Effect of limb and cuff positioning on measurement of arterial blood pressure with an oscillometry device (PetMAP) in anaesthetized cats.

Arterial blood pressure (ABP) is often measured with oscillometry during anaesthesia. Changing the height of the measuring cuff with respect to the level of the heart is known to affect oscillometry accuracy in some species; however, this effect has not been investigated in cats. The objective of this study was to determine the effects of raising and lowering the measuring cuff from standard position (level of the heart) on ABP, measured with PetMAP, in anaesthetised cats. ABP readings were obtained from 29 cats with the cuff at standard position (baseline), and 5 cm above and below the heart. The end-tidal isoflurane concentrations were maintained constant during data acquisition. There were no differences between baseline values and those measured below the heart, while ABP measured above the heart was consistently lower than baseline for both the thoracic and pelvic limbs (P < 0.001), with absolute differences of 8.2 (2.5 - 14) mmHg and 6.5 (3.0 - 15.0) mmHg, respectively. Systolic ABP readings at the pelvic limb were consistently higher than those at the thoracic limb at standard position (112 ± 26 versus 103 ± 21 mmHg, p = 0.010), above (106 ± 22 versus 95 ± 20 mmHg, p = 0.003), and below the heart (116 ± 26 versus 107 ± 22 mmHg, p = 0.011). This study shows that raising the cuff by 5 cm above the heart, which may become necessary during procedural positioning, results in clinically significant underestimation of ABP measured with PetMAP.

High throughput and quantitative enzymology in the genomic era.

Accurate predictions from models based on physical principles are the ultimate metric of our biophysical understanding. Although there has been stunning progress toward structure prediction, quantitative prediction of enzyme function has remained challenging. Realizing this goal will require large numbers of quantitative measurements of rate and binding constants and the use of these ground-truth data sets to guide the development and testing of these quantitative models. Ground truth data more closely linked to the underlying physical forces are also desired. Here, we describe technological advances that enable both types of ground truth measurements. These advances allow classic models to be tested, provide novel mechanistic insights, and place us on the path toward a predictive understanding of enzyme structure and function.

Revealing enzyme functional architecture via high-throughput microfluidic enzyme kinetics.

Systematic and extensive investigation of enzymes is needed to understand their extraordinary efficiency and meet current challenges in medicine and engineering. We present HT-MEK (High-Throughput Microfluidic Enzyme Kinetics), a microfluidic platform for high-throughput expression, purification, and characterization of more than 1500 enzyme variants per experiment. For 1036 mutants of the alkaline phosphatase PafA (phosphate-irrepressible alkaline phosphatase of Flavobacterium), we performed more than 670,000 reactions and determined more than 5000 kinetic and physical constants for multiple substrates and inhibitors. We uncovered extensive kinetic partitioning to a misfolded state and isolated catalytic effects, revealing spatially contiguous regions of residues linked to particular aspects of function. Regions included active-site proximal residues but extended to the enzyme surface, providing a map of underlying architecture not possible to derive from existing approaches. HT-MEK has applications that range from understanding molecular mechanisms to medicine, engineering, and design.

Programmable Microfluidic Synthesis of Over One Thousand Uniquely Identifiable Spectral Codes.

Encoded microparticles have become a powerful tool for a wide array of applications, including high-throughput sample tracking and massively parallel biological multiplexing. Spectral encoding, where particles are encoded with distinct luminescence spectra, provides a particularly appealing encoding strategy because of the ease of reading codes and assay flexibility. To date, spectral encoding has been limited in the number of codes that can be accurately resolved. Here, we demonstrate an automated 5-dimensional spectral encoding scheme using lanthanide nanophosphors that is capable of producing isotropic spherical microparticles with up to 1,100 unique codes, which we term MRBLEs (Microspheres with Ratiometric Barcode Lanthanide Encoding). We further develop a quantitative framework for evaluating global ability to distinguish codes and demonstrate that for six different sets of MRBLEs ranging from 106 to 1,101 codes in size, > 98% of MRBLEs can be assigned to a code with 99.99% confidence. These > 1,000 code sets represent the largest spectral code libraries built to date. We expect that these MRBLEs will enable a wide variety of novel multiplexed assays.

Nature and governance of veterinary clinical research conducted in the UK.

In order to quantify the amount of clinical research conducted on client-owned animals under the Veterinary Surgeons Act 1966, and the nature and extent of any ethical review of that research, a questionnaire was sent to 6 UK veterinary schools, 1 charity veterinary clinic and 12 private referral clinics. The questionnaire examined whether and how much clinical research respondents undertook, and the composition of any ethical review panels examining research proposals. The questionnaire revealed a substantial amount of clinical research was conducted in the UK, with over 200 veterinary surgeons involved in the year of the survey, with at least 170 academic papers involving clinical research published by respondents in the same year. However, it proved impossible to quantify the full extent of clinical research in the UK. All UK veterinary schools required ethical review of clinical research. The composition and working practices of their ethical review panels generally reflected skill sets in ethical review panels set-up under statute to consider the ethics of non-clinical biomedical research on animals and clinical research conducted on human patients. The process for review of clinical research in the private sector was less clear.

Correction: Programmable microfluidic synthesis of spectrally encoded microspheres.

Correction for 'Programmable microfluidic synthesis of spectrally encoded microspheres' by R. E. Gerver et al., Lab Chip, 2012, 12, 4716-4723.

Comparison of surgical duration of canine ovariectomy and ovariohysterectomy in a veterinary teaching hospital.

OBJECTIVE: To prospectively evaluate ovariectomy and ovariohysterectomy via midline coeliotomy when being employed by supervised final year veterinary students for the purpose of routine canine neutering. METHODS: One hundred and eight female dogs of various breeds, presented to a veterinary teaching hospital for neutering, were randomly allocated to one of two surgery groups, ovariectomy or ovariohysterectomy. The specified procedure was performed by a supervised final year veterinary student. If the duration of surgery exceeded 2 hours or if major surgical or anaesthetic complications occurred, the supervising surgeon intervened to complete the procedure. RESULTS: Data analysed included age, weight, time from first incision to start of closure, duration of closure, total surgical time and length of incision. Fifty-four dogs underwent each procedure. There was no significant difference between the two surgery groups for any of the measured variables. CLINICAL SIGNIFICANCE: Ovariectomy is not associated with shorter surgical times or smaller abdominal incisions than ovariohysterectomy when employed by inexperienced surgeons. As no major complications novel to ovariectomy occurred in this cohort of dogs, this study adds support to the existing literature indicating that ovariectomy is an acceptable alternative to ovariohysterectomy for canine neutering.

Programmable microfluidic synthesis of spectrally encoded microspheres.

Spectrally encoded fluorescent beads are an attractive platform for assay miniaturization and multiplexing in the biological sciences. Here, we synthesize hydrophilic PEG-acrylate polymer beads encoded with lanthanide nanophosphors using a fully automated microfluidic synthesis device. These beads are encoded by including varying amounts of two lanthanide nanophosphors relative to a third reference nanophosphor to generate 24 distinct ratios. These codes differ by less than 3% from their target values and can be distinguished from each other with an error rate of <0.1%. The encoded bead synthesis strategy we have used is readily extensible to larger numbers of codes, potentially up to millions, providing a new platform technology for assay multiplexing.

Systematic characterization of feature dimensions and closing pressures for microfluidic valves produced via photoresist reflow.

Multilayer soft lithography (MSL) provides a convenient and low-cost method for fabricating poly(dimethyl siloxane) (PDMS) microfluidic devices with on-chip valves for automated and precise control of fluid flow. MSL casting molds for flow channels typically incorporate small patches of rounded positive photoresist at valve locations to achieve the rounded cross-sectional profile required for these valves to function properly. Despite the importance of these rounded features for device performance, a comprehensive characterization of how the rounding process affects feature dimensions and closing pressures has been lacking. Here, we measure valve dimensions both before and after rounding and closing pressures for 120 different valve widths and lengths at post-rounding heights between 15 and 84 µm, for a total of 1200 different geometries spanning a wide range of useful sizes. We find that valve height and width after rounding depend strongly on valve aspect ratios, with these effects becoming more pronounced for taller and narrower features. Based on the measured data, we provide a simple fitted model and an online tool for estimating the pre-rounding dimensions needed to achieve desired post-rounding dimensions. We also find that valve closing pressures are well explained by modelling valve membranes in a manner analogous to a suspension bridge, shedding new light on device physics and providing a practical model for estimating closing pressures during device design.

Search for the decay KL --> pi(0)e+e-.

We report on a search for the decay KL-->pi(0)e+e- carried out by the KTeV/E799 experiment at Fermilab. This decay is expected to have a significant CP violating contribution and the measurement of its branching ratio could support the Cabibbo-Kobayashi-Maskawa mechanism for CP violation or could point to new physics. Two events were observed in the 1997 data with an expected background of 1.06+/-0.41 events, and we set an upper limit B(KL-->pi(0)e+e-)<5.1 x 10(-10) at the 90% confidence level.

Neuritis of the cauda equina in the horse.

Ultrastructural lesions of the cranial nerves and their ganglia and the autonomic nervous system from 5 cases of neuritis of the cauda equina in the horse are described. They include lysosomal inclusions within the semilunar, geniculate and sympathetic chain ganglia, granulomatous involvement of the coeliaco-mesenteric ganglion and accumulation of axonal organelles in unmyelinated fibres of the great splanchnic nerve, sympathetic chain and oesophageal vagus.

Use of an ELISA in the differential diagnosis of cauda equina neuritis and other equine neuropathies.

In 27 potential neuropathies an enzyme-linked immunosorbent assay, using P2 preparations from either bovine or equine myelin, detected all cases of cauda equina neuritis in which there was caudal involvement. The test was of limited value in differentiating neuropathies involving only cranial or other peripheral nerves.

Rhinovirus plaque formation in WI-38 cells with methylcellulose overlay.

A sensitive, reliable plaque assay system is described for five rhinoviruses using freshly prepared methylcellulose overlay and human embryonic diploid cells. Circular plaques with irregular edges, 2 mm in size, were formed by rhinoviruses 1A, 2, 6, and 13 after 6 or 7 days of incubation. A fifth rhinovirus, 17, formed a 1- to 2-mm feather plaque after 14 days of incubation. Plaque counts of rhinoviruses 1A and 13 were not affected by varying the pH of the overlay from 6.9 to 7.5. Plaque sizes and plaque-forming unit values of high passage rhinoviruses 1A and 13 were equivalent when tested at 26, 31, or 36 C. The rhinoviruses tested were sensitive to incubation at 40 C or heating at 50 C. Enhancement of plaques was observed when Mg(++) was incorporated into agar overlays, but enhancement did not occur when Mg(++) was added to methylcellulose overlays.

Propagation of Rhinovirus on WI-38 Cell Monolayers in Rolling Bottles.

Techniques are described for the propagation of rhinoviruses on WI-38 monolayers in rolling bottles. High yields of viruses were obtained, as indicated by infectivity titers and electron microscopy. When crude harvests were subjected to low-speed centrifugation and then filtered through a 0.45-mu membrane filter, little or no loss in infectivity titer was observed. However, electron microscopic examination indicated that the concentration of viral physical particles was reduced below detectable levels after filtration. The guinea pig potency test on the lot of unfiltered rhinovirus 14 vaccine prepared in rolling bottles indicated that this vaccine stimulated higher reciprocal serum-neutralizing titers than a rhinovirus 14 vaccine prepared in stationary monolayers.