Biomedical subjects

K C Graham

Publications and source records attributed to K C Graham.

14 recordsLinked to original sources

PubMed2006

Volume measurement variability in three-dimensional high-frequency ultrasound images of murine liver metastases.

The identification and quantification of tumour volume measurement variability is imperative for proper study design of longitudinal non-invasive imaging of pre-clinical mouse models of cancer. Measurement variability will dictate the minimum detectable volume change, which in turn influences the scheduling of imaging sessions and the interpretation of observed changes in tumour volume. In this paper, variability is quantified for tumour volume measurements from 3D high-frequency ultrasound images of murine liver metastases. Experimental B16F1 liver metastases were analysed in different size ranges including less than 1 mm3, 1-4 mm3, 4-8 mm3 and 8-70 mm3. The intra- and inter-observer repeatability was high over a large range of tumour volumes, but the coefficients of variation (COV) varied over the volume ranges. The minimum and maximum intra-observer COV were 4% and 14% for the 1-4 mm3 and <1 mm3 tumours, respectively. For tumour volumes measured by segmenting parallel planes, the maximum inter-slice distance that maintained acceptable measurement variability increased from 100 to 600 microm as tumour volume increased. Comparison of free breathing versus ventilated animals demonstrated that respiratory motion did not significantly change the measured volume. These results enable design of more efficient imaging studies by using the measured variability to estimate the time required to observe a significant change in tumour volume.

Algorithms↗

PubMed2005

Leg wound infections following cardiac surgery: a scoring system for assessment and management.

OBJECTIVE: Well-defined criteria are needed to provide guidance for the appropriate management of leg wounds following saphenous vein harvest in coronary artery bypass graft surgery (CABG). METHOD: A score named DISINFECT was devised to carefully define the variables to be considered for assessing saphenous vein harvest wounds. RESULTS: This preliminary study included 100 consecutive patients undergoing first-time isolated CABG requiring the saphenous vein as a conduit. Wounds were assessed and the points combined to create a daily score (D) according to the presence of increased C-reactive protein/white blood cells (I), surrounding tissue (S), quality of the incision (I), new skin (N), foreign material (F), exudate (E), positive cultures (C) and temperature (T). CONCLUSION: Taking into account the stages of wound healing, severity of infection and appropriate use of antibiotics, this method of wound management would improve the consistency with which leg wounds are managed, reduce hospital stays and increase resistance to hospital-acquired infection.

Aged↗

PubMed2000

Identification and characterization of CKIP-1, a novel pleckstrin homology domain-containing protein that interacts with protein kinase CK2.

The catalytic subunits of protein kinase CK2, CK2alpha and CK2alpha', are closely related to each other but exhibit functional specialization. To test the hypothesis that specific functions of CK2alpha and CK2alpha' are mediated by specific interaction partners, we used the yeast two-hybrid system to identify CK2alpha- or CK2alpha'-binding proteins. We report the identification and characterization of a novel CK2-interacting protein, designated CKIP-1, that interacts with CK2alpha, but not CK2alpha', in the yeast two-hybrid system. CKIP-1 also interacts with CK2alpha in vitro and is co-immunoprecipitated from cell extracts with epitope-tagged CK2alpha and an enhanced green fluorescent protein fusion protein encoding CKIP-1 (i.e. EGFP-CKIP-1) when they are co-expressed. CK2 activity is detected in anti-CKIP-1 immunoprecipitates performed with extracts from non-transfected cells indicating that CKIP-1 and CK2 interact under physiological conditions. The CKIP-1 cDNA is broadly expressed and encodes a protein with a predicted molecular weight of 46,000. EGFP-CKIP-1 is localized within the nucleus and at the plasma membrane. The plasma membrane localization is dependent on the presence of an amino-terminal pleckstrin homology domain. We postulate that CKIP-1 is a non-enzymatic regulator of one isoform of CK2 (i.e. CK2alpha) with a potential role in targeting CK2alpha to a particular cellular location.

Amino Acid Sequence↗

PubMed2000

The regulatory beta subunit of protein kinase CK2 mediates formation of tetrameric CK2 complexes.

Protein kinase CK2 is a tetrameric enzyme composed of two catalytic (alpha and/or alpha') subunits and two regulatory (beta) subunits. Because CK2beta is synthesized in excess of CK2alpha, we hypothesized that formation of CK2beta homodimers precedes the incorporation of the catalytic subunits of CK2 into complexes. To test this hypothesis, we cotransfected cells with two epitope-tagged variants of CK2beta. The results of these cotransfection studies demonstrate that interactions between two CK2beta subunits take place in the absence of CK2alpha. Together with results from previous biosynthetic labeling studies, these results suggest that formation of CK2beta homodimers occurs before incorporation of catalytic subunits of CK2 into CK2 complexes. We also cotransfected Cos-7 cells with a deletion fragment of CK2beta (i.e. Myc-beta1-166) together with full-length hemagglutinin (HA)-tagged CK2beta and/or CK2alpha'. Although complexes between Myc-beta1-166 and HA-beta were readily detected, we obtained no evidence of direct interactions between Myc-beta1-166 and HA-CK2alpha'. These results suggest that residues within the N-terminal 166 amino acids of CK2beta are sufficient for interactions between CK2beta subunits, whereas the C-terminal domain of CK2beta is required for complex formation with the catalytic subunits of CK2. Finally, we observed that expression of full-length HA-beta promotes phosphorylation of Myc-beta1-166 by HA-CK2alpha'.

Animals↗

PubMed1997

Identification of proteins that interact with a protein of interest: applications of the yeast two-hybrid system.

The yeast two-hybrid system is a molecular genetic test for protein interaction. Here we describe a step by step procedure to screen for proteins that interact with a protein of interest using the two-hybrid system. This process includes, construction and testing of the bait plasmid, screening a plasmid library for interacting fusion proteins, elimination of false positives and deletion analysis of true positives. This procedure is designed to allow investigators to identify proteins and their encoding cDNAs that have a biologically significant interaction with your protein of interest.

Cloning, Molecular↗

PubMed1996

Huntingtin is ubiquitinated and interacts with a specific ubiquitin-conjugating enzyme.

Using the yeast two-hybrid system, we have identified a human ubiquitin-conjugating enzyme (hE2-25K) as a protein that interacts with the gene product for Huntington disease (HD) (Huntingtin). This protein has complete amino acid identity with the bovine E2-25K protein and has striking similarity to the UBC-1, -4 and -5 enzymes of Saccharomyces cerevisiae. This protein is highly expressed in brain and a slightly larger protein recognized by an anti-E2-25K polyclonal antibody is selectively expressed in brain regions affected in HD. The huntingtin-E2-25K interaction is not obviously modulated by CAG length. We also demonstrate that huntingtin is ubiquitinated. These findings have implications for the regulated catabolism of the gene product for HD.

Amino Acid Sequence↗

PubMed1996

Knowledge representation and problem solution in expert and novice youth baseball players.

The purpose of this study was to examine differences in knowledge representation and problem solutions in expert and novice youth baseball players. Ninety-four players in two age divisions, 7-8 years of age and 9-10 years of age, were assigned to three levels of expertise: high; average; and low skilled. Each subject participated in an interview session to elicit knowledge representation and solutions to five different defensive game situations. Interviews were transcribed and analyzed for content, solution to the problem, errors in problem solution, and qualitative trends. The frequency of advanced solutions to each of the five situations were analyzed in separate chi-square tests for age and expertise. Differences among the levels of expertise were found for the accuracy of solutions to three complex situations. Age was significant for only one situation. Patterns of knowledge content accessed during advanced and less advanced responses indicated both experts and novices were in a beginning stage of developing baseball knowledge structures. Errors in problem solutions indicated children had difficulty monitoring critical conditions and making correct inferences. Players' and teammates' ability to execute baseball skills seemed to influence the content and structure of tactical knowledge accessed during problem solution.

Age Factors↗

PubMed1995

Interactions between the subunits of casein kinase II.

Casein kinase II (CKII) is a protein serine/threonine kinase known to control the activity of a variety of regulatory nuclear proteins. This enzyme has a tetrameric structure composed of two catalytic (alpha and/or alpha ') subunits and two beta subunits. We have examined the subunit composition of tetrameric complexes of purified bovine CKII by immunoprecipitation using alpha, alpha ', or beta subunit-specific antibodies. These experiments indicate that the enzyme can exist as homotetramers (i.e., alpha 2 beta 2 or alpha 2' beta 2) as well as heterotetramers (i.e. alpha alpha ' beta 2). To further examine subunit interactions between the alpha, alpha ', or beta subunits of CKII, we have utilized the yeast two-hybrid system (Fields, S. and Song, O. (1989) Nature 340: 245-246). For these studies, each subunit of human CKII was expressed in yeast as a fusion with the DNA binding domain or with the transcriptional activation domain of the yeast GAL4 transcriptional activator. These studies demonstrate that the alpha or alpha ' subunits of CKII can interact with the beta subunits of CKII, but not with other alpha or alpha ' subunits. By comparison, the beta subunits of CKII can interact with alpha, alpha ', or beta subunits. These results indicate that the CKII holoenzyme forms because of the ability of beta subunits to dimerize, bringing two heterodimers (alpha beta or alpha ' beta) into a tetrameric complex.

Animals↗

PubMed1989

Paradigms for the study of teacher-student behavior: an alternative perspective.

In conclusion, this commentary--like Schempp's (1987) article--advances the view that a multiple perspective approach to the study of classroom processes is indeed critical to an increased understanding of teacher-student behavior. Contrary to the work of Schempp (1987), however, the rationale extended here as underscoring the need for this approach lies in the limited capacity of any single research approach--be it quantitative or naturalistic in nature--to adequately capture the complexity of classroom interaction. Finally, the view is offered that while each of these traditions of research offers its own contribution to the grand map of teaching (Shulman, 1986), the cumulative outcomes can complement one another. The challenge facing us as an educational research community is to engage in dialogue regarding the design of future investigations so that multiple perspective analyses can indeed become reality. The merit of such a posture is summarized by Weinstein (1983): It is difficult to maintain that one view of classroom events is more accurate than another. Rather, we must learn from each perspective, identify matches and mismatches among perspectives, and examine relationships between perceptions and behavior.... By investigating...[the various perspectives], we will improve our understanding of the social reality of classrooms. (p. 306).

Faculty↗

PubMed1987