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Biomedical subjects

R Woods

Publications and source records attributed to R Woods.

At least 37 records · Page 2Linked to original sources

Quantification of HER2/neu gene amplification by competitive pcr using fluorescent melting curve analysis.

BACKGROUND: Molecular detection methods for HER2/neu gene amplification include fluorescence in situ hybridization (FISH) and competitive PCR. We designed a quantitative PCR system utilizing fluorescent hybridization probes and a competitor that differed from the HER2/neu sequence by a single base change. METHODS: Increasing twofold concentrations of competitor were coamplified with DNA from cell lines with various HER2/neu copy numbers at the HER2/neu locus. Competitor DNA was distinguished from the HER2/neu sequence by a fluorescent hybridization probe and melting curve analysis on a fluorescence-monitoring thermal cycler. The percentages of competitor to target peak areas on derivative fluorescence vs temperature curves were used to calculate copy number. RESULTS: Real-time monitoring of the PCR reaction showed comparable relative areas throughout the log phase and during the PCR plateau, indicating that only end-point detection is necessary. The dynamic range was over two logs (2000-250 000 competitor copies) with CVs < 20%. Three cell lines (MRC-5, T-47D, and SK-BR-3) were determined to have gene doses of 1, 3, and 11, respectively. Gene amplification was detected in 3 of 13 tumor samples and was correlated with conventional real-time PCR and FISH analysis. CONCLUSION: Use of relative peak areas allows gene copy numbers to be quantified against an internal competitive control in < 1 h.

DNA↗

Laparoscopic management of intestinal endometriosis.

Intestinal involvement by endometriosis traditionally required open laparotomy for bowel resection and anastomosis. Operative laparoscopy may offer the most effective form of treatment for these women. Two women with endometriosis of the rectum and right hemicolon, respectively, underwent transvaginal resection of the rectum and laparotomy for hemicolectomy, assisted by laparoscopy. The only morbidity was postoperative ileus in the former patient. Both women were asymptomatic at the 6-week postoperative visit.

Adult↗

Intestinal sleeve anastomosis: a comparative study with end-to-end anastomosis.

This study examines the utility of a sleeve anastomosis with comparison to conventional end to end anastomosis. Thirty New Zealand white rabbits were randomized to sleeve (n = 15) or end-to-end (n = 15) small bowel anastomosis. Five rabbits of each group were sacrificed at 3 days, 7 days, and 6 weeks. Anastomoses were assessed for integrity, bursting strength, and stenosis and examined histologically. Ten control specimens of small bowel were tested for bursting pressure. Three rabbits died postoperatively (1 sleeve and 2 end-to-end). A fourth rabbit (sleeve) was sacrificed early at 3 weeks and had a total stenosis at the anastomosis. The remaining 26 rabbits were reoperated at the prescribed times. There was no evidence of infection or dehiscence in any of these rabbits. Both end-to-end and sleeve anastomoses were equivalent for bursting pressure at all times and, at 7 days and 6 weeks, were similar to controls. The stenotic index revealed no evidence of proximal dilation suggestive of obstruction in the 26 rabbits. For sleeve anastomoses the length of the projected bowel into the lumen persisted at the 6-week stage. Histologically there was good evidence of healing in both the sleeve and end-to-end anstomoses and the serosal surface of the sleeved bowel had epithelialized. Sleeve anastomosis has been demonstrated to heal well and to be as strong as conventional end-to-end anastomosis. Further studies are warranted to determine its role in intestinal anastomosis and potential as a valve.

Anastomosis, Surgical↗

Pharmacokinetics of intravenously administered ketamine in southern elephant seals (Mirounga leonina).

Southern elephant seals (Mirounga leonina) are large, potentially dangerous animals which must be restrained before study or treatment. However, chemical restraint is unpredictable, possibly because of differences in body composition during fasting ashore, and circulatory adaptations to enable diving. The pharmacokinetics of ketamine (1.1 mg/kg i.v.) was studied in 15 southern elephant seals which had come ashore on Macquarie Island. The animals were first sedated with pethidine (5 mg/kg i.m.) to allow intravenous access. There was great variability in the calculated pharmacokinetic parameters, possibly due to circulatory changes associated with periods of apnoea which characterise this animal's response to anaesthetics. The median values were similar to those reported for other species: distribution t1/2 = 2.5 min (range 1-11 min); elimination t1/2 = 43 min (range 17-108 min); apparent volume of distribution, 1474 ml/kg (range 830-9301 ml/kg); clearance, 33 ml/min/kg (range 12-57 ml/min/kg). This was the first investigation of drug disposition in any seal species. and the first in any free-ranging wild animal. It is important to obtain pharmacological data in animals in whom drugs are used, but pre-catheterised captive animals may provide less variable data.

Anesthetics, Dissociative↗

An audit of error rates in a UK district hospital transfusion laboratory.

We have audited the error rates of our transfusion laboratory and compared these with error rates reported in the transfusion literature. Error rates were calculated using workload data from the department. The majority of errors that were detected were preanalytical and related to inadequate or incomplete data provided on the sample or request form. These errors were all corrected prior to any further action being taken on that request. The main analytical errors were transcription errors in entering patient identification information into the laboratory computer by transfusion staff together with the incorrect performance of blood group testing. For postanalytical errors the main errors were failure of nursing staff to follow procedures for the collection of blood components prior to transfusion. There were no serious consequences identified of the errors detected in this study. It was difficult to compare these results with those published in the literature in view of the different methodologies that have been reported when error rates have been determined. A standard method should be developed in the UK for calculating error rates so that laboratories can benchmark their performance against comparable organizations.

Blood Grouping and Crossmatching↗

Total colectomy for Crohn's disease.

BACKGROUND: Total colectomy for Crohn's disease of the colon may be restorative with ileorectal anastomosis or with an ileostomy and rectal stump. The present paper retrospectively audits the results of total colectomy and in particular assesses the number of patients who had a permanent ileostomy and whether this was related to disease in the rectum at the time of the original operation. METHODS: A retrospective case note review was undertaken of patients operated upon between 1968 and 1994. RESULTS: Thirty-eight patients were identified (mean age 35 years; range 17-65 years). One patient died perioperatively from an anastomotic leak. Median follow-up for the remaining patients was 7 years (range 1-29 years). Ileorectal anastomosis was performed in 17 patients and total colectomy and ileostomy in 20 patients. Indications for surgery were failure of medical treatment (61%); toxic colitis (18%); abscess (8%); perforation (5%); large bowel obstruction (5%); and colovesical fistula (3%). Subsequent proctectomy (14 patients, 38%) was more likely with subtotal colectomy and ileostomy (nine patients, 45%) than ileorectal anastomosis (five patients, 29%). This was not statistically significant (P = 0.33). Additionally, seven patients had diversion of the rectum making 21 with an ileostomy (57%). Rectal involvement at the time of the original procedure significantly increased the likelihood of permanent ileostomy (P = 0.001). The presence of anal disease did not increase the prospect of ileostomy. One patient died with advanced adeno carcinoma in a defunctioned rectum. CONCLUSIONS: A permanent ileostomy after total colectomy for Crohn's disease is common and significantly more likely with rectal involvement.

Adolescent↗

A direct comparison of the activities of two humanized respiratory syncytial virus monoclonal antibodies: MEDI-493 and RSHZl9.

Two humanized monoclonal antibodies, MEDI-493 and RSHZ19, were developed independently as potential improvements over RSV-IGIV for prevention of respiratory syncytial virus (RSV) infection. RSV-IGIV is a polyclonal human antibody preparation for intravenous infusion enriched for RSV neutralizing activity. A phase III clinical trial showed that MEDI-493 significantly reduced hospitalizations due to RSV infection. In a separate trial, RSHZ19 failed to show significant efficacy. In new studies, the in vitro and in vivo activities of MEDI-493 and RSHZ19 were compared to determine whether the different clinical results are related to differences in biologic activity. MEDI-493 was consistently 4- to 5-fold more potent than RSHZ19 in antigen binding, RSV neutralization, and fusion inhibition assays. Although both MEDI-493 and RSHZ19 were effective against A and B subtypes of RSV in the cotton rat model of RSV infection, 2- to 4-fold higher doses of RSHZ19 were required for similar protection. The enhanced activity of MEDI-493 compared with RSHZ19 may, in part, explain its better clinical effect.

Animals↗

Fungal pneumonia in a captive black rhinoceros.

A captive black rhinoceros (Diceros bicornis) with a hoof abscess was treated with long-term antibiotic therapy. After 9 months of treatment, there was rapid deterioration, marked weight loss and reluctance to stand. Profuse, bilateral epistaxis developed accompanied by collapse and the animal was euthanased. Necropsy revealed pulmonary aspergillosis with concurrent Pseudomonas aeruginosa infection. Though a well-recognized disease of black rhinoceros, fungal pneumonia has not been reported in this species in Australia. The cost and efficacy of treatment have been questioned, however, prophylactic antifungal drug administration will be considered in any further cases of chronic, debilitating illness in black rhinoceros at Western Plains Zoo.

Abscess↗

A new questionnaire for the repeat of the first stage of the European Community Respiratory Health Survey: a pilot study.

In the early 1990s a multicentre survey on asthma was performed on the young adult population (European Community Respiratory Health Survey - ECRHS). This study is to be repeated in order to estimate changes in the prevalence of asthma-like symptoms during the last decade and to assess the social and economic costs of the disease and their variations among countries. The self-administered questionnaire devised for this purpose is a two-page questionnaire. The first page contains the same items as those used in the first survey with four additional questions related to: 1) the frequency and severity of asthma attacks; 2) the presence of chronic bronchitis; 3) smoking habits; and 4) a visual analogue scale assessing perception of outdoor pollution. The second page aims to collect information regarding the direct and indirect costs of asthma. The influence of the length of the questionnaire on the response rate was assessed in a pilot study in Italy. Two random samples of 150 subjects received either the one-page questionnaire (first page) or the two-page questionnaire. The response rate was compared with that obtained from the first postal wave in the 1991-1992 survey. Although the response rate was unchanged when using the one-page questionnaire (45% versus 45%), it decreased by 7% when the two-page questionnaire was used (38% versus 45%). On the basis of these results, no problem should arise if four more questions are added to the one-page questionnaire. The slight reduction in the response rate of the two-page questionnaire is worrying but could be corrected by the use of telephone interviews.

Adult↗

Can staff training reduce behavioural problems in residential care for the elderly mentally ill?

The effect of a brief in-service training programme on the psychosocial management of behavioural disturbance in residential care is described. Although the incidence of problematic behaviour did not change 3 months after training, staff in the experimental homes reported a significant improvement in their management of problematic behaviour, as compared to the control home. However, these effects were not maintained a year later. The importance of post-training supervision and the sociocultural context of the home in the maintenance of psychosocial intervention in residential settings is discussed.

Aged↗

Klinefelter's syndrome as a model of anomalous cerebral laterality: testing gene dosage in the X chromosome pseudoautosomal region using a DNA microarray.

Consistent handedness and language laterality are two of the most striking behavioral and cognitive asymmetries observed in humans. Alterations in the typical pattern of cerebral laterality, termed "anomalous dominance," is observed in left-handers and some patients with verbal learning disabilities. We undertook the study of a genetically distinct group of subjects, XXY males (Klinefelter's syndrome; KS), who demonstrate anomalous dominance in a variety of testing paradigms in order to begin to elucidate the molecular basis of anomalous dominance in this population. KS subjects manifest specific verbal learning disability, evidence of altered functional laterality for phonologic processing, and an increase in left-handedness when measured by skill. It is proposed that an alteration in gene dosage in the pseudoautosomal region (PAR) of the sex chromosomes is the most likely explanation for anomalous dominance in these patients. This is especially intriguing in light of previously described genetic models of cerebral laterality that suggest a contributing locus in the PAR, or adjacent high homology regions of the X chromosome. We have developed an ordered DNA microarray covering the X chromosome PAR at high resolution for hybridization with two-color fluorescently labeled probes. We demonstrate the ability to detect changes in hybridization signal that will facilitate efficient large-scale screening of this region for alterations in gene dosage associated with features of anomalous dominance and other cognitive or behavioral phenotypes.

Case-Control Studies↗

Visual-motor, visual-perceptual, and fine motor outcomes in very-low-birthweight children at 5 years.

The prevalence and severity of visual-motor deficits and the relation of visual-motor function to visual perception and fine motor skills was examined in a group of 83 neurologically and intellectually normal (IQ >84) very-low-birthweight (VLBW) children at age 5. Fifteen children (17%) had below average visual motor scores (<1SD below mean). While relatively few children (N=9) had below average scores in visual perception (11%), 58 (71%) had below average scores for fine motor skills. Nineteen (23%) were considered impaired (<1.5SD); of these, eight were severely impaired (<2SD). Fine motor scores were significantly lower in children who had been born <28 weeks' gestation, with hyaline membrane disease, or had required a longer period of ventilation. There was significant correlation between visual-motor and fine motor scores (r = 0.50, P < 0.001) and between visual-motor and visual perception scores (r = 0.42, P < 0.001). The implications of these findings and management of these 'normal' children need further research. Previous reports of visual-motor dysfunction in school-age VLBW children could be related to fine motor difficulties.

Apgar Score↗

Development of a humanized monoclonal antibody (MEDI-493) with potent in vitro and in vivo activity against respiratory syncytial virus.

Neutralizing polyclonal antibody to respiratory syncytial virus (RSV) has been shown to be an effective prophylactic agent when administered intravenously in high-risk infants. This study describes the generation of a humanized monoclonal antibody, MEDI-493, that recognizes a conserved neutralizing epitope on the F glycoprotein of RSV. The affinity of MEDI-493 was found to be equal to or slightly better than an isotype-matched chimeric derivative of the parent antibody. In plaque reduction, microneutralization, and fusion-inhibition assays, MEDI-493 was significantly more potent than the polyclonal preparation. Broad neutralization of a panel of 57 clinical isolates of the RSV A and B subtypes was demonstrated. Pretreatment of cotton rats with MEDI-493 resulted in 99% reduction of lung RSV titers at a dose of 2.5 mg/kg, corresponding to a serum concentration of 25-30 microg/mL. Further, MEDI-493 did not induce increased RSV infection or pathology in either a primary or a secondary challenge.

Amino Acid Sequence↗

'Sickness is a baffling matter'. A reply to James C. Riley.

This paper replies to the comments made by James C. Riley. It provides a defence of the assumptions adopted in 'Physician, heal thyself' (Social History of Medicine, 9(1996), 1-30) to estimate the average duration of work-preventing sickness experienced by members of the medical profession in England in the 1860s as well as offering some new estimates. It also provides further criticisms of Riley's contention that although the rate of mortality declined in England between the 1860s and 1890s that of morbidity increased, which is based on surveys of friendly society members. In doing so it reiterates the warning given by Jacques Bertillon in 1892 concerning the use of friendly society surveys for the measurement of variations and trends in morbidity patterns by age.

History, 19th Century↗