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D L Weeks

Publications and source records attributed to D L Weeks.

52 records · Page 3Linked to original sources
PubMed1992

Evaluation of a system for rating edema in dairy cattle.

Our objective was to evaluate a subjective edema rating system to quantitate edema severity. Edema was evaluated with 20 nulliparous Holstein heifers from 3 wk before their expected calving date until 3 wk postpartum. Severity of udder edema was evaluated independently daily by five people throughout the 68-d experiment; a 10-point rating system (0 = no edema, 10 = severe edema) was used. One of the people developing the scoring system served as the official scorer. The accuracy of the rating system, defined as the closeness of test evaluator scores to official scores, was highest for scores 2 to 8. The precision of the system, identified by the standard error of the mean, was similar for each score in the rating system. The largest disagreement between official and test evaluator scores was evident postpartum. The large number of defined points in this rating system and the similarity between official and test scores in this study indicate the usefulness of this tool to evaluate precisely the severity of edema in dairy cattle.

Animals↗
PubMed1991

The Xenopus localized messenger RNA An3 may encode an ATP-dependent RNA helicase.

The maternal messenger RNA An3 was originally identified localized to the animal hemisphere of Xenopus laevis oocytes, eggs and early embryos. Xenopus embryos depend on mRNA and protein present in the egg before fertilization (maternal molecules) to provide the information needed for early development. Localization of maternal mRNA gives cells derived from different regions of the egg distinctive capacities for protein synthesis. We show here that An3 mRNA encodes a protein with 74% identity to a protein encoded by the testes-specific mRNA PL10 found in mouse, which is proposed to have RNA helicase activity. Because the gene encoding An3 mRNA is reactivated after gastrulation and remains active throughout embryogenesis, we have examined its distribution in embryonic and adult tissues. Unlike PL10 mRNA, which is primarily restricted to the testes, An3 mRNA is broadly distributed in later development.

Amino Acid Sequence↗
PubMed1991

Epithelial-mesenchymal transformation of embryonic cardiac endothelial cells is inhibited by a modified antisense oligodeoxynucleotide to transforming growth factor beta 3.

During early cardiac development, the progenitor cells of the heart valves and membranous septa undergo an epithelial-mesenchymal transformation. Previous studies have shown that this transformation depends on the activity of a transforming growth factor beta (TGF beta) molecule produced by the heart. In the present study, we have used modified antisense oligodeoxynucleotides generated to nonconserved regions of TGF beta 1, -2, -3, and -4 to examine the possible roles of these members in this transformation. A phosphoramidate-modified oligonucleotide complementary to TGF beta 3 mRNA was capable of inhibiting normal epithelial-mesenchymal transformation by 80%. Unmodified oligonucleotides to TGF beta 3, modified oligonucleotides to TGF beta 1, -2, and -4, and two modified control oligonucleotides were unable to inhibit the transformation. These data demonstrate that a specific member of the TGF beta family, TGF beta 3, is essential for the epithelial-mesenchymal cell transformation.

Animals↗
PubMed1991

Pathways of degradation and mechanism of action of antisense oligonucleotides in Xenopus laevis embryos.

Recently, we described a new class of antisense oligonucleotides that can be used to direct the cleavage of mRNAs in Xenopus laevis embryos by RNase H (Dagle et al., Nucleic Acids Res. 18, 4751-4757). In this study, we have examined several factors that determine the activity of these derivatives. In embryos, oligodeoxyribonucleotides were found to be rapidly degraded by a 3' exonuclease. Modification of 3'-terminal phosphodiester linkages as phosphoramidates blocks this activity. The predominant sites of endonucleolytic cleavage within the embryo are localized close to the 5' termini demonstrating the necessity of multiply modifying phosphodiester linkages at each end of the molecule. A stretch of at least six consecutive phosphodiester linkages is required to form an effective substrate for Xenopus RNase H; mRNA degradation with an oligonucleotide containing fewer than six contiguous unmodified internucleoside linkages is greatly diminished. Injection of an anti-cyclin B oligonucleotide containing eight unmodified residues results in degradation of cyclin B mRNAs and subsequent inhibition of embryonic cell division. An oligonucleotide with the same sequence but containing four consecutive phosphodiesters has no observable effect on the cell cycle. This last observation suggests that, in Xenopus embryos, hybridization alone has a limited role, if any, in oligonucleotide-mediated inhibition of gene expression.

Animals↗
PubMed1991

Effect of ivermectin on the volume of blood ingested by two species of ticks (Acari: Ixodidae) feeding on cattle.

Females of the lone star tick, Amblyomma americanum (L.), and the American dog tick, Dermacentor variabilis (Say), were fed on ivermectin (Ivomec)-treated and untreated bovines to determine the effect of the acaricide on volume of blood ingested and to compare the weight differences between the treatment and control groups at various time intervals after attachment. Adult females from each genus were collected from Bos tarus hosts and subjected to hematin assays on three collection dates to estimate the volume of blood ingested. Before feeding, lone star ticks contained an average of 2.0 microliters of blood and had an average weight of 5.2 mg. Unengorged American dog tick females had an average blood volume of 3.3 microliters and a mean weight of 5.8 mg. Ticks of both species reacted to ivermectin by expressing lower mean weights, and they consumed smaller quantities of blood. Lone star tick females were significantly affected in terms of amount of blood consumed and body weight changes when compared with control ticks. After feeding on treated cattle, lone star tick females contained smaller quantities of blood than pretreatment females, but there were no significant differences observed until day 12 between the control and the treated groups. American dog tick females on treated hosts had measurable quantities of blood that were significantly different among the experimental groups.

Animals↗
PubMed1991

Cyclin B mRNA depletion only transiently inhibits the Xenopus embryonic cell cycle.

The control of the cell cycle is dependent on the ability to synthesize and degrade proteins called cyclins. When antisense oligonucleotides are used to deplete Xenopus embryos of mRNA encoding cyclin B protein, embryonic cleavage is inhibited. Surprisingly, after missing several rounds of cleavage, the cell cycle and cell division resumes. These studies indicate that the early embryonic cell cycle can proceed with undetectable levels of cyclin B encoding mRNA. In contrast, other events of normal development, including the activation of embryonic transcription and gastrulation, are inhibited.

Animals↗
PubMed1990

Targeted degradation of mRNA in Xenopus oocytes and embryos directed by modified oligonucleotides: studies of An2 and cyclin in embryogenesis.

We have designed antisense oligodeoxyribonucleotides which are both highly resistant to nucleolytic degradation and also serve as substrates for ribonuclease H. Using these compounds we have targeted the specific degradation of several maternal mRNAs present in Xenopus laevis oocytes and early embryos. Several internucleoside linkages at both the 3' and 5' ends of the oligonucleotides were modified as phosphoramidates to provide complete protection against exonucleases, the predominant nucleolytic activity found in both oocytes and embryos. Eight Internal linkages were left unmodified to provide a substrate for RNase H. Degradation of specific embryonic mRNAs was accomplished using subtoxic amounts of the modified oligonucleotides. Specific depletion of An2, a localized mRNA encoding the alpha subunit of the mitochondrial ATPase, produced embryos that gastrulated later than control embryos and arrested in development prior to neurulation. A modified oligonucleotide targeting Xenopus cyclin B1 and cyclin B2 mRNA was also synthesized. Following the injection of one blastomere of a two-cell embryo with the anti-cyclin oligonucleotide, cell division in that half of the embryo was inhibited, demonstrating the in vivo importance of these cyclins in mitosis. The oligonucleotide analogs described here should be useful in studying developmentally significant proteins in Xenopus.

Adenosine Triphosphatases↗
PubMed1988

Temporal constraints in the control of prehensile movement.

Three experiments were conducted to investigate the control of the manipulation (i.e., finger-thumb aperture) and transportation (i.e., wrist velocity) components in prehensile movement (Jeannerod, 1981, 1984). In all experiments, subjects were seated and instructed to grasp a dowel mounted on a joystick following a discrete movement over a set distance. Thus, the amount of dowel movement following the grasp could be determined. In Experiment 1, the tolerance (i.e., the amount of allowable dowel movement) was manipulated using a computer-generated boundary around the dowel. The results indicated that the transportation component changed dependent on the tolerance condition, and there were trends that maximum aperture was also affected. Experiment 2 manipulated both tolerance and dowel size (i.e., diameter) factorially in a within-subject design. Dowel size affected only the manipulation component, supporting Jeannerod's (1981) earlier work, but tolerance clearly influenced both components. Experiment 3 investigated Wing, Turton, and Fraser's (1986) proposition that speed of movement influences aperture size. Distance and movement time were combined factorially to produce conditions with different average velocities. Maximum aperture was dependent on the movement time rather than the speed of movement. The relation between the control of the components was examined by using a new method of calculating within-trial correlations between aperture size and wrist velocity in Experiments 2 and 3. The correlations were related to the temporal aspects of the movement with higher correlations in the rapid movement time conditions. Also, the temporal occurrence of maximum aperture remained invariant across the different movement conditions. In general, the results suggest a strong functional linkage between the two components, which may be dependent on the temporal characteristics of the movement.

Journal Article↗
PubMed1987

A maternal mRNA localized to the vegetal hemisphere in Xenopus eggs codes for a growth factor related to TGF-beta.

We report that Vg1, a maternal mRNA localized to the vegetal hemisphere of frog eggs, encodes a member of the transforming growth factor-beta (TGF-beta) family of proteins. Furthermore, we show that Vg1 mRNA is distributed to presumptive endodermal cells after fertilization. Previous studies had shown that the vegetal end of a frog egg produces a signal that induces the overlying animal pole cells to form mesodermal tissue. More recently it has been shown that fibroblast growth factor (FGF) and TGF-beta can participate in the induction of muscle. Together, these results lead us to propose that the formation of mesoderm during frog development is specified by the products of localized maternal mRNAs, including Vg1.

Amino Acid Sequence↗
PubMed1987

A maternal mRNA localized to the animal pole of Xenopus eggs encodes a subunit of mitochondrial ATPase.

We have previously isolated several cDNA clones of mRNAs that have the unusual property of being localized to either the animal pole or the vegetal pole of frog eggs. To gain insight into the function of these maternal mRNAs we have determined their DNA sequence and deduced the sequence of the proteins they encode. Here we report that An2, an mRNA localized to the animal pole of Xenopus oocytes and eggs, codes for the alpha chain of mitochondrial ATPase. Furthermore, we compare the intracellular localization of the An2 mRNA and mitochondria in oocytes and eggs and find that they do not have the same degree of localization. In the light of these results we discuss possible reasons for the maternal localization of the An2 mRNA.

Adenosine Triphosphatases↗
PubMed1985

Adenovirus E3-early promoter: sequences required for activation by E1A.

To identify sequences within the adenovirus-5 E3 promoter necessary for E1A trans-activation, a series of promoter deletion mutants were constructed and analysed. A region between positions -82 and -105 was shown to be critical both for E1A induced expression as well as uninduced expression. The importance of this region was confirmed by constructing hybrid promoters consisting of E3 and Herpes simplex virus thymidine kinase sequences. The E1A insensitive tk promoter could be converted to an E1A sensitive promoter by replacing sequences upstream of position -79 with the corresponding region of the E3 promoter. This critical region of the E3 promoter contains a sequence 5' AGATGACTA3' which is also present in important upstream regions of the E2A and E4 promoters.

Adenoviruses, Human↗
PubMed1985

Identification and cloning of localized maternal RNAs from Xenopus eggs.

A central question in developmental biology is to explain how cells in different regions of an embryo acquire different developmental fates. We have begun to address this question by investigating whether specific RNAs are localized within a frog egg. Differential screening of a cDNA library shows that most maternal RNAs are uniformly distributed along the animal-vegetal axis. However, we find that a rare class of maternal RNAs is localized. cDNA clones of four localized RNAs have been characterized. Three of these cDNAs are derived from maternal RNAs that are concentrated in the animal hemisphere of unfertilized eggs and remain localized through the early blastula stage. One cDNA is derived from a maternal RNA found almost exclusively in the vegetal hemisphere at both stages. These studies show that some informational molecules, specifically RNAs, are localized in eggs and are inherited by particular blastomeres.

Animals↗
PubMed1983

Regulation of adenovirus transcription by an E1a gene in microinjected Xenopus laevis oocytes.

The regulation of adenovirus type 5 gene expression by the E1a gene product was examined in microinjected Xenopus laevis oocytes. Chimeric genes were constructed which included the promoter region of early adenovirus type 5 gene 3 and the structural sequence which codes for the bacterial enzyme chloramphenicol-3-O-acetyltransferase (CAT). A plasmid containing this chimeric gene as well as plasmids containing the E1a gene were coinjected into oocyte nuclei. The presence of the E1a gene was shown to increase CAT activity by up to 8.5-fold over basal levels. Synthesis of the functional product from the E1a gene requires the removal of intron sequences by RNA splicing. The E1a gene and a derivative that precisely lacks the intron were equally effective in increasing CAT activity, suggesting that splicing of the primary E1a transcript is efficiently accomplished in the oocyte nucleus. This was confirmed by directly examining the E1a mRNAs by the S1 mapping procedure. A protein extract from adenovirus type 5-infected HeLa cells enriched for the E1a protein may supplant the E1a plasmid in enhancing CAT activity. Synthesis of the CAT enzyme after gene injection is invariant in oocytes from the same frog, but oocytes from different frogs show a high degree of variability in their ability to synthesize the CAT enzyme. Microinjected X. laevis oocytes appear to be an extremely useful system to study the effects of protein elements on transcription.

Acetyltransferases↗
PubMed1983

E1A control of gene expression is mediated by sequences 5' to the transcriptional starts of the early viral genes.

A product of the adenovirus E1A gene is a positive regulator of early viral gene expression. In this report we show that E1A regulates at the transcriptional level and that sequences located 5' to the early viral regions contain sites which confer regulation by the E1A gene product. We constructed chimeric genes in which the sequences at the 5' end of the E2A, E3, and E4 regions were fused to the structural sequences of either the herpes simplex virus thymidine kinase gene, the bacterial gene encoding the enzyme neomycin phosphotransferase, or the chloramphenicol acetyltransferase gene. In all cases, expression of the chimeric genes was induced by a product of the E1A region. It was also found that the insertion of a fragment from the left-hand end of the adenovirus type 5 genome into a plasmid harboring the thymidine kinase gene resulted in elevated frequencies of transformation of TK- cells to TK+. The elevated transformation frequencies were only detected when the insert and tk gene were covalently joined. This effect occurred even when the insert was several kilobase upstream from, and regardless of its orientation to, the transcriptional initiation site of the tk gene. We propose that this region of the adenovirus type 5 genome harbors a cis-acting enhancer of transcription.

Adenoviridae↗
PubMedDate not supplied

Interaction between attention demanding motor and cognitive tasks and static postural stability.

BACKGROUND: Due to the often-reported decrease in postural stability in the elderly, it is important to understand factors that may contribute to reduced postural stability. It is possible that attention-demanding focal tasks performed concurrent with postural regulation influence postural stability. OBJECTIVE: This study utilized dual-task methodology to determine if motor or cognitive focal tasks interact with center of pressure (COP) excursion during static bipedal stance in healthy young and healthy elderly subjects (n = 18). METHODS: The cognitive task involved silently solving an orally-presented multi-step arithmetic problem over a 30-second period. The motor task was a 30-second bilateral static finger-thumb pinch task performed at 10% of maximal voluntary contraction with a pair of pinch-force transducers. Each focal task was performed separately, and in a condition in which both tasks were performed simultaneously. COP excursion was compared in quiet standing (no focal task) and during performance of the focal tasks with full vision and with vision occluded. RESULTS: Performance on the focal tasks was unaffected by increased postural demands during stance as compared to a seated baseline condition. This was the case for both age groups, and for the full vision and occluded vision conditions. Medio-lateral COP excursion was reduced over the quiet standing pretest condition when attentional focus was on the cognitive task, suggesting that COP was influenced centrally during cognition. In contrast, COP excursion increased over the quiet standing pretest condition when performing the motor focal task, suggesting a reduced ability to suppress sway when the motor system was concurrently occupied with a voluntary task that shared the same input-output resources. CONCLUSION: The ability to share attentional resources among focal and postural tasks was similar in healthy young and elderly subjects.

Adult↗