Effect of prepartum exercise on lying behavior, labor length, and cortisol concentrations.

The objective of this study was to assess the effect of exercise and pasture turnout on lying behavior, labor length, and cortisol concentrations around the time of parturition in dairy cows. Twenty-nine primiparous and 31 multiparous, pregnant, nonlactating Holstein (n = 58) and Jersey × Holstein cross (n = 2) dairy cows were assigned to control (n = 20), exercise (n = 20), or pasture (n = 20) treatments at dry-off using rolling enrollment. Control cows remained in the dry cow group pen. Exercise cows were removed from the dry cow group pen 5 times per week and walked for 1.4 ± 0.1 h at 1.88 ± 0.58 km/h. Pasture cows were moved to an outdoor paddock 5 times per week for 1.8 ± 0.3 h/d. Cows were housed in deep-bedded sand freestalls in a naturally ventilated, 4-row freestall barn. Cows were moved into maternity pens on the day of projected calving or when cows displayed signs that calving was imminent (restlessness, raised or lifted tail, ruptured amniotic sac, or swollen vulva), and treatments were discontinued. Cameras continuously recorded cows from entry into the pen until farm staff noted a calf, and one observer continuously watched video for two visually observable periods throughout the calving process: time from initial observation of amniotic sac to initial observation of calf's feet, and time from initial observation of calf's feet to full expulsion of calf. Assisted calvings were excluded. Accelerometers were attached to the rear fetlocks of cows 3 d before dry-off and removed 14 d postpartum. Activity was summarized by day for the 7 d before and after delivery time recorded from video observation into lying time (hours per day), lying bout frequency (bouts per day), lying bout duration (minutes per bout), and steps (number per day). Plasma total cortisol concentration was measured on d 0 and 3 postpartum and determined by a radioimmunoassay procedure using a commercially available kit. Data were analyzed using mixed linear model. During calving, time from appearance of the amniotic sac to appearance of the calf's feet was longer for pasture cows compared with control. Control cows engaged in fewer lying bouts and less overall lying time compared with pasture and exercise cows. Cortisol concentrations were higher on the day of calving compared with 3 d later, regardless of treatment. Understanding the effects of lying alterations around calving and increases in labor period length may allow for physical activity recommendations for late-gestation dairy cows.

Effect of prepartum exercise, pasture turnout, or total confinement on hoof health.

Lameness is a major welfare concern in the dairy industry, and access to physical activity during the dry period may improve hoof health. The objective of this study was to determine the effects of forced exercise, pasture turnout, or total confinement of dry cows on horn growth and wear and sole thickness. Twenty-nine primiparous and 31 multiparous, pregnant, nonlactating Holstein (n = 58) and Jersey-Holstein crossbred (n = 2) dairy cows were assigned to either total confinement (n = 20), exercise (n = 20), or pasture (n = 20) treatments at dry-off using rolling enrollment from January to November 2015. Cows were managed with a 60-d dry period (58.5 ± 5.4 d) divided into far-off (dry-off to 2 wk before parturition) and close-up periods (2 wk before projected parturition). Cows were housed in a naturally ventilated, 4-row freestall barn at the University of Tennessee's Little River Animal and Environmental Unit (Walland, TN) with concrete flooring and deep-bedded sand freestalls. Cows assigned to confinement remained in the housing pen. Exercise cows were walked for a targeted 1.5 h at 3.25 km/h, 5 times/wk until calving. Pasture cows were turned out for a targeted 1.5 h, 5 times/wk until calving. Hoof growth and wear and sole thickness of the rear hooves were measured on d 2 and 44, relative to dry-off. Data were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Cranial and caudal horn wear was greater for exercise cows than confinement and pasture cows. Exercise cows experienced more equal rates of horn growth and wear cranially. Confined cows tended to increase sole thickness from d 2 to 44, relative to dry-off. Frequent, short duration exercise on concrete did not impair the hoof health of late-gestation dry cows. Further, exercise may improve overall hoof health, potentially improving cow welfare.

Effect of maternal exercise on calf dry matter intake, weight gain, behavior, and cortisol concentrations at disbudding and weaning.

The objective of this study was to determine the effect of maternal physical activity during late gestation on calf dry matter intake, weight gain, behavior, and cortisol concentration during disbudding and weaning. Fifty-five Holstein and 5 Jersey × Holstein crossbred calves were enrolled into the study during gestation. Calves were born from pregnant, nonlactating Holstein (n = 58) and Jersey × Holstein crossbred (n = 2) dairy cows. Cows were assigned to either confinement (n = 20 cows; 13 female calves, 7 male calves), exercise (n = 20 cows; 8 female calves, 12 male calves), or pasture (n = 20 cows; 11 female calves, 9 male calves) treatments at dry-off from January to November 2015. Enrollment in treatment was balanced by parity (1.8 ± 0.9), projected mature-equivalent fat-corrected milk (13,831 ± 2,028 kg/lactation), dam breed, and projected calving date. Cows assigned to confinement remained in the pen throughout the dry period. Cows assigned to exercise were walked 5 times/wk at a targeted 1.5 h at 3.25 km/h. Cows assigned to pasture were turned out 5 times/wk for a targeted 1.5 h/d. Treatments were terminated on the expected due date or at signs of calving. Calves were removed from cows immediately once observed by farm staff and subsequently weighed and moved into a straw deep-bedded hutch. Data loggers were attached to the rear fetlock of each calf -3 d to +6 d relative to disbudding and weaning to monitor changes in lying behavior. Calves were weighed on d -7, -5, -3, -1, 0, 1, 3, 5, and 7, and grain was weighed the 7 d preceding and following disbudding and weaning. Blood was collected 24 h before and 0, 1, and 4 h after disbudding and d -1, 0, 1, and 2 relative to weaning to determine cortisol concentrations. Data were analyzed using mixed linear model in SAS (SAS Institute Inc., Cary, NC). Calf weight gain decreased the day after disbudding and calves tended to have elevated cortisol concentrations 1 h after disbudding, regardless of maternal treatment. Calf weight gain decreased the day of and after weaning; calves had elevated cortisol concentrations the day after weaning, regardless of treatment. Behavior did not differ by treatment at disbudding, but calves from pasture cows lay down for less time compared with confinement and exercise maternal treatments and less frequently than exercise maternal treatments at weaning. More research investigating the significance of lying time and restlessness around stressful events is needed to further understand the implications of such behavioral responses.

Effects of bedding with recycled sand on lying behaviors, udder hygiene, and preference of lactating Holstein dairy cows.

Effects of bedding with recycled sand and season on lying behaviors, hygiene, and preferences of late-lactation Holstein cows were studied. It was hypothesized that recycled sand will decrease lying time and increase hygiene scores due to increased moisture content and organic matter, and thus a preference for the control sand will be evident. Cows (n = 64) were divided into 4 groups (n = 8 per group) per season. In summer (August to September), cows were balanced by days in milk (268.1 ± 11.9 d) and parity (2.0 ± 0.2). In winter (January to February), mean DIM was 265.5 ± 34.1 d. Cows were assigned to 1 of 2 treatments using a crossover design with each treatment lasting 7 d (no-choice phase): bedding with recycled sand (RS; n = 32) or control (CO; clean sand; n = 32). Stocking density was maintained at 100%. The choice phase allowed cows to have access to either treatment with stocking density at 50%. Accelerometers recorded daily lying time, number of lying bouts per day, lying bout duration (min/bout), and total steps per day. Teat swabs, milk, sand samples, and udder hygiene scores were collected on d 0, 3, and 7 of each experimental week. Samples were cultured for streptococci, staphylococci, and gram-negative bacteria. Video data were used to assess bedding preferences. All data were analyzed using the MIXED and GLIMMIX procedures of SAS 9.4 (SAS Institute Inc., Cary, NC). Lying time was not affected by treatment, but cows did take more steps during winter. Bacterial counts were elevated for cows on recycled sand. A preference was observed for clean sand during the summer, but no preference was observed for sand during the winter. Regardless of bedding, the most commonly observed behavior was lying in the stalls, which suggested either bedding might be suitable. Caution should be used with this interpretation of preference, as sand was recycled only once. This limited reclamation was still sufficient to potentially alter the composition of sand, driving the observed preference. If these changes in composition continue, then the strength of the preference may also change. However, considering all variables within the current study, recycled sand is a viable bedding source to use for dairy cows.

Short communication: Short-term changes in stocking density did not alter meal characteristics of lactating Holstein dairy cattle.

The study objectives were to determine the effect of short-term increases in stocking density and milking on meal duration, meal frequency, and time between meals and to determine the bioequivalence of different meal criterions in a competitive environment. Forty-eight Holstein dairy cows were allotted to 1 of 4 groups (n=12 per group). Stocking density treatments of 100 (one cow per freestall and headlock), 113, 131, and 142% were assigned to groups using a 4×4 Latin square with treatments imposed for 14-d periods. On d 11 of each period, feeding time was recorded for 24h using 10-min scan samples from direct observation. Meals were defined as repeated observations of eating with a maximum of 20, 30, or 40min of not eating between observations constituting the same meal. A new meal was established when a cow was observed feeding and then not feeding for greater than 2 (20min), 3 (30min), or 4 (40min) observations. To evaluate diurnal effects, the 24-h period of data was divided into 8-h intervals (based on milking time); morning (0400-1200h), afternoon (1200-2000h), and night (2000-0400h). Feed delivery occurred daily at 0430h, with feed pushed up throughout the day. A mixed linear model was used to determine the effect of stocking density and time of day on meals per day, meals per hour, meal duration, time between meals, and meal duration 2h before and after milking. Regardless of stocking density, meal duration, meal frequency, meals per hour, and time between meals did not differ. Regardless of stocking density, mean meal duration was longer during the morning and afternoon compared with night. Meal duration was also greater after milking compared with before milking, regardless of stocking density. These results suggest meal length decreased throughout the day, relative to feed delivery, with periodic increases in length due to return from milking. Meals per hour, meal duration before and after milking, and meal frequency established bioequivalence for the 20-, 30-, and 40-min meal criteria. Bioequivalence was not met for meal duration when the meal criterion was increased from 20 to 40min. Short-term increases in stocking density of 14-d duration did not affect the feeding pattern of lactating dairy cows, indicating that mid-lactation dairy cows can compensate for reduced feed bunk access during short-term overstocking. When calculating feeding behaviors, including meal frequency and time between meals, using a meal criterion of 20, 30, or 40min resulted in similar outcomes when using 10-min scan samples. Future studies should investigate changes in other behaviors, such as resting, which may be altered to compensate for reduced access to the feed bunk.

The relationship between compost bedded pack performance, management, and bacterial counts.

The objective of this study was to assess the relationships among temperature, moisture, carbon-to-nitrogen (C:N) ratio, space per cow, and bacterial counts from bedding material collected from compost bedded pack (CBP) barns. A field survey of 42 routinely aerated CBP barns was conducted in Kentucky between October 2010 and March 2011. Two bedding material samples of 1,064.7 cm(3) each were collected during a single site visit from 9 evenly distributed locations throughout each barn and thoroughly mixed to create a composite sample representative of the entire CBP. Bacterial counts were determined for coliforms, Escherichia coli, streptococci, staphylococci, and Bacillus spp. University of Kentucky Regulatory Services (Lexington) laboratory personnel performed nutrient analyses to determine moisture, carbon, and nitrogen contents. Surface and 10.2-cm pack depth temperatures were collected for each of the 9 evenly distributed locations and the mean calculated to produce a composite temperature. Space per cow was calculated as the total CBP area divided by number of cows housed on the CBP. The GLM procedure of SAS (SAS Institute Inc., Cary, NC) generated models to describe factors affecting bacterial counts. Bacterial counts were 6.3 ± 0.6, 6.0 ± 0.6, 7.2 ± 0.7, 7.9 ± 0.5, and 7.6 ± 0.5 log 10 cfu/g of dry matter for coliform, Escherichia coli, streptococci, staphylococci, and Bacillus spp., respectively. Composite temperature, CBP moisture, C:N ratio, and space per cow had no effect on coliform counts. Escherichia coli reached a peak concentration when the C:N ratio was between 30:1 and 35:1. Staphylococci counts increased as ambient temperature increased. Streptococci counts decreased with increased space per cow and composite temperature and increased with increasing ambient temperature and moisture. Streptococci counts peaked at a C:N ratio ranging from 16:1 to 18:1. Bacillus spp. counts were reduced with increasing moisture, C:N ratio, and ambient temperature. Mastitis-causing bacteria thrive in similar conditions to that of composting bacteria and microbes, making elimination of these at higher temperatures (55 to 65°C) difficult in an active composting environment. Producers must use recommended milking procedures and other preventative practices to maintain low somatic cell count in herds with a CBP barn.

Compost bedded pack dairy barn management, performance, and producer satisfaction.

The objective of the research was to characterize herd performance, producer satisfaction and recommendations, and management practices used by compost bedded pack (CBP) managers in Kentucky (42 farms and 47 CBP facilities). Farms were visited between October 2010 and March 2011. A random selection of cows housed solely in the CBP were scored for locomotion and hygiene. Changes in monthly Dairy Herd Improvement Association performance records, including milk production, SCC, reproductive performance, and daily bulk-tank somatic cell count after moving into the CBP were analyzed using the MIXED procedure of SAS (SAS 9.3; SAS Institute Inc., Cary, NC). The GLM procedure of SAS (SAS 9.3) was used to develop models to describe CBP moisture, CBP temperature at 20.3 cm, and mean herd hygiene. Producers provided 9.0 ± 2.2 m2 of pack space per cow (n = 44). Barns constructed with an attached feed alley cost $1,051 ± 407 per cow (n = 40). Barns constructed without an attached feed alley cost $493 ± 196 per cow (n = 13). Kiln-dried shavings required 0.05 ± 0.04 m3 of bedding per cow per day (n = 15). Green shavings required 0.07 ± 0.06 m3 of bedding per cow per day (n = 12). The most-frequently cited benefits of the CBP included cow comfort (n = 28), cow cleanliness (n = 14), and the low-maintenance nature of the system (n = 10). Increased stirring frequency, stirring depth, and ambient temperature increased pack temperature, measured at 20.3 cm below the CBP surface. Increased stirring depth, pasture-adjusted space per cow, and drying rate decreased CBP moisture. Mean herd locomotion and hygiene scores were 1.5 ± 0.3 (n = 34) and 2.2 ± 0.4 (n = 34), respectively. Increased 20.3-cm depth CBP temperature and ambient temperatures improved mean herd hygiene. Bulk-tank somatic cell count decreased from the year before to the year after moving into the CBP barn (323,692 ± 7,301 vs. 252,859 ± 7,112 cells/mL, respectively) for farms using the CBP barn as the primary housing facility (n = 9). Daily milk production, collected from monthly Dairy Herd Improvement Association tests, increased from before moving into the CBP barn to the second year after (29.3 ± 0.3 vs. 30.7 ± 0.3 kg, respectively) for farms using the CBP barn as the primary housing facility (n = 8). Calving interval decreased from the year before to the second year after (14.3 ± 0.1 vs. 13.7 ± 0.1 mo) moving into the CBP barn for farms using the CBP as primary housing (n = 8).

ADAMs: focus on the protease domain.

ADAMs are proteins containing a disintegrin and metalloproteinase domain. Several important insights were provided in the past year regarding ADAM metalloproteinases. ADAM 10 was implicated in the Notch signaling pathway. ADAM 17 was shown to be the long sought after tumor necrosis factor-alpha convertase and the crystal structure of its metalloproteinase domain was determined.

Rapid and specific conversion of precursor interleukin 1 beta (IL-1 beta) to an active IL-1 species by human mast cell chymase.

Secretory granules of human dermal mast cells contain a chymotrypsin-like serine proteinase called chymase. In this study, we demonstrate that the inactive cytokine, 31 kD interleukin 1 beta (IL-1 beta), can be converted rapidly to an 18 kD biologically active species by human mast cell chymase. The product formed is three amino acids longer at the amino terminus than the mature IL-1 beta produced by peripheral blood mononuclear cells and has comparable biological activity. Because chymase is a secretory granule constituent, it is likely to be released into the surrounding tissue when mast cells degranulate. It is also known that non-bone marrow derived cells resident in skin (keratinocytes, fibroblasts) produce but do not process 31 kD IL-1 beta. In this context, chymase may be a potent activator of locally produced 31 kD IL-1 beta. Mast cells lie in close apposition to blood vessels in dermis; therefore, chymase mediated conversion of 31 kD IL-1 beta might be expected to have a critical role in the initiation of the inflammatory response in skin.

A metalloprotease inhibitor blocks shedding of the 80-kD TNF receptor and TNF processing in T lymphocytes.

TNF is synthesized as a 26-kD membrane-anchored precursor and is proteolytically processed at the cell surface to yield the mature secreted 17-kD polypeptide. The 80-kD tumor necrosis factor (TNF) receptor (TNFR80) is also proteolytically cleaved at the cell surface (shed), releasing a soluble ligand-binding receptor fragment. Since processing of TNF and TNFR80 occurs concurrently in activated T cells, we asked whether a common protease may be involved. Here, we present evidence that a recently described inhibitor of TNF processing N-(D,L-[2-(hydroxyaminocarbonyl)methyl]-4-methylpentanoyl)L- 3-(2'naphthyl)- alanyl-L-alanine, 2-aminoethyl amide (TAPI) also blocks shedding of TNFR80, suggesting that these processes may be coordinately regulated during T cell activation. In addition, studies of murine fibroblasts transfected with human TNFR80, or a cytoplasmic deletion form of TNFR80, reveal that inhibition of TNFR80 shedding by TAPI is independent of receptor phosphorylation and does not require the receptor cytoplasmic domain.

Development of a Bioactive Polymeric Drug Eluting Coronary Stent Coating Using Electrospraying.

Drug-eluting stents are now routinely used in the treatment of acute coronary syndromes caused by coronary artery disease. Whilst the sustained release of anti-proliferative drugs from these devices has greatly reduced the need for repeat revascularisation procedures, this approach is not suitable for all patients and appears to delay regrowth of the endothelium, necessitating the use of prolonged dual anti-platelet therapy. Although the development of more advanced stent platforms and drug coatings has produced modest improvements in performance, these devices have not fully addressed the limitations experienced with their first-generation counterparts. In the present study, we developed a novel stent coating that provides controlled sirolimus release from a bioactive polymer (accelerate™ AT) that has previously been shown to support endothelial cell growth in vitro. A bespoke electrospray deposition process provided control over the coating thickness, surface roughness, drug load, and release kinetics. The resultant optimised coating combines rapid release of an anti-proliferative agent from a bioactive polymer coating that promotes re-endothelialisation, thereby offering potential protection against in-stent restenosis and thrombosis. This novel, dual-action coating therefore has significant therapeutic potential, with the enhanced control of drug load and release kinetics offered by electrospray deposition also opening up opportunities for more personalised treatment approaches. Further development and evaluation of these technologies in vitro and in vivo is therefore warranted.

Acetylcholine receptor in a C2 muscle cell variant is retained in the endoplasmic reticulum.

We have examined the properties and intracellular localization of acetylcholine receptors in the C2 muscle cell line and in a variant (T-) that accumulates AChR intracellularly. On immunoblots, the subunit structures of the AChR from wild-type and T- cells were similar except that the gamma and delta subunits of the variant AChR had altered mobilities. Digestion with endoglycosidases H and F demonstrated that this difference results from a failure of high-mannose N-linked oligosaccharides on AChR subunits to be processed to complex forms in the variant. N-linked glycosylation of other proteins in the variant was normal. When examined by immunocytochemistry, the distribution of internal AChR in wild-type cells was consistent with a location both in the endoplasmic reticulum and in the Golgi. Variant cells, however, showed no evidence of Golgi staining. Subcellular fractionation experiments also demonstrated AChR in the Golgi fractions of wild-type cells, but not in those derived from T- cells. We conclude that in T- myotubes most of the AChR fails to be transported out of the endoplasmic reticulum.

An essential role for ectodomain shedding in mammalian development.

The ectodomains of numerous proteins are released from cells by proteolysis to yield soluble intercellular regulators. The responsible protease, tumor necrosis factor-alpha converting enzyme (TACE), has been identified only in the case when tumor necrosis factor-alpha (TNFalpha) is released. Analyses of cells lacking this metalloproteinase-disintegrin revealed an expanded role for TACE in the processing of other cell surface proteins, including a TNF receptor, the L-selectin adhesion molecule, and transforming growth factor-alpha (TGFalpha). The phenotype of mice lacking TACE suggests an essential role for soluble TGFalpha in normal development and emphasizes the importance of protein ectodomain shedding in vivo.

Complex patterns of genetic and phenotypic divergence in an island bird and the consequences for delimiting conservation units.

Substantial phenotypic and genetic variation is often found below the species level and this may be useful in quantifying biodiversity and predicting future diversification. However, relatively few studies have tested whether different aspects of intraspecific variation show congruent patterns across populations. Here, we quantify several aspects of divergence between 13 insular populations of an island endemic bird, the Vanuatu white-eye (Zosterops flavifrons). The components of divergence studied are mitochondrial DNA (mtDNA), nuclear DNA microsatellites and morphology. These different aspects of divergence present subtly different scenarios. For instance, an mtDNA phylogenetic tree reveals a potential cryptic species on the most southerly island in Vanuatu and considerable divergence between at least two other major phylogroups. Microsatellite loci suggest that population genetic divergence between insular populations, both between and within phylogroups, is substantial, a result that is consistent with a low level of interisland gene flow. Finally, most populations were found to be strongly morphologically divergent, but no single population was morphologically diagnosable from all others. Taken together, our results show that, although many measures of divergence are concordant in this system, the number of divergent units identified varies widely depending on the characters considered and approach used. A continuum of divergence and a degree of discordance between different characters are both to be expected under simple models of evolution, but they present problems in terms of delimiting conservation units.

Avian haematozoan parasites and their associations with mosquitoes across Southwest Pacific Islands.

The degree to which haematozoan parasites can exploit a range of vectors and hosts has both ecological and evolutionary implications for their transmission and biogeography. Here we explore the extent to which closely related mosquito species share the same or closely related haematozoan parasites, and examine the overlap in parasite lineages with those isolated from avian hosts, Zosterops species, sampled across the same study sites. Mosquito samples were collected and analysed (14 species, n = 804) from four islands in Vanuatu and the main island of New Caledonia. Using polymerase chain reaction, 15.5% (14/90) of pooled mosquito (thoracic) samples showed positive amplifications. Subsequent phylogenetic analysis of the cytochrome b gene identified four genetically distinct Plasmodium and two Haemoproteus lineages from these samples, five of which were identical to parasite lineages (n = 21) retrieved from the avian hosts. We found that three Plasmodium lineages differing by a maximum of 0.9% sequence divergence were recovered from different species and genera of mosquitoes and two Haemoproteus lineages differing by 4.6% sequence divergence were carried by 10 distantly related (11-21% divergent) mosquito species. These data suggest a lack of both cospeciation and invertebrate host conservatism. Without experimental demonstration of the transmission cycle, it is not possible to establish whether these mosquitoes are the biological vectors of isolated parasite lineages, reflecting a limitation of a purely polymerase chain reaction-based approach. Nonetheless, our results raise the possibility of a new transmission pathway and highlight extensive invertebrate host shifts in an insular mosquito-parasite system.

Measurement of pulsatile haemodynamic forces in a model of a bifurcated stent graft for abdominal aortic aneurysm repair.

The longitudinal haemodynamic force (LF) acting on a bifurcated stent graft for abdominal aortic aneurysm repair has been estimated previously using a simple one-dimensional analytical model based on the momentum equation which assumes steady flow of an inviscid fluid. Using an instrumented stent-graft model an experimental technique was developed to measure the LF under pulsatile flow conditions. The physical stent-graft model, with main trunk diameter of 30mm and limb diameters of 12 mm, was fabricated from aluminium. Strain gauges were bonded on to the main trunk to determine the longitudinal strain which is related to the LF. After calibration, the model was placed in a pulsatile flow system with 40 per cent aqueous glycerol solution as the circulating fluid. The LF was determined using a Wheatstone bridge signal-conditioning circuit. The signals were averaged over 590 cardiac cycles and saved to a personal computer for subsequent processing. The LF was strongly dependent on the pressure but less so on the flowrate. The measured forces were higher than those predicted by the simplified mathematical model by about 6-18 per cent during the cardiac cycle. The excess measured forces are due to the viscous drag and the effect of pulsatile flow. The peak measured LF in this model of 30 mm diameter may exceed the fixation force of some current clinical endovascular stent grafts.

Anatomical flow phantoms of the nonplanar carotid bifurcation, part II: experimental validation with Doppler ultrasound.

A nonplanar wall-less anatomical flow phantom of a healthy human carotid artery is described, the construction of which is based on a lost-core technique described in the companion paper (Part I) by . The core was made by rapid prototyping of an idealized three-dimensional computer model of the carotid artery. Flow phantoms were built using these idealized non planar carotid artery bifurcations. Physiologically realistic flow waveforms were produced with resistance index values of 0.75, 0.72 and 0.63 in the common, external and internal carotid artery branches, respectively. Distension of the common carotid using M-mode imaging was found to be at 10% of diameter. Although differences in vessel diameter between the phantom and that of the original computer model were statistically significant (p < 0.05), there was no difference (p > 0.05) in measurements made on the lost-cores and those obtained by B-mode ultrasound on the resulting flow phantoms. In conclusion, it was possible to reliably reproduce geometrically similar anatomical flow phantoms that are capable of producing realistic physiological flow patterns and distensions.

Anatomical flow phantoms of the nonplanar carotid bifurcation, part I: computer-aided design and fabrication.

Doppler ultrasound is widely used in the diagnosis and monitoring of arterial disease. Current clinical measurement systems make use of continuous and pulsed ultrasound to measure blood flow velocity; however, the uncertainty associated with these measurements is great, which has serious implications for the screening of patients for treatment. Because local blood flow dynamics depend to a great extent on the geometry of the affected vessels, there is a need to develop anatomically accurate arterial flow phantoms with which to assess the accuracy of Doppler blood flow measurements made in diseased vessels. In this paper, we describe the computer-aided design and manufacturing (CAD-CAM) techniques that we used to fabricate anatomical flow phantoms based on images acquired by time-of-flight magnetic resonance imaging (TOF-MRI). Three-dimensional CAD models of the carotid bifurcation were generated from data acquired from sequential MRI slice scans, from which solid master patterns were made by means of stereolithography. Thereafter, an investment casting procedure was used to fabricate identical flow phantoms for use in parallel experiments involving both laser and Doppler ultrasound measurement techniques.

A new device for in vitro evaluation of thrombogenicity.

A device to measure the time of coagulation of whole blood has been designed in order to facilitate testing of thrombogenicity of biomaterials. The principle of operation of the apparatus is to record the time taken for a sphere to fall through a sample of blood. The coagulation time is defined here as the time from collection of the blood sample to coagulation, coagulation having deemed to have occurred when the sphere is prevented from falling by the presence of the fibrin-cell network. The device was tested with homogeneous fluids of different viscosity, milk containing different amounts of rennin and calcium chloride and non anti-coagulated whole blood obtained from five volunteers. Repeat measurements made with each homogeneous fluid show that the variance in the transit time of the sphere is small and consistent with small dispersion. In contrast, the onset of coagulation in milk and blood samples was readily detected. The clotting of milk was highly dependant on the concentration of rennin and calcium chloride. The coagulation time of blood samples from five individuals ranged from 23 to 33 min but simultaneous measurements of samples from the same individual in two identical devices agreed to within +/-1 min in all cases. This device may be easily adapted for use in studies to determine the thromboresistance of biomaterials where the onset of coagulation of whole blood in contact with different materials may be readily compared.

Attachment, morphology and adherence of human endothelial cells to vascular prosthesis materials under the action of shear stress.

In an effort to improve the long-term patency of vascular prostheses several groups now advocate seeding autologous endothelial cells (ECs) onto the lumen of the vessel prior to implantation, a procedure that involves pre-treating the prosthesis material with fibrin, collagen and/or other matrix molecules to promote cell attachment and retention. In this study, we examined the degree to which human umbilical venous endothelial cells (HUVECs) adhered to three materials commonly used polymeric vascular prosthesis that had been coated with the same commercial extra cellular matrix proteins, and after exposure to fluid shear stresses representative of femoro-distal bypass in a cone-and-plate shearing device. We quantified cell number, area of coverage and degree of cell spreading using image analysis techniques. The response of cells that adhered to the surface of each material, and following exposure to fluid shear stress, depended on surface treatment, topology and cell type. Whereas collagen coating improved primary cellular adhesion and coverage significantly, the degree of spreading depended on the underlying surface structure and on the application of the shear stress. In some cases, fewer than 30% of cells remained on the surface after only 1-h exposure to physiological levels of shear stress. The proportion of the surface that was covered by cells also decreased, despite an increase in the degree to which individual cells spread on exposure to shear stress. Moreover, the behaviour of HUVECs was distinct from that of fibroblasts, in that the human ECs were able to adapt to their environment by spreading to a much greater extent in response to shear. The quality of HUVEC attachment, as measured by extent of cell coverage and resistance to fluid shear stress, was greatest on expanded polytetrafluoroethylene samples that had been impregnated with Type I/III collagen.