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Biomedical subjects

J McKinney

Publications and source records attributed to J McKinney.

At least 19 recordsLinked to original sources
PubMed2001

Conformation of the substrate and pterin cofactor bound to human tryptophan hydroxylase. Important role of Phe313 in substrate specificity.

Tryptophan hydroxylase (TPH) carries out the 5-hydroxylation of L-Trp, which is the rate-limiting step in the synthesis of serotonin. We have prepared and characterized a stable N-terminally truncated form of human TPH that includes the catalytic domain (Delta90TPH). We have also determined the conformation and distances to the catalytic non-heme iron of both L-Trp and the tetrahydrobiopterin cofactor analogue L-erythro-7,8-dihydrobiopterin (BH2) bound to Delta90TPH by using 1H NMR spectroscopy. The bound conformers of the substrate and the pterin were then docked into the modeled three-dimensional structure of TPH. The resulting ternary TPH-BH2-L-Trp structure is very similar to that previously determined by the same methods for the complex of phenylalanine hydroxylase (PAH) with BH2 and L-Phe [Teigen, K., et al. (1999) J. Mol. Biol. 294, 807-823]. In the model, L-Trp binds to the enzyme through interactions with Arg257, Ser336, His272, Phe318, and Phe313, and the ring of BH2 interacts mainly with Phe241 and Glu273. The distances between the hydroxylation sites at C5 in L-Trp and C4a in the pterin, i.e., 6.1 +/- 0.4 A, and from each of these sites to the iron, i.e., 4.1 +/- 0.3 and 4.4 +/- 0.3 A, respectively, are also in agreement with the formation of a transient iron-4a-peroxytetrahydropterin in the reaction, as proposed for the other hydroxylases. The different conformation of the dihydroxypropyl chain of BH2 in PAH and TPH seems to be related to the presence of nonconserved residues, i.e., Tyr235 and Pro238 in TPH, at the cofactor binding site. Moreover, Phe313, which seems to interact with the substrate through ring stacking, corresponds to a Trp residue in both tyrosine hydroxylase and PAH (Trp326) and appears to be an important residue for influencing the substrate specificity in this family of enzymes. We show that the W326F mutation in PAH increases the relative preference for L-Trp as the substrate, while the F313W mutation in TPH increases the preference for L-Phe, possibly by a conserved active site volume effect.

Binding Sites↗
PubMed2001

Inhibition of Escherichia coli viability by external guide sequences complementary to two essential genes.

Narrow spectrum antimicrobial activity has been designed to reduce the expression of two essential genes, one coding for the protein subunit of RNase P (C5 protein) and one for gyrase (gyrase A). In both cases, external guide sequences (EGS) have been designed to complex with either mRNA. Using the EGS technology, the level of microbial viability is reduced to less than 10% of the wild-type strain. The EGSs are additive when used together and depend on the number of nucleotides paired when attacking gyrase A mRNA. In the case of gyrase A, three nucleotides unpaired out of a 15-mer EGS still favor complete inhibition by the EGS but five unpaired nucleotides do not.

Bacterial Proteins↗
PubMed2000

Recombinant bacillus Calmette-Guérin as a potential vector for preventive HIV type 1 vaccines.

In August 1997, the World Health Organization (WHO) and the Joint United Nations Programme on HIV/AIDS (UNAIDS) convened an expert working group to discuss current strategies for the development of HIV type 1 vaccines. Based on the recent findings of investigators from Japan's National Institute of Infectious Diseases (NIID) in Tokyo using recombinant bacillus Calmette-Guérin (rBCG) as a potential vectored vaccine for HIV, a recommendation was made that further work in this area is a priority. As a result, the working group reconvened in September 1998 to discuss the progress to date with this vaccine approach, as well as areas of related research to assess the feasibility of a BCG-vectored HIV vaccine. This report summarizes the discussions addressing the available scientific data on the potential use of rBCG as a vector for preventive HIV vaccines, the work necessary to move such candidate vaccines into Phase 1 clinical trials, and recommendations targeted at facilitating the long-term development of rBCG-vectored HIV vaccines.

AIDS Vaccines↗
PubMed2000

Clathrin in mitotic spindles.

Subconfluent cultures of Madin-Darby canine kidney (MDCK) and CV-1 cells were immunostained with two monoclonal antibodies (MAbs), MAb X-22 and MAb 23, against clathrin heavy chain and with polyclonal antiserum against a conserved region of all mammalian clathrin light chains. In interphase MDCK and CV-1 cells, staining by all three antibodies resulted in the characteristic intracellular punctate vesicular and perinuclear staining pattern. In mitotic cells, all three anti-clathrin antibodies strongly stained the mitotic spindle. Staining of clathrin in the mitotic spindle was colocalized with anti-tubulin staining of microtubular arrays in the spindle. Staining of the mitotic spindle was evident in mitotic cells from prometaphase to telophase and in spindles in mitotic cells released from a thymidine-nocodazole block. In CV-1 cells, staining of clathrin in the mitotic spindle was not affected by brefeldin A. On Western blots, clathrin was detected, but not enriched, in isolated spindles. The immunodetection of clathrin in the mitotic spindle may suggest a novel role for clathrin in mitosis. Alternatively, the recruitment of clathrin to the spindle may suggest a novel regulatory mechanism for localization of clathrin in mitotic cells.

Animals↗
PubMed2000

Clathrin in gastric acid secretory (parietal) cells: biochemical characterization and subcellular localization.

Clathrin from H-K-ATPase-rich membranes derived from the tubulovesicular compartment of rabbit and hog gastric acid secretory (parietal) cells was characterized biochemically, and the subcellular localization of membrane-associated clathrin in parietal cells was characterized by immunofluorescence, electron microscopy, and immunoelectron microscopy. Clathrin from H-K- ATPase-rich membranes was determined to be comprised of conventional clathrin heavy chain and a predominance of clathrin light chain A. Clathrin and adaptors could be induced to polymerize quantitatively in vitro, forming 120-nm-diameter basketlike structures. In digitonin-permeabilized resting parietal cells, the intracellular distribution of immunofluorescently labeled clathrin was suggestive of labeling of the tubulovesicular compartment. Clathrin was also unexpectedly localized to canalicular (apical) membranes, as were alpha-adaptin and dynamin, suggesting that this membrane domain of resting parietal cells is endocytotically active. At the ultrastructural level, clathrin was immunolocalized to canalicular and tubulovesicular membranes. H-K-ATPase was immunolocalized to the same membrane domains as clathrin but did not appear to be enriched at the specific subdomains that were enriched in clathrin. Finally, in immunofluorescently labeled primary cultures of parietal cells, in contrast to the H-K-ATPase, intracellular clathrin was found not to translocate to the apical membrane on secretagogue stimulation. Taken together, these biochemical and morphological data provide a framework for characterizing the role of clathrin in the regulation of membrane trafficking from tubulovesicles and at the canalicular membrane in parietal cells.

Animals↗
PubMed1999

Prevalence of antibodies to Sarcocystis neurona, Toxoplasma gondii and Neospora caninum in horses from Argentina.

Sera from 76 horses from Argentina were examined for antibodies to Sarcocystis neurona, Toxoplasma gondii and Neospora caninum. Antibodies to S. neurona were found in 27 (35.5%) of 76 horses using immunoblots with culture derived merozoites as antigen. Antibodies to T. gondii were found in 10 (13.1%) of 76 horses by using the modified agglutination test with formalin-fixed tachyzoites and mercaptoethanol; titers were 1:25 (two horses), 1:50 (six horses), 1:100 (two horses), and 1:200 (one horse). Antibodies to N. caninum were not found in any of the 76 horses by the use of N. caninum agglutination test. This is the first report of S. neurona infection in horses in Argentina.

Agglutination Tests↗
PubMed1999

Identification of a STAT4 binding site in the interleukin-12 receptor required for signaling.

The specificity of the various STAT SH2 domains for different tyrosine-containing peptides enables cytokines to activate different signaling pathways and to induce distinct patterns of gene expression. We show that STAT4 has a unique peptide specificity and binds to the peptide sequence pYLPSNID (where pY represents phosphotyrosine). This motif is found at tyrosine residue 800 in the beta2 subunit of the interleukin-12 receptor and is required for DNA binding and transcriptional activity of STAT4. Our data demonstrate that transfection of interleukin-12 receptor beta1 and beta2 subunits is sufficient for STAT4 activation but not for STAT1 or STAT3 activation.

Base Sequence↗
PubMed1997

Battered women's perceptions of loss and health.

The article reports a study investigating whether a positive relationship exists between a battered women's perceptions of loss and her perceptions of health. The study further investigated whether a period of time away from the abusive situation would change the battered women's perceptions of loss and her perceptions of health. A convenience sample of 20 women who had experienced battering and abuse and had sought help at a protective shelter was studied. Questionnaires on perception of loss, health response, and demographics were used for the collection of data. The findings indicated that a significant relationship exists between a battered women's perceptions of loss and her perception of health. Perceptions of loss and health were shown to decline after a period of 5 days away from the abusive situation. Additional qualitative data obtained from in-depth interviews were also found to be consistent with the quantitative results and supported the literature documented in this study.

Adult↗
PubMed1997

The MHSIP mental health report card. A consumer-oriented approach to monitoring the quality of mental health plans. Mental Health Statistics Improvement Program.

Health care report cards have been endorsed as a mechanism for efficiently comparing key quantifiable aspects of performance across a range of health systems or plans. There are challenges in determining what to measure; how to gather and analyze data; and how to report, interpret, and use findings. Mental health has received little attention, and a consumer perspective is typically not included. The proposed MHSIP mental health report card (MMHRC) addresses these concerns. General issues for report cards are discussed, and the MMHRC is described in terms of content, data sources and quality, and analysis and reporting.

Community Participation↗
PubMed1992

Is START a switch?

The cell cycle in Saccharomyces cerevisiae is controlled by regulation of START in late G1. The CLN1, CLN2 and CLN3 family of cyclin homologues is required for cells to pass START. They probably act by activating the CDC28 protein kinase. Expression of CLN1 or CLN3 under the control of an inducible promoter shows that transcription of either gene is sufficient for cyclin-deficient strains arrested in G1 to traverse START. A model of START regulation involves activation of CDC28 kinase by any CLN protein, leading to activation of CLN1 and CLN2 transcription in a positive feedback loop and passage through START. The cell cycle-dependent transcriptional regulators SWI4 and SWI6 may be components of the feedback loop. Cell cycle commitment entails resistance to the inhibitory action of mating factor, which correlates with peak levels of CLN1 and CLN2 mRNAs. FAR1 encodes an alpha-factor-dependent inhibitor of CLN function whose expression is markedly reduced at the time of START. The interplay of all these factors may sharpen the START transition such that it is close to an all-or-nothing switch event. This may be important for several START-dependent events to be activated at the same time, leading to coordinated cell cycle progression.

Cell Cycle↗
PubMed1992

Occupational therapists in private practice.

Although increasing numbers of occupational therapists are choosing to work in private practice, little data exist describing this sector of the profession. In the present study, experienced occupational therapists were asked about their moves into private practice, including (a) their motivation, (b) their preparation, and (c) their perceptions of the move's risks and benefits before and after the move. A survey was sent to a national random sample of 105 occupational therapists, 74 of whom responded. According to the survey, autonomy was the most important motivating factor for occupational therapists moving into private practice. However, once they were in private practice, the occupational therapists noted that increased income was a major benefit. These occupational therapists had planned for the risks of reimbursement, referral sources, and overhead but had not anticipated problems with staffing shortages. Incomes increased for occupational therapists who moved into private practice. The survey compared the incomes of occupational therapists before and after they entered private practice. It also compared their income and educational levels. Other comparisons included income and work experience, income and work role, and income and geographic location. Autonomy and financial considerations appear to be the overriding issues for occupational therapists choosing careers in private practice. Almost unanimously, the survey respondents said that private practice was a good career choice.

Adult↗
PubMed1991

Disabling cerebral transient ischemic attacks.

The surgical management of an emerging clinical entity, namely disabling transient cerebral ischemic attacks, is described. A series of 19 patients treated in a 2-year period (12 with anterior circulation dysfunction and seven with posterior insufficiency) met the following criteria: 1) stereotypical recurrent episodes of transient neurological dysfunction related to the anterior or posterior circulation distribution; 2) failure of maximum medical therapy to control the transient neurological dysfunction; 3) four-vessel cerebral angiography demonstrating an isolated vascular territory corresponding to patient symptoms; 4) inhalation xenon cerebral blood flow studies with at least three of eight probe-pairs showing significant asymmetries in the initial slope index, localizing an area of relative oligemia to the symptomatic hemisphere (anterior circulation only); and 5) severe restriction of lifestyle due to transient ischemic attacks (TIA's). Seventeen patients underwent surgical bypass therapy: deep sylvian superficial temporal artery (STA)-middle cerebral artery (MCA) bypass in nine; surface STA-MCA bypass in three; STA-superior cerebellar artery bypass in three; STA-posterior cerebral artery bypass in one; and aorta-carotid artery bypass in one. There was one perioperative death and four perioperative strokes (two ipsilateral and two contralateral to the operated side). The average follow-up period was 14 months. Of the 16 surviving surgically treated patients, 13 (81%) have had an excellent to good outcome with complete resolution of TIA's and minimal neurological deficits. Three patients had a poor outcome with either a significant persistent neurological deficit or continued TIA's. The two patients not treated surgically continue to have vertebrobasilar insufficiency episodes while receiving oral anticoagulation medication. The overall mortality rate (5.5%) and stroke morbidity rate (22.2%) of surgical therapy for disabling TIA's are high in this neurologically unstable group of patients, but are associated with an 81% excellent to good response. Although the natural history of disabling TIA's is not known, these patients present with significant to total disability due to their symptoms. It is concluded that disabling TIA's respond to surgical revascularization and may represent an indication for cerebral revascularization surgery.

Cerebral Revascularization↗
PubMed1991

Implementation of a local area network for nursing management.

This paper describes the planning and implementation of a Local Area Network (LAN) for the nursing service of a 504-bed urban teaching hospital. The major goals of the network were: support for nurse executives, nurse managers, and the departmental assistants assigned to administrative offices; increased efficiency and effectiveness of the nursing administrative areas; and improved communication systems. Collaboration between the nursing service and the Computer Information Center (CIC) resulted in a network of over 70 workstations, spanning 11 buildings. The network provides access to multiple programs that support clinical, managerial, and research activities. Gateways provide access to the hospital's two mainframes.

Boston↗
PubMed1990

Comparison between the counter-rotational toothbrush and multi-action toothbrush.

Previous studies have demonstrated that two types of powered toothbrushes (Interplak and Braun) are more effective than a manual toothbrush in the removal of bacterial plaque from tooth surfaces. The purpose of this study was to compare the effectiveness of the Interplak Home Plaque Removal Instrument (A) and the Braun Electric Toothbrush (B) on supragingival plaque removal and gingival bleeding following (1) one time instruction; (2) reinstruction and one week of home use; (3) a third instruction and three weeks of home use. The sample population for this study consisted of twelve subjects: six (6) were randomly assigned to use the Interplak, and the remaining six (6) were assigned to use the Braun. Subjects met specific qualification criteria to participate in this examiner-blind study. The Turesky Modification of the Quigley-Hein Plaque Index and an Objective-Subjective Gingival Abrasion Index were scored before and after brushing at the initial appointment, one week later, and at the final appointment in three weeks. A timed bleeding index (Barnett-Muhleman) and probing depths were recorded at the beginning of all three appointments. Subjects received instructions on the use of the assigned toothbrush in a clinical demonstration and in a printed format to take home. Subjects were then supervised in their brushing technique. Data was analyzed using a Wilcoxon Signed Rank Test and Analysis of Variance with Repeated Measures where appropriate. Although the number of patients was small, a statistically significant difference (p less than .05) in plaque scores between the groups was obtained in favor of the Interplak.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1989

Histone H1 subtype-specific consensus elements mediate cell cycle-regulated transcription in vitro.

In this study we used nuclear extracts from centrifugally elutriated cell populations to study histone H1 transcriptional regulation during the cell cycle. Analysis of mutations within the H1 promoter establish that both of the H1 subtype-specific consensus elements participate in induction of transcription upon entry into S phase. The DNA binding activity of H1TF2, which specifically interacts with the H1 proximal subtype-specific element, is increased in S-phase nuclear extracts, whereas no increase in DNA binding is observed for the H1 distal subtype-specific DNA transcription factor H1TF1 or the H2b subtype-specific factor OTF1. These data strongly support the idea that histone gene subtype-specific transcription factors are important for S-phase-dependent expression of histone genes. Further studies of these factors will be important for increased understanding of the transition from G1 to S phase of the mammalian cell cycle.

Animals↗
PubMed1988

An evaluation of labeling-then-doing with moderately handicapped persons: acquisition and generalization with complex tasks.

We conducted two experiments in which moderately mentally retarded persons were trained first to label and then to enter characters into a computer, calculator, or checkbook (label-then-do) within a multiple baseline design. In Experiment 1, 5 young adults were trained to enter statistical programs into computers in an office setting. Following training, all subjects' use of verbal labels and key-entry skills generalized across tasks (programs) and settings (offices and computer terminals). In Experiment 2, 3 junior high school students were trained with self-labeling procedures to complete a key-entry task and to balance a checkbook. The performance of all students generalized across tasks and settings, and the use of labels generalized for 2 of the students. Results are discussed relative to mediated generalization and to establishing verbal control over behavior.

Adolescent↗