Determination of cell division axes in the early embryogenesis of Caenorhabditis elegans

The establishment of cell division axes was examined in the early embryonic divisions of Caenorhabditis elegans. It has been shown previously that there are two different patterns of cleavage during early embryogenesis. In one set of cells, which undergo predominantly determinative divisions, the division axes are established successively in the same orientation, while division axes in the other set, which divide mainly proliferatively, have an orthogonal pattern of division. We have investigated the establishment of these axes by following the movement of the centrosomes. Centrosome separation follows a reproducible pattern in all cells, and this pattern by itself results in an orthogonal pattern of cleavage. In those cells that divide on the same axis, there is an additional directed rotation of pairs of centrosomes together with the nucleus through well-defined angles. Intact microtubules are required for rotation; rotation is prevented by inhibitors of polymerization and depolymerization of microtubules. We have examined the distribution of microtubules in fixed embryos during rotation. From these and other data we infer that microtubules running from the centrosome to the cortex have a central role in aligning the centrosome-nuclear complex.     

Polarized-light sensitivity in rainbow trout (Oncorhynchus mykiss): characterization from multi-unit responses in the optic nerve

Integrated spike activity of axons from the optic nerve was measured in an investigation of the e-vector sensitive mechanism underlying the ability of rainbow trout (Oncorhynchus mykiss) for orientation in downwelling, linearly-polarized light. In anaesthetized, immobilized fish, one eye was exposed to incremental light flashes which were superimposed over closely controlled background conditions. Under scotopic and various photopic conditions, intensity/response curves were generated from the on-response of the optic nerve. Relative sensitivity curves were then obtained as a function of e-vector direction for the 5 kinds of receptor cells in this trout's retina: rods, ultraviolet cones (UV), short wavelength cones (S), medium wavelength cones (M), and long wavelength cones (L).Under scotopic conditions, no sensitivity to e-vector was apparent: thus, rods do not mediate polarization sensitivity. Under photopic conditions, parr weighing 8–10 g exhibited e-vector sensitivity in two orthogonal channels. A UV stimulus (380 nm) on a white background evoked a three-peaked response (0°, 90°, and 180°) to the e-vector orientations presented in 30° increments between 0° and 180°. In contrast, when the background was illuminated with appropriate short and long wavelength cut-off filters, M-and L-cones showed maximum responses only to the horizontal (90°) plane whether they were stimulated at their -absorption band or their -absorption band in the near UV. Isolated UV-cones gave maximum responses to the vertical (0° and 180°) e-vector, thus corresponding to a second channel. The blue sensitive, S-cones, did not show evidence of polarization sensitivity. As well, no evidence of the polarization sensitivity was observed under UV isolating background conditions in larger individuals, 50–78 g smolts, although the other cone mechanisms responded as in smaller individuals.     

A new non-orthogonal decomposition method to determine effective torques for three-dimensional joint rotation

This paper describes a new non-orthogonal decomposition method to determine effective torques for three-dimensional (3D) joint rotation. A rotation about a joint coordinate axis (e.g. shoulder internal/external rotation) cannot be explained only by the torque about the joint coordinate axis because the joint coordinate axes usually deviate from the principal axes of inertia of the entire kinematic chain distal to the joint. Instead of decomposing torques into three orthogonal joint coordinate axes, our new method decomposes torques into three "non-orthogonal effective axes" that are determined in such a way that a torque about each effective axis produces a joint rotation only about one of the joint coordinate axes. To demonstrate the validity of this new method, a simple internal/external rotation of the upper arm with the elbow flexed at 90 degrees was analyzed by both orthogonal and non-orthogonal decomposition methods. The results showed that only the non-orthogonal decomposition method could explain the cause-effect mechanism whereby three angular accelerations at the shoulder joint are produced by the gravity torque, resultant joint torque, and interaction torque. The proposed method would be helpful for biomechanics and motor control researchers to investigate the manner in which the central nervous system coordinates the gravity torque, resultant joint torque, and interaction torque to control 3D joint rotations.     

Comparison of different calculations of three-dimensional joint kinematics from video-based system data

When skin-fixed marker trajectories are used to calculate 3D joint kinematics, the measurement errors (i.e. the difference between the trajectories of the external markers and those of the skeleton) influence to some extent the accuracy of the results, depending both on the calculation method and on the axes about which the rotations are expressed. The purpose of this paper is to compare several expressions of joint angular variations. Two kinematic concepts are used to calculate the changes in the orientation of the distal segment versus the proximal one: the first method consists of computing the components of the spatial attitude vector, the second one deals with the determination of elementary rotations about successive axes. For each of these methods, two sets of three axes are tested to express the results: the axes forming the reference frame affixed to the body segment adjacent to the joint (named fixed axes), and a set consisting of a first axis belonging to the proximal segment, a third axis belonging to the distal segment and a second (floating) axis defined as the cross-product between the two other ones (named mobile axes). To compare these four distinct expressions on the knee joint, numerical simulations of perturbed skin marker trajectories are performed, based on experimental data recorded by a Motion Analysis system during a normal gait cycle. A significant difference is pointed out only for the internal–external rotation angle, for which the best expression — from the viewpoint of sensitivity to experimental errors — is obtained using the components of the attitude vector in a segment-embedded reference frame.     

Production of L-aspartic acid from fumaric acid by a fumaric acid-assimilating obligate thermophile,Bacillus stearothermophilus KP 1041

Summary A fumaric acid-assimilating obligate thermophile having a high aspartase activity was isolated from soil. The isolate (KP 1041) that grew at 45 to 68 °C was assigned to a strain of Bacillus stearothermophilus. The cell suspensions produced L-aspartate from fumarate and ammonium ion, with the rapidest initial rate at 65 °C and pH 9.5. The Michaelis constant for fumarate was 0.2 M. The cellular aspartase was relatively stable for 18 h at and below 50 °C over a pH range 6.7–8.3 in the presence of ammonium fumarate; this substance protected the enzyme from heat inactivation. The best yield in L-aspartic acid production was achieved at 6 h incubation at 53 °C and pH 8.5, using 0.88 M fumarate, 3.1 M ammonium ion, and the cells at 53 mg dry weight per ml. In this case, 85% of fumarate added was converted into aspartic acid. The structure of the product was determined from its infrared spectrum, specific rotation, melting point and ultimate analysis.Presented at the Annual Meeting of the Agricultural Chemical Society of Japan, Yokohama, April 2, 1977     

Loss and restoration of viability of E. coli due to combinations of mutations affecting DNA polymerase I and repair activities

Summary We have found that the cells possessing the polA6 mutation affecting DNA polymerase I are unable to accept another mutation (uvr502) leading to UV-sensitivity. The introduction of the polA12 mutation determining the synthesis of a temperature sensitive DNA polymerase I into the uvr502 mutant results in the temperature sensitivity of colony forming ability of the double mutant. These data show that the uvr502 derivatives lacking DNA polymerase I are inviable. Reversions to temperature resistance in the population of the double mutant uvr502 polA12 may occur because of reverse mutations at one of the mutated sites or because of mutations suppressing DNA polymerase I deficiency but not UV- or MMS-sensitivity of revertants. DNA and protein synthesis in uvr502 polA12 cells continues after a shift to 45°C with rates almost indistinguishable from those in single mutants or wild type cells. No differences in DNA degradation were observed during incubation of single and double mutants at 45°C. The single strand molecular weight distribution of parent DNA from the double mutant as well as that from wild type cells is not affected by the shift to 45°C and 3 hours incubation at this temperature. We suggest that DNA polymerase I and/or the product altered by the uvr502 mutation are required for some step(s) of discontinuous DNA replication nonessential for the formation of acid insoluble DNA. The DNA polymerase I and the uvr gene product seem to be able to substitute for each other in accomplishing this process.     

Cellular basis for polarized light perception in the spider crab,Libinia

Summary Differential increases in the numbers of pinocytotic vesicles, multivesicular bodies and total complex bodies occurred in the cytoplasm of specific photoreceptor cells in the compound eye of the crab Libinia exposed for six hours to polarized light with various e-vector orientations. These data coupled with previous results on the same species proved that the seven retinular cells in each ommatidium formed two functional groups selectively light adaptable by e-vectors oriented 90° apart. One group (Channel I, comprising Cells 1, 4 and 5) was more affected by horizontal polarization; the other (Channel II, comprising Cells 2, 3, 6 and 7) was more affected by vertical polarization.This confirmed by a quite independent technique the conclusion reached from electrophysiological experiments on the crab Cardisoma that decapod compound eyes have two orthogonal polarization analyzer channels. In addition the present data showed that both channels occur in each ommatidium as hypothesized on previous electron microscopic evidence and that the axes of maximum absoprtion in the two retinal channels were parallel to the long axes of their cells' rhabdom microvilli, horizontal in Channel I and vertical in Channel II. The latter relations in turn supported the hypothesis that the dichroism of rhodopsin was fundamental to the analyzer mechanism.This research has been supported by U. S. Air Force Grant AFOSR 1064 and NASA Grant NGR 07-004-055. The authors wish to thank Professor Joseph G. Gall for generously sharing his electron microscope facilities.     

NeuroTerrain – a client-server system for browsing 3D biomedical image data sets

Background  

Three dimensional biomedical image sets are becoming ubiquitous, along with the canonical atlases providing the necessary spatial context for analysis. To make full use of these 3D image sets, one must be able to present views for 2D display, either surface renderings or 2D cross-sections through the data. Typical display software is limited to presentations along one of the three orthogonal anatomical axes (coronal, horizontal, or sagittal). However, data sets precisely oriented along the major axes are rare. To make fullest use of these datasets, one must reasonably match the atlas' orientation; this involves resampling the atlas in planes matched to the data set. Traditionally, this requires the atlas and browser reside on the user's desktop; unfortunately, in addition to being monolithic programs, these tools often require substantial local resources. In this article, we describe a network-capable, client-server framework to slice and visualize 3D atlases at off-axis angles, along with an open client architecture and development kit to support integration into complex data analysis environments.     

Effects of ethanol on thermal death and on the maximum temperature for growth of the yeast Kluiveromyces fragilis

Summary The maximum temperature for growth of a strain of Kluyveromyces fragilis was depressed by increasing ethanol concentration from its normal value around 45 °C to 26.5 °C at 6.4% (w/v) of ethanol. Ethanol enhanced thermal death by increasing S, the entropy of activation of thermal death. Entropy coefficients of 7.6 entropy units per mol of ethanol per liter of medium for unadapted cells and of 5.7 for adapted cells ware obtained.     

Pyrolytic characteristics of heterotrophic Chlorella protothecoides for renewable bio-fuel production

Heterotrophic Chlorella protothecoides cells were pyrolyzed in athermogravimetric analyzer to investigate the pyrolytic characteristics anddetermine the kinetic parameters. Heating rates of 15, 40, 60 and 80 ^°C^-1 up to a final temperature of 800 ^°C wereused. The pyrolysis reactions mainly took place between 160–520^°C with a volatile yield of about 80%. The devolatilization stageconsisted of two main temperature zones (I and II) with a transition at300–320 ^°C. Crude lipid in cells decomposed at Zone II whileother main components at Zone I. The increase of heating rate caused alateral shift to higher temperatures in the thermograms, a decrease ofactivation energies for the devolatilization stage and an increase of both theinstantaneous maximum and average reaction rates. The difference ofactivation energies between two zones implied that more energy input forlipid pyrolysis seems needed in comparison with other main components.These data are useful for the design, operation, and modeling of thepyrolysis systems for microalgae.     

Fusion of radiostereometric analysis data into computed tomography space: application to the elbow joint

Improvement of joint prostheses is dependent upon information concerning the biomechanical properties of the joint. Radiostereometric analysis (RSA) and electromagnetic techniques have been applied in previous cadaver and in vivo studies on the elbow joint to provide valuable information concerning joint motion axes. However, such information is limited to mathematically calculated positions of the axes according to an orthogonal coordinate system and is difficult to relate to individual skeletal anatomy. The aim of this study was to evaluate the in vivo application of a new fusion method to provide three-dimensional (3D) visualization of flexion axes according to bony landmarks. In vivo RSA data of the elbow joint's flexion axes was combined with data obtained by 3D computed tomography (CT). Results were obtained from five healthy subjects after one was excluded due to an instable RSA marker. The median error between imported and transformed RSA marker coordinates and those obtained in the CT volume was 0.22 mm. Median maximal rotation error after transformation of the rigid RSA body to the CT volume was 0.003 degrees . Points of interception with a plane calculated in the RSA orthogonal coordinate system were imported into the CT volume, facilitating the 3D visualization of the flexion axes. This study demonstrates a successful fusion of RSA and CT data, without significant loss of RSA accuracy. The method could be used for relating individual motion axes to a 3D representation of relevant joint anatomy, thus providing important information for clinical applications such as the development of joint prostheses.     

Convergence of neck and macular vestibular inputs on vestibulospinal neurons projecting to the lumbosacral segments of the spinal cord

The activity of LVN neurons was recorded in decerebrate cats and analyzed during separate stimulation of macular vestibular and neck receptors elicited by sinusoidal rotation about the longitudinal axis at 0.026 Hz, 10 degrees peak amplitude. Of 119 LVN units examined, the great majority, i.e. 106, were vestibulospinal neurons antidromically identified following stimulation of the spinal cord at T12-L1, thus projecting to the lumbosacral segments of the spinal cord (IVS neurons); the remaining 13 units were nonantidromically activated. Among the 119 LVN neurons, 77 (64.7%) responded with a periodic modulation of their firing rate to roll tilt of the animal and 81 (68.1%) responded to neck rotation. Convergence of macular and neck inputs was found in 58/119 (48.7%) lateral vestibular neurons; in these units, the gain as well as the sensitivity of the first harmonic of responses corresponded on the average to 0.58 +/- 0.45, S.D. imp./sec/deg and 4.39 +/- 3.58, S.D.%/deg for the neck responses and 0.52 +/- 0.49, S.D. imp./sec/deg and 3.85 +/- 3.35, S.D.%/deg for the macular responses, respectively. In addition to these convergent units, 19/119 (16.0%) and 23/119 (19.3%) lateral vestibular units responded to selective stimulation either of macular receptors or of neck receptors only. These units, which showed on the average an higher firing rate and a lower conduction velocity of the corresponding vestibulospinal axons than the convergent units, displayed a significantly lower response gain and sensitivity to animal tilt and neck rotation with respect to those obtained from convergent units. Most of the convergent lateral vestibular units were maximally excited by the direction of stimulus orientation, the first harmonic of responses showing an average phase lead of +51.4 degrees with respect to neck position and +21.9 degrees with respect to animal position. Two populations of convergent neurons were observed. The first group of units (53/58, i.e. 91.4%) showed reciprocal ("out-of-phase") responses to the two inputs in that they were mainly excited during side-down animal tilt and side-up neck rotation. The remaining group of units (5/58, i.e. 8.6%) showed parallel ("in phase") responses to the two inputs and they were mainly excited by side-up neck rotation and animal tilt. Interestingly, the former group of units displayed an average gain and sensitivity to the labyrinth and neck inputs which were more than twice higher than the values obtained from the latter group of units.(ABSTRACT TRUNCATED AT 400 WORDS)     

Thermal sensitivity of heart rate and insensitivity of blood pressure in the Antarctic nototheniid fish <Emphasis Type="Italic">Pagothenia borchgrevinki</Emphasis>

The Antarctic notothenioids are among the most stenothermal of fishes, well adapted to their stable, cold and icy environment. The current study set out to investigate the thermal sensitivity/insensitivity of heart rate and ventral aortic blood pressure of the Antarctic nototheniid fish Pagothenia borchgrevinki over a range of temperatures. The heart rate increased rapidly from –1 to 6°C (Q₁₀=2.0–3.3), but was relatively insensitive to temperature above the ~6°C lethal limit of the species (Q₁₀=1.2). The increase in heart rate from –1 to 6°C was the result of a 45% increase in excitatory adrenergic tone, masking a 37% increase in inhibitory cholinergic tone. Ventral aortic pressure was regulated well above the lethal limit, up to at least 10°C. With the return of the fish to environmental temperatures, the heart rate rapidly decreased back to control levels, while ventral aortic pressure increased and remained elevated for over an hour following a 6°C exposure.     

Nemo is required in a subset of photoreceptors to regulate the speed of ommatidial rotation

Both dramatic and subtle morphogenetic movements are of paramount importance in molding cells and tissues into functional form. Cells move either independently or as populations and the distance traversed by cells varies greatly, but in all cases, the output is common: to organize cells into or within organs and epithelia. In the developing Drosophila eye, a highly specialized, 90° rotational movement of subsets of cells imposes order by polarizing the retinal epithelium across its dorsoventral axis. This process was proposed to take place in two 45° steps, with the second under control of the gene nemo (nmo), a serine/threonine kinase. While our analysis confirms that these subsets of cells, the ommatidial precursors, do stall at 45°, we demonstrate that nmo is also required through most of the first 45° of rotation to regulate the speed at which the ommatidial precursors move. In addition, although the precursors reach only the halfway point by the end of larval life, this work demonstrates that patterning events that occur during pupal life move the ommatidial units an additional 15°. A re-analysis of nmo mosaic clones indicates that nmo is required in photoreceptors R1, R6 and R7 for normal orientation. This work also demonstrates that two major isoforms of nmo rescue the nmo^P1 phenotype. Finally, a dominant modifier screen of a nmo misexpression background identified genomic regions that potentially regulate rotation. The results presented here suggest a model in which a motor for rotation is established in a nemo-dependent fashion in a subset of cells.     

Variation in patellofemoral kinematics due to changes in quadriceps loading configuration during in vitro testing

This study investigated changes in patellofemoral (PF) kinematics for different loading configurations of the quadriceps muscle: single line of action (SL), physiological-based multiple lines of action (ML), weak vastus medialis (WVM), and weak vastus lateralis (WVL). Fourteen cadaveric knees were flexed from 15° to 120° knee flexion using a loading rig with the ability to load different heads of the quadriceps and hamstring muscles in their anatomical orientation. PF rotation in the sagittal plane) and medial lateral translation were significantly different (p<0.05) for SL and ML, with maximum differences of 2.8° and 0.9 mm at 15° and 45° knee flexion, respectively. Compared to the ML, the WVM induced an average lateral shift of 1.5 mm and an abduction rotation of 0.8°, whereas a 0.9 mm medial shift and 0.6° adduction rotation was seen when simulating a WVL. The difference in the sagittal plane resultant force orientation of 26° between SL and ML was the major contributor to the change in PF rotation in the sagittal plane, while the difference in the frontal plane resultant force orientation of both the WVM and WVL from the ML (17° medial and 8° lateral, respectively) were the primary reasons for the change in PF frontal plane rotation and medial lateral translation. The two PF kinematic were significantly different from the ML for WVM and WVL (p<0.05). The results suggest that quadriceps muscle loading configuration can have a large influence on PF kinematics during full extension but less in deeper flexion. Therefore, using quadriceps single line loading for simulating activities with low flexion angles might not be sufficient to accurately replicate the physiological condition.     

Selection of yeast able to produce ethanol from glucose at 40° C

Summary A total of 55 yeast strains selected from 7 genera known to ferment carbohydrates to ethanol were screened for their ability to ferment glucose to ethanol in shaken flask culture at 37°, 40° and 45°C. Yields of more than 50% of the theoretical maximum were obtained with 28 strains at 37°C, but only 12 at 40°C. Only 6 could grow at 45°C, but they produced poor yields. In general Kluyveromyces strains were more thermotolerant than Saccharomyces and Candida strains, but Saccharomyces strains produced higher ethanol yields. The 8 strains with the highest yields at 40°C were evaluated in batch fermentations. Three of these, two Saccharomyces and one Candida, were able to meet minimum commercial targets set at 8% (v/v) ethanol from 14% (w/v) glucose at 40°C.     

Excretion of metabolites by hydrogen bacteria

Summary D(–)-3-hydroxybutanoic acid was produced by a double mutant of Alcaligenes eutrophus, unable to synthesize poly-3-hydroxybutanoic acid and to utilize 3-hydroxybutanoate as a substrate. About 3.4 g of 3-hydroxybutanoate/l were produced under optimum conditions at pH 7.0 to 7.3, at a temperature of 30°C after exhaustion of ammonia and at restricted aeration rates allowing 10–12% of the maximum respiratory rate of the cells to occur. D(–)-3-hydroxybutanoic acid was identified by means of gas and ion exchange chromatography, IR-spectrometry and specific rotation.     

Functional knee axis based on isokinetic dynamometry data: Comparison of two methods, MRI validation, and effect on knee joint kinematics

This paper compares geometry-based knee axes of rotation (transepicondylar axis and geometric center axis) and motion-based functional knee axes of rotation (fAoR). Two algorithms are evaluated to calculate fAoRs: Gamage and Lasenby's sphere fitting algorithm (GL) and Ehrig et al.'s axis transformation algorithm (SARA). Calculations are based on 3D motion data acquired during isokinetic dynamometry. AoRs are validated with the equivalent axis based on static MR-images. We quantified the difference in orientation between two knee axes of rotation as the angle between the projection of the axes in the transversal and frontal planes, and the difference in location as the distance between the intersection points of the axes with the sagittal plane. Maximum differences between fAoRs resulting from GL and SARA were 5.7° and 15.4mm, respectively. Maximum differences between fAoRs resulting from GL or SARA and the equivalent axis were 5.4°/11.5mm and 8.6°/12.8mm, respectively. Differences between geometry-based axes and EA are larger than differences between fAoR and EA both in orientation (maximum 10.6°).and location (maximum 20.8mm). Knee joint angle trajectories and the corresponding accelerations for the different knee axes of rotation were estimated using Kalman smoothing. For the joint angles, the maximum RMS difference with the MRI-based equivalent axis, which was used as a reference, was 3°. For the knee joint accelerations, the maximum RMS difference with the equivalent axis was 20°/s(2). Functional knee axes of rotation describe knee motion better than geometry-based axes. GL performs better than SARA for calculations based on experimental dynamometry.     

Netrins and Wnts Function Redundantly to Regulate Antero-Posterior and Dorso-Ventral Guidance in C. elegans

Guided migrations of cells and developing axons along the dorso-ventral (D/V) and antero-posterior (A/P) body axes govern tissue patterning and neuronal connections. In C. elegans, as in vertebrates, D/V and A/P graded distributions of UNC-6/Netrin and Wnts, respectively, provide instructive polarity information to guide cells and axons migrating along these axes. By means of a comprehensive genetic analysis, we found that simultaneous loss of Wnt and Netrin signaling components reveals previously unknown and unexpected redundant roles for Wnt and Netrin signaling pathways in both D/V and A/P guidance of migrating cells and axons in C. elegans, as well as in processes essential for organ function and viability. Thus, in addition to providing polarity information for migration along the axis of their gradation, Wnts and Netrin are each able to guide migrations orthogonal to the axis of their gradation. Netrin signaling not only functions redundantly with some Wnts, but also counterbalances the effects of others to guide A/P migrations, while the involvement of Wnt signaling in D/V guidance identifies Wnt signaling as one of the long sought mechanisms that functions in parallel to Netrin signaling to promote D/V guidance of cells and axons. These findings provide new avenues for deciphering how A/P and D/V guidance signals are integrated within the cell to establish polarity in multiple biological processes, and implicate broader roles for Netrin and Wnt signaling - roles that are currently masked due to prevalent redundancy.