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Biomedical subjects

V Allen

Publications and source records attributed to V Allen.

At least 19 recordsLinked to original sources
PubMed2004

Defaecation and weight of the gastrointestinal tract contents after feed and water withdrawal in broilers.

1. Broilers were held under constant light for up to 24 h with access to feed and water, access to water only, or access to neither feed nor water. 2. Birds given continuous access to feed ate evenly over the 24-h period. Feed deprivation reduced drinking behaviour to about half of that when feed was available. Deprivation of feed, or of feed and water, had similar effects on defaecation and the reduction in the weight of the gut contents. It reduced the overall number of defaecations over the 24-h period to about 40% of that in birds given food and water, and the weight of the contents by about 75%. The maximum effect of deprivation on defaecation was seen after about 4 h, and on gut contents weight after about 12 h. 3. With longer fasting the contents of most parts of the gut, except the caeca, became wetter. 4. The findings may have implications for the practice of preslaughter food withdrawal to reduce the potential for soiling of the live birds and contamination of the carcase after slaughter. However, the likelihood is that relatively short overall feed deprivation times of not more than 8 to 12 h before killing may be desirable, but specifying firm recommendations on optimal feed withdrawal times requires further work.

Animals↗
PubMed2001

Alveolar metabolism of natural vs. synthetic surfactants in preterm newborn rabbits.

We compared the recoveries of four surfactant preparations: two natural [term fetal rabbit surfactant (FRS) and adult rabbit surfactant (ARS)] and two commercially available preparations [apoprotein-based Survanta (S) and synthetic Exosurf (E)] from 27-day gestation rabbit pups treated at birth and ventilated up to 120 min. At 5, 60, and 120 min, we measured the recovery of the heavy-aggregate, metabolically active form (H) and the light-aggregate, nonsurface active metabolic breakdown form (L) of alveolar surfactant and determined the phospholipid content and composition of the intracellularly stored lamellar body (LB) pool. Pups treated with FRS had <15% loss of H by 2 h. ARS-treated pups had a >50% loss of H by 1 h, and E- and S-treated pups had approximately 50% loss by 5 min, with a slower rate of continuing loss of up to 80% by 2 h. The major losses of H phospholipid were not explained by the L-form recovery. LB phospholipid significantly increased only in the E-treated pups and only at 2 h. FRS provides a biologically active form (H) of surfactant that appeared to remain in the airway for a significantly longer time than the other surfactant preparations. The unique properties of FRS merit further study.

Animals↗
PubMed2000

Inhibition of Ca(2+) signalling by p130, a phospholipase-C-related catalytically inactive protein: critical role of the p130 pleckstrin homology domain.

p130 was originally identified as an Ins(1,4,5)P(3)-binding protein similar to phospholipase C-delta but lacking any phospholipase activity. In the present study we have further analysed the interactions of p130 with inositol compounds in vitro. To determine which of the potential ligands interacts with p130 in cells, we performed an analysis of the cellular localization of this protein, the isolation of a protein-ligand complex from cell lysates and studied the effects of p130 on Ins(1,4,5)P(3)-mediated Ca(2+) signalling by using permeabilized and transiently or stably transfected COS-1 cells (COS-1(p130)). In vitro, p130 bound Ins(1,4,5)P(3) with a higher affinity than that for phosphoinositides. When the protein was isolated from COS-1(p130) cells by immunoprecipitation, it was found to be associated with Ins(1,4,5)P(3). Localization studies demonstrated the presence of the full-length p130 in the cytoplasm of living cells, not at the plasma membrane. In cell-based assays, p130 had an inhibitory effect on Ca(2+) signalling. When fura-2-loaded COS-1(p130) cells were stimulated with bradykinin, epidermal growth factor or ATP, it was found that the agonist-induced increase in free Ca(2+) concentration, observed in control cells, was inhibited in COS-1(p130). This inhibition was not accompanied by the decreased production of Ins(1,4,5)P(3); the intact p130 pleckstrin homology domain, known to be the ligand-binding site in vitro, was required for this effect in cells. These results suggest that Ins(1,4,5)P(3) could be the main p130 ligand in cells and that this binding has the potential to inhibit Ins(1,4,5)P(3)-mediated Ca(2+) signalling.

3T3 Cells↗
PubMed1999

Involvement of EF hand motifs in the Ca(2+)-dependent binding of the pleckstrin homology domain to phosphoinositides.

The pleckstrin homology (PH) domains of phospholipase C (PLC)-delta1 and a related catalytically inactive protein, p130, both bind inositol phosphates and inositol lipids. The binding to phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] by PLC-delta1 is proposed to be the critical interaction required for membrane localization to where the substrate resides; it is also required for the Ca(2+)-dependent activation of PLC-delta1 observed in the permeabilized cells. In the proximity of the PH domain, both PLC-delta1 and p130 possess the EF-hand domain, containing classical motifs implicated in calcium binding. Therefore, in the present study we examined whether the binding of the PH domain to PtdIns(4,5)P2 is regulated by changes in free Ca2+ concentration within the physiological range. A Ca2+ dependent increase in the binding to PtdIns(4,5)P2 was observed with a full-length PLC-delta1, while the isolated PH domain did not show any Ca2+ dependence. However, the connection of the EF-hand motifs to the PH domain restored the Ca2+ dependent increase in binding, even in the absence of the C2 domain. The p130 protein showed similar properties to PLC-delta1, and the EF-hand motifs were again required for the PH domain to exhibit a Ca2+ dependent increase in the binding to PtdIns(4,5)P2. The isolated PH domains from several other proteins which have been demonstrated to bind PtdIns(4,5)P2 showed no Ca2+ dependent enhancement of binding. However, when present within a chimera also containing PLC-delta1 EF-hand motifs, the Ca2+ dependent binding was again observed. These results suggest that the binding of Ca2+ to the EF-hand motifs can modulate binding to PtdIns(4,5)P2 mediated by the PH domain.

Blood Proteins↗
PubMed1999

Molecular detection of prostate cells in ejaculate and urethral washings in men with suspected prostate cancer.

PURPOSE: To determine whether prostatic cells were normally present in ejaculate and if the sensitivity and specificity of the detection of malignant prostate cells in ejaculate and urethral washings from men with suspected prostate cancer could be improved using the more sensitive molecular technique of reverse transcriptase-polymerase chain reaction (RT-PCR). MATERIALS AND METHODS: RT-PCR for prostate-specific antigen (PSA), prostate-specific membrane antigen (PSM) and Apoliprotein D (3 putative prostate-specific and/or cancer-specific markers) was performed on RNA extracts of ejaculate (80) and urethral washings (52) from 77 men with suspected prostate cancer and 12 young controls (<30 years of age) and urines from 5 men who had radical prostatectomies and 10 women. RESULTS: PSA, PSM and Apolipoprotein D expression was detected in ejaculates and urethral washings from both patient and control groups. No differences were observed in the results obtained for 58 men with suspected or 19 men with confirmed prostate cancer or the 18 vasectomized men within the patient group. Urines from the 5 men who had radical prostatectomies and 10 women were all negative for PSA, but PSM was detected in 2 female urines and in 3 radical prostatectomy samples. As few as 10 LNCaP prostate tumor cells could be detected by PSA RT-PCR when added to female urine. CONCLUSION: We have established a sensitive method of detecting prostatic cells in ejaculate and urethral washings and shown that PSA RT-PCR is a reliable indicator of prostate cells in these samples. However, RT-PCR for PSA, PSM and Apoliprotein D were not useful for discriminating malignant from non-malignant prostate cells.

Adult↗
PubMed1998

Replacements of single basic amino acids in the pleckstrin homology domain of phospholipase C-delta1 alter the ligand binding, phospholipase activity, and interaction with the plasma membrane.

The pleckstrin homology (PH) domain of phosphatidylinositol-specific phospholipase C-delta1 (PLC-delta1) binds to both D-myo-inositol 1,4, 5-trisphosphate (Ins(1,4,5)P3) and phosphatidylinositol 4, 5-bisphosphate (PtdIns(4,5)P2) with high affinities. We have previously identified a region rich in basic amino acids within the PH domain critical for ligand binding (Yagisawa, H., Hirata, M., Kanematsu, T., Watanabe, Y., Ozaki, S., Sakuma, K., Tanaka, H., Yabuta, N., Kamata, H., Hirata, H., and Nojima, H. (1994) J. Biol. Chem. 269, 20179-20188; Hirata, M., Kanematsu, T., Sakuma, K., Koga, T., Watanabe, Y., Ozaki, S., and Yagisawa, H. (1994) Biochem. Biophys. Res. Commun. 205, 1563-1571). To investigate the role of these basic residues, we have performed site-directed mutagenesis replacing each of the basic amino acid in the N-terminal 60 residues of PLC-delta1 (Lys24, Lys30, Lys32, Arg37, Arg38, Arg40, Lys43, Lys49, Arg56, Lys57, and Arg60) with a neutral or an acidic amino acid. The effects of these mutations on the PH domain ligand binding properties and their consequence for substrate hydrolysis and membrane interactions of PLC-delta1 were analyzed using several assay systems. Analysis of [3H]-Ins(1,4,5)P3 binding, measurement of the binding affinities, and measurements of phospholipase activity using PtdIns(4,5)P2-containing phospholipid vesicles, demonstrated that residues Lys30, Lys32, Arg37, Arg38, Arg40, and Lys57 were required for these PLC-delta1 functions; in comparison, other mutations resulted in a moderate reduction. A subset of selected mutations was further analyzed for the enzyme activity toward substrate present in cellular membranes of permeabilized cells and for interaction with the plasma membrane after microinjection. These experiments demonstrated that mutations affecting ligand binding and PtdIns(4,5)P2 hydrolysis in phospholipid vesicles also resulted in reduction in the hydrolysis of cellular polyphosphoinositides and loss of membrane attachment. All residues (with the exception of the K43E substitution) found to be critical for the analyzed PLC-delta1 functions are present at the surface of the PH domain shown to contain the Ins(1,4,5)P3 binding pocket.

Amino Acid Sequence↗
PubMed1998

Validation of an automated technique for determining the mechanical characteristics of coronary arteries during balloon angioplasty: laboratory assessment with necropsy segments.

OBJECTIVES: To develop a technique for automatic inflation of a percutaneous transluminal coronary angioplasty (PTCA) balloon, with continuous measurement of the balloon pressure and volume; to validate the technique for determining the mechanical characteristics of coronary arteries. METHODS: During necropsy examination of the hearts of nine patients, 17 coronary artery samples were obtained for histological examination. A PTCA balloon was inserted into each artery, and the balloon pressure and volume were measured continuously during four repeat automatic inflations of the balloon. RESULTS: Of the 17 arteries, eight showed elastic, six plastic, and three fracture pressure-volume deformation characteristics. For the plastic deformations, the first inflation required a higher pressure than subsequent repeat inflations of 82 (61) kPa (mean (SD), range 25 to 175 kPa). For the three in the fracture group, the pressure drop because of the fracture occurred between 210 and 540 kPa. Two of these three showed a tear on visual inspection, and the other showed disruption of the intimal plaque on blinded histological examination. Of the six with plastic deformation characteristics alone, one showed a tear, and on histological examination two others showed splitting of the internal and external elastic lamina and one showed separation of intima and media. None in the elastic group showed any of these characteristics. CONCLUSIONS: Plastic and fracture deformation characteristics could be differentiated from elastic characteristics. Visual or histological evidence of fracturing was present in all three arteries identified during angioplasty as having pressure-volume fracture characteristics.

Angioplasty, Balloon, Coronary↗
PubMed1997

Regulation of inositol lipid-specific phospholipase cdelta by changes in Ca2+ ion concentrations.

Studies of inositol lipid-specific phospholipase C (PLC) have elucidated the main regulatory pathways for PLCbeta and PLCgamma but the regulation of PLCdelta isoenzymes still remains obscure. Here we demonstrate that an increase in Ca2+ ion concentration within the physiological range (0.1-10 microM) is sufficient to stimulate PLCdelta1, but not PLCgamma1 and PLCbeta1, to hydrolyse cellular inositol lipids present in permeabilized cells. The activity of PLCdelta1 is further enhanced in the presence of phosphatidylinositol transfer protein (PI-TP). Both full activation by Ca2+ ions and stimulation in the presence of PI-TP require an intact PH domain involved in the membrane attachment of PLCdelta1. The physiological implication of this study is that PLCdelta1 could correspond to a previously uncharacterized PLC responsible for Ca2+ ion-stimulated inositol lipid hydrolysis observed in many cellular systems.

Animals↗
PubMed1997

Measurement of angioplasty lumen volume and wall compliance: a laboratory study.

We have developed a device and technique for measuring the pressure-volume characteristics of an arterial segment from a standard angioplasty balloon, enabling us in this laboratory study to investigate the ability of the technique to measure changes in lumen volume and wall compliance. Percutaneous transluminal coronary angioplasty (PTCA) is a valuable clinical procedure for alleviating arterial stenosis, but the long-term success of the technique is less than ideal. The ability to monitor the effect of the balloon on the artery during inflations could lead to improvements in the technique. In this study, 10 different lumen diameters (1.6 to 3.4 mm) were produced in a Perspex block, and six different wall compliances produced with silicone tubes of constant inner diameter (2.0 mm) but different outer diameters (3.5 to 6.0 mm) A standard PTCA balloon was inflated under automatic control at a constant rate of 0.05 ml s-1 to 800 kPa (8 bar). Pressure and volume data were recorded to computer continuously throughout the procedure. Differences in lumen volume of +/-0.01 ml and wall compliance of +/-5% could be differentiated with greater than 95% confidence. We conclude that our technique allows small changes in lumen volume and wall compliance to be differentiated using a standard angioplasty catheter and balloon.

Angioplasty, Balloon, Coronary↗
PubMed1995

Angioplasty pressure-volume measurement.

An instrument has been developed for controlled inflation and deflation of an angioplasty balloon. The mean difference between inflation to 800 kPa in air and a simulated coronary artery is 71.2 kPa. The repeatability of four inflation/deflation cycles is 6.4 kPa with the balloon in air and 9.0 kPa in the simulated artery.

Angioplasty, Balloon, Coronary↗
PubMed1994

A 12-year experience with enterovesical fistulas.

OBJECTIVES: To review our experience with enterovesical fistulas in order to determine the most accurate diagnostic studies and most effective method of treatment. METHODS: A retrospective record review of 76 patients who were diagnosed and treated for enterovesical fistulas over a 12-year period was performed. Data collection focused on presenting symptoms, urinary disease process, diagnostic studies, and methods of management. RESULTS: Diverticular disease was the primary etiologic factor in the majority of patients (59%), with colonic malignancy, granulomatous bowel disease, and radiation therapy accounting for the majority of the remainder. Cystoscopy (60%) and cystography (44%) were the most sensitive diagnostic studies. There was no statistical difference in the complication rate between groups treated with single or multistage repair. CONCLUSIONS: One-stage repair of enterovesical fistulas can be safely performed when the cause is diverticular or granulomatous bowel disease. Staged repairs may be more judicious in patients with large intervening pelvic abscesses or those in whom advanced malignancy or radiation changes are present.

Adult↗
PubMed1994

Simplified body-surface electrocardiographic maps with depolarization magnitude and direction.

A new technique is presented for extracting the magnitude and direction of ventricular depolarization at the body surface from surface electrocardiographic (ECG) map data. Bipolar electrocardiograms were obtained from 36 sites on the chest surface in five normal subjects. The direction and magnitude of depolarization as seen from the chest surface were calculated for 18 body-surface areas centred between electrode positions V1 and V6. Each area was bounded by three electrodes with an electrode spacing of 5 cm. A major depolarization component could be calculated for all triangular areas, with 48% of areas having a smaller second component. The area with the greatest magnitude in each subject had a depolarization vector pointing downwards and to the left, with an average angle to the horizontal of 55 degrees. This was consistent with an average angle of 51 degrees obtained from the subjects' 12-lead electrocardiograms. There was more variability in vector angle between adjacent areas on the right-hand side. At the V5/V6 areas, close to the cardiac apex, the vector component had an upwards orientation in all subjects, opposing the overall downward component of ventricular depolarization. The technique was able to determine local depolarization directions which were in agreement with the normal cardiac vector derived from standard electrocardiography. Reversal of the vector direction close to the cardiac apex and the collision of depolarization components from different directions could be detected. This simple form of body-surface mapping can reduce the essential features of depolarization to a single map, and provide information not directly available from a 12-lead electrocardiogram.

Adult↗
PubMed1994

Immunohistological expression of p53 in primary pT1 transitional cell bladder cancer in relation to tumour progression.

OBJECTIVE: To determine whether p53 expression is a marker of tumour progression in superficially invasive (pT1) transitional cell carcinoma of the bladder. PATIENTS AND METHODS: The immunohistochemical status of the p53 protein in 28 pT1 primary bladder cancers was determined on frozen tissue and archival paraffin block sections using three primary antibodies (CM-1, PAB1801 and D07). The findings were compared with the patients' progress. All the patients, except for those who died during the course of the study, were followed up by check cystoscopy for a minimum of 2 years. RESULTS: Immediately adjacent frozen sections stained identically in 10 of 16 cases for CM-1 and PAB1801. For paraffin sections, identical staining patterns were seen for PAB1801 and D07 in 21 of 26 sections. However, inter- and intra-tumour staining for p53 was very variable, even with the same antibody. The heterogeneity of p53 positive cell distribution in the tumours indicates potential for significant sampling errors if random sections are chosen as representative of p53 status. CONCLUSION: The p53 status of the primary tumours did not relate to patient outcome. The results obtained do not support the use of immunohistological p53 expression as a discriminating prognostic indicator in pT1 transitional cell carcinoma of the bladder.

Biomarkers, Tumor↗
PubMed1993

Repeatability of subject/bed interface pressure measurements.

The repeatability of a technique for measuring interface pressures has been assessed. Pressure was measured using a Talley SA500 Pressure Evaluator under six anatomical sites (occiput, scapula, elbow, sacrum, buttock, and heel) of six healthy subjects lying supine on a Clinifloat mattress (trademark, SSI). For each site of each subject, four repeat readings were taken per day on four separate days. Mean pressures varied significantly between subjects (p < 0.02), though differences in mean pressures between sites were greater. Pressure was not significantly related to subject mass. The overall repeatability of the technique was +/- 0.77 kPa (+/- 5.8 mm Hg) which was much smaller than the range of pressures found under different sites (2.72 kPa or 20.4 mm Hg at the sacrum to 9.00 kPa or 67.5 mm Hg at the heel). Repeatability varied from site to site, from +/- 0.47 kPa (+/- 3.5 mm Hg) at the buttocks to +/- 1.20 kPa (+/- 9.0 mm Hg) at the heel. Measurements were found to vary significantly more between days than between repeats on the same day (p < 0.02).

Adult↗
PubMed1993

Accuracy of interface pressure measurement systems.

Interface pressure measurement is needed to assess beds designed to prevent pressure sores, so it is therefore important to establish the accuracy of interface pressure measuring systems. In this study, the Talley SA500 pressure evaluator (with 28 mm and 100 mm sensor pads), the DIPE (with 100 mm sensor pad), and a water-filled bladder system (with 0.1 ml and 0.3 ml water) were assessed. Measurement errors were evaluated using a loading system with pressures up to 7.4 kPa (55 mm Hg) in steps of 0.9 kPa (6.9 mm Hg). All systems tested over-measured interface pressure, the error being approximately linearly proportional to the loading pressure. The repeatability for a given system was approximately constant. The mean error (+/- SD) (%) and repeatability (kPa) for the systems were: 28 mm Talley 12 +/- 1%, +/- 0.07 kPa; 100 mm Talley 15 +/- 1%, +/- 0.07 kPa; DIPE 27 +/- 3%, +/- 0.12 kPa; 0.1 ml water bladder 17 +/- 1%, +/- 0.13 kPa; 0.3 ml water bladder 26 +/- 3%, +/- 0.07 kPa. Different interfaces affected accuracy markedly, and repeatability was affected when an inhomogeneous interface was used. The study shows that the errors associated with interface pressure measurement systems can be substantial, and can vary from one system to another.

Beds↗
PubMed1993

Air-fluidized beds and their ability to distribute interface pressures generated between the subject and the bed surface.

Pressures were measured under five anatomical sites prone to pressures sores for ten subjects, supine and sitting on two different air-fluidized beds. The beds were the Clinitron (trademark, SSI) and the Fluidair Plus (trademark, KCI Mediscus). Mean supine pressures were less than 4 kPa under four sites. The average supine buttock pressure was 2.65 kPa. This increased to 3.71 kPa upon sitting up, though pressures did not rise above the accepted capillary closing pressure, on either bed. Low interface pressures at these sites were due to good moulding between subject and bed. Heel pressures averaging 7.08 kPa, were a factor of 2.67 times greater than buttock pressure, and were higher than expected considering the depth the heels sunk to in both beds. This exceeded the accepted capillary closing pressure and was attributed to covering sheets preventing true floatation at the heels.

Adult↗
PubMed1992

The influence of temperature and organic matter on the bactericidal activity of short-chain organic acids on salmonellas.

Acetic and lactic acids and BioAdd, a commercial preparation of formic and propionic acid, were tested at a concentration of 0.1% (w/w) at 20, 30, 40 and 50 degrees C and in the presence of organic material for bactericidal activity against Salmonella serotype Kedougou. BioAdd was the most active of the solutions at all temperatures, followed by lactic acid and acetic acid. The presence of horse blood at all four temperatures, and milk and serum at 50 degrees C, did not greatly affect the antibacterial activity of the acids although yeast extract (50 degrees C) provided some protection for the salmonella. Acid activity was related to low pH values although the bactericidal activity of acetic acid with blood and milk was greater than the unadulterated acid even though the pH was 0.4 units higher.

Acetates↗