Biomedical subjects

D L Weeks

Publications and source records attributed to D L Weeks.

At least 37 records · Page 2Linked to original sources

PubMed1998

Cationic oligonucleotides can mediate specific inhibition of gene expression in Xenopus oocytes.

Base-specific hydrogen bonding between an oligonucleotide and the purines in the major groove of a DNA duplex provide an approach to selective inhibition of gene expression. Oligonucleotide-mediated triplex formation in vivo may be enhanced by a number of different chemical modifications. We have previously described an in vitro analysis of triplex formation using oligonucleotides containing internucleoside phosphate linkages modified with the cation N , N -diethyl-ethylenediamine (DEED). When compared with unmodified oligonucleotides of identical base composition, DEED-modified oligonucleotides were better able to form DNA triplexes under conditions that approximate the pH, magnesium and potassium levels found in vivo . Here we report the ability of DEED-modified oligonucleotides to inhibit the expression of plasmid DNA injected into Xenopus oocytes. Inhibition is specific to plasmids containing a triplex formation target and sensitive to sequence alteration in the triplex forming target site. Inhibition of gene expression was nearly complete when oligonucleotide and plasmid were mixed together prior to injection. Inhibition was partial when oligonucleotide was injected first and not evident when plasmid was injected and allowed to form chromatin prior to oligonucleotide injection. Thus, access to DNA is a determining factor in effective triplex inhibition of gene expression.

Animals↗

PubMed1998

Production of the transforming growth factor-beta binding protein endoglin is regulated during chick heart development.

The early embryonic heart consists of two cell types. The cells form an inner epithelial tube of endocardium within an outer tube of myocardium separated by a cell-free extracellular matrix. A crucial process in heart development is the production of cushion mesenchyme in the atrioventricular (AV) canal and outflow tract (OT). Cushion mesenchyme differentiates from the endocardium in response to signaling molecules produced by the adjacent myocardium. In chicken hearts, both transforming growth factor-beta3 (TGF-beta3) and TGF-beta2 are present and have been identified as being important in the production of cushion mesenchyme. We were interested in how the signals from these two similar molecules may be differentiated during early heart development. To this end, we examined the expression of endoglin, a TGF-beta receptor molecule, in the developing chick heart. Endoglin is typically located on endothelial cell layers and binds tightly to TGF-beta1 and TGF-beta3 but not well to TGF-beta2. We show that during the formation of the primitive heart tube, endoglin is found at relatively high levels in both presumptive myocardium and endocardium. However, as myocardium differentiates and development proceeds, endoglin expression is progressively reduced. At stage 20 in the heart, endoglin expression is most readily seen in the AV canal and the OT. This pattern of expression is similar to the reported TGF-beta3 expression patterns in the heart.

Animals↗

PubMed1998

Relative frequency of knowledge of performance and motor skill learning.

This study examined the effects of variations in relative frequency of knowledge of performance (KP) on acquisition, retention, and transfer of form for a multilimb closed sport skill. Two groups received either 100% relative frequency of KP or 33% relative frequency of KP while learning the soccer throw-in skill. Participants were boys between the ages of 11 and 14 years who were unfamiliar with the skill. Participants performed a 30-trial acquisition phase in which KP was provided about one of eight aspects of form. Following acquisition, five trial retention and transfer (to a target at a different distance than experienced in acquisition) tests were administered at 5 min, 24 hr, and 72 hr. Although no group differences were found for accuracy scores, the 33% group had higher form scores in acquisition and all retention and transfer tests. It was concluded that reducing the relative frequency of KP eliminated a dependency on KP to guide performance in acquisition, which was beneficial for maintaining form in conditions in which KP was absent.

Adolescent↗

PubMed1997

An3 protein encoded by a localized maternal mRNA in Xenopus laevis is an ATPase with substrate-specific RNA helicase activity.

ATP-dependent RNA helicases from the DEAD box family of proteins are involved in a number of RNA processing and utilization events. An3 protein from Xenopus laevis is an RNA helicase of the DEAD box family of proteins. An3 is synthesized by a mRNA that is localized to one end of Xenopus laevis oocytes. An3 protein is found in the nucleus of ooctes, and more specifically, during the middle stages of oocyte development, with extra nucleoli that contain amplified copies of rRNA genes in the nucleolus. By expressing glutathione-S-transferase:An3 fusion proteins in E. coli, sufficient amounts of An3 protein were isolated to examine its enzymatic activities. ATPase activity, NTP substrate range and RNA helicase activity were tested. An3 protein ATPase activity was evident but not stimulated by any of a variety of RNA tested. An3 protein was able to resolve the duplex formed by an in vitro substrate, in the presence of ATP or dATP. An3 required both 3' and 5' single-stranded regions of RNA flanking the RNA duplex it resolves.

Adenosine Triphosphatases↗

PubMed1996

Positively charged oligonucleotides overcome potassium-mediated inhibition of triplex DNA formation.

The formation of triplex DNA using unmodified, purine-rich oligonucleotides (ODNs) is inhibited by physiologic levels of potassium. Changing negative phosphodiester bonds in a triplex forming oligonucleotide (TFO) to neutral linkages causes a small increase in triplex formation. When phosphodiester bonds in a TFO are converted to positively-charged linkages the formation of triplex DNA increases dramatically. In the absence of KCl, a 17mer TFO containing 11 positively-charged linkages at a concentration of 0.2 microM converts essentially all of a 30 bp target duplex to a triplex. Less than 15% of the target duplex is shifted by 2 microMolar of the unmodified TFO. In 130 mM KCl, triplex formation is undetectable using the unmodified TFO, while triplex formation is nearly complete with 2 microM positively-charged TFO. With increasing potassium, TFOs containing a higher proportion of modified linkages show enhanced triplex formation compared with those less modified. In contrast with unmodified TFOs, triplex formation with more heavily modified TFOs can occur in the absence of divalent cations. We conclude that replacement of phosphodiester bonds with positively-charged phosphoramidate linkages results in more efficient triplex formation, suggesting that these compounds may prove useful for in vivo applications.

Base Composition↗

PubMed1996

Changes in nuclear localization of An3, a RNA helicase, during oogenesis and embryogenesis in Xenopus laevis.

The immunolocalization of An3 protein, an ATP-dependent RNA helicase and a member of the DEAD box family, was compared with the localization of fibrillarin, a protein essential for rRNA processing, and snRNPs, which are involved in mRNA splicing reactions, during oogenesis and embryogenesis in Xenopus laevis. Although An3 protein was detected in the cytoplasm of all stages of oocytes, in most stages An3 protein was also present in the nucleus. Prior to stage I An3 protein was uniformly dispersed throughout the entire germinal vesicle; from stages I to V it was in nucleoli. By stage VI nucleolar labeling with anti-An3 disappeared and the protein was no longer present within nuclei. An3 reactivity was also present throughout the nuclei of follicle cells surrounding prestage I to stage VI oocytes. Both cytoplasmic and nuclear An3 staining were present in cells of stages 8 to 35 embryos; however, nuclear staining was punctate and uniformly distributed throughout the nucleoplasm. Fibrillarin was diffusely distributed throughout the entire germinal vesicle prior to stage I, localized exclusively to nucleoli of oocytes between stages I and VI and in nucleoli of stages 12 and 35 embryonic cells. Reactivity for snRNPs (anti-Sm) in germinal vesicles of prestage I oocytes was diffuse, and similar to the distribution of An3 and fibrillarin; in later stage oocytes anti-Sm staining was restricted to a population of granules, much fewer in number and more heterogeneous in size than nucleoli. Anti-Sm activity was apparent in nuclei of embryonic cells of stages 8 to 35 embryos. Although colocalization of the Sm epitope and An3 was not observed in developing oocytes and in embryonic cells, Sm reactive material was frequently found in close association with An3-positive nucleoli (oocytes) and nuclear deposits (embryonic cells). In stage IV and V oocytes treated with actinomycin D (4 micrograms/ml) to inhibit rRNA synthesis, nucleoli, which continued to possess fibrillarin, lacked An3; staining of follicle cell nuclei for An3 was unchanged. Treatment with 200 micrograms/ml actinomycin D to block mRNA synthesis, inhibited An3 but not fibrillarin staining in nuclei of prestage I oocytes and follicle cells. The changing patterns of An3 reactivity and the differential effects of actinomycin D on such localizations observed here are consistent with a role for An3 in the processing/production of RNA.

Animals↗

PubMed1996

A comparison of imitation strategies in observational learning of action patterns.

The effects of different arrangements of demonstration and imitation of modeled actions on the learning of the 26 handshapes of the American manual alphabet were investigated. A concurrent group (N =16), which imitated handshapes concurrently with their demonstration, was compared with a delayed group (N = 16), which delayed imitation until 3 handshapes had been displayed, and with a combination group (N = 16), which practiced under a combination of concurrent conditions early in acquisition and delayed conditions later in acquisition. Following acquisition, learning was assessed by means of immediate and long-term recall and recognition tests. The delayed group was superior to the concurrent group in long-term serial recall and in immediate and long-term recognition of 3-letter sequences (in nonserial order); the performance of the combination group was between those of the delayed and concurrent groups. Therefore, delaying imitation in acquisition required subjects to expend more cognitive effort to retain and produce handshapes when requested than did concurrent imitation. This was beneficial to development of task knowledge that could be relied on for postacquisition recall and recognition of handshapes.

Journal Article↗

PubMed1996

One-trial adaptation of movement to changes in load.

1. We analyzed the rapid adaptation of elbow movement to unexpected changes in external load conditions at the elbow joint. The experimental approach was based on the lambda model, which defines control variables (CVs) setting the positional frames of reference for recruitment of flexor and extensor motoneurons. CVs may be specified by the nervous system independently of the current values of output variable such as electromyographic (EMG) activity, muscle torques, and kinematics. The CV R specifies the referent joint angle (R) at which the transition of flexor to extensor activity or vice versa can be observed during changes in the actual joint angle, theta, elicited by an external force. The other CV, the coactivation (C) command, instead of a single transition angle, defines an angular range in which flexor and extensor muscles may be simultaneously active (if C > 0) or silent (if C < 0). Changes in the R command result in shifts in the equilibrium state of the system, a dynamic process leading to EMG modifications resulting in movement or isometric force production if movement is obstructed. Fast movements are likely produced by combining the R command with a positive C command, which provides movement stability and effective energy dissipation, diminishing oscillations at the end of movement. 2. According to the model, changes in the load characteristic (e.g., from a 0 to a springlike load) influence the system's equilibrium state, leading to a positional error. This error may be corrected by a secondary movement produced by additional changes in R and C commands. In subsequent trials, the system may reproduce the CVs specified after correction in the previous trial. This behavior is called the recurrent strategy. It allows the system to adapt to the new load condition in the subsequent trials without corrections (1-trial adaptation). Alternatively, the system may reproduce the CVs specified before correction (invariant strategy). If the movement was perturbed only in a single trial, the invariant strategy allows the system to reach the target in the subsequent trials without corrections. 3. To test the assumption on the dominant role of the recurrent strategy in rapid adaptation of movement to new load conditions, we performed experiments in which subjects (n = 6) used a pivoting manipulandum and made fast 60 degrees movements to a target. After a random number of trials (5-10) with no load, we introduced opposing (experiment 1), assisting (experiment 2), or randomly varied opposing or assisting loads (experiment 3) for 5-10 trials before unexpectedly switching loads again (14-18 switches in total). The opposing or assisting torque was created by position feedback to a torque motor and was a linear function of the displacement of the manipulandum form the initial position (springlike load). Subjects were instructed to correct positional errors as soon as possible to reach the target. The EMG activity of two elbow flexors (biceps brachii and brachioradialis) and two elbow extensors (triceps brachii and anconeus), elbow position, velocity, and torque were recorded. Kinematic and EMG patterns were compared with those obtained in similar experiments in which subjects were instructed not to correct errors. 4. In 94% of the trials in which a change in the load occurred, the primary movement was in error and was followed by a corrective secondary movement. In primary movements, both the phasic and tonic levels of EMG activity as well as the kinematics were load dependent, implicating reflex and intramuscular mechanisms in the adaptation of muscle forces counteracting external loads. These mechanisms, however, were not sufficient to eliminate positional errors. 5. An undershoot error occurred in trials with an opposing load after those with no load or in trials with no load after those with an assisting load. After adaptation to a new load condition, a sudden return to the previous load condition resulted in an error of the oppo

Adult↗

PubMed1995

Mechanisms of cell transformation in the embryonic heart.

The process of cell transformation in the heart is a complex one. By use of the invasion bioassay, we have been able to identify several critical components of the cell transformation process in the heart. TGF beta 3 can be visualized as a switch in the environment that contributes to the initial process of cell transformation. Our data show that it is a critical switch in the transformation process. Even so, it is apparently only one of the factors involved. Others may include other TGF beta family members, the ES antigens described by Markwald and co-workers and additional unknown substances. Observing the sensitivity of the process to pertussis toxin, there is likely to be a G-protein-linked receptor involved, yet we have not identified a known ligand for this type of receptor. Clearly, there are several different signal transduction processes involved. The existence of multiple pathways is consistent with the idea that the target endothelial cells receive a variety of environmental imputs, the sum of which will produce cell transformation at the correct time and place. Adjacent endothelial cells of the ventricle that do not undergo cell transformation are apparently refractory to one or more of the stimuli. Figure 4 depicts a summary diagram of this invasion process with localization of most of the molecules mentioned in this narrative. As hypothesized here, elements of the transformation process may recapitulate aspects of gastrulation. Since some conservation of mechanism is expected in cells, it is not surprising that cells undergoing phenotypic change might reutilize mechanisms used previously to produce mesenchyme from the blastodisk. Though we have preliminary data to suggest this point, confirmation of the hypothesis by perturbation of genes such as brachyury, msx-1, etc. will be required to establish this point. The advantage of this hypothesis is that it provides, from the work of others in the area of gastrulation, a ready source of molecules and mechanisms that can be tested in the transforming heart. Whereas, perturbation of such mechanisms at gastrulation may be lethal to the embryo, such molecules and mechanisms may be responsible for the high incidence of birth defects in the heart.

Animals↗

PubMed1994

An3 mRNA encodes an RNA helicase that colocalizes with nucleoli in Xenopus oocytes in a stage-specific manner.

An3 is a maternal mRNA localized to the animal hemisphere of oocytes and early embryos. We have analyzed the enzymatic activity and the subcellular localization of the protein encoded by An3 mRNA during Xenopus oogenesis. Antibodies raised using recombinant full-length and truncated An3 protein recognized a single protein in Xenopus and single proteins from HeLa cells, Drosophila, mouse testes, and Saccharomyces cerevisiae. An3 protein immunoprecipitated from stage IV and stage VI oocytes had ATP-dependent RNA helicase activity. The subcellular location of An3 protein changed during oocyte development. In previtellogenic oocytes, An3 was present throughout the nucleus; cytoplasmic localization was relatively sparse. Nuclear localization in mid-vitellogenic oocytes was primarily nucleolar; cytoplasmic staining increased relative to earlier stages. In stage VI oocytes, An3 protein was detected only in the cytoplasm. The temporal change in An3 protein localization is consistent with a role in the production of large maternal pools of rRNA during oogenesis.

Animals↗

PubMed1994

Age-related differences in temporal scaling of postural EMG activity.

Electromyographical (EMG) burst onset times for postural muscles were compared in young and elderly male and female subjects performing a forceful abrupt pull with the right arm against a load cell apparatus while standing. EMG activity was collected via surface electrodes from the posterior deltoid, and right and left hamstring and quadriceps muscles. Pulls were performed at varying force levels from 100% to approximately 50% of a maximum effort. Regression equations were developed for each postural muscle for each group, using force production values as the independent variable, and postural muscle onset latencies (relative to deltoid onset) as the dependent variable. Regression slopes for postural muscles of elderly subjects ranged from 0.59 to 0.33, while slopes for the young subjects ranged from 0.72 to 0.63. Less elevated slopes for the elderly may indicate a reduced ability to temporally scale postural muscle onset latencies to variations in the force of the upper-limb task; a reduction in this ability may constitute a possible contributing factor to instability in the elderly.

Adult↗

PubMed1994

A comparison of knowledge of results scheduling methods for promoting motor skill acquisition and retention.

Providing knowledge of results (KR) to a learner about each trial in a series of trials only after the series is completed is termed summary KR. A variation of summary KR is average KR, in which KR is presented as an average value about the series of trials. This study compared groups receiving either average KR, summary KR, or KR following every attempt in acquisition of a force production task. Following acquisition, groups performed no-KR immediate and long-term retention tests. Analysis of variable error revealed group main effects and group by block interactions in acquisition and long-term retention. Post hoc tests indicated that differences were due primarily to inconsistent performance of the group receiving KR following each acquisition attempt. Post hoc tests did not, however, indicate differences between the average KR and summary KR groups in acquisition or retention. It was concluded that average and summary KR promote consistency in performance in acquisition, with this stability in responding carried over to retention performance.

Adult↗

PubMed1994

Attentional style in ratings of perceived exertion during physical exercise.

This study assessed the effects of associative and dissociative psychological strategies of attention on heart rate and self-report ratings of perceived exertion (RPE) during cycling performance. Seven trained cyclists performed a control ride, a dissociation ride, and an association ride on a bicycle ergometer at a work rate corresponding to 75% of their maximal heart rate. For the dissociation ride, subjects watched a videotape unrelated to cycling and responded to a key word each time it occurred on tape. For the association ride, subjects focused attention on heart-rate feedback available throughout the ride. During the control ride, attentional focus was not intentionally manipulated. Analysis indicated that the deliberate application of an attentional strategy did not significantly affect heart rate or RPE scores; however, the dissociation condition yielded somewhat higher RPE scores. From a postexperimental interview, four subjects responded that the association ride was the easier to complete, while three subjects responded the control ride was the easier one, matching a possible trend in the data.

Adolescent↗

PubMed1993

Two related localized mRNAs from Xenopus laevis encode ubiquitin-like fusion proteins.

The uneven distribution of maternal mRNAs in unfertilized eggs and the unequal inheritance of these molecules by dividing blastomeres may be one mechanism for determining cell fate during embryogenesis. Complementary DNA (cDNA) clones corresponding to maternal mRNAs localized to specific regions of the Xenopus laevis egg have been previously identified and cloned [Rebagliati et al., Cell 42(1985) 769-777]. The maternal mRNA, An1, was originally identified as being localized to the animal hemisphere of X. laevis eggs and early embryos. We describe here the two proteins encoded by two An1 mRNA isoforms which we designate An1a and An1b. These mRNAs are both approximately 3.0 kb long and are concentrated in the animal hemisphere of unfertilized eggs. The predicted amino acid (aa) sequences encoded by An1a and An1b correspond to 76.9 and 78.6 kDa, respectively, and are 88% identical. Both proteins contain a single N-terminal ubiquitin (Ub)-like domain (50% identical to X. laevis Ub) and a putative Zn(2+)-binding region near the C terminus. Unlike Ub polyproteins and most Ub fusion proteins, the N-terminal Ub-like domain found in the An1 proteins does not undergo proteolytic processing. In contrast to earlier studies showing that the An1 mRNA represents a strictly maternal transcript, we report that both related An1 transcripts are found in later embryonic stages and in all adult tissues tested.

Amino Acid Sequence↗

PubMed1993

Effects of self-modeling on self-efficacy and balance beam performance.

The purpose of this investigation was to assess the effect of self-modeling on self-efficacy and performance of balance beam routines. Subjects were intermediate-level female gymnasts who were randomly assigned to one of two groups, a self-modeling or a control group. For the self-modeling group, self-modeling videotapes were made of each subject performing her balance beam routine. During a 6-wk. period, self-modeling group subjects viewed the videotape of themselves three times a week prior to practice. During this time, the control group and self-modeling group participated in their normal instructional program. All subjects completed self-efficacy inventories and balance beam skill tests at four intervals, a pretest, a 2-wk. test, a 4-wk. test, and a 6-wk. posttest. Although no significant differences in ratings of self-efficacy or balance beam performance between the groups were found, the correlation between subjects' self-rated performance scores and actual performance scores for the self-modeling group was significant (r = .92). This correlation was not significant for the control group (r = .02). This significant correlation suggests that self-modeling may enhance performers' ability to assess their own performance realistically, improving their understanding and use of instructional feedback to enhance performance.

Adolescent↗

PubMed1992

Sense and antisense TGF beta 3 mRNA levels correlate with cardiac valve induction.

The formation of the valves in the heart is a spatially and temporally controlled process. A tissue interaction between the endothelium and its adjacent myocardium initiates the transformation of the endothelium into the mesenchymal precursors of the heart valve. One or more of the molecules implicated as critical for valve formation are members of the transforming growth factor beta family of molecules. Presented here is a spatial and temporal analysis of TGF beta 2 and TGF beta 3 in the chick heart during valve formation. We show that TGF beta 3 mRNA is concentrated in AV canal tissue where valve formation will occur, consistent with previous observations that TGF beta 3 production is critical during valve formation. Additionally, an RNA complementary to TGF beta 3 encoding mRNA is present in the heart. The temporally controlled appearance of RNA complementary to TGF beta 3 suggests that this molecule may play a role in the regulation of TGF beta 3 production in the heart.

Animals↗

PubMed1992

TGF-beta 3-mediated tissue interaction during embryonic heart development.

A critical process during early heart development is the formation of mesenchymal cells which will contribute to valves and septa of the mature heart. These cells arise by an epithelial-mesenchymal transformation of endothelial cells in the atrioventricular (AV) canal and outflow tract areas of the heart. Adjacent endothelial cells in the atrium and ventricle remain epithelial. A three-dimensional collagen gel culture system has been exploited to examine the interactions that mediate this transformation. The AV canal myocardium produces a stimulus that is transmitted through an intervening extracellular matrix to the AV canal endothelium. This interaction is regionally specific, such that ventricular myocardium does not provide an adequate stimulus and ventricular endothelium does not respond to the AV canal myocardial stimulus. Exogenous TGF-beta 1 (or TGF-beta 2) can complement ventricular myocardium to produce transformation by AV canal endothelium. A blocking antibody, effective against several TGF-beta, prevents cell transformation. To identify the specific member of the TGF-beta family that functions in situ, antisense oligonucleotides for each of the numbered TGF-beta were topically added to AV canal explant cultures. Only the oligonucleotide targeted to TGF-beta 3 was an effective inhibitor of mesenchymal cell formation. Studies have been undertaken to localize specific mRNas by in situ hybridization and RNase protection assays. These assays have concentrated on the regional and temporal appearance of TGF-beta 2 and 3. Surprisingly, RNase protection assays with a TGF-beta 3 sense probe showed the presence of a transcript complementary to TGF-beta 3.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

A comparison of modeling modalities in the observational learning of an externally paced skill.

This study contrasted prepractice modeling with either the perceptual component (perceptual modeling) or the motor component (movement pattern) of a coincident-timing task to determine whether experiencing the modalities singly or in combination enhanced timing performance on initiation of active practice. The motor component was a 60-cm right-to-left arm movement coincident with the illumination of lights on a Bassin timer runway to displace a barrier as the final runway light was illuminated. Four groups were compared (n = 12 per group). A perceptual modeling group passively viewed stimulus runway lights prior to attempting the task. A motoric modeling group viewed a videotape prior to practice of a model performing the motor component of the skill with zero timing error. A perceptual modeling plus motoric modeling group experienced both modeling modalities prior to performance. Finally, a no modeling group simply initiated practice on the task without modeling. Results indicated that the groups experiencing perceptual modeling initiated practice with significantly less average timing error and variability. Thus, perceptual modeling appeared to be at least as important as motoric modeling as a source of prepractice information to make available to a learner to optimize coincident-timing skill acquisition.

Analysis of Variance↗