Concerns over confidentiality.
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Dementia↗
Biomedical subjects
J Sanford
Publications and source records attributed to J Sanford.
Expression of an antimicrobial peptide via the chloroplast genome to control phytopathogenic bacteria and fungi.
The antimicrobial peptide MSI-99, an analog of magainin 2, was expressed via the chloroplast genome to obtain high levels of expression in transgenic tobacco (Nicotiana tabacum var. Petit Havana) plants. Polymerase chain reaction products and Southern blots confirmed integration of MSI-99 into the chloroplast genome and achievement of homoplasmy, whereas northern blots confirmed transcription. Contrary to previous predictions, accumulation of MSI-99 in transgenic chloroplasts did not affect normal growth and development of the transgenic plants. This may be due to differences in the lipid composition of plastid membranes compared with the membranes of susceptible target microbes. In vitro assays with protein extracts from T(1) and T(2) plants confirmed that MSI-99 was expressed at high levels to provide 88% (T(1)) and 96% (T(2)) inhibition of growth against Pseudomonas syringae pv tabaci, a major plant pathogen. When germinated in the absence of spectinomycin selection, leaf extracts from T(2) generation plants showed 96% inhibition of growth against P. syringae pv tabaci. In addition, leaf extracts from transgenic plants (T(1)) inhibited the growth of pregerminated spores of three fungal species, Aspergillus flavus, Fusarium moniliforme, and Verticillium dahliae, by more than 95% compared with non-transformed control plant extracts. In planta assays with the bacterial pathogen P. syringae pv tabaci resulted in areas of necrosis around the point of inoculation in control leaves, whereas transformed leaves showed no signs of necrosis, demonstrating high-dose release of the peptide at the site of infection by chloroplast lysis. In planta assays with the fungal pathogen, Colletotrichum destructivum, showed necrotic anthracnose lesions in non-transformed control leaves, whereas transformed leaves showed no lesions. Genetically engineering crop plants for disease resistance via the chloroplast genome instead of the nuclear genome is desirable to achieve high levels of expression and to prevent pollen-mediated escape of transgenes.
Adaptation, Physiological↗
Effect of the E200K mutation on prion protein metabolism. Comparative study of a cell model and human brain.
The hallmark of prion diseases is the cerebral accumulation of a conformationally altered isoform (PrP(Sc)) of a normal cellular protein, the prion protein (PrP(C)). In the inherited form, mutations in the prion protein gene are thought to cause the disease by altering the metabolism of the mutant PrP (PrP(M)) engendering its conversion into PrP(Sc). We used a cell model to study biosynthesis and processing of PrP(M) carrying the glutamic acid to lysine substitution at residue 200 (E200K), which is linked to the most common inherited human prion disease. PrP(M) contained an aberrant glycan at residue 197 and generated an increased quantity of truncated fragments. In addition, PrP(M) showed impaired transport of the unglycosylated isoform to the cell surface. Similar changes were found in the PrP isolated from brains of patients affected by the E200K variant of Creutzfeldt-Jakob disease. Although the cellular PrP(M) displayed some characteristics of PrP(Sc), the PrP(Sc) found in the E200K brains was quantitatively and qualitatively different. We propose that the E200K mutation cause the same metabolic changes of PrP(M) in the cell model and in the brain. However, in the brain, PrP(M) undergoes additional modifications, by an age-dependent mechanism that leads to the formation of PrP(Sc) and the development of the disease.
Amino Acid Substitution↗
Use of metronidazole in horses.
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Animals↗
Infection surveillance in home care: device-related incidence rates.
BACKGROUND: Four home health agencies of different sizes, including a rural agency with many branch offices, participated in a surveillance study. METHODS: These agencies used the same definitions for symptomatic urinary tract infections in patients with urinary catheters and for bloodstream infections in patients receiving intravenous therapy. Identical data and methods were used for calculating infection rates, with device-days consistently used as the denominator. Each agency's primary goal was to develop baseline information about its own infection rates and to use these statistics to detect problems and trends. Chart audits were performed if trends or problems were identified. A secondary goal was to compare or benchmark infection rates, because there is almost no published information in this area of home care. RESULTS: We found that our mean infection rates for symptomatic urinary tract infection among patients with urinary catheters and for bloodstream infection among patients receiving intravenous therapy were similar among the four agencies. The mean rates for all four agencies were 4.5 symptomatic urinary tract infections/1000 device-days and 1.1 bloodstream infections/1000 device-days. CONCLUSION: The methods used here can be implemented in other home care agencies. Using the same definitions, collecting data for the same type of infections, and using the same rate calculations make infection control benchmarking possible between home health agencies.
Home Care Services↗
Detection of aneuploidy in mesothelioma cell lines by fluorescence in situ hybridization (FISH)
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Aneuploidy↗
Herbert Evans and the collecting of medicine and science.
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Anatomy↗
Mercy Healthcare's CARE 2000: an evolution in progress.
In a learning environment of shared governance, continuous quality improvement, and redesign principle application, disciplines of Mercy Healthcare San Diego produced their patient care delivery redesign model, Creative Actions Reflecting Excellence. Nurses, pharmacists, medical technologists, respiratory care practitioners, physicians, educators, managers, and many other professional and technical partners converted change and transition into opportunities. As disciplines understood and appreciated each other's unique and shared contributions, quality of care, stakeholder satisfaction, and process efficiencies increased.
Decision Making, Organizational↗
Breast cancer in males: DNA content and sex chromosome constitution.
The infrequent occurrence of breast cancer in males may reflect different etiologic factors and decreased hormonal dependence in comparison with the disease in females. We have attempted to define genetic differences in tumors of males that might reflect alterations in pathogenesis, by examining 22 tumors and 11 examples of gynecomastia for ploidy in relation to X- and Y- chromosome copy number by fluorescence in situ hybridization. Ploidy values were obtained for 15 infiltrating ductal and four papillary tumors (three invasive and one intraductal), two ductal carcinomas in situ, one papilloma, and 11 cases of gynecomastia by flow cytometry of disaggregated cells from the paraffin blocks. The malignant papillary lesions and 14 of the ductal tumors were examined by fluorescence in situ hybridization. DNA probes for pericentromeric regions of the X and Y chromosomes were reacted with formalin-fixed paraffin-embedded sections using a combination of avidin-biotin, digoxigenin-antidigoxigenin, and direct-labeling techniques. Seven multimodal tumors, including one carcinoma in situ, were clearly aneuploid by flow cytometry; the remaining 14 malignant tumors, the papilloma, and 11 examples of gynecomastia were within the diploid range. In two cases, aneuploid-tetraploid clones in tumors that were not recognized by flow cytometry were detected as subpopulations because of extra copies of the X and Y chromosomes. Two cases with aneuploid subpopulations by fluorescence in situ hybridization showed evidence of excess X-chromosome copy number, suggestive of preferential increase of this chromosome within the tumor, and the fluorescence in situ hybridization results also supported exclusion of constitutional Klinefelter's syndrome, although mosaicism could not be excluded.
Adult↗
Low back pain: program description and outcome in a case series.
Studies are needed to enhance our understanding of functional outcomes. The purpose of this paper is to describe a community clinic program for injured workers with low back pain and to report outcomes of the first 50 consecutive patients to enter the program who were evaluated using a standardized assessment procedure. Data for this report were collected from a retrospective chart review as part of an evaluation of the program. The patients referred to the clinic entered a 4-week treatment program. They were assessed at entry and discharge using the Toronto-Hamilton Lumbar Database. The database assessment is a standardized evaluation for documenting subjective and objective clinical data, and the protocol includes a diagnostic classification system and pain and function ratings. The results of this investigation include a statistically significant (p < 0.05) decrease in pain and increase in function as measured by the Jan van Breemen pain and disability scales and the Sickness Impact Profile. Seventy-four percent of the treatment group had returned to work by 6 weeks postdischarge from the program. This study suggests that a significant improvement in functional capabilities, a decrease in pain and disability indices, and higher return-to-work rates can be achieved through a 4-week, community-based multiprofessional rehabilitation program.
Adult↗
A human chromosome 11 NotI end clone library.
A NotI end clone library has been constructed from a human-hamster hybrid cell line containing only human chromosome 11. Fifty-one NotI clones were chosen to characterize the library. The majority of NotI clones hybridize to small 15- to 200-kb fragments and have proven to be valuable for chromosome 11 physical mapping by detecting fragments not previously recognized by random probes. These NotI end clones have been used to isolate corresponding NotI linking cosmids which were then used to identify adjacent NotI fragments on pulsed-field gels. The clones were mapped using fluorescence in situ hybridization and a somatic cell hybrid panel. Although these clones were localized over the entirety of chromosome 11, a nonrandom distribution was observed. Northern blot analysis indicated that 57% (17/30) of the NotI clones examined detected poly(A)+ transcripts in HeLa cell RNA.
Animals↗
Reliability of the Fugl-Meyer assessment for testing motor performance in patients following stroke.
BACKGROUND AND PURPOSE: The purpose of this study was to establish the interrater reliability of assessments made with the Fugl-Meyer evaluation of physical performance in a rehabilitation setting. SUBJECTS: Twelve patients (7 male, 5 female), aged 49 to 86 years (mean = 66), who had sustained a cerebrovascular accident participated in the study. All patients were admitted consecutively to a rehabilitation center and were between 6 days and 6 months poststroke. METHODS: Three physical therapists, each with more than 10 years of experience, assessed the patients in a randomized and balanced order using this assessment. The therapists standardized the assessment approach prior to the study but did not discuss the procedure once the study began. RESULTS: The overall reliability was high (overall intraclass correlation coefficient = .96), and the intraclass correlation coefficients for the subsections of the assessment varied from .61 for pain to .97 for the upper extremity. CONCLUSION AND DISCUSSION: The relative merits of using the Fugl-Meyer assessment as a research tool versus a clinical assessment for stroke are discussed.
Aged↗
Measuring physical impairment and disability with the Chedoke-McMaster Stroke Assessment.
BACKGROUND AND PURPOSE: The Chedoke-McMaster Stroke Assessment measures the physical impairments and disabilities that impact on the lives of individuals with stroke. This measure has three overall purposes: 1) to stage motor recovery to classify individuals in terms of clinical characteristics, 2) to predict rehabilitation outcomes, and 3) to measure clinically important change in physical function. This study was carried out to evaluate the ability of this measure to yield reliable and valid results. METHODS: Thirty-two subjects from a stroke rehabilitation treatment unit were assessed by research and treating physical therapists using multiple measures on multiple occasions. The measure's three purposes dictated the study objectives and design. RESULTS: Intrarater, interrater, and test-retest reliabilities of the impairment and disability inventories were estimated. Reliability coefficients for the total scores ranged from 0.97 to 0.99. Construct and concurrent validities were studied by examining the correlations between this and other measures. A priori hypothetical constructs stated that these correlations should exceed 0.60. These constructs were confirmed; the impairment inventory total score was found to correlate with the Fugl-Meyer Test (r = 0.95, p < 0.001) and the disability inventory with the Functional Independence Measure (r = 0.79, p < 0.05). Additional study hypotheses were also substantiated. CONCLUSIONS: This study confirms that the Chedoke-McMaster Stroke Assessment yields both reliable and valid results. With the evaluation study now completed, the Chedoke-McMaster Stroke Assessment can be used with confidence as both a clinical and a research tool that can discriminate among subjects and evaluate patient outcomes.
Adolescent↗
Clinical evaluation: physiotherapists' ranking of competencies.
The competencies to be assessed as part of the process of clinical evaluation of students need to be determined. This paper describes a method establishing the emphasis or weights assigned to each area of competency. Clinicians and clinical educators were asked to rate a number of dummy student evaluation forms and the individual competencies on each form. Category weights were derived and the magnitude of these weights were used to rank the categories. The derived ranks were compared to the rater's overt ranking of the categories. The weightings are compared to those reported in other studies which examine the value placed on clinical competencies by clinicians.
Attitude of Health Personnel↗
Dissection of a pollen-specific promoter from maize by transient transformation assays.
We have previously reported the isolation and characterization of a gene (Zm13) from Zea mays which shows a pollen-specific pattern of expression. Stably transformed tobacco plants containing a reporter gene linked to portions of the Zm13 5' flanking region show correct temporal and spatial expression of the gene. Here we present a more detailed analysis of the 5' regions responsible for expression in pollen by utilizing a transient expression system. Constructs containing the beta-glucuronidase (GUS) gene under the control of various sized fragments of the Zm 13 5' flanking region were introduced into Tradescantia and Zea mays pollen via high-velocity microprojectile bombardment, and monitored both visually and with a fluorescence assay. The results suggest that sequences necessary for expression in pollen are present in a region from -100 to -54, while other sequences which amplify that expression reside between -260 and -100. The replacement of the normal terminator with a portion of the Zm13 3' region containing the putative polyadenylation signal and site also increased GUS expression. While the -260 to -100 region contains sequences similar to other protein-binding domains reported for plants, the -100 to -54 region appears to contain no significant homology to other known promoter fragments which direct pollen-specific expression. The microprojectile bombardment of Tradescantia pollen appears to be a good test system for assaying maize and possibly other monocot promoter constructs for pollen expression.
Gene Expression↗
Antiendotoxin monoclonal antibodies for gram-negative sepsis: guidelines from the IDSA.
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Antibodies, Monoclonal↗
Gamma-subunits of G proteins, but not their alpha- or beta-subunits, are polyisoprenylated. Studies on post-translational modifications using in vitro translation with rabbit reticulocyte lysates.
Lipid modifications that may be introduced into several subunits of G proteins were explored by in vitro translation of recombinant mRNAs in reticulocyte lysates. In agreement with studies by others, myristic acid was incorporated into alpha i's and alpha o, but not alpha s, beta, or gamma's. In contrast, mevalonate (Mev) was incorporated only into gamma-subunits. Both, the gamma-subunit of transducin (gamma T) and that of other G proteins (gamma G) were modified by the lysates but with different characteristics. Labeled gamma T was unstable and was rapidly proteolyzed. Labeled gamma G was stable. The Mev-derivative in gamma G was sensitive to methyliodide and, after cleavage and chromatographic analysis, comigrated with the C20 polyisoprenol geranylgeraniol. This indicated that gamma G had been geranylgeranylated and that this polyisoprenoid was attached to the protein through a thioether linkage. It is thought that polyisoprenylation is defined by the COOH-terminal sequence Cys-A-A-X, where A is an aliphatic acid and X is any amino acid. Replacement by mutation of the Cys of the COOH-terminal -Cys-Ala-Ile-Leu sequence of gamma G with Ser abolished Mev incorporation, suggesting this Cys as the site of attachment of the geranylgeranyl moiety. Yet, Mev incorporation was less than 10% as much into gamma G with the Cys-A-A-X sequence -Cys-Ala-Ile-Trp. Consistent with geranylgeranylation, the C15 farnesyl moiety of farnesyl pyrophosphate was not incorporated into gamma G unless the incubations were fortified with Mev. In contrast, the farnesyl moiety was incorporated in an Mev-independent manner into gamma T (COOH terminus: -Cys-Val-Ile-Ser) and c-Ha-ras (COOH terminus: -Cys-Val-Leu-Ser) which are both farnesylated rather than geranylgeranylated. Thus, 1) separate enzymes appear to be involved in transferring farnesyl and geranylgeranyl groups to proteins, 2) structural factors other than the CAAX box contribute to the activity of the polyisoprenylating enzymes, and 3) this type of lipidation may be part of a proteolytic signaling system. Polyisoprenylation, which increases hydrophobicity of the derivatized protein, may play a role in anchoring not only ras but also G proteins to membranes.
Animals↗