GM3 derivatives, especially NeuGcGM3 per gram (dry weight) intestinal brush border decreases rapidly from newborn through 4 weeks of age. These results may help explain the age-sensitivity of piglets to severe rotavirus diarrhea.

Age Factors↗

Immunohistochemical detection of proliferating cell nuclear antigen in middle ear cholesteatoma.

Cholesteatoma epithelium is characterized by a keratinocyte dysregulation accompanied by destruction of the ossicles and temporal bone. Immunohistochemical methods using antibodies to cell-cycle-related antigens can be used as a means for assessing various aspects of proliferation in cholesteatoma tissue. They also have the important advantage of preserving the spatial orientation of proliferating cells in histological sections. Proliferating cell nuclear antigen (PCNA) is a 36 kDa DNA-delta-polymerase-associated protein that is directly involved in the mechanisms of DNA synthesis. In the present study the expression of PCNA was investigated in formalin-fixed, paraffin-embedded biopsy specimens of cholesteatomas and normal skin. Normal skin revealed nuclear staining in a small number of keratinocytes (PCNA grade, 1.5) located in the basal cell layer. In contrast, an increased number of PCNA-labeled basal and suprabasal epidermal cells (PCNA grade, 9.3) were found in cholesteatoma samples. Our findings indicate that PCNA represents a reliable marker for epithelial proliferation, showing that cholesteatoma epithelium proliferates at a higher rate than normal epidermis. These findings also support the concept of keratinocyte dysregulation in middle ear cholesteatoma.

Cell Division↗

Afferent nerve endings in the tracheal muscle of guinea-pigs and rats.

The trachea of guinea-pigs was stained as a whole-mount preparation with the zinc iodide-osmium technique. A distinct class of nerve endings was observed associated with the tracheal muscle. The endings, issued from myelinated fibres of the vagus nerve via the recurrent laryngeal nerve, are distributed on either side of the midline and ventral to the tips of cartilages. They are interpreted as afferent nerve endings that may correspond to slow adapting stretch receptors identified by physiological studies. Each nerve contributes predominantly, but not exclusively, to the receptors of the ipsilateral side. There are 120-180 receptors along the full length of the guinea-pig trachea, their density being higher at the cranial end. The receptors are variable in size and structural complexity, and, to some extent, also in spatial orientation, but distinct subtypes are not recognizable. Receptors of similar morphology and distribution are found also in the rat trachea. The receptors can also be visualized with a cytochrome oxidase method for nerve endings, but they do not stain with immunohistochemistry for the neuropeptides substance P, calcitonin gene-related peptide, vasointestinal polypeptide and neurotensin.

Animals↗

Temporal analysis of adaptation in moth (Trichoplusia ni) pheromone receptor neurons.

The temporal pattern of response in chemoreceptor neurons reflects both the temporal distribution of stimuli and the timing of signal transduction, action potential generation and propagation. Here we analyze the temporal characteristics of the responses elicited in pheromone receptor neurons by computer-controlled rectangular pulses of odorant. Extracellular recordings from the HS sensilla trichodea on the antenna of male Trichoplusia ni reveal the activity of two neurons: the "A" neuron, which responds to the major component of the female pheromone blend, (Z)7-dodecenyl acetate and the "B" neuron, which responds to (Z)7-dodecenol. "B" neurons were divided into two classes (HR, LR), based on the magnitude and temporal pattern of their response to (Z)7-dodecenol. Most "A" and HR "B" neurons responded to rectangular pulses of various durations (0.1-40 s) with an initial phasic burst (approximately 100 ms), followed by a slowly declining tonic component. At moderate and elevated pheromone doses, prolonged stimulation resulted in significant reductions in the tonic response levels (adaptation); stimuli of increasing duration effected greater adaptation. Most LR "B" neurons lacked a phasic response component and showed virtually no adaptation with prolonged stimulation. Pheromone receptor neurons may differ in both their spectral and temporal response properties which may provide the animal with additional sensory information for blend discrimination and spatial orientation in complex natural pheromone plumes. The potential functional value of adaptation in the moth pheromone communication system is discussed.

Action Potentials↗

Negative gravitropism in Chara protonemata: a model integrating the opposite gravitropic responses of protonemata and rhizoids.

The unicellular protonema of Chara fragilis Desv. was investigated in order to establish a reaction chain for negative gravitropism in tip-growing cells. The time course of gravitropic bending after stimulation at angles of 45 degrees or 90 degrees showed three distinct phases of graviresponse. During the first hour after onset of stimulation a strong upward shift of the tip took place. This initial response was followed by an interval of almost straight growth. Complete reorientation was achieved in a third phase with very low bending rates. Gravitropic reorientation could be completely abolished by basipetal centrifugation of the cells, which lastingly removed conspicuous dark organelles from the protonema tip, thus identifying them as statoliths. Within minutes after onset of gravistimulation most or all statoliths were transported acropetally from their resting position 20-100 micrometers from the cell apex to the lower side of the apical dome. This transport is actin-dependent since it could be inhibited with cytochalasin B. Within minutes after arrival of the statoliths, the apical dome flattened on its lower side and bulged on the upper one. After this massive initial response the statoliths remained firmly sedimented, but the distance between this sedimented complex and the cell vertex increased from 7 micrometers to 22 micrometers during the first hour of stimulation and bending rates sharply declined. From this it is concluded that only statoliths inside the apical dome convey information about the spatial orientation of the cell in the gravitropic reaction chain. After inversion of the protonema the statoliths transiently arranged into a disk-shaped complex about 8 micrometers above the vertex. When this statolith complex tilted towards one side of the apical dome, growth was shifted in the opposite direction and bending started. It is argued that the statoliths intruding into the apical dome may displace a growth-organizing structure from its symmetrical position in the apex and may thus cause bending by bulging. In the positively gravitropic Chara rhizoids only a more stable anchorage of the growth-organizing structure is required. As a consequence, sedimented statoliths cannot dislocate this structure from the vertex. Instead they obstruct a symmetrical distribution of cell-wall-forming vesicles around the structure and thus cause bending by bowing.

Actin Cytoskeleton↗

Outgrowth in vitro of cervical epithelium separated from the uterovaginal anlage of newborn mice. An ultrastructural study.

After gently trypsinization, the pseudostratified columnar Müllerian epithelium that lines the uterine cervix of newborn mice could be separated from the enclosing stromal tissue Pure epithelial tubes explanted in vitro and were allowed to grow in a standard medium for 3-4 days forming a confluent colony of rather closely-fitting cells. The cell sheet was studied by a preparatory technique that allows examination of a large number of cells with preserved intercellular spatial orientation. Attempts were made to identify cultured cells according to the morphology of cell types in the cervicovaginal epithelium in vivo. Electron micrographs revealed that, close to the explant, the cultured cell sheet exhibited several features similar to the Müllerian epithelium in vivo. Outside these central areas of the colony was a broad transitional zone consisting of thin platelike cells distinguished by an abundance of microfilaments. At the periphery of the colonies, bulky cells possessing microvilli and a vacuolated cytoplasm tended to overlpa adjoining platelike cells. These bulky cells had a morphology resembling that of the superficial cells seen in the upper vagina and common cervical canal of immature and diestrous animals. The epithelial development in the cultures apparently simulated the transformation iatified epithelium resembling that of the uppermost vagina and common cervical canal of immature animals. Judged by morphological and cytochemical criteria, the Müllerian cells in the out-growth obviously had many changed features. It thus seems questionable wheter the cells grown in vitro are comparable with the corresponding cells in vivo when used for experiments requiring the corresponding of the culture environment.

Animals↗

Development of the filiform hairs on the cerci of Gryllus bimaculatus deg. (saltatoria, gryllidae).

The filiform hairs, mechanoreceptors of Gryllus, pass through six developmental stages during the last larval stage. The cytoplasm of their sense cells suggests intensive synthesis of protein for cellular metabolism and intercytoplasmic exchange of material via glial evaginations. Ultrahistochemical tests demonstrated acid phosphatase in the lysosomes as well as in components of the Golgi apparatus. There was no significant change in the appearance of the sense cell cytoplasm, indicating a maintained functional state also during molting. The new cuticular apparatus is formed after apolysis by the three enveloping cells. Formation of the replacement hairs is initiated by a cytoplasmic outgrowth of the trichogen cell. During morphogenesis of the new hair, the microtubules serve as a cytoskeleton and probably control the flow of vesicles, which contain phenol oxidase, also demonstrated in the Golgi apparatus, and are incorporated into the new cuticle. Bundles of microfibrils are involved in the surface sculpturing of the replacement hair. The trichogen cell also forms a number of structural elements, e.g. the "cup" and "strut" marked geometric peculiarities of which indicate that they are important in the spatial orientation of the dendrite and thus also in transduction. Reduction of the apical cell membrane of the tormogen cell after apolysis permits unrestricted growth of the new hair into the exuvial space. The tormogen cell participates in the formation of the joint membrane, parts of the socket and the articulation of the hair.

Acid Phosphatase↗

Catalytic and ligand binding properties of bovine trypsinogen and its complex with the effector dipeptide Ile-Val. A comparative study.

Steady-state and pre-steady-state kinetic data for the trypsinogen catalyzed hydrolysis of a series of synthetic substrates (i.e. p-nitrophenyl esters of N-alpha-carbobenzoxy-L-amino acids) have been obtained as a function of pH (3.4-8). Moreover, the effect of ethylamine on the hydrolysis of a neutral substrate and benzamidine binding have been extensively studied. In order to obtain direct information on the transition of trypsinogen to a beta-trypsin-like structure, the role of the effector dipeptide Ile-Val on the catalytic and ligand binding properties of the zymogen has been investigated. Kinetic and thermodynamic data for beta-trypsin and alpha-chymotrypsin are also reported for the purpose of an homogeneous comparison of the various (pro)enzymes. Under all the experimental conditions, kinetic data for (pro)enzyme catalysis are consistent with the minimum three-step mechanism: (formula; see text) involving the acyl intermediate E X P. In the presence of Ile-Val dipeptide, trypsinogen assumes catalytic and ligand binding properties that are reminiscent of activated beta-trypsin. This is at variance with free trypsinogen, which shows a alpha-chymotrypsin-like behavior. The large differences in the results of kinetic and thermodynamic measurements for free trypsinogen, as compared to its binary adduct with Ile-Val, can be ascribed to the substantial differences in the two molecular species, which include the spatial orientation of Asp189.

Animals↗

A neuronal correlate of spatial stability during periods of self-induced visual motion.

Motion of background visual images across the retina during slow tracking eye movements is usually not consciously perceived so long as the retinal image motion results entirely from the voluntary slow eye movement (otherwise the surround would appear to move during pursuit eye movements). To address the question of where in the brain such filtering might occur, the responses of cells in 3 visuo-cortical areas of macaque monkeys were compared when retinal image motion of background images was caused by object motion as opposed to a pursuit eye movement. While almost all cells in areas V4 and MT responded indiscriminately to retinal image motion arising from any source, most of those recorded in the dorsal zone of area MST (MSTd), as well as a smaller proportion in lateral MST (MST1), responded preferentially to externally-induced motion and only weakly or not at all to self-induced visual motion. Such cells preserve visuo-spatial stability during low-velocity voluntary eye movements and could contribute to the process of providing consistent spatial orientation regardless of whether the eyes are moving or stationary.

Animals↗

Vestibular control of swimming in lamprey. II. Characteristics of spatial sensitivity of reticulospinal neurons.

1. Experiments were carried out on an in vitro preparation of the lamprey brainstem isolated together with intact labyrinths. Responses of reticulospinal neurons from different brainstem reticular nuclei (mesencephalic, MRN; anterior rhombencephalic, ARRN; middle rhombencephalic, MRRN; and posterior rhombencephalic, PRRN) to rotation of the preparation (0 degrees-360 degrees) either in the sagittal plane (pitch tilt, or nose up-down movement) or in the transverse plane (roll tilt, or left-right inclination) were recorded. 2. Responses to roll tilt were qualitatively similar in all nuclei: contralateral side down tilt (in relation to the location of the neuron in the brain) caused an activation of reticulospinal neurons. The angular thresholds for activation differed, however, between nuclei as well as the angle at which the maximal activity occurred. The maximal response for MRN was at 45 degrees, for MRRN and PRRN at 90 degrees, for ARRN at 180 degrees. Thus, the zones of spatial sensitivity differed in different nuclei, and they covered the whole range of possible inclinations in the transverse plane. 3. Responses to pitch tilt were not uniform in the different nuclei. MRN neurons responded preferentially in the range of 45 degrees-90 degrees nose-up inclinations, but a proportion of the cells responded in the range of 45 degrees-90 degrees nose-down inclinations. The ARRN neurons had their maximal response when the brain was turned to a dorsal side-down position (180 degrees). In the MRRN, three subgroups of neurons could be distinguished, the first responding at around 90 degrees nose-down, the second responding at around 90 degrees nose-up and the third responding in both zones. However, the activation in the nose-up zone was less robust: responses in this zone were present only in approximately one half of the experiments. Finally, the PRRN neurons were found to be very heterogeneous, with their zones of sensitivity being distributed throughout the whole space (0 degrees-360 degrees). Thus, also in the sagittal plane, the zones of spatial sensitivity in the different nuclei covered the whole range of possible inclinations. 4. Long-term recording of MRRN neurons having the zone of sensitivity around 90 degrees nose-up showed that this response was rather unstable. Its amplitude varied considerably and could disappear with time to reappear later. These results, together with the fact that in a part of the experiments the MRRN neurons responded only in the 90 degrees nose-down zone (see above), leads us to suggest that the system of spatial orientation can dynamically re-organize.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

The role of haptic cues from rough and slippery surfaces in human postural control.

Haptic information is critically important in complex sensory-motor tasks such as manipulating objects. Its comparable importance in spatial orientation is only beginning to be recognized. We have shown that postural sway in humans is significantly reduced by lightly touching a stable surface with a fingertip at contact force levels far below those physically necessary to stabilize the body. To investigate further the functional relationship between contact forces at the hand and postural equilibrium, we had subjects stand in the tandem Romberg stance while being allowed physically supportive (force contact) and non-physically supportive (touch contact) amounts of index fingertip force on surfaces with different frictional characteristics. Mean sway amplitude (MSA) was reduced by over 50% with both touch and force contact of the fingertip, compared to standing without fingertip contact. No differences in MSA were observed when touching rough or slippery surfaces. The amplitude of EMG activity in the peroneal muscles and the timing relationships between fingertip forces, body sway and EMG activity suggested that with touch contact of the finger or with force contact on a slippery surface long-loop "reflexes" involving postural muscles were stabilizing sway. With force contact of the fingertip on a rough surface, MSA reduction was achieved primarily through physical support of the body. This pattern of results indicates that light touch contact cues from the fingertip in conjunction with proprioceptive signals about arm configuration are providing information about body sway that can be used to reduce MSA through postural muscle activation.

Adult↗