Biomedical subjects

S Howe

Publications and source records attributed to S Howe.

At least 19 recordsLinked to original sources

PubMed2005

Long-term preservation of retinal function in the RCS rat model of retinitis pigmentosa following lentivirus-mediated gene therapy.

The Royal College of Surgeons (RCS) rat is a well-characterized model of autosomal recessive retinitis pigmentosa (RP) due to a defect in the retinal pigment epithelium (RPE). It is homozygous for a null mutation in the gene encoding , a receptor tyrosine kinase found in RPE cells, that is required for phagocytosis of shed photoreceptor outer segments. The absence of Mertk results in accumulation of outer segment debris. This subsequently leads to progressive loss of photoreceptor cells. In order to evaluate the efficacy of lentiviral-mediated gene replacement therapy in the RCS rat, we produced recombinant VSV-G pseudotyped HIV-1-based lentiviruses containing a murine Mertk cDNA driven by a spleen focus forming virus (SFFV) promoter. The vector was subretinally injected into the right eye of 10-day-old RCS rats; the left eye was left untreated as an internal control. Here, we present a detailed assessment of the duration and extent of the morphological rescue and the resulting functional benefits. We examined animals at various time points over a period of 7 months by light and electron microscopy, and electroretinography. We observed correction of the phagocytic defect, slowing of photoreceptor cell loss and preservation of retinal function for up to 7 months. This study demonstrates the potential of gene therapy approaches for the treatment of retinal degenerations caused by defects specific to the RPE and supports the use of lentiviral vectors for the treatment of such disorders.

Animals↗

PubMed2003

Gene therapy progress and prospects: gene therapy for severe combined immunodeficiency.

Severe combined immunodeficiencies have long been targeted as a group of disorders amenable to gene therapy because of their defined molecular biology and pathophysiology, and the prediction that corrected cells would have profound growth and survival advantage. Recently, several clinical studies have shown that conventional gene transfer technology can produce major beneficial therapeutic effects in these patients, but, as for all cellular and pharmacological treatment approaches, with a finite potential for toxicity.

Adenosine Deaminase↗

PubMed2003

A comparison of the performance of commercially available chromogenic agars for the isolation and presumptive identification of organisms from urine.

AIMS: To compare four media-UTI medium, BBL CHROMagar, CPS ID2, and Harlequin CLED-using a collection of fully characterised organisms and subsequent "field trial". METHODS: Seven hundred and eighty seven fully characterised isolates (730 Gram negative bacteria, 47 Gram positive bacteria, and 10 yeasts) were used to test for accuracy of organism identification. To assess isolation rates and ability to detect mixed cultures, 1435 urine samples were cultured in the three best performing chromogenic media (UTI medium, BBL CHROMagar, and CPS ID2) and CLED. RESULTS: The chromogenic agars differed in their accuracy of identification, with BBL CHROMagar performing best and Harlequin CLED performing least well. Similarly, BBL CHROMagar achieved a higher overall isolation rate than UTI medium and CPS ID2. When mixed growth was defined as greater than two organism types, BBL CHROMagar detected more mixed cultures than did UTI medium and CPS ID2, although the differences were not significant. When mixed growth was defined as greater than one organism type the increased number of mixed growths detected by BBL CHROMagar became significant, largely because of differences in enterococcal isolation rates. CONCLUSION: The use of BBL CHROMagar, UTI medium, or CPS ID2 chromogenic agar as a replacement for CLED agar would improve the detection rate of contaminated urine samples. Enhanced identification helps to distinguish different species, facilitating the monitoring of bacterial resistance in support of the national antibiotic strategy. BBL CHROMagar gave the highest overall organism recovery rates, greatest ability to detect mixed cultures, and the most accurate identification of organisms.

Agar↗

PubMed2003

Groundwater abstraction pollution risk assessment.

A generic groundwater pollution risk assessment methodology has been developed to enable the evaluation and ranking of the potential risk of pollution to groundwater abstractions. The ranking can then be used to prioritise risk management or mitigation procedures in a robust and quantifiable framework and thus inform business investment decisions. The risk assessment consider the three components of the pollution transport model: source-pathway-receptor. For groundwater abstractions these correspond to land use (with associated pollutants and shallow subsurface characteristics), aquifer and the abstraction borehole. An hierarchical approach was chosen to allow the risk assessment to be successfully carried out with different quality data for different parts of the model. The 400-day groundwater protection zone defines the catchment boundary that form the spatial limit of the land use audit for each receptor. A risk score is obtained for each land use (potential pollution source) within the catchment. These scores are derived by considering the characteristics (such as load, persistence and toxicity) of all pollutants pertaining to each land use, their on-site management and the potential for the unsaturated subsurface to attenuate their effects in the event of a release. Risk scores are also applied to the aquifer characteristics (as pollutant pathway) and to the abstraction borehole (as pollutant receptor). Each risk score is accompanied by an uncertainty score which provides a guide to the confidence in the data used to compile the risk assessment. The application of the methodology has highlighted a number of problems in this type of work and results of initial case studies are being used to trial alternative scoring methods and a more simplified approach to accelerate the process of pollution risk assessment.

Models, Theoretical↗

PubMed2002

Recruitment strategies for cervical cancer prevention study.

OBJECTIVE: The aim of this study was to describe recruitment strategies for a single-visit cervical cancer prevention study. METHODS: From January through December 1999, low-income, predominantly Latino women were recruited to participate in a single-visit cervical cancer prevention study. For the first 6 months, all women who had ever visited one of two community-based study clinics were invited to participate (clinic registry recruitment). For the remainder of the year, recruitment was modified to be primarily inclusive of advertisements in English- and Spanish-language community newspapers and fliers left in local businesses and organizations (media campaign recruitment). Eligible volunteers were randomized to one of two study arms, usual-care program or single-visit program. All study subjects completed demographic and medical questionnaires delivered by bilingual staff. Women who declined to participate in this study were asked to provide reasons for this preference. Statistical analyses included the use of chi-square, logistic regression, and Student's t test. RESULTS: The proportion of women who agreed to participate was higher in the media recruitment group than in the clinic registry group [51% (535/1041) compared to 26% (405/1542), P < 0.001]. The no-show rate among participants solicited from the media strategy was significantly less than that from the clinic registry. There were no significant differences in the median age, number of months since the last Papanicolaou smear, incidence of abnormal Papanicolaou smear, education, or income of the subjects based on the recruitment strategy. CONCLUSION: A media-based recruitment strategy was effective for this single-visit cervical prevention study. This approach may be effective for recruitment of other low-income groups to clinical trials.

Adult↗

PubMed2002

An evaluation of the performance of XLD, DCA, MLCB, and ABC agars as direct plating media for the isolation of Salmonella enterica from faeces.

AIMS: To compare the performance of four media, singly and in combination, as direct plating media for the isolation of Salmonella enterica from human faeces. METHODS: Two thousand four hundred and nine routine, faecal samples received by four laboratories were inoculated on to xylose lysine desoxycholate (XLD), desoxycholate citrate (DCA), mannitol lysine crystal violet brilliant green (MLCB), and alpha-beta chromogenic (ABC) agars using standardised protocols, reagents, and data collection. Isolates of presumptive salmonellae were identified using standard laboratory techniques and the results were analysed statistically. RESULTS: Direct plating recovered 46 of the 60 possible isolates of Salmonella spp recovered via enrichment broth. No isolates were recovered from direct plating that were not recovered via selenite enrichment. MLCB gave the highest isolation rate individually (84.8%) and amounts of competing flora (CF) did not affect the recognition of colonies. ABC proved highly specific, but insensitive, and isolation rates were adversely affected by any amount of CF. Isolation rates from XLD and DCA were only affected when the CF load was heavy. DCA was least specific, with only 9.01% of picks positive and greatest number of confirmatory tests. XLD and MLCB, in combination, gave the highest isolation rate. CONCLUSIONS: Where the earlier results of direct plating may be advantageous, XLD and MLCB provide the optimal combination. For non-typhi salmonellae, MLCB is the best, single direct plating medium. For routine diagnostic work, XLD is most effective.

Bacteriological Techniques↗

PubMed2002

A comparison of the performance of cystine lactose electrolyte deficient (CLED) agar with Oxoid chromogenic urinary tract infection (CUTI) medium for the isolation and presumptive identification of organisms from urine.

AIMS: As part of the UK antimicrobial resistance strategy and action plan, the Public Health Laboratory Service (PHLS) is required to collect antibiotic susceptibility data so that resistance trends and patterns can be monitored. Most laboratories report urine Gram negative isolates, as "coliforms" according to morphological appearance, but without an acceptable identification system the antimicrobial surveillance data will be meaningless. Commercially available identification systems tend to be expensive and time consuming. Chromogenic agars, which claim to improve the detection of mixed cultures and identification of organisms from urine, have now become available and may provide a cost effective alternative. The primary aim of this study was to compare the performance of cystine lactose electrolyte deficient (CLED) agar with a chromogenic agar (Oxoid urinary tract infection medium; CUTI) in terms of isolation rates and ability to detect mixed cultures. Secondary aims were to evaluate the correlation of "presumptive" identification of isolates from chromogenic media with that of two commercial identification systems and to appraise the sensitivity of the semiquantitative loop and filter paper strip culture techniques. METHOD: One thousand, four hundred and sixty six urine samples were examined in four laboratories using the semiquantitative culture methods of 1 microl loop and filter paper strip. The degree of accuracy of organism identification was measured by comparing the presumptive identification using colony colour supplemented with simple bench tests, with identification obtained from two more complex commercial systems. RESULTS: There was no significant difference between the performance of the loop and filter paper strip methods on the CLED agar, but the CUTI agar performed significantly better than the CLED agar for the detection of significant isolates and mixed cultures. This difference was greater using the loop method. Identification of the organisms using the commercial systems gave > 99% agreement and was therefore considered suitable as a standard against which to compare the presumptive CUTI identification. Using the manufacturer's colony colour criteria in combination with a bench indole test, the CUTI medium was 99% specific for Escherichia coli, although this was reduced to 97% if the indole test was omitted. Citrobacter spp were the most commonly misidentified organisms, giving false presumptive identification as E coli. By testing oxidase activity to differentiate Pseudomonas spp and the absence of indole production to support the identification of Proteus mirabilis, the CUTI medium provided a suitable identification for 86.8% of Gram negative isolates. The remaining 13.2% would require further identification. CONCLUSION: CUTI medium improves the detection of mixed cultures, thereby improving the reliability of reporting of significant isolates when compared with CLED agar. When supplemented with simple bench tests it provides an identification system capable of speciating 86.8% of Gram negative isolates and providing a valuable cost effective mechanism for antimicrobial resistance surveillance.

Agar↗

PubMed2002

Effect of incubation temperature on the isolation of thermophilic species of Campylobacter from faeces.

Although no large-scale clinical study has been performed, it has been reported that incubation at 37 degrees C gives better isolation rates for all common species of Campylobacter than incubation at 42 degrees C, while also improving the recovery of the more unusual species. In this study, 2,570 faecal samples were examined in four laboratories, using a standardised protocol. Isolation rates of Campylobacter spp. were compared after incubation on modified campylobacter blood-free selective agar at 37 degrees C and 42 degrees C. Campylobacter spp. isolates were made from 185 samples (7.2%); 25 were recovered only at 42 degrees C and three only at 37 degrees C (p < 0.001). There were significantly more colonies at 42 degrees C (p = 0.001). Competing flora were heavier at 37 degrees C, but this did not account for the difference in isolation rates or quantity of growth. It is recommended that cultures for Campylobacter spp. be routinely incubated at 42 degrees C. More specific techniques are required to seek for fastidious campylobacters.

Agar↗

PubMed2002

Comparison of the performance of lactose and mannitol selenite enriched broths, subcultured to DCA and XLD agars, in the isolation of Salmonella spp. from faeces.

Selenite-based enrichment broths using either lactose or mannitol as a carbohydrate source are generally used as selective enrichment media for the isolation of Salmonella spp. from human faeces in the UK, but few studies have compared the relative efficacy of the available formulations. A variety of solid media is used for the routine subculture from these selective broths, but similarly we have been unable to find published evidence as to which medium performs best. Four thousand and nineteen faecal samples were examined in four laboratories and the isolation rates of Salmonella spp. from lactose (LS) or mannitol selenite (MS) broths, plated onto either xylose lysine desoxycholate agar (XLD) or desoxycholate citrate agar (DCA) were compared. MS performed significantly better than LS (p = 0.02), recovering 95 salmonellae compared with 87. No significant difference in isolation rates was found between XLD and DCA, although colonial appearances of suspected salmonellae on XLD were much more specific, resulting in significantly fewer colonies having to be selected for supplementary testing (p < 0.001) and so reducing confirmatory work. An opportunity exists to simplify holdings of media by choosing to use the MS/XLD combination.

Bacteriological Techniques↗

PubMed2001

An element in intron 1 of the CFTR gene augments intestinal expression in vivo.

The elements controlling the complex developmental and tissue-specific expression of the cystic fibrosis transmembrane conductance regulator (CFTR) gene lie outside the basal promoter region and have not been characterized. We previously identified a tissue-specific DNase I hypersensitive site (DHS) in intron 1 (185 + 10 kb) of the CFTR gene. Here we show that removal of the core element abolishes the activity of this DHS in transient transfection assays of reporter/enhancer gene constructs. We then compared expression from a 310 kb yeast artificial chromosome (YAC) that contains the entire CFTR gene with expression from the same YAC from which the DHS element had been deleted. Stable transfection of a human colon carcinoma cell line showed that transcription from the deleted YAC was reduced by approximately 60%. In transgenic mice, deletion of the intron 1 DHS had no effect on expression in the lung, but reduced expression in the intestine by approximately 60%. Thus, the regulatory element associated with the intron 1 DHS is tissue-specific and is required for normal CFTR expression levels in the intestinal epithelium in vivo.

Animals↗

PubMed2001

A comparison of the performance of bacitracin-incorporated chocolate blood agar with chocolate blood agar plus a bacitracin disk in the isolation of Haemophilus influenzae from sputum.

The lack of selectivity of chocolated blood agar (CBA), routinely used for the isolation of Haemophilus influenzae, may lead to masking of the growth of H. influenzae due to overgrowth of competing flora. Bacitracin can be used as a selective agent, either incorporated into the medium or applied to the medium in a filter paper. However, neither method has been evaluated or compared in a large study. Sputum samples (1990) were examined in four laboratories and the isolation rates of H. influenzae on chocolated blood agar with bacitracin added to the medium (BCA) and chocolated blood agar (CBA) with a bacitracin disk were compared. A plain blood agar plate was also inoculated to facilitate the isolation of Streptococcus pneumoniae so that its effects on the isolation of H. influenzae could be assessed. No significant difference was found between the isolation rates of H. influenzae on BCA and CBA with a bacitracin disk, although competing flora was greatly reduced and quantity of growth of H. influenzae increased on BCA. The presence of S. pneumoniae did not affect the isolation of H. influenzae in this study.

Anti-Bacterial Agents↗

PubMed2000

Modular bacterial artificial chromosome vectors for transfer of large inserts into mammalian cells.

To facilitate the use of large-insert bacterial clones for functional analysis, we have constructed new bacterial artificial chromosome vectors, pPAC4 and pBACe4. These vectors contain two genetic elements that enable stable maintenance of the clones in mammalian cells: (1) The Epstein-Barr virus replicon, oriP, is included to ensure stable episomal propagation of the large insert clones upon transfection into mammalian cells. (2) The blasticidin deaminase gene is placed in a eukaryotic expression cassette to enable selection for the desired mammalian clones by using the nucleoside antibiotic blasticidin. Sequences important to select for loxP-specific genome targeting in mammalian chromosomes are also present. In addition, we demonstrate that the attTn7 sequence present on the vectors permits specific addition of selected features to the library clones. Unique sites have also been included in the vector to enable linearization of the large-insert clones, e. g., for optical mapping studies. The pPAC4 vector has been used to generate libraries from the human, mouse, and rat genomes. We believe that clones from these libraries would serve as an important reagent in functional experiments, including the identification or validation of candidate disease genes, by transferring a particular clone containing the relevant wildtype gene into mutant cells or transgenic or knock-out animals.

Animals↗

PubMed2000

Expression of the human CFTR gene from episomal oriP-EBNA1-YACs in mouse cells.

Plasmids carrying the origin of plasmid replication ( oriP ) and expressing the EBNA-1 protein from the Epstein-Barr virus replicate and segregate in human cells and are thus potentially useful vectors for gene therapy. As very large circular molecules, up to 660 kb in size, can be maintained episomally using this system, it is possible to include intact human genes with all their long-range controlling elements which might give high levels of tissue-specific and controlled gene expression. We have shown previously that a 320 kb yeast artificial chromosome (YAC) carrying the intact human CFTR gene can complement the Cambridge null cystic fibrosis mice as a transgene. We have now modified this YAC to a circular molecule carrying both oriP and the EBNA-1 gene. We show that this oriP-EBNA1-YAC can be stably maintained as unrearranged episomes in mouse LA-9 cells, which do not express endogenous cftr, and in mouse CMT-93 cells, which do express endogenous cftr. The human CFTR gene is expressed in some of the cell lines, but the level of expression is very variable between cell lines and is not related to the copy number of the elements.

Alanine↗

PubMed2000

Ondansetron versus placebo for prophylaxis of nausea and vomiting in patients undergoing ambulatory laparoscopic cholecystectomy.

BACKGROUND: Postoperative nausea and vomiting is a common problem in patients undergoing laparoscopic cholecystectomy (LC). The purpose of this study was to evaluate the efficacy of ondansetron given at the induction of anesthesia in patients scheduled for ambulatory LC. METHODS: A total of 84 patients undergoing ambulatory LC were enrolled in a randomized, prospective, double-blinded study in which the subjects received either placebo or 4 mg ondansetron intravenously at induction of anesthesia. A nausea scoring system was employed utilizing a 5-point linear scale, with 1 point given for no nausea and a maximum of 5 points for an episode of emesis. Each patient received a total of four scores postoperatively. RESULTS: The patients receiving placebo had significantly more episodes of nausea (53 versus 32; P <0.009) and emesis (11 versus 2; P <0.02), higher mean total nausea scores (7.2 versus 5.4; P <0.006), and need for additional postoperative antiemetics (23 versus 14; P <0.05) than those receiving ondansetron. CONCLUSIONS: In patients undergoing ambulatory laparoscopic cholecystectomy, ondansetron at induction was highly effective in decreasing postoperative nausea and vomiting and should become the standard.

Adult↗

PubMed2000

Carboxyhemoglobin half-life in carbon monoxide-poisoned patients treated with 100% oxygen at atmospheric pressure.

STUDY OBJECTIVES: There are large reported differences for the carboxyhemoglobin (COHb) half-life (COHb t(1/2)) in humans breathing 100% atmospheric O(2) following CO inhalation in tightly controlled experiments compared to the COHb t(1/2) observed in clinical CO poisoning (range, 36 to 131 min, respectively). Other reports have suggested that the COHb t(1/2) may be affected by gender differences, age, and lung function. We wished to test the hypothesis that the COHb t(1/2) might also be influenced by CO poisoning vs experimental CO exposure, by a history of loss of consciousness (LOC), concurrent tobacco smoking, and by PaO(2). The purpose of the present study was to measure the COHb t(1/2) in a cohort of CO-poisoned patients and to determine if those listed factors influenced the COHb t(1/2). DESIGN: Retrospective chart review from 1985 to 1995. We calculated the COHb t(1/2) of CO-poisoned patients who were treated with high-flow supplemental atmospheric pressure O(2) delivered by nonrebreather face mask or endotracheal tube. SETTING: Hyperbaric medicine department of a tertiary-care teaching hospital. PATIENTS: Of 240 CO-poisoned patients, 93 had at least two COHb measurements > 2% (upper limit of normal) with recorded times of the measurements, permitting calculation of the COHb t(1/2). RESULTS: The COHb t(1/2) was 74 +/- 25 min (mean +/- 1 SD) with a range from 26 to 148 min. By stepwise multiple linear regression analysis, the PaO(2) influenced the COHb t(1/2) (R(2) = 0.19; p < 0.001), whereas the COHb t(1/2) was not influenced by gender, age, smoke inhalation, history of LOC, concurrent tobacco smoking, degree of initial metabolic acidosis (base excess), or initial COHb level. CONCLUSIONS: The COHb t(1/2) of 93 CO-poisoned patients treated with 100% O(2) at atmospheric pressure was 74 +/- 25 min, considerably shorter than the COHb t(1/2) reported in prior clinical reports (approximately 130 +/- 130 min) and was influenced only by the patient's PaO(2).

Adolescent↗

PubMed1998

Family rigidity, adolescent problem-solving deficits, and suicidal ideation: a mediational model.

Family rigidity and adolescent problem-solving deficits have each been linked to adolescent suicidal behaviors. The objective of the present study was to evaluate the hypothesis that family rigidity has primarily an indirect effect on adolescent suicidal ideation through its effect on adolescent problem-solving deficits. College students and their parents completed the Family Adaptability and Cohesion Evaluation Scale II. Students also completed the Problem-Solving Inventory and the Adult Suicidal ideation Questionnaire. Structural equation models confirmed that family rigidity has an indirect effect on adolescent suicidal ideation through its effect on adolescent problem-solving deficits. Directions for treatment efforts are discussed.

Adaptation, Psychological↗

PubMed1998

Demographic differences in prostate cancer incidence and stage: an examination of population diversity in California.

INTRODUCTION: Geographic and racial/ethnic variability in prostate cancer incidence rates and stage distribution may be partly attributed to differences in screening and early detection. METHODS: Using California Cancer Registry data we aimed to characterize variability in prostate cancer rates statewide and to examine differences in the stage at diagnosis of prostate cancer by racial/ethnic group statewide and by census tract per capita income in San Diego County. We calculated annual average (1988-1991) age-adjusted incidence rates per 100,000 (AAIR) of prostate cancer for 49,880 men over age 34 years. Racial/ethnic groups were compared using incidence rate ratios (IRR) (AAIR localized plus regional stages divided by AAIR distant stage). RESULTS: Statewide, Caucasians showed a higher IRR [6.16, 95% confidence interval (CI), 6.00-6.30] than did African Americans (2.34, 95% CI, 1.89-2.89), Hispanics (3.84, 95% CI, 3.63-4.05), or Asian/others (3.61, 95% CI, 1.80-7.22). Within San Diego County, Caucasians living in higher per capita income census tracts (> or = 65th percentile) had a significantly higher IRR (8.80, 95% CI 7.84-9.89) than did lower-income tracts (5.68, 95% CI, 5.13-6.30). CONCLUSION: Findings from the present and similar studies suggest that outcomes research is needed to determine the impact of these demographic differences on prostate cancer mortality and quality of life. This is particularly important given the current controversy regarding the treatment of clinically localized prostate cancers, increasingly found through early detection, which often involve difficult choices between aggressive therapies including prostatectomy or watchful waiting.

Adult↗

PubMed1998

Differences in breast cancer stage at diagnosis between non-Hispanic white and Hispanic populations, San Diego County 1988-1993.

The incidence of breast cancer in the U.S. is lower among Hispanic women than non-Hispanic white women. However, population-based studies show that Hispanic women are more likely to be diagnosed at a later stage than non-Hispanic whites. We aimed to determine whether: 1) a lower proportion of breast cancer was diagnosed at early vs. late stages in Hispanic compared to non-Hispanic white women from 1988-93 in San Diego County, and 2) lower income is related to later stage at diagnosis for both groups. All incident cases of breast cancer in San Diego County from the California Cancer Registry (10,161 cases) were stratified by 'early' (in situ or localized) or 'late' (regional or distant) stage, and by race/ethnicity. Annual average age-adjusted incidence rates/100,000 (AAIR) were calculated. Incidence rate ratios (IRR) (AAIR for early stages divided by AAIR for late stages) were used as a surrogate of early detection. AAIRs for early and late stage disease were significantly higher for non-Hispanic whites (89.3, 42.3) than Hispanic women (46.7, 27.2). The IRR was significantly higher for non-Hispanic whites than Hispanics, (2.11 vs 1.72, p = 0.01). This difference was greatest among women under 50 years old (IRR difference 0.63), and not apparent for women 65 or older (IRR difference 0.06). There was also an association between increasing census tract per capita income and higher rates of early stage disease among non-Hispanic whites but not Hispanics. Results suggest that Hispanic women and lower income women should be targeted for early detection.

Adolescent↗