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M Kelly

Publications and source records attributed to M Kelly.

At least 91 records · Page 5Linked to original sources
PubMed1999

Construction of gene targeting vectors from lambda KOS genomic libraries.

We describe a highly redundant murine genomic library in a new lambda phage, lambda knockout shuttle (lambda KOS) that facilitates the very rapid construction of replacement-type gene targeting vectors. The library consists of 94 individually amplified subpools, each containing an average of 40,000 independent genomic clones. The subpools are arrayed into a 96-well format that allows a PCR-based efficient recovery of independent genomic clones. The lambda KOS vector backbone permits the CRE-mediated conversion into high-copy number pKOS plasmids, wherein the genomic inserts are automatically flanked by negative-selection cassettes. The lambda KOS vector system exploits the yeast homologous recombination machinery to simplify the construction of replacement-type gene targeting vectors independent of restriction sites within the genomic insert. We outline procedures that allow the generation of simple and more sophisticated conditional gene targeting vectors within 3-4 weeks, beginning with the screening of the lambda KOS genomic library.

Animals↗
PubMed1999

Prat falls, missteps, & solutions in implementing change.

Agencies across the country are preparing for OASIS, PPS, and other changes. The Visiting Nurse Association of Northern California shares its story of preparing for changes while simultaneously shifting gears to achieve compliance with regulations amid competing priorities and financial pressures.

California↗
PubMed1999

Comparison of costing tools in paediatric intensive care.

The cost of service provision is an issue of concern to all professionals working in health care. Department of Health reports have highlighted the need for more effective costing mechanisms for provision of paediatric intensive care (PIC). This study aimed to determine whether nurse-patient dependency scoring would be as effective as the Therapeutic Intervention Scoring System (TISS) at providing a differential costing tool for PIC. Data were collected on 251 patient admissions over 1,741 nursing shifts and analysis of variance undertaken. Findings suggest that a modified nurse-patient dependency score would be as good as TISS as a potential costing tool in PICU.

Activities of Daily Living↗
PubMed1998

A genome-wide search for schizophrenia susceptibility genes.

We completed a systematic genome-wide search for evidence of loci linked to schizophrenia using a collection of 70 pedigrees containing multiple affected individuals according to three phenotype classifications: schizophrenia only (48 pedigrees; 70 sib-pairs); schizophrenia plus schizoaffective disorder (70 pedigrees; 101 sib-pairs); and a broad category consisting of schizophrenia, schizoaffective disorder, paranoid or schizotypal personality disorder, psychosis not otherwise specified (NOS), delusional disorder, and brief reactive psychosis (70 pedigrees; 111 sib-pairs). All 70 families contained at least one individual affected with chronic schizophrenia according to DSM-III-R criteria. Three hundred and thirty-eight markers spanning the genome were typed in all pedigrees for an average resolution of 10.5 cM (range, 0-31 cM) and an average heterozygosity of 74.3% per marker. The data were analyzed using multipoint nonparametric allele-sharing and traditional two-point lod score analyses using dominant and recessive, affecteds-only models. Twelve chromosomes (1, 2, 4, 5, 8, 10, 11, 12, 13, 14, 16, and 22) had at least one region with a nominal P value <0.05, and two of these chromosomes had a nominal P value <0.01 (chromosomes 13 and 16), using allele-sharing tests in GENEHUNTER. Five chromosomes (1, 2, 4, 11, and 13) had at least one marker with a lod score >2.0, allowing for heterogeneity. These regions will be saturated with additional markers and investigated in a new, larger set of families to test for replication.

Chromosome Mapping↗
PubMed1998

Improved method for the on-line metal chelate affinity chromatography-high-performance liquid chromatographic determination of tetracycline antibiotics in animal products.

An improved on-line metal chelate affinity chromatography-high-performance liquid chromatography (MCAC-HPLC) method for the determination of tetracycline antibiotics in animal tissues and egg has been developed. Extraction was carried out with ethyl acetate. The extract was then evaporated to dryness and reconstituted in methanol prior to on-line MCAC clean-up and HPLC-UV determination. Recoveries of tetracycline, oxytetracycline, demeclocycline and chlortetracycline in the range 42% to 101% were obtained from egg, poultry, fish and venison tissues spiked at 25 micrograms kg-1. Limits of detection less than 10 microgram kg-1 were estimated for all four analytes. This method has higher throughput, higher recovery and lower limits of detection than a previously reported on-line MCAC-HPLC method which involved aqueous extraction and solid-phase extraction clean-up.

Animals↗
PubMed1998

The effects of iron and vitamin C co-supplementation on oxidative damage to DNA in healthy volunteers.

The effects of co-supplementing healthy volunteers with iron (14 mg/day ferrous sulphate) and vitamin C (either 60 mg/day or 260 mg/day as ascorbic acid) on levels of oxidative DNA damage in white blood cells were studied. The subjects were divided into two groups: one group of 20 volunteers with a higher mean initial level of plasma vitamin C (71.9 +/- 14.0 mumol/l) and a second group of 18 volunteers with a lower mean level (50.4 +/- 25.8 mumol/l). In the first group there was a significant rise in several oxidative DNA base damage products and in total oxidative DNA damage in DNA extracted from white blood cells, but not in 8-hydroxyguanine, after 6 weeks of supplementation. However, after 12 weeks levels returned approximately to normal. In the group with the lower initial level of plasma ascorbate, presupplemental levels of oxidative DNA damage were higher and decreased on supplementation with iron and ascorbate. Since oxidative DNA damage has been suggested as a risk factor for the development of cancer, the implications of increased levels in well-nourished subjects after iron/ascorbate supplementation are disturbing in view of the frequent use of dietary supplements containing both iron salts and ascorbate.

Adult↗
PubMed1998

A preliminary study to determine the effect of an infusion of cryopreserved autologous lymphocytes on immunocompetence and viral load in HIV-infected patients.

Therapeutic measures aimed at boosting the immunity of HIV-infected patients are a critical component of strategies for effective therapy of HIV and AIDS. To improve immunocompetence in patients with progressive disease, autologous lymphocytes that were collected and cryopreserved earlier in the course of HIV-infection were reinfused. None of the 12 patients receiving cell infusions experienced any adverse effects. Improvements in immunologic parameters (CD4+ counts, CD8+ counts, or both; HIV-specific cytotoxic T-lymphocyte (CTL) activity; or viral load) were seen in seven patients. Restoration of the CD4+ count to the level recorded at the time of cell harvest was achieved in two patients with less advanced disease. Plasma HIV RNA was reduced by >0.5 logs in two of the four patients tested. These preliminary results suggest that cellular immunotherapy using cryopreserved autologous lymphocytes has the potential to improve some measures of immunity in patients with HIV/AIDS and warrants further investigation.

Adult↗
PubMed1998

Comparison of responses of tattoos to picosecond and nanosecond Q-switched neodymium: YAG lasers.

OBJECTIVE: To test the hypothesis that picosecond laser pulses are more effective than nanosecond domain pulses in clearing of tattoos. DESIGN: Intratattoo comparison trial of 2 laser treatment modalities. SETTING: A large interdisciplinary biomedical laser laboratory on the campus of a tertiary medical center. PATIENTS: Consecutive patients with black tattoos were enrolled; all 16 patients completed the study. INTERVENTION: We treated designated parts of the same tattoo with 35-picosecond and 10-nanosecond pulses from 2 neodymium:YAG lasers. Patients received a total of 4 treatments at 4-week intervals. All laser pulse parameters were held constant except pulse duration. Radiation exposure was 0.65 J/cm2 at the skin surface. Biopsies were performed for routine microscopic and electron microscopic analysis at the initial treatment session and 4 weeks after the final treatment in 8 consenting patients. Also, ink samples were irradiated in vitro. MAIN OUTCOME MEASURES: In vivo, on the completion of treatment, a panel of dermatologists not associated with the study (and blinded to the treatment type) evaluated photographs to assess tattoo lightening. Formalin-fixed specimens were examined for qualitative epidermal and dermal changes as well as depth of pigment alteration. Electron micrographs were examined for particle electron density and size changes (in vivo and in vitro). The gross in vitro optical density changes were measured. RESULTS: In 12 of 16 tattoos, there was significant lightening in the picosecond-treated areas compared with those treated with nanosecond pulses. Mean depth of pigment alteration was greater for picosecond pulses, but the difference was not significant. In vivo biopsy specimens showed similar electron-lucent changes for both pulse durations. In vitro results were similar for both pulse durations, showing increases in particle sizes and decreased electron density as well as gross ink lightening. CONCLUSIONS: Picosecond pulses are more efficient than nanosecond pulses in clearing black tattoos. Black tattoos clear principally by laser-induced changes in the intrinsic optical properties of the ink.

Aluminum Silicates↗
PubMed1998

Multiplicity-Selective Coherence Transfer Steps for the Design of Amino Acid-Selective Experiments-A Triple-Resonance Experiment Selective for Asn and Gln

A multiplicity-selective coherence transfer step is discussed, that can replace the normal INEPT transfer in triple-resonance experiments. Depending on the pulse sequence in which they are implemented, amino acid-selective experiments will be created. Two experiments selective for Asn and Gln are proposed. Copyright 1998 Academic Press.

Journal Article↗
PubMed1998

Gestational carrier pregnancy.

OBJECTIVE: To evaluate the results of a gestational carrier program in terms of pregnancy rates in fresh and cryopreserved cycles. DESIGN: Retrospective analysis. SETTING: Private IVF program. PATIENT(S): Women with uterine or medical reasons for unsuccessful gestation. INTERVENTION(S): In vitro fertilization of oocytes with fresh or cryopreserved ET to gestational carriers. MAIN OUTCOME MEASURE(S): Pregnancy rates and deliveries. RESULT(S): A PR of 56.3% and a cycle rate of 30.8% was achieved in each patient <40 years of age in 117 cycles of fresh or cryopreserved ET. There were no pregnancies for nine patients >40 years of age in 27 cycles. CONCLUSION(S): Carrier gestation offers a satisfactory solution to childlessness caused by uterine or major medical factors.

Adult↗
PubMed1998

Pelvic fractures: the golden hour.

Major accident units should be able to apply standard treatment methods to control life threatening haemorrhage within the 'Golden Hour', as recommended by the ATLS guidelines. In this study all 31 major accident units in Scotland were examined to determine their ability to stabilize a pelvic fracture within this period. We found that only eight units could potentially stabilize a pelvis within 1 h. In eight other units no appropriate emergency treatment was available. We recommend that all units designated to receive multiple injuries should have a pelvic stabilization device as well as the training and expertise to use it.

Acute Disease↗
PubMed1998

Differential regulation of ventricular adrenomedullin and atrial natriuretic peptide gene expression in pressure and volume overload in the rat.

1. Adrenomedullin is a recently discovered vasodilating and natriuretic peptide whose physiological and pathophysiological roles remain to be established. Like atrial natiuretic peptide adrenomedullin is expressed in the left ventricle. Ventricular expression of atrial natriuretic peptide is known to be markedly increased by volume or pressure overload. In this study we investigated whether ventricular expression of adrenomedullin is similarly stimulated under such conditions. 2. Ventricular adrenomedullin and atrial natriuretic peptide mRNA levels as well as those of a loading control mRNA (glyceraldehyde-3-phosphate dehydrogenase) were quantified by Northern blot analysis in (a) rats with severe post-infarction heart failure induced by left coronary ligation at 30 days post-surgery and (b) in rats with pressure-related cardiac hypertrophy induced by aortic banding at several time points (0.5, 1 and 4 h, and 1, 4, 7 and 28 days) after surgery. Levels were compared with those in matched sham-operated controls. 3. The mRNA level of atrial natriuretic peptide was markedly increased (8-10-fold) in the left ventricle of animals with post-infarction heart failure. In contrast, there was only a modest (40%) increase in the level of adrenomedullin mRNA. In rats with pressure-induced cardiac hypertrophy the ventricular level of atrial natriuretic peptide mRNA was again markedly increased (maximum 10-fold). The increase was first noticeable at 24 h post-banding and persisted until 28 days. In contrast, there was no change in adrenomedullin mRNA level compared with sham-operated rats at any time point. 4. Despite having similar systemic effects, the expression of adrenomedullin and atrial natriuretic peptide in the left ventricle is differently regulated. The findings imply distinct roles for the two peptides. The results do not support an important role for ventricular adrenomedullin expression in the remodelling process that occurs during the development of cardiac hypertrophy but suggest that ventricular adrenomedullin participates in the local and/or systemic response to heart failure.

Adrenomedullin↗
PubMed1998

A high-resolution microsatellite map of the mouse genome.

The European Collaborative Interspecific Backcross (EUCIB) resource was constructed for the purposes of high-resolution genetic mapping of the mouse genome (). The large Mus spretus/C57BL/6 backcross of 982 progeny has a genetic resolution of 0.3 cM at the 95% confidence level ( approximately 500 kb in the mouse genome). We have used the EUCIB mapping resource to develop a genome-wide high-resolution genetic map incorporating 3368 microsatellites. The microsatellites are distributed among 2302 genetically separated bins with 1.46 markers per bin on average. Average bin separation is 0.61 cM. This high-resolution genetic map will aid the construction of a robust physical map of the mouse genome.

Animals↗
PubMed1998

CCR5 expression correlates with susceptibility of maturing monocytes to human immunodeficiency virus type 1 infection.

The chemokine receptor CCR5 and to a lesser extent CCR3 and CCR2b have been shown to serve as coreceptors for human immunodeficiency virus type 1 (HIV-1) entry into blood- or tissue-derived macrophages. Therefore, we examined the expression of the chemokine receptors CCR1, CCR2b, CCR3, CCR5, and CXCR4 as RNAs or as membrane-expressed antigens in monocytes maturing into macrophages and correlated these results with the susceptibility of macrophages to HIV-1 infection, as measured by their concentrations of extracellular p24 antigen and levels of intracellular HIV DNA by quantitative PCR. There was little change in levels of CCR1, CCR2b, and CCR5 RNAs. CCR3 RNA and surface antigen were undetectable throughout maturation of adherent monocytes over 10 days. CXCR4 RNA and membrane antigen were strongly expressed in newly adherent monocytes, but their levels declined at day 7. The amounts of CCR5 RNA remained stable, but the amounts of CCR5 antigen increased from undetectable to peak levels at day 7 and then declined slightly at day 10. Levels of susceptibility to laboratory (HIV-1BaL) and clinical strains of HIV-1 showed parallel kinetics, peaking at day 7 and then decreasing at days 10 to 14. The concordance of levels of HIV DNA and p24 antigen suggested that the changes in susceptibility with monocyte maturation were at or immediately after entry and correlated well with CCR5 expression and inversely with CXCR4 expression.

Base Sequence↗
PubMed1998

A third-generation lentivirus vector with a conditional packaging system.

Vectors derived from human immunodeficiency virus (HIV) are highly efficient vehicles for in vivo gene delivery. However, their biosafety is of major concern. Here we exploit the complexity of the HIV genome to provide lentivirus vectors with novel biosafety features. In addition to the structural genes, HIV contains two regulatory genes, tat and rev, that are essential for HIV replication, and four accessory genes that encode critical virulence factors. We previously reported that the HIV type 1 accessory open reading frames are dispensable for efficient gene transduction by a lentivirus vector. We now demonstrate that the requirement for the tat gene can be offset by placing constitutive promoters upstream of the vector transcript. Vectors generated from constructs containing such a chimeric long terminal repeat (LTR) transduced neurons in vivo at very high efficiency, whether or not they were produced in the presence of Tat. When the rev gene was also deleted from the packaging construct, expression of gag and pol was strictly dependent on Rev complementation in trans. By the combined use of a separate nonoverlapping Rev expression plasmid and a 5' LTR chimeric transfer construct, we achieved optimal yields of vector of high transducing efficiency (up to 10(7) transducing units [TU]/ml and 10(4) TU/ng of p24). This third-generation lentivirus vector uses only a fractional set of HIV genes: gag, pol, and rev. Moreover, the HIV-derived constructs, and any recombinant between them, are contingent on upstream elements and trans complementation for expression and thus are nonfunctional outside of the vector producer cells. This split-genome, conditional packaging system is based on existing viral sequences and acts as a built-in device against the generation of productive recombinants. While the actual biosafety of the vector will ultimately be proven in vivo, the improved design presented here should facilitate testing of lentivirus vectors.

Animals↗