The force exerted by a molecular motor.

The stochastic driving force exerted by a single molecular motor (e. g., a kinesin, or myosin) moving on a periodic molecular track (microtubule, actin filament, etc.) is discussed from a general viewpoint open to experimental test. An elementary "barometric" relation for the driving force is introduced that (i) applies to a range of kinetic and stochastic models, (ii) is consistent with more elaborate expressions entailing explicit representations of externally applied loads, and (iii) sufficiently close to thermal equlibrium, satisfies an Einstein-type relation in terms of the velocity and diffusion coefficient of the (load-free) motor. Even in the simplest two-state models, the velocity-vs.-load plots exhibit a variety of contrasting shapes (including nonmonotonic behavior). Previously suggested bounds on the driving force are shown to be inapplicable in general by analyzing discrete jump models with waiting time distributions.     

Partial tuning of motor cortex neurons to final posture in a free-moving paradigm

Motor cortex neurons in the monkey brain were tested with a diverse and naturalistic arm movement set. Over this global set of movements, the neurons showed a limited but significant degree of tuning to the multijoint posture attained by the arm at the end of each movement. Further supporting the hypothesis that the neurons are partially tuned to end posture, the postures preferred by the neurons significantly matched the postures evoked by electrical stimulation of the same cortical sites. However, much of the variance in neuronal activity remained unexplained even by the end-posture model, and thus other variables must have contributed to the response profile of the neurons. One possibility is that motor cortex neurons become tuned to the wide variety of movement parameters that are relevant to the animal's normal behavioral repertoire, and, therefore, any one parameter accounts for only a limited amount of neuronal variance.     

Thermal modulation of birefringence observed in a crystalline molecular gyrotop

Recently, functional organic materials have been put into practical use. The application of molecular motions has the potential to create new molecule-based materials. For this reason, considerable attention has been focused on the chemistry and properties of molecular machines in which mechanical motions of parts of the molecules are observed. In particular, phenylene rotation in the crystalline state has been investigated using framed molecular gyrotops having a phenylene rotor encased in three long alkyl spokes. In this study, we show thermal modulation of birefringence in a crystal due to the states of dynamic equilibrium of a novel molecular gyrotop. A macrocage molecule having a bridged phenylene rotor was synthesized as a novel molecular gyrotop. Rapid rotation of the phenylene rotor of the molecular gyrotop was confirmed by solid-state (2)H NMR spectroscopy that showed changes in the optical properties of a single crystal, i.e., the thermal modulation of birefringence. These results are the first application of the dynamic states in a crystal causing an optical change. These phenomena were also confirmed by control experiments using a molecular gyrotop with a nonrotating xylene rotor. We anticipate our finding to be a starting point for the creation of a new field of material chemistry that will make use of the dynamic states of molecules.     

Cholesteryl myristate conformation in liquid crystalline mesophases determined by neutron scattering.

The possible involvement of cholesteryl ester states in the development and persistence of atherosclerosis and the transport and storage of cholesteryl esters has led to questions concerning the organization and conformation of cholesteryl ester molecules in both pure phases and membranes. The experiments we report here were designed to measure the distance between the center of mass of the fatty acyl terminal methyl group and the center of mass of the three-carbon branched terminus of the cholesterol moiety at the opposite end of the molecule. The distance obtained is thus a gauge of cholesteryl ester conformation through the conformational range from a completely extended conformation to a U-shaped conformation. Neutron scattering experiments on partially deuterated samples of pure cholesteryl myristate in the crystalline, smectic, cholesteric, and isotropic phases indicate that the molecule is extended in each of these states. A discussion of specific molecular models consistent with these results and extension of these conclusions to other cholesteryl esters is included.     

Resonant tectorial membrane motion in the inner ear: its crucial role in frequency tuning.

The tectorial membrane has long been postulated as playing a role in the exquisite sensitivity of the cochlea. In particular, it has been proposed that the tectorial membrane provides a second resonant system, in addition to that of the basilar membrane, which contributes to the amplification of the motion of the cochlear partition. Until now, technical difficulties had prevented vibration measurements of the tectorial membrane and, therefore, precluded direct evidence of a mechanical resonance. In the study reported here, the vibration of the tectorial membrane was measured in two orthogonal directions by using a novel method of combining laser interferometry with a photodiode technique. It is shown experimentally that the motion of the tectorial membrane is resonant at a frequency of 0.5 octave (oct) below the resonant frequency of the basilar membrane and polarized parallel to the reticular lamina. It is concluded that the resonant motion of the tectorial membrane is due to a parallel resonance between the mass of the tectorial membrane and the compliance of the stereocilia of the outer hair cells. Moreover, in combination with the contractile force of outer hair cells, it is proposed that inertial motion of the tectorial membrane provides the necessary conditions to allow positive feedback of mechanical energy into the cochlear partition, thereby amplifying and tuning the cochlear response.     

Myosin-X is a molecular motor that functions in filopodia formation

Despite recent progress in understanding lamellipodia extension, the molecular mechanisms regulating filopodia formation remain largely unknown. Myo10 is a MyTH4-FERM myosin that localizes to the tips of filopodia and is hypothesized to function in filopodia formation. To determine whether endogenous Myo10 is required for filopodia formation, we have used scanning EM to assay the numerous filopodia normally present on the dorsal surfaces of HeLa cells. We show here that siRNA-mediated knockdown of Myo10 in HeLa cells leads to a dramatic loss of dorsal filopodia. Overexpressing the coiled coil region from Myo10 as a dominant- negative also leads to a loss of dorsal filopodia, thus providing independent evidence that Myo10 functions in filopodia formation. We also show that expressing Myo10 in COS-7 cells, a cell line that normally lacks dorsal filopodia, leads to a massive induction of dorsal filopodia. Because the dorsal filopodia induced by Myo10 are not attached to the substrate, Myo10 can promote filopodia by a mechanism that is independent of substrate attachment. Consistent with this observation, a Myo10 construct that lacks the FERM domain, the region that binds to integrin, retains the ability to induce dorsal filopodia. Deletion of the MyTH4-FERM region, however, completely abolishes Myo10's filopodia-promoting activity, as does deletion of the motor domain. Additional experiments on the mechanism of Myo10 action indicate that it acts downstream of Cdc42 and can promote filopodia in the absence of VASP proteins. Together, these data demonstrate that Myo10 is a molecular motor that functions in filopodia formation.     

Visual form discrimination from color or motion cues: functional anatomy by positron emission tomography.

To explore the extent to which various cortical functional pathways are involved in processing and analyzing different types of information that yield the same perceptual entity, we mapped anatomical structures in the human brain participating in the discrimination of visual forms mediated either by motion or color cues. Changes in regional cerebral blood flow were measured in 10 young male volunteers with positron emission tomography and with [15O]butanol. During the measurements, the subjects performed four visual discrimination tasks (form-from-motion, motion alone, form-from-color, and color alone discrimination). The individual regional cerebral blood flow images were standardized in shape and size with the help of a computerized brain atlas. Subtraction images were determined and averaged across data from all subjects. The resulting images were analyzed for statistically significant changes between specific and reference tasks. The discrimination of form by means of motion cues activated functional fields bilaterally in the inferior and lateral occipital gyri, in the lingual, anterior cingulate, middle frontal and orbitofrontal gyri, and in the left fusiform and right inferior temporal gyri. Form discrimination by color cues resulted in activation bilaterally in the inferior temporal, lateral occipital, and orbitofrontal gyri, the left precuneus and intraparietal sulcus, and the right precentral gyrus. The regions engaged in the two kinds of form discrimination did not overlap, demonstrating that differences in visual forms mediated by color or motion cues are processed and analyzed by disparate networks of functional fields in human cerebral cortex.     

Pre-steady-state DNA unwinding by bacteriophage T4 Dda helicase reveals a monomeric molecular motor

Helicases are molecular motor enzymes that unwind and translocate nucleic acids. One of the central questions regarding helicase activity is whether the process of coupling ATP hydrolysis to DNA unwinding requires an oligomeric form of the enzyme. We have applied a pre-steady-state kinetics approach to address this question with the bacteriophage T4 Dda helicase. If a helicase can function as a monomer, then the burst amplitude in the pre-steady state might be similar to the concentration of enzyme, whereas if the helicase required oligomerization, then the amplitude would be significantly less than the enzyme concentration. DNA unwinding of an oligonucleotide substrate was conducted by using a Kintek rapid quench-flow instrument. The substrate consisted of 12 bp adjacent to 12 nucleotides of single-stranded DNA. Dda (4 nM) was incubated with substrate (16 nM) in buffer, and the unwinding reaction was initiated by the addition of ATP (5 mM) and Mg(2+) (10 mM). The reaction was stopped by the addition of 400 mM EDTA. Product formation exhibited biphasic kinetics, and the data were fit to the equation for a single exponential followed by a steady state. The amplitude of the first phase was 3.5 +/- 0.2 nM, consistent with a monomeric helicase. The burst amplitude of product formation was measured over a range of enzyme and substrate concentrations and remained consistent with a functional monomer. Thus, Dda can rapidly unwind oligonucleotide substrates as a monomer, indicating that the functional molecular motor component of a helicase can reside within a single polypeptide.     

On the motion of a sphere in a perfect fluid, with application to liquid helium

Momentum relationships involved in the motion of a sphere through a perfect fluid are considered. The mechanism, by which the momentum given to a sphere by an external impulsive force is transferred to the container as a whole, is traced through in some detail, and energy and momentum relationships are discussed. Comparison is made with a ³He atom or a roton moving in superfluid ⁴He.     

Unidirectional retrograde atrioventricular nodal block in man: determinants of reversibility by vagal antagonism

The mechanism of unidirectional retrograde atrioventricular (AV) nodal block remains largely unknown. In this study, factors determining the reversal of the unidirectional block by atropine were evaluated in 12 patients who had no demonstrable ventriculoatrial (VA) conduction during ventricular pacing. Six patients demonstrated 1:1 VA conduction after atropine (group I), while the remaining six patients continued to show VA block (group II). During the control study there was no significant difference in the sinus cycle length and AH interval between the two groups. The percent decrease in sinus cycle length after atropine was also similar in groups I and II (i.e., 23 +/- 12 and 26 +/- 6, respectively). The effect on antegrade AV nodal conduction (i.e., the percent decrease in AH interval), however, was significantly greater in group I (24 +/- 9) as compared to group II (9 +/- 5) (p less than 0.004). The onset of VA conduction appeared to correlate with the improvement of antegrade conduction. The ratio of these two effects of atropine (i.e., percent decrease in AH interval to percent decrease in sinus cycle length) was higher when VA conduction was first demonstrated in group I (2.3 +/- 1.1) than at the maximal effect of atropine (1.2 +/- 0.3), reflecting a relatively greater decrease in sinus cycle length. Three of six group I patients redeveloped VA block at maximal effect of atropine. The results suggest a functional and dynamic nature of the unidirectional AV nodal block, possibly caused by vagal influence exaggerating the well-known directional asymmetry of AV nodal conduction in man.(ABSTRACT TRUNCATED AT 250 WORDS)     

Low molecular weight heparin prevents activation of coagulation in a hypobaric environment.

It is commonly thought that people are at increased risk of venous thrombosis during air flights, but the magnitude of the risk is unknown. Suggested risk factors are hypobaric hypoxia, stasis, and dehydration. In a previous experimental study, we found immediate activation of coagulation as determined by the levels of prothrombin fragments 1 + 2 (F(1 + 2)) and thrombin-antithrombin complex (TAT) after rapid exposure to a hypobaric and hypoxic environment (76 kPa). The aim of the present study was to examine the ability of low molecular weight heparin (LMWH) to prevent such activation. Twelve healthy male volunteers were given 40 mg enoxaparin as a single subcutaneous injection 1 h prior to exposure from 96.3 to 76 kPa. We found no activation of coagulation as judged by F(1 + 2) or TAT. Anti-activated factor X activity levels and release of tissue factor pathway inhibitor was normal. We conclude that high prophylactic doses of a LMWH most probably prevent activation of coagulation in a hypobaric environment.     

Anhydrous cholesterol: a new crystalline form in gallstones

A new crystalline form of anhydrous cholesterol called cholesterol II has been found in gallstones. This material can change within a few weeks into the usual form of anhydrous cholesterol but pure synthetic cholesterol II is stable.     

Illusory motion in Enigma: a psychophysical investigation

Experiments to determine the optimum conditions for perceiving illusory motion in Engima-like patterns have also demonstrated that the illusory motion is not the result of unintended motion of the image on the retina due to microsaccades or accommodative changes in the lens of the eye but instead has a cortical origin. The perceived illusory activity is believed to be a consequence of neural signals emanating from high-contrast bars and edges in the image that emit randomly fluctuating signals, as expected from spiking cortical neurons. These fluctuations may induce illusory motion in the channels by a mechanism similar to that responsible for the Omega effect, in which sequences of random patterns of black dots presented in an annular channel produce the perception of illusory rotation of these dots within the annulus.     

Dynamic chemical devices: modulation of contraction/extension molecular motion by coupled-ion binding/pH change-induced structural switching

Dynamic chemical devices involve morphological or constitutional modifications in molecular or supramolecular systems, induced by internal or external physical or chemical triggers. Reversible changes in shape result in molecular motions and define motional dynamic devices presenting mechanical-like actions of various types. Suitably designed polyheterocyclic strands such as compounds 1-5 wrap into helical conformations. The binding of lead(II) ions to the coordination subunits contained in the strand leads to complete uncoiling and yields a polymetallic complex presenting a fully extended shape. The addition of a cryptand complexing agent that strongly binds lead(II) ions and releases them under protonation allows a reversible pH-modulation of lead(II) levels in the medium, which in turn induces coiling/uncoiling of the molecular strand. This system thus represents a motional dynamic device which performs a mechano-chemical process, realizing alternating extension/contraction motions triggered by ion binding. It achieves a linear motor-type of action of very large stroke amplitude fueled by ionic processes.     

Intra-cytoplasmic crystalline inclusions in acute myeloid leukemia: a rare event

Unusual intracytoplasmic crystal-like inclusions within granulocytic precursor cells from a patient with acute myeloid leukemia (AML) are described. Based upon their cytochemical and immunochemical properties, the possible composition and origin of these inclusions are discussed. They represent a rare event in leukemic cell metabolism since, to our knowledge, only 1 case with identical characteristics has previously been reported.     

Unidirectional coronary collateral blood flow in a patient with congenital anomaly of a coronary artery: an angiographic artefact

A 53-year-old female underwent coronary arteriography for evaluation of chest discomfort. A congenital anomaly of a large interconnecting vessel between the right coronary and circumflex arteries was documented. Selective injection of the right coronary artery resulted in visualization of the circumflex branch of the left coronary through a large interconnecting vessel. Through careful analysis of the left ventriculogram and coronary arteriograms, it appears that retrograde filling of the circumflex artery during right coronary injection is an angiographic artefact.     

Detection of specific DNA sequences by fluorescence amplification: a color complementation assay.

We have developed a color complementation assay that allows rapid screening of specific genomic DNA sequences. It is based on the simultaneous amplification of two or more DNA segments with fluorescent oligonucleotide primers such that the generation of a color, or combination of colors, can be visualized and used for diagnosis. Color complementation assay obviates the need for gel electrophoresis and has been applied to the detection of a large and small gene deletion, a chromosomal translocation, an infectious agent, and a single-base substitution. DNA amplification with fluorescent oligonucleotide primers has also been used to multiplex and discriminate five different amplified DNA loci simultaneously. Each primer set is conjugated to a different dye, and the fluorescence of each dye respective to its amplified DNA locus is scored on a fluorometer. This method is valuable for DNA diagnostics of genetic, acquired, and infectious diseases, as well as in DNA forensics. It also lends itself to complete automation.