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'Fixed-axis' magnetic orientation by an amphibian: non-shoreward-directed compass orientation, misdirected homing or positioning a magnetite-based map detector in a consistent alignment relative to the magnetic field?

Experiments were carried out to investigate the earlier prediction that prolonged exposure to long-wavelength (>500 nm) light would eliminate homing orientation by male Eastern red-spotted newts Notophthalmus viridescens. As in previous experiments, controls held in outdoor tanks under natural lighting conditions and tested in a visually uniform indoor arena under full-spectrum light were homeward oriented. As predicted, however, newts held under long-wavelength light and tested under either full-spectrum or long-wavelength light (>500 nm) failed to show consistent homeward orientation. The newts also did not orient with respect to the shore directions in the outdoor tanks in which they were held prior to testing. Unexpectedly, however, the newts exhibited bimodal orientation along a more-or-less 'fixed' north-northeast-south-southwest magnetic axis. The orientation exhibited by newts tested under full-spectrum light was indistinguishable from that of newts tested under long-wavelength light, although these two wavelength conditions have previously been shown to differentially affect both shoreward compass orientation and homing orientation. To investigate the possibility that the 'fixed-axis' response of the newts was mediated by a magnetoreception mechanism involving single-domain particles of magnetite, natural remanent magnetism (NRM) was measured from a subset of the newts. The distribution of NRM alignments with respect to the head-body axis of the newts was indistinguishable from random. Furthermore, there was no consistent relationship between the NRM of individual newts and their directional response in the overall sample. However, under full-spectrum, but not long-wavelength, light, the alignment of the NRM when the newts reached the 20 cm radius criterion circle in the indoor testing arena (estimated by adding the NRM alignment measured from each newt to its magnetic bearing) was non-randomly distributed. These findings are consistent with the earlier suggestion that homing newts use the light-dependent magnetic compass to align a magnetite-based 'map detector' when obtaining the precise measurements necessary to derive map information from the magnetic field. However, aligning the putative map detector does not explain the fixed-axis response of newts tested under long-wavelength light. Preliminary evidence suggests that, in the absence of reliable directional information from the magnetic compass (caused by the 90 degrees rotation of the response of the magnetic compass under long-wavelength light), newts may resort to a systematic sampling strategy to identify alignment(s) of the map detector that yields reliable magnetic field measurements.

Animals↗

Chromophore orientation in bacteriorhodopsin determined from the angular dependence of deuterium nuclear magnetic resonance spectra of oriented purple membranes.

The orientation of prosthetic groups in membrane proteins is of considerable importance in understanding their functional role in energy conversion, signal transduction, and ion transport. In this work, the orientation of the retinylidene chromophore of bacteriorhodopsin (bR) was investigated using 2H NMR spectroscopy. Bacteriorhodopsin was regenerated with all-trans-retinal stereospecifically deuterated in one of the geminal methyl groups on C1 of the cyclohexene ring. A highly oriented sample, which is needed to obtain individual bond orientations from 2H NMR, was prepared by forming hydrated lamellar films of purple membranes on glass slides. A Monte Carlo method was developed to accurately simulate the 2H NMR line shape due to the distribution of bond angles and the orientational disorder of the membranes. The number of free parameters in the line shape simulation was reduced by independent measurements of the intrinsic line width (1.6 kHz from T2e experiments) and the effective quadrupolar coupling constant (38. 8-39.8 kHz from analysis of the line shape of a powder-type sample). The angle between the C1-(1R)-1-CD3 bond and the purple membrane normal was determined with high accuracy from the simultaneous analysis of a series of 2H NMR spectra recorded at different inclinations of the uniaxially oriented sample in the magnetic field at 20 and -50 degrees C. The value of 68.7 +/- 2.0 degrees in dark-adapted bR was used, together with the previously determined angle of the C5-CD3 bond, to calculate the possible orientations of the cyclohexene ring in the membrane. The solutions obtained from 2H NMR were then combined with additional constraints from linear dichroism and electron cryomicroscopy to obtain the allowed orientations of retinal in the noncentrosymmetric membrane structure. The combined data indicate that the methyl groups on the polyene chain point toward the cytoplasmic side of the membrane and the N-H bond of the Schiff base to the extracellular side, i.e., toward the side of proton release in the pump pathway.

Bacteriorhodopsins↗

Comparison of theoretically oriented and patient-oriented behavioral science courses.

Effective teaching of medical behavioral science is an important challenge for psychiatrists. Two behavioral science courses taught at different times at the Yale University School of Medicine are described: a theoretically oriented course emphasizing symbolic function, primary source reading, and conceptual thinking and a patient-oriented course emphasizing the relevance of behavioral science to the practice of medicine, taught primarily by consultation-liaison psychiatrists. Students' evaluations of the two courses during the fifth year each course was taught were compared. The students' acceptance of the patient-oriented course was dramatically greater than that of the theoretically oriented course. The class mean scores on the behavioral science subtest of the Part I examination of the National Board of Medical Examiners during the last four years that the courses were taught were compared. The mean scores of the classes that had taken the patient-oriented course were significantly higher than those of the classes that had taken the theoretically oriented course. The mean score for the class that took the fifth year of the patient-oriented course was 52 points higher than the mean score for the class that took the fifth year of the theoretically oriented course. The authors concluded that the skills that consultation-liaison psychiatrists use every day in their work may be exactly those that are useful in teaching a successful behavioral science course for medical students.

Behavioral Sciences↗

A methodology for the systematic and quantitative study of cell contact guidance in oriented collagen gels. Correlation of fibroblast orientation and gel birefringence.

Despite the likely role of contact guidance in every physiological process involving cell migration, its study in a three-dimensional tissue-equivalent environment has been precluded, heretofore, by inherent difficulties in systematically preparing well-defined contact guidance fields and quantifying the resultant contact guidance. Here, we describe a novel use of a magnetic field to orient collagen fibrils during fibrillogenesis, entrapping cells dispersed in the collagen solution. Using computer-controlled staging and image analysis, we show from automated birefringence measurements of the resultant slab of cell-populated gel contained in a specially designed observation chamber that the fibril orientation is biased along the long axis of the chamber uniformly throughout the chamber. Further, we show that the degree of fibril orientation, and consequently the elicited contact guidance, can be controlled by independently varying the magnetic field strength or temperature during fibrillogenesis. We characterize the contact guidance response to the imposed contact guidance field by measuring cell orientation relative to the axis of fibril orientation from still images obtained in time-lapse via automated image analysis. We present the first quantitative correlation of contact guidance (based on cell orientation) with collagen fibril orientation (based on birefringence) for human foreskin fibroblasts cultured in a collagen gel, by using gels of varying orientation resulting from different magnetic field strengths and temperatures during fibrillogenesis, and by using sufficiently low cell concentrations and early observation times.

Birefringence↗

Merotelic kinetochore orientation versus chromosome mono-orientation in the origin of lagging chromosomes in human primary cells.

Defects in chromosome segregation play a critical role in producing genomic instability and aneuploidy, which are associated with congenital diseases and carcinogenesis. We recently provided evidence from immunofluorescence and electron microscopy studies that merotelic kinetochore orientation is a major mechanism for lagging chromosomes during mitosis in PtK1 cells. Here we investigate whether human primary fibroblasts exhibit similar errors in chromosome segregation and if at least part of lagging chromosomes may arise in cells entering anaphase in the presence of mono-oriented chromosomes. By using in situ hybridization with alphoid probes to chromosome 7 and 11 we showed that loss of a single sister is much more frequent than loss of both sisters from the same chromosome in anatelophases from human primary fibroblasts released from a nocodazole-induced mitotic arrest, as predicted from merotelic orientation of single kinetochores. Furthermore, the lagging of pairs of separated sisters was higher than expected from random chance indicating that merotelic orientation of one sister may promote merotelic orientation of the other. Kinetochores of lagging chromosomes in anaphase human cells were found to be devoid of the mitotic checkpoint phosphoepitopes recognized by the 3F3/2 antibody, suggesting that they attached kinetochore microtubules prior to anaphase onset. Live cell imaging of H2B histone-GFP-transfected cells showed that cells with mono-oriented chromosomes never enter anaphase and that lagging chromosomes appear during anaphase after chromosome alignment occurs during metaphase. Thus, our results demonstrate that the mitotic checkpoint efficiently prevents the possible aneuploid burden due to mono-oriented chromosomes and that merotelic kinetochore orientation is a major limitation for accurate chromosome segregation and a potentially important mechanism of aneuploidy in human cells.

Cells, Cultured↗

Exploratory factor analysis of the Sport Orientation Questionnaire and the Task and Ego Orientation in Sport Questionnaire in a Japanese sport setting.

This study examined sport achievement orientation within the japanese sport setting. 1,836 male and 425 female athletes (M age = 18.6 yr.) from 47 sports completed the Japanese version of the Sport Orientation Questionnaire (24 items), and 1,781 males and 421 females (M age = 18.6 yr.) from 47 sports completed that of the Task and Ego Orientation in Sport Questionnaire (14 items). While the original English versions of the questionnaires are composed of three and two factors, respectively, present exploratory factor analyses identified four factors in the Sport Orientation Questionnaire and three in the Task and Ego Orientation in Sport Questionnaire. In this study, two types of the SOQ Competitiveness and the TEOSQ Ego Orientation emerged even though the original versions included only one type. A one-way analysis of variance indicated that sex differences were significant for all subscales. Sex differences in the Sport Orientation Questionnaire of Japanese resembled those of Americans. The overall factor reliability and validity based on the sample suggested that the modified Japanese versions of the two questionnaires can be valuable in the investigation of sport achievement orientation in Japanese sport and exercise settings.

Achievement↗

The orientation coach: an important player in competency-based orientation.

Competency-based orientation (CBO) is mandated by JCAHO's Agenda for Change and can improve quality patient care and employee retention. JCAHO redefined its requirements for staff orientation, training and education in 1993, emphasizing the use of competency assessment systems to foster continuous improvements in organizational quality. Proper training for staff members who conduct orientation for new employees--"orientation coaches"--is critical to the success of CBO programs. Our institution designed a training workshop to establish a common knowledge base for orientation coaches. The workshop clarifies the coaches' role and responsibilities, supplies practical information for the coaches' use and promotes a consistent approach to competency assessment. Coaches are familiarized with transitions or adjustment phases new employees typically experience and with adult learning principles, teaching behaviors and learning style differences. The orientation coaches' workshop has met organizational expectations by preparing qualified orientation leaders. New employees benefit from systematic, customized orientation plans facilitated by coaches who are comfortable with their roles and responsibilities, knowledgeable about adult education principles and competency assessment.

Competency-Based Education↗

[Action-oriented versus state-oriented reactions to experimenter-induced failures].

The present study assessed different effects of action-oriented versus state-oriented styles of coping with failure on achievement-related performance and cognition. In a learned helplessness experiment, students were exposed to an academic failure situation and were then tested on a series of problem-solving tasks, either immediately after the pretreatment or after a delay of 24 hours. Performance and cognitive concomitants were measured during both experimental periods. Results demonstrated that action orientation was associated with self-immunizing cognitions during helplessness training. Action-oriented participants improved their performance level even after repeated failure feedbacks. Moreover, action-oriented students assigned to the delayed test condition responded with increased striving for success and showed performance increments, even in comparison with control subjects. In contrast, state-oriented participants developed symptoms of helplessness and showed impaired performance during failure inductions. In later tests on problem-solving tasks, state-oriented groups responded with increased fear of failure. Independent of immediate or delayed test conditions, they soon lapsed into new performance decrements.

Achievement↗

Determination of orientational order parameters from 2H NMR spectra of magnetically partially oriented lipid bilayers.

The partial orientation of multilamellar vesicles (MLVs) in high magnetic fields is known to affect the shape of 2H NMR spectra. There are numerical methods for extracting either the orientational order parameters of lipid molecules for a random distribution of domain orientations in the sample, or the distribution of orientations for a known set of spectral anisotropies. A first attempt at determining the orientational order parameters in the presence of an unknown nonrandom distribution of orientations is presented. The numerical method is based on the Tikhonov regularization algorithm. It is tested using simulated partially oriented spectra. An experimental spectrum of a phospholipid-ether mixture in water is analyzed as an example. The experimental spectrum is consistent with an ellipsoidal shape of MLVs with a ratio of semiaxes of approximately 3.4.

Algorithms↗

Orientation of the tetranuclear manganese cluster and tyrosine Z in the O(2)-evolving complex of photosystem II: An EPR study of the S(2)Y(Z)(*) state in oriented acetate-inhibited photosystem II membranes.

Inhibitory treatment by acetate, followed by illumination and rapid freezing, is known to trap the S(2)Y(Z)(*) state of the O(2)-evolving complex (OEC) in photosystem II (PS II). An EPR spectrum of this state exhibits broad split signals due to the interaction of the tyrosyl radical, Y(Z)(*), with the S = 1/2 S(2) state of the Mn(4) cluster. We present a novel approach to analyze S(2)Y(Z)(*) spectra of one-dimensionally (1-D) oriented acetate-inhibited PS II membranes to determine the magnitude and relative orientation of the S(2)Y(Z)(*) dipolar vector within the membrane. Although there exists a vast body of EPR data on isolated spins in oriented membrane sheets, the present study is the first of its kind on dipolar-coupled electron spin pairs in such systems. We demonstrate the feasibility of the technique and establish a rigorous treatment to account for the disorder present in partially oriented 1-D membrane preparations. We find that (i) the point-dipole distance between Y(Z)(*) and the Mn(4) cluster is 7.9 +/- 0.2 A, (ii) the angle between the interspin vector and the thylakoid membrane normal is 75 degrees, (iii) the g(z)()-axis of the Mn(4) cluster is 70 degrees away from the membrane normal and 35 degrees away from the interspin vector, and (iv) the exchange interaction between the two spins is -275 x 10(-)(4) cm(-)(1), which is antiferromagnetic. Due to the sensitivity of EPR line shapes of oriented spin-coupled pairs to the interspin distance, the present study imposes a tighter constraint on the Y(Z)-Mn(4) point-dipole distance than obtained from randomly oriented samples. The geometric constraints obtained from the 1-D oriented sample are combined with published models of the structure of Mn-depleted PS II to propose a location of the Mn(4) cluster. A structure in which Y(Z) is hydrogen bonded to a manganese-bound hydroxide ligand is consistent with available data and favors maximal orbital overlap between the two redox center that would facilitate direct electron- and proton-transfer steps.

Electron Spin Resonance Spectroscopy↗

Multiple groups of orientation-selective visual mechanisms underlying rapid orientated-line detection.

Visual search for an edge or line element differing in orientation from a background of other edge or line elements can be performed rapidly and effortlessly. In this study, based on psychophysical measurements with ten human observers, threshold values of the angle between a target and background line elements were obtained as functions of background-element orientation, in brief masked displays. A repeated-loess analysis of the threshold functions suggested the existence of several groups of orientation-selective mechanisms contributing to rapid orientated-line detection; specifically, coarse, intermediate and fine mechanisms with preferred orientations spaced at angles of approximately 90 degrees, 35 degrees, and 10 degrees-25 degrees, respectively. The preferred orientations of coarse and some intermediate mechanisms coincided with the vertical or horizontal of the frontoparallel plane, but the preferred orientations of fine mechanisms varied randomly from observer to observer, possibly reflecting individual variations in neuronal sampling characteristics.

Adult↗

Relation of cortical cell orientation selectivity to alignment of receptive fields of the geniculocortical afferents that arborize within a single orientation column in ferret visual cortex.

Neurons in the primary visual cortex of higher mammals are arranged in columns, and the neurons in each column respond best to light-dark borders of particular orientations. The basis of cortical cell orientation selectivity is not known. One possible mechanism would be for cortical cells to receive input from several lateral geniculate nucleus (LGN) neurons with receptive fields that are aligned in the visual field (Hubel and Wiesel, 1962). We have investigated the relationship between the arrangement of the receptive fields of geniculocortical afferents and the orientation preferences of cortical cells in the orientation columns to which the afferents provide visual input. Radial microelectrode penetrations were made into primary visual cortex of anesthetized adult sable ferrets. Cortical cells were recorded throughout the depth of the cortex, and their orientation preferences were determined. Cortical cell responses were then eliminated by superfusion of the cortex with either kainic acid (Zahs and Stryker, 1988) or muscimol. After the drug treatment, responses from many single units with distinct receptive fields were recorded. These responses were presumed to be those of geniculocortical afferents, because they had the response properties characteristic of LGN neurons, and because they could be recorded only in cortical layers that receive geniculate input. In 16 of 18 cases, the afferent receptive fields recorded in a single penetration covered an elongated region of visual space. In these penetrations, the best-fit line through the centers of the afferent receptive fields generally paralleled the preferred orientation of cortical cells recorded at the same site in cortex. These results are consistent with the Hubel and Wiesel (1962) model for the construction of oriented visual cortical receptive fields from geniculate inputs with aligned receptive fields.

Afferent Pathways↗

Code 50: an orientation matrix to track orientation cost.

Estimates of staff orientation costs can range from $20,000-50,000 per person. The Director of Finance and the Director of Nursing Education and Research developed a system to track the orientation process to determine the actual costs. Head nurses and educators approached the orientation process differently; therefore, there was no consistency as to the time allotted for orientation. Although competency-based orientation tools were used for all new orientees, the average amount of time for new orientees to become productive ranged from 3-9 months. By creating the orientation matrix, a unique pay class (code 50) was assigned for all new orientees, and a computer program was developed to track costs. The overtime hours for orientees dropped to 159 from 941, resulting in an 83% reduction in overtime hours. The matrix ensured a more cost effective approach to orientation.

Clinical Competence↗

Does orientation-independent object recognition precede orientation-dependent recognition? Evidence from a cuing paradigm.

Object recognition may entail an incremental normalization process before access to canonical orientation representations, but is this process guided by prior access to object-centered representations? In Experiment 1, the authors showed observers figure-ground stimuli known to reflect access to, and output from, stored shape representations. The stimuli appeared in each of 6 different orientations, preceded by cues providing either (a) no information, (b) upright shape information only, (c) upright shape information plus orientation information (separately), or (d) shape information in the same orientation as the upcoming figure-ground test stimulus. Contrary to predictions by a postaccess account, the cues failed to eliminate orientation dependency in shape recognition. The results favor a preaccess account of the normalization process within the context of canonical orientation representations.

Adult↗

The orientation anisotropy and orientation constancy: a visual evoked potential study.

Two experiments were conducted on the orientation anisotropy in which averaged visual evoked potentials (VEPs) were recorded from the occipital scalp. The first experiment confirmed the findings of Maffei and Campbell (1970) that obliquely oriented gratings alternated back and forth produced smaller-amplitude VEPs than when the gratings were oriented horizontally or vertically. Since no asymmetry was found in VEPs produced by a Julesz figure presented under identical conditions, it was concluded that direction of displacement could not have been contributing to the effect. In a second experiment head tilt of the subject was manipulated together with grating orientation and the results indicated that the orientation anisotropy is retinally rather than gravitationally referenced. It was concluded that the site of orientation constancy is located either at higher levels of the primary visual system or in the second visual system.

Adult↗

Human perceptual learning in identifying the oblique orientation: retinotopy, orientation specificity and monocularity.

1. Human perceptual learning in discrimination of the oblique orientation was studied using psychophysical methods. Subjects were trained daily to improve their ability to identify the orientation of a circular 2.5 deg diameter unidimensional noise field. Dramatic improvements in sensitivity to contour orientation occurred over a period of 15-20 days. The improved performance persisted for several months. Improvement was more evident between daily sessions than within sessions. This was partly due to fatigue interfering with the learning effect. Moreover, a consolidation period seemed to be required. 2. Improvement was restricted to the position of the stimulus being trained. This position dependency of the learning effect proved very precise. After training at a specific stimulus position, merely displacing the stimulus to an adjacent position caused a marked increase in thresholds. 3. No transfer of the training effect was observed between orientations. Following a shift of 90 deg away from the trained orientation, performance fell, even below the initial level. 4. We observed complete to almost complete transfer between the two eyes. 5. Our results suggest plastic changes at a level of the visual processing stream where input from both eyes has come together, but where generalization for spatial localization and orientation has not yet occurred.

Discrimination, Psychological↗

Orientation of the hemes of high potential cytochromes relative to photosynthetic membranes, as shown by the linear dichroism of oriented preparations.

The orientations of high potential cytochromes with respect to photosynthetic membranes was investigated in spinach chloroplasts and in Rhodopseudomonas viridis. The general approach consists in detection with polarized light of photoinduced absorbance changes related to the oxidation of the cytochromes. The orientation of cytochrome c-558 was measured at room temperature in chromatophores and whole cells of Rps. viridis, oriented on glass slides and in a magnetic field, respectively. The orientation of cytochrome b-559 of green plants was detected at 77 K in magnetically oriented chloroplasts. In both cases the dichroic ratio for the alpha band shows that the heme plane makes an angle greater than 35 degrees C with the membrane plane. Moreover, the dichroic ratio is not constant throughout the alpha and beta bands, for both cytochrome c-558 and b-559. Linear dichroism spectra of oriented pure horse heart cytochrome c and cytochrome c2 of Rhodopseudomonas sphaeroides in stretched polyvinyl alcohol films show that the variations of the dichroic ratio in the alpha and beta bands can be explained by the occurrence of x- and y-polarized transitions absorbing at slightly different wavelengths.

Chloroplasts↗

Measuring orientation of actin filaments within a cell: orientation of actin in intestinal microvilli.

Orientational distribution of actin filaments within a cell is an important determinant of cellular shape and motility. To map this distribution we developed a method of measuring local orientation of actin filaments. In this method actin filaments within cells are labeled with fluorescent phalloidin and are viewed at high magnification in a fluorescent microscope. Emitted fluorescence is split by a birefringent crystal giving rise to two images created by light rays polarized orthogonally with respect to each other. The two images are recorded by a high-sensitivity video camera, and polarization of fluorescence at any point is calculated from the relative intensity of both images at this point. From the value of polarization, the orientation of the absorption dipole of the dye, and thus orientation of F-actin, can be calculated. To illustrate the utility of the method, we measured orientation of actin cores in microvilli of chicken intestinal epithelial cells. F-actin in microvillar cores was labeled with rhodamine-phalloidin; measurements showed that the orientation was the same when microvillus formed a part of a brush border and when it was separated from it suggesting that "shaving" of brush borders did not distort microvillar structure. In the absence of nucleotide, polarization of fluorescence of actin cores in isolated microvilli was best fitted by assuming that a majority of fluorophores were arranged with a perfect helical symmetry along the axis of microvillus and that the absorption dipoles of fluorophores were inclined at 52 degrees with respect to the axis. When ATP was added, the shape of isolated microvilli did not change but polarization of fluorescence decreased, indicating statistically significant increase in disorder and a change of average angle to 54 degrees. We argue that these changes were due to mechanochemical interactions between actin and myosin-I.

Actins↗