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Biomedical subjects

M Kirkland

Publications and source records attributed to M Kirkland.

8 recordsLinked to original sources

Clinical agreement between automated and calculated ultrasound measurements of bladder volume.

Non-invasive urine volume measurement is an important tool in the management of dysfunctional and neuropathic bladders in children. Ultrasound imaging devices have been used for many years for this purpose. An automated scanner (Bladderscan) is now available and has been recommended by a number of authors, but there is conflicting evidence in the literature regarding the accuracy and appropriate clinical application of the device. We aimed to assess the level of clinical agreement between the two methods. 36 urine volume measurements were made on 11 children using both instruments. Although there was a good correlation between the methods (r=0.97), the clinical agreement was poor (limits of agreement +/-77 ml). 13 voided volumes were directly measured and compared with the difference between pre- and post-void ultrasound measurements. The systematic errors were small but the mean absolute errors were 54 ml and 23 ml, respectively, for the automated and ultrasound imaging methods. If used correctly, ultrasound imaging provides more accurate results and can compete with the cost, convenience and ease of use of the automated method. Low cost, highly portable ultrasound imaging devices are now available and should be used in preference to the Bladderscan.

Automation↗

High-density immobilization of an antibody fragment to a carboxymethylated dextran-linked biosensor surface.

There are numerous chemical methods published that enable protein coupling to carboxymethyl (CM) dextran. Here we have taken traditional amine coupling using N-hydroxysuccinimide (NHS) and N'-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and coupled an antibody fragment (scFv) to CM dextran at a very high density. Using an upgraded BIAlite from Biacore AB, more than 7000 RU of scFv was coupled to a CM dextran biosensor chip. In addition, scanning electron microscopy was performed on CM dextran biosensor chips following amine coupling of 30 nm gold anti-IgG particles. This showed that amine coupling was uniform across the biosensor chip surface. Calculations show that 7620 RU of an scFv coupled to such a surface results in a mean distance between binding sites of 8.8 nm. This equates to a packing volume of approximately 20% of the available space occupied by the antibody fragment. Comparisons made with densities of covalently coupled IgG show that a greater number of antibody fragment molecules can be coupled per unit area. This is most likely due to the smaller size of an antibody fragment (scFv), which has a volume of less than 20% of an IgG molecule. The significance of these findings is discussed.

Animals↗

Protein interactions with DNA elements in variant equine infectious anemia virus enhancers and their impact on transcriptional activity.

The long terminal repeats (LTRs) from various cloned equine infectious anemia virus (EIAV) proviruses differ significantly, but all contain cis-acting DNA elements identical to MDBP-, PEA2-, AP-1-, and PU.1 (ets)-binding sites. A prototype EIAV LTR would contain one of each of these conserved elements. The LTR variations originate from the insertion of novel sequences between the PEA2 and AP-1 elements in the transcriptional enhancer unit. Viewed in this way, the LTR from provirus clone lambda 12 has an 11-bp insertion containing a PEA2 site and the LTR of the lambda 6 provirus has a 31-bp insertion/duplication containing PEA2, AP-1, and PU.1 sites. Two other LTRs were cloned by amplification of cDNAs from the persistently infected cell line, EIAV-FEA. A third LTR was generated by site-directed mutagenesis of one of the LTRs from EIAV-FEA cells. The latter three had a single base change in the element next to the TATA box that abolished PU.1 binding; however, the variable regions of these LTRs were shown by gel mobility shift assays to contain one or two PU.1 sites. One variable region was shown to have an octamer site overlapping its tandem PU.1 elements. Basal, PMA-activated, and Tat trans-activated transcriptional activities of the LTRs were compared in several different cell lines by transient transfection. The various promoters displayed different relative levels of activity depending on the cell line used and the condition of activation. This natural set of variant promoters may help define how changes in the components of the transcription complex influence transactivation by Tat. The diverse LTRs could endow their respective proviruses with a unique pattern of expression and activation in vivo.

Base Sequence↗

An in vitro technique to assess oxygenator potential for respiratory failure therapies.

To assess microporous membrane oxygenators (MMO) for long-term respiratory support, an in vitro test of 4 days duration has been developed. The objective is to screen oxygenators before they undergo expensive animal trials. As a sensitive indicator of performance change, CO2 transfer rates (VCO2) are determined with fresh bovine blood at the start, after 48 hours and after 96 hours of perfusion. Blood is used only for gas exchange evaluation and is replaced with plasma in the intervening periods. Data are obtained at blood flow rates (QB) of 1.0, 1.5 and 2.0 l/min for gas: blood flow ratios (QG:QB) of 2:1 and 5:1. The test has been conducted on Bentley BOS-CM50 oxygenators (n = 10). With heated dry air ventilation there was no significant change in VCO2 with time for all QB and QG:QB combinations. Plasma leakage did not occur into the gas compartment of the MMO. Similar results were obtained with heated humidified air ventilation. The findings are in agreement with existing animal studies of 4-7 days duration.

Animals↗

The effects of submaximal endurance exercise upon LH pulsatility.

The acute effects of submaximal endurance exercise (three consecutive 20-min runs on a treadmill at 50, 60 and 70% of the subjects' maximum oxygen uptake) upon the pulsatile release of LH were compared with those accompanying leisurely strolling for a similar period in seven normally menstruating young women. All trials were conducted during the early to mid-luteal phase, as determined by body temperature patterns, ultrasonic scans of the ovaries, detection of the LH surge in first morning urine specimens, and serial measurements of plasma progesterone. Blood was sampled every 10 min via an indwelling cannula for 8 h before and 12 h after exercise and serum LH measured by radioimmunoassay. LH pulsations were analysed by a time series method. Following cannulation, mean LH levels declined but then rose to reach a maximum 2 h before the beginning of the exercise bout. LH concentrations remained virtually unchanged during exercise itself, and exhibited a declining trend throughout the post-exercise period. The findings in the two groups were similar in all respects, except that in the control study the rate of LH pulsatility was significantly diminished (P less than 0.05) during the first 2 h of sampling as compared with the subsequent 2-h period. The approximate half-life of LH varied from 27 to 57 min, with a mean of 41 min.

Adult↗

The detection of peaks in luteinizing hormone secretion.

Fluctations in luteinizing hormone are believed to consist of irregularly spaced sharp increases separated by periods of exponential decay. A simple method is presented for analysing such fluctuations when the data consist of uniformly sampled observations of hormone. Specific allowance for the exponential decay in the absence of pulses is made via a time series model before assessing the number and extent of pulses. All calculations are done using MINITAB regression programs. The results have been compared with those obtained by three established models and are in general agreement.

Activity Cycles↗