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Biomedical subjects

K Taylor

Publications and source records attributed to K Taylor.

At least 181 records · Page 10Linked to original sources
PubMed1995

Plasmid and host functions required for lambda plasmid replication carried out by the inherited replication complex.

We have shown previously that in amino acid-starved, relaxed (rel-) mutants of Escherichia coli replication of the lambda plasmid occurs via the lambda O-containing replication complex (RC) that was assembled prior to the onset of amino acid starvation and is inherited by one of the two daughter plasmid circles in each replication cycle. This replication is regulated neither by binding of the lambda O initiator to ori lambda, nor by the lambda Cro-mediated repression. Here we show that it is dependent on both RNA polymerase and DnaA functions, which is consistent with our recent finding that transcriptional activation of ori lambda is under the control of DnaA. In the system studied, DnaA-regulated transcriptional activation of ori lambda seems to be the only rate-limiting process. The lambda plasmid replication mediated by the inherited RC appeared to be independent of the functions of lambda P and DnaJ required in RC assembly In vitro experiments performed by others suggest that DnaJ first binds to the ori lambda-bound lambda O-lambda P-DnaB pre-primosome and subsequently lambda P complexed with DnaJ is preferentially recognized by DnaK-GrpE; chaperone-mediated rearrangement of this structure relieves DnaB helicase of lambda P inhibition. Recently we proposed that this process is directly coupled to the insertion of the pre-primosome between DNA strands transiently separated by transcription. This last-mentioned process may be required in lambda plasmid replication mediated by the inherited RC, which appeared in turn to be dependent on DnaK and GrpE functions.

Bacterial Proteins↗
PubMed1995

Transcriptional activation of the origin of coliphage lambda DNA replication is regulated by the host DnaA initiator function.

The initiator of phage lambda DNA replication, the lambda O protein, is considered to be an analogue of the initiator of DNA replication (DnaA) of its host, Escherichia coli. Both specifically recognize their origins of replication, ori lambda and oriC, respectively, and organize the assembly of specific replication complexes. However, DnaA has an additional activation function, acting on oriC-proximal DnaA-boxes, and regulating transcription initiated at promoters in and around oriC. Here, we demonstrate that lambda plasmid replication can be synchronized by a temperature shift-down that caused renaturation of the previously denatured DnaAts protein. Moreover, we show that elimination of the activating DnaA function affects transcriptional activation at ori lambda. DnaA may act by binding to DnaA-boxes, situated around the lambda pR promoter; there are no such sequences in ori lambda. Our results being to explain in molecular terms why lambda plasmid replication is DnaA-dependent [Kur et al., J. Mol. Biol. 198 (1987) 203-210] and why the initiation of phage lambda DNA replication is blocked (in E. coli devoid of prophage Rac) after inactivation of DnaA [Wegrzyn et al., Genetics (1995) in press].

Bacterial Proteins↗
PubMed1995

Protection of coliphage lambda O initiator protein from proteolysis in the assembly of the replication complex in vivo.

We have shown previously that, in contrast to the free coliphage lambda O initiator protein rapidly degraded by ClpP/ClpX protease, the lambda present in the replication complex (RC) is protected from proteolysis. Now we asked at which step of the pathway of RC assembly in vivo does the stabilization of lambda O occur. In accordance with the in vitro established order we found that lambda P and DnaB helicase functions are, but those of DnaJ and GrpE chaperones are not, required for the protection of lambda O from proteolysis. Therefore, our results suggest that the first lambda O protecting structure of the pathway of RC assembly is the lambda O-lambda P-DnaB preprimosome. The next step of the pathway, the chaperone-mediated rearrangement of the preprimosome, is not essential for lambda O stabilization. However, in contrast to other chaperones, the DnaK function was required for the protection of lambda O from proteolysis, suggesting an earlier access of DnaK to the pathway of RC assembly in vivo, in accordance with current models by which molecular chaperones facilitate protein assembly.

Bacterial Proteins↗
PubMed1995

Preservation of hemostatic and structural properties of rehydrated lyophilized platelets: potential for long-term storage of dried platelets for transfusion.

Currently, therapeutic platelet concentrates can be stored for only 5 days. We have developed a procedure that permits long-term storage of fixed and lyophilized platelets that retain hemostatic properties after rehydration. These rehydrated lyophilized platelets (RL platelets) restore hemostasis in thrombocytopenic rats and become incorporated in the hemostatic plug of bleeding time wounds of normal dogs as well as von Willebrand disease dogs with partially replenished plasma von Willebrand factor. Ultrastructurally, these platelets are well preserved and are comparable to control normal washed platelets. Flow cytometry analysis shows that RL platelets react with antibodies to the major surface receptors, glycoprotein (GP)Ib and GPIIb/IIIa. These receptors are involved in platelet agglutination, aggregation, and adhesion. In vitro functional tests document the ability of RL platelets to adhere to denuded subendothelium and to spread on a foreign surface. Circulating RL platelets participated in carotid arterial thrombus formation induced in normal canine subjects. The participation of RL platelets in these vital hemostatic properties suggests that with further development they could become a stable platelet product for transfusion.

Animals↗
PubMed1995

Comparative mapping of YRRM- and TSPY-related cosmids in man and hominoid apes.

Using chromosomal in situ hybridization it has been demonstrated that specific members of the YRRM and the TSPY families are multicopy and Y chromosome specific in hominoids. After hybridization with the YRRM-related cosmid A5F and the TSPY-related cosmids cos36 and cY91, a reverse and complementary pattern of main and secondary signals is detected on the Y chromosomes of the human, the pygmy chimpanzee and the gorilla, while the location of signals coincides on the Y chromosomes of the chimpanzee, both orang-utan subspecies and the white hand gibbon. This complementary distribution of YRRM and TSPY sequences on the hominoid Y chromosomes possibly originates from a similar sequence motif that is shared by and evolutionarily conserved between certain members of both gene families and/or repeated elements flanking those genes. Otherwise this complementary distribution could go back to a common organization of these genes next to each other on an ancient Y chromosome which was disrupted by chromosomal rearrangements and amplification of one or other of the genes at each of the locations.

Animals↗
PubMed1995

Trends in oral cancer by subsite in New Zealand.

A significant increase in incidence rates for all subsites of oral cancer in men and all but cancer of the salivary glands in women has occurred in New Zealand over the past 35 years. However, only male mortality rates of cancers of the nasopharynx and pharynx (excluding the nasopharynx) have significantly increased during this time period. Significant birth-cohort effects existed in the trends in incidence and mortality for cancers of the tongue, mouth, pharynx (excluding the nasopharynx), and nasopharynx among men. For cancers of the mouth and pharynx (excluding the nasopharynx) an increased risk for men born 1912-1932 existed, while an increased risk of cancer of the tongue occurred for men born 1922 onwards. For nasopharyngeal cancer, men born around 1912 and those born from about 1932 onwards were found to have an increased risk. For women, no consistent trends by birth-cohort were found for any oral subsite of cancer.

Adult↗
PubMed1995

Allelotype of squamous cell carcinoma of the head and neck: fractional allele loss correlates with survival.

Allelic imbalance or loss of heterozygosity (LOH) studies have been used extensively to identify regions on chromosomes that may contain putative tumour-suppressor genes. We have undertaken an extensive allelotype of 80 specimens of squamous cell carcinoma of the head and neck (SCCHN) using 145 polymorphic microsatellite markers on 39 chromosome arms. Allelic imbalances were found most frequently on chromosome arms 3p, 9p, 17p and 18q with over 45% LOH and imbalances on 1p, 1q, 2p, 5q, 6p, 6q, 8p, 8q, 9q, 11q, 13q, 17q and 19q were found in more than 20% of SCCHN. These LOH data were analysed against a range of clinicopathological parameters which included previously untreated and previously treated tumours; correlations were found between LOH on 9q and nodes at pathology (P = 0.02) and between histopathological grade and LOH on 12q (P = 0.02) and 13q (P = 0.01). In the group of previously untreated tumours, a correlation was found between site of tumour and LOH on 3p (P = 0.019), and 8p (P = 0.029), while TNM staging correlated with LOH on 3p (P = 0.019) and 17p (P = 0.016). Fractional allele loss (FAL) was calculated for 52 tumours with LOH data on nine or more chromosomal arms and found to have a median value of 0.22 (range 0.0-0.80). Correlations were found between FAL > median value and nodes at pathology (P = 0.01) and tumour grade (P = 0.06), demonstrating that advanced tumours with lymph node metastasis often had LOH at multiple sites. FAL > median value was found to correlate with a poor survival (P < 0.03) and, furthermore, FAL > median value correlated with poor survival in the previously untreated patients (P < 0.019). These results indicate that assessment of the accumulation of genetic damage, as provided by allelotype data, provides a useful molecular indicator of the tumour behaviour and clinical outcome.

Alleles↗
PubMed1995

Involvement of the host initiator function dnaA in the replication of coliphage lambda.

We demonstrate that the initiation of coliphage lambda DNA replication is dependent on the host initiator function dnaA, provided that the lambdoid prophage Rac is absent. Presence of Rac compensated the absence of dnaA function, causing initiation of replication. In dnaAts rac+ cells at 43 degrees, most of parental phage DNA molecules, after one round of theta replication, switched to a replication with features of the sigma mode and produced progeny at high yield. Initiation of replication of the lambda Pts1 mutant at 43 degrees was blocked by dnaA function; however, under dnaA-rac+ conditions all parental phage DNA molecules, after one round of theta replication, switched to the sigma mode and produced progeny at high yield. Taking into account our recent finding that transcriptional activation of ori lambda seems to be dnaA-regulated (to be published elsewhere), we suggest that the DnaA-lambda Pts1 incompatibility occurs at the insertion of the ori lambda-bound lambda O-lambda P-DnaB preprimosome between the complementary lambda DNA strands. The role of Rac and the mechanism of the switch from theta to sigma mode of lambda phage DNA replication are discussed.

Bacterial Proteins↗
PubMed1995

Replication of coliphage lambda DNA.

A general scheme of lambda phage and plasmid DNA replication in Escherichia coli is presented, and results of in vivo experiments from the authors' laboratory are superimposed. The initiator lambda O functions in the assembly of the replication complex (RC) at ori lambda, making it a stable component of this structure. ClpP/ClpX protease-specific action on lambda O does not affect the regulation of replication; it only degrades the surplus of synthesized lambda O. The initiator lambda O becomes protected from proteolysis at a distinct step of the pathway of RC assembly. The host DnaA initiator-regulated transcriptional activation of ori lambda seems to be coupled with RC assembly at the step of chaperone-mediated rearrangement of the pre-primosome. The once-assembled RC is inherited by one of two lambda plasmid daughter copies at each round of circle-to-circle (theta) replication. The inherited, old RC-driven replication is also dependent on RNA polymerase and DnaA functions. It seems that DnaA licenses lambda plasmid DNA for only one replication round, resembling the putative eukaryotic licensing factor in this respect. The lambda O binding to ori lambda does not seem to play any role in regulation of lambda plasmid replication, and the Cro-autoregulatory loop may be deleted. The emerging picture shows lambda plasmid circles with RCs bound to their ori, awaiting a signal triggering initiation of replication. The host DnaA initiator-regulated transcriptional activation of ori lambda may be involved in signal transmission. Inactivation of DnaA function blocks initiation of lambda phage DNA replication, but the lambdoid prophage Rac compensates this defect and all parental phage DNA molecules, after one round of theta replication switch to the sigma mode and produce progeny in high yield. We suspect that DnaA-regulated transcriptional activation is involved in installation and adequate positioning of two RCs, required for bidirectional replication, but in the Rac-promoted process only one RC may be installed, leading to unidirectional replication continued in the sigma mode. In wild-type cells consumption of DnaA function by the rapidly replicating lambda phage DNA may switch replication from bidirectional theta to unidirectional theta, and later to the sigma mode; the lambda circles produced earlier may play the role of Rac, which is required only when DnaA function has been inactivated prior to phage infection.

Bacteriophage lambda↗
PubMed1995

Interleukin-1 beta enhances spinal cord blood flow after intrathecal administration in the normal rat.

The effects of acute intrathecal recombinant human interleukin-1 beta (rhIL-1 beta) administration on spinal cord blood flow (SCBF), volume, and velocity were determined by laser-Doppler flowmetry in normal anesthetized rats with the use of a randomized and blinded protocol. The intrathecal administration of rhIL-1 beta (0.16-16 ng) produced a dose-dependent increase in SCBF that was not related to changes in blood pressure; arterial pH, PO2, PCO2; or spinal cord temperature. The IL-1 beta-induced enhancement of SCBF was directly proportional to the resultant elevation of spinal cord rhIL-1 beta content and was significantly correlated with an elevated blood velocity. The IL-1 receptor antagonist (IL-1ra) in concentrations 50- and 200-fold higher than IL-1 beta completely blocked the IL-1 beta-induced increase in SCBF when both compounds were administered concomitantly, but when administered alone, IL-1ra did not affect SCBF or other parameters. This suggests that IL-1 beta action was mediated by a specific interaction with an IL-1 membrane receptor site. The results suggest a role of IL-1 beta in the regulation of spinal cord hemodynamics. A potential pharmacological approach using IL-1 agonists for the treatment of the delayed appearance of posttraumatic spinal ischemia is proposed.

Animals↗
PubMed1995

Estimation of stature from the length of the cervical, thoracic, and lumbar segments of the spine in American whites and blacks.

In order to estimate stature from the length of cervical, thoracic, lumbar, thoraco-lumbar (T-L) and cervico-thoraco-lumbar (C-T-L) segments of the spine, measurements were made on white and black Americans, both male and female, autopsied during 1977-1993. Sample sizes were as follows: white males = 167; white females = 58; black males = 43; black females = 31. Separate measurements were made of the vertebral segments along the anterior surface of the spine. Regression formulae were calculated for each segment in each of the four groups. Standard errors of estimate ranged from 2.60 to 7.11 cm. Comparison was made with previous work published for Japanese. The Japanese formulae could not predict stature of the American populations using our data. The method is useful for estimating the stature of severely burned or mutilated bodies.

Adolescent↗
PubMed1995

Computer-assisted decision support systems for patient management in an intensive care unit.

The application of the intelligent monitoring techniques of case-based reasoning and neural network analysis to physician decision making concerning patient care in an Intensive Car Unit (ICU) is described. Case-based reasoning offers a model for quickly matching--using a predetermined hierarchical structure--a single patient's parameters (text or numeric) to similar parameters contained in a clinical database. The output produces a group of patients which may be set to match exactly on certain characteristics and may also be set to match "as closely as possible" on a gradient of patient properties. Clinicians may thus use the system to find the group of the closest matching cases to their current patient. Aspects of the ICU history of the selected group may then be displayed graphically (e.g., mortality, length of stay, hours of ventilation, procedures utilized, and complications encountered). Neural network analysis is a pattern recognition technique which uses a training set of patient data (text or numeric) to seek mathematical relationships between various subsets of patient parameters. The discovered relationships from the training set are then applied to estimate the outcomes (e.g., mortality, length of stay, hours of ventilation) of new patients. The effects of these intelligent monitoring techniques are scheduled to be tested in a field trial held in a regional referral center ICU.

Algorithms↗
PubMed1994

Energy-dependent protein-triacylglycerol interaction in a cell-free system from 3T3-L1 adipocytes.

Triacylglycerol is synthesized from the precursors sn-1,2-diacylglycerol and and palmitoyl-CoA in a reaction catalyzed by the microsomal enzyme diacylglycerol acyltransferase (EC 2.3.1.20). Isolated 3T3-L1 adipocyte microsomal vesicles from cells pulse-labeled with L-[35S]methionine were found to release microsomal proteins into a low density form during the synthesis of triacylglycerol. The proteins released, which represent a subset of those present in the labeled microsomes, include a 62-kDa protein found in high concentration in mature fat droplets. The formation of the triacylglycerol-protein complexes was dependent on time and temperature, was not stimulated by cytosol, and required ATP as well as diacylglycerol and palmitoyl-CoA. Only nucleoside triphosphates and not non-hydrolyzable analogues could replace ATP in the reaction. Unlike the enzyme reaction that measures the synthesis of triacylglycerol, the formation of low density membrane is thus dependent on ATP hydrolysis as well as enzyme substrates. The newly formed, low density particles are selectively enriched in triacylglycerol synthesized during the reaction as well as that synthesized prior to the reaction. The cell-free system described thus appears to represent an early adipogenic event leading to the lipid vacuoles found in mature adipocytes.

3T3 Cells↗
PubMed1994

Delineation of extended lengths of coronary arteries by multiplane transesophageal echocardiography.

OBJECTIVES: The purpose of this study was to evaluate the utility of multiplane transesophageal echocardiography in assessing the coronary artery tree. BACKGROUND: Evaluation of coronary disease with single-plane and biplane transesophageal echocardiography is limited to the very proximal vessels. The numerous views provided by multiplane imaging may enhance visualization of coronary arteries and detection of their abnormalities. METHODS: Intraoperative multiplane transesophageal echocardiography was performed in 45 consecutive adults who had recently undergone angiography. Recordings were reviewed in blinded manner. RESULTS: We describe the coronary segments visualized with the different imaging planes and define new views. The left main coronary artery with its bifurcation was visualized in all 45 patients. Sensitivity and specificity for detection of coronary narrowings were 100% when results were compared with angiographic data. Visualization of proximal, mid and distal segments of the left anterior descending coronary artery was possible in 69%, 31% and 16% of patients, respectively. Among patients in whom the proximal segment was visualized, sensitivity and specificity for detection of significant narrowings were 80% and 100%. Proximal, mid and distal portions of the left circumflex coronary artery were visualized in 80%, 51% and 20% of patients. Among patients in whom the proximal portion was well seen, sensitivity and specificity were 89% and 100%. The proximal, mid and distal portions of the right coronary artery were visualized in 84%, 16% and 11% of patients. Among patients in whom the proximal segment was visualized, sensitivity and specificity were 82% and 100%. Color Doppler examination was less useful because it detected only 52% of all patients with proximal stenosis. CONCLUSIONS: Multiplane transesophageal echocardiography allows enhanced visualization of extended lengths of coronary arteries and the reliable identification of coronary artery abnormalities.

Adult↗
PubMed1994

Neither absence nor excess of lambda O initiator-digesting ClpXP protease affects lambda plasmid or phage replication in Escherichia coli.

Owing to rapid proteolysis of the coliphage lambda-coded initiator protein, lambda O, this protein is considered to carry a rate-limiting step in lambda DNA replication. The discovery of ClpXP protease responsible for lambda O protein turnover allowed an opportunity to verify this hypothesis. However, neither absence nor excess of this protease significantly affected the transformation efficiency and copy number of lambda plasmid, or the kinetics of the lambda phage growth. These results are also incompatible with the hypothesis that the stabilization of lambda O plays a role in the switch from early (circle-to-circle) to late (rolling-circle) lambda phage DNA replication. Transcriptional activation of ori lambda, probably assisted by the Escherichia coli DnaA function, remains as the possible rate-limiting step in lambda DNA replication.

ATP-Dependent Proteases↗