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Biomedical subjects

A E Hill

Publications and source records attributed to A E Hill.

At least 19 recordsLinked to original sources
PubMed2006

A new approach to epithelial isotonic fluid transport: an osmosensor feedback model.

A model for control of the transport rate and osmolarity of epithelial fluid (isotonic transport) is presented by using an analogy with the control of temperature and flow rate in a shower. The model brings recent findings and theory concerning the role of aquaporins in epithelia together with measurements of epithelial paracellular flow into a single scheme. It is not based upon osmotic equilibration across the epithelium but rather on the function of aquaporins as osmotic sensors that control the tonicity of the transported fluid by mixing cellular and paracellular flows, which may be regarded individually as hyper- and hypo-tonic fluids, to achieve near-isotonicity. The system is built on a simple feedback loop and the quasi-isotonic behavior is robust to the precise values of most parameters. Although the two flows are separate, the overall fluid transport rate is governed by the rate of salt pumping through the cell. The model explains many things: how cell pumping and paracellular flow can be coupled via control of the tight junctions; how osmolarity is controlled without depending upon the precise magnitude of membrane osmotic permeability; and why many epithelia have different aquaporins at the two membranes. The model reproduces all the salient features of epithelial fluid transport seen over many years but also indicates novel behavior that may provide a subject for future research and serve to distinguish it from other schemes such as simple osmotic equilibration. Isotonic transport is freed from constraints due to limited permeability of the membranes and the precise geometry of the system. It achieves near-isotonicity in epithelia in which partial water transport by co-transporters may be present, and shows apparent electro-osmotic effects. The model has been developed with a minimum of parameters, some of which require measurement, but the model is flexible enough for the basic idea to be extended both to complex systems of water and salt transport that still await a clear explanation, such as intestine and airway, and to systems that may lack aquaporins or use other sensors.

Animals↗
PubMed2006

AQP and the control of fluid transport in a salivary gland.

Experiments were performed with the perfused rat submandibular gland in vitro to investigate the nature of the coupling between transported salt and water by varying the osmolarity of the source bath and observing the changes in secretory volume flow. Glands were submitted to hypertonic step changes by changing the saline perfusate to one containing different levels of sucrose. The flow rate responded by falling to a lower value, establishing a new steady-state flow. The rate changes did not correspond to those expected from a system in which fluid production is due to simple osmotic equilibration, but were much larger. The changes were fitted to a model in which fluid production is largely paracellular, the rate of which is controlled by an osmosensor system in the basal membrane. The same experiments were done with glands from rats that had been bred to have very low levels of AQP5 (the principal aquaporin of the salivary acinar cell) in which little AQP5 is expressed at the basal membrane. In these rats, salivary secretion rates after hypertonic challenges were small and best modelled by simple osmotic equilibration. In rats which had intermediate AQP5 levels the changes in flow rate were similar to those of normal rats although their AQP5 levels were reduced.Finally, perfused normal glands were subject to retrograde ductal injection of salines containing different levels of Hg(2+) ions (0, 10 and 100 microM: ) which would act as inhibitors of AQP5 at the apical acinar membrane. The overall flow rates were progressively diminished with rising Hg(2+) concentration, but after hypertonic challenge the changes in flow rates were unchanged and similar to those of normal rats. All these results are difficult to explain by a cellular osmotic model but can be explained by a model in which paracellular flow is controlled by an osmosensor (presumably AQP5) present on the basal membrane.

Animals↗
PubMed2005

Prevalence of and risk factors for Escherichia coli O157 in market-ready beef cattle from 12 U.S. feedlots.

Determination of Escherichia coli O157 prevalence immediately prior to shipment and harvest is an important facet of the ecology of this organism, which requires further elucidation. As part of a larger study to measure the effects of within-pen prevalence of E. coli O157 on subsequent carcass contamination, fecal samples from 15 pens of cattle in each of 12 different feedlots in three states (Colorado, Nebraska, and Montana) were collected from June through September 2002. Thirty fresh fecal samples were collected from each pen floor within 36 h of shipment to slaughter. Fecal samples underwent standard enrichment, immunomagnetic separation, and isolation procedures for E. coli O157. Multivariable logistic regression was used to determine which factors best predicted pen-level positive culture results, and to estimate the magnitude of association between each factor and the outcome, while adjusting for other factors in the model. Thirteen (86.7%) of the 15 pens had at least one positive sample, and the within-pen prevalence of E. coli O157 in positive pens ranged from 3.3% to 77.8%. The odds of E. coli O157 positive fecal samples from cattle fed brewers grains were six times that for cattle not fed brewers grains. The odds of E. coli O157 positive fecal samples from pens of cattle from Central Nebraska was nine times that for pens of cattle from Eastern Colorado. These data demonstrate that the presence of E. coli O157 in fecal samples from finished feedlot cattle is associated with feeding of brewers grain and geographic location. Additional studies to further characterize geographic distribution of E. coli O157 and to investigate pen-level intervention strategies should be conducted.

Animal Feed↗
PubMed2004

What are aquaporins for?

The prime function of aquaporins (AQPs) is generally believed to be that of increasing water flow rates across membranes by raising their osmotic or hydraulic permeability. In addition, this applies to other small solutes of physiological importance. Notable applications of this 'simple permeability hypothesis' (SPH) have been epithelial fluid transport in animals, water exchanges associated with transpiration, growth and stress in plants, and osmoregulation in microbes. We first analyze the need for such increased permeabilities and conclude that in a range of situations at the cellular, subcellular and tissue levels the SPH cannot satisfactorily account for the presence of AQPs. The analysis includes an examination of the effects of the genetic elimination or reduction of AQPs (knockouts, antisense transgenics and null mutants). These either have no effect, or a partial effect that is difficult to explain, and we argue that they do not support the hypothesis beyond showing that AQPs are involved in the process under examination. We assume that since AQPs are ubiquitous, they must have an important function and suggest that this is the detection of osmotic and turgor pressure gradients. A mechanistic model is proposed--in terms of monomer structure and changes in the tetrameric configuration of AQPs in the membrane--for how AQPs might function as sensors. Sensors then signal within the cell to control diverse processes, probably as part of feedback loops. Finally, we examine how AQPs as sensors may serve animal, plant and microbial cells and show that this sensor hypothesis can provide an explanation of many basic processes in which AQPs are already implicated. Aquaporins are molecules in search of a function; osmotic and turgor sensors are functions in search of a molecule.

Aquaporins↗
PubMed2003

Survey of restriction-modification systems and transformation in Mannheimia haemolytica and Pasteurella trehalosi.

A significant obstacle to molecular studies of Mannheimia (Pasteurella) haemolytica, has been its resistance to genetic transformation. The lack of competence of many M. haemolytica strains has been attributed to the presence of restriction modification systems. In this study, representative strains of 12 M. haemolytica serotypes and four Pasteurella trehalosi serotypes were successfully transformed by electroporation using a recombinant vector derived from the native M. haemolytica A1 serotype plasmid pNSF2176. Transformation was achieved despite PCR-based evidence for the presence of genes encoding a type I restriction enzyme, phaI, and a type II restriction enzyme hsdM, in each of the M. haemolytica strains.

DNA Restriction-Modification Enzymes↗
PubMed2003

Visual impairment in childhood: insights from a community-based survey.

BACKGROUND: The concept of visual impairment (VI) in childhood has changed over the last 30 years. There has been a decrease in the number of children with an isolated visual problem and an increase in the numbers with VI and coexisting neurological disability. This study aimed to produce a profile of VI in childhood with a view to informing future services and to raise awareness of the need for comprehensive assessment including developmental remediation and educational advice. METHODS: Children with a VI were identified from multiple sources including hospital- and community-based paediatricians and statutory blind registers. RESULTS: Seventy-six children with a VI were identified giving a childhood prevalence of 1.61 per 1000. Thirty-two per cent had a normal pattern of development. Global delays/severe learning difficulty were found in 43%. Only 21% of the children had an isolated VI. Additional medical problems were present in 79% of which cerebral palsy, occurring in 33%, was the most common. Nine per cent of the children were classified as totally blind. Cortical visual impairment was diagnosed in 45%. Twenty-two per cent of the children were registered blind or partially sighted. CONCLUSIONS: Most cases of VI in children did not appear on the statutory blind or partially sighted registers, thus these have limited value for service development. The implications for practice highlight the need for early assessment and advice from a co-ordinated team to optimize visual potential in childhood.

Adolescent↗
PubMed2001

The paracellular component of water flow in the rat submandibular salivary gland.

1. The pathway of water flow during salivary secretion by the isolated, perfused rat submandibular gland was examined using a family of homologous radiodextran molecules as probes of paracellular fluid transfer. 2. The secretion/perfusate ratio (S/P) of the secreted probes versus molecular radius during fluid secretion evoked by ACh could be resolved into two components: one that fitted a free-diffusion (Stokes-Einstein) curve and indicated diffusion through large channels, and a convective component that was linearly related to radius. 3. The convective component had a cut-off point at 0.5 nm (5 A) radius and an S/P intercept of near 1.0 at the radius of water, which indicates that most of the volume flow was paracellular. 4. The nature of such a paracellular flow is discussed together with the possible integration of this volume flow with the cellular transport of ions, resulting in an isotonic primary secretion from the gland.

Animals↗
PubMed2001

Single water channels of aquaporin-1 do not obey the Kedem-Katchalsky equations.

The Kedem-Katchalsky (KK) equations are often used to obtain information about the osmotic properties and conductance of channels to water. Using human red cell membranes, in which the osmotic flow is dominated by Aquaporin-1, we show here that compared to NaCl the reflexion coefficient of the channel for methylurea, when corrected for solute volume exchange and for the water permeability of the lipid membrane, is 0.54. The channels are impermeable to these two solutes which would seem to rule out flow interaction and require a reflexion coefficient close to 1.0 for both. Thus, two solutes can give very different osmotic flow rates through a semi-permeable pore, a result at variance with both classical theory and the KK formulation. The use of KK equations to analyze osmotic volume changes, which results in a single hybrid reflexion coefficient for each solute, may explain the discrepancy in the literature between such results and those where the equations have not been employed. Osmotic reflexion coefficients substantially different from 1.0 cannot be ascribed to the participation of other 'hidden' parallel aqueous channels consistently with known properties of the membrane. Furthermore, we show that this difference cannot be due to second-order effects, such as a solute-specific interaction with water in only part of the channel, because the osmosis is linear with driving force down to zero solute concentration, a finding which also rules out the involvement of unstirred-layer effects. Reflexion coefficients smaller than 1.0 do not necessitate water-solute flow interaction in permeable aqueous channels; rather, the osmotic behaviour of impermeable molecular-sized pores can be explained by differences in the fundamental nature of water flow in regions either accessible or inaccessible to solute, created by a varying cross-section of the channel.

Aquaporin 1↗
PubMed2001

Risk factors for and outcomes of noncatastrophic suspensory apparatus injury in Thoroughbred racehorses.

OBJECTIVE: To evaluate effects of toe grabs, exercise intensity, and distance traveled as risk factors for subclinical to mild suspensory apparatus injury (SMSAI) in Thoroughbred racehorses and to compare incidence of severe musculoskeletal injury (MSI) in horses with and without SMSAI. DESIGN: Nested case-control study. ANIMALS: 219 Thoroughbred racehorses racing or in race training. PROCEDURE: Racehorses were examined weekly for 90 days to determine incidence of suspensory ligament injury and monitor horseshoe characteristics. Every horse's exercise speeds and distances were recorded daily. Conditional logistic regression was used to compare exposure variables between incident case (n = 25) and selected control (125) horses. Survival analysis was used to compare time to MSI for horses with (n = 41) and without (76) SMSAI. RESULTS: The best-fitting logistic model for the data included age (< 5 vs > or = 5 years old), toe grab height the week of injury (none vs very low, low, regular, or Quarter Horse height), and weekly distance the week preceding injury (miles). Although the 95% confidence intervals for all odds ratios included 1, the odds for SMSAI appeared to increase with the presence of a toe grab, higher weekly distance, and age > or = 5 years. Horses that had SMSAI were significantly more likely to have a severe MSI or severe suspensory apparatus injury than were horses that did not. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that pre-existing SMSAI is associated with development of severe MSI and severe suspensory apparatus injury. Modifying training intensity and toe grab height for horses with SMSAI may decrease the incidence of severe MSI.

Age Factors↗
PubMed2001

Relation between objectively measured feeding competence and nutrition in children with cerebral palsy.

This study aimed to investigate the prevalence of undernutrition in children with cerebral palsy (CP) and to determine the relation with feeding ability. Ninety children with CP from special needs schools were examined. Undernutrition was diagnosed on one or more of the following criteria: weight <2nd centile, triceps or subscapular skinfold measurement <3rd centile, mid-arm circumference <5th centile. Feeding competence was scored with respect to seven specific oromotor tasks using the Multidisciplinary Feeding Profile. Thirty-six participants (40%) had diplegia, 29 (33%) quadriplegia, 13 (14%) hemiplegia, and 12 participants (13%) had dyskinetic CP. Age ranged from 2.6 to 18.7 years (mean 10.8 years). Forty-six percent (41 of 90) were undernourished. In all aspects of feeding, those undernourished had lower feeding competence scores compared to adequately nourished children (p<0.002). Each modality of feeding competence correlated significantly to the centiles of weight, triceps or subscapular skinfold measurement and mid-arm circumference (p<0.02). A positive association of weight, triceps skinfold measurement, and mid-arm circumference with chewing ability was present independent of other feeding modalities (p<0.05). Undernutrition was common in this group and was associated with poorer feeding ability.

Adolescent↗
PubMed2000

Identification and characterization of a novel cytochrome c(3) from Shewanella frigidimarina that is involved in Fe(III) respiration.

Shewanella frigidimarina NCIMB400 is a non-fermenting, facultative anaerobe from the gamma group of proteobacteria. When grown anaerobically this organism produces a wide variety of periplasmic c-type cytochromes, mostly of unknown function. We have purified a small, acidic, low-potential tetrahaem cytochrome with similarities to the cytochromes c(3) from sulphate-reducing bacteria. The N-terminal sequence was used to design PCR primers and the cctA gene encoding cytochrome c(3) was isolated and sequenced. The EPR spectrum of purified cytochrome c(3) indicates that all four haem irons are ligated by two histidine residues, a conclusion supported by the presence of eight histidine residues in the polypeptide sequence, each of which is conserved in a related cytochrome c(3) and in the cytochrome domains of flavocytochromes c(3). All four haems exhibit low midpoint redox potentials that range from -207 to -58 mV at pH 7; these values are not significantly influenced by pH changes. Shewanella cytochrome c(3) consists of a mere 86 amino acid residues with a predicted molecular mass of 11780 Da, including the four attached haem groups. This corresponds closely to the value of 11778 Da estimated by electrospray MS. To examine the function of this novel cytochrome c(3) we constructed a null mutant by gene disruption. S. frigidimarina lacking cytochrome c(3) grows well aerobically and its growth rate under anaerobiosis with a variety of electron acceptors is indistinguishable from that of the wild-type parent strain, except that respiration with Fe(III) as sole acceptor is severely, although not completely, impaired.

Amino Acid Sequence↗
PubMed1997

Ion, fluid and charge transport across Necturus intestinal epithelium in response to alanine.

Fluxes of Na, Cl and volume were followed across Necturus small intestine under zero voltage clamp. 20 mM L-alanine doubles the net Na and fluid transfer. Although there is a ouabain-sensitive Na pump present in Necturus a major fraction of the net Na flux can be measured for an hour after application of 10(-3) M ouabain. Collected fluid transferred by the epithelium is quasi-isotonic over a range of luminal osmolarities from 100 to 250 milliosmolar in alanine saline. The net Na fluxes account for the Na found in this transported fluid. Fluid transfer also shows a large ouabain-insensitive fraction after the addition of alanine. Compartmental analysis of 22Na-loaded epithelium was used to separate cellular and paracellular fluxes. The estimated Na concentration in the cell derived from its Na content is 9-10 mM, in agreement with that determined with microelectrodes. The Na efflux from cell to serosa is stimulated by alanine, but this increase accounts for only a quarter of the simultaneous rises in Na, fluid and current flow across the epithelium. The increase of Na efflux from the cell induced by alanine is apparently insensitive to ouabain although the cell Na content rises to circa 20 mM but no higher even after 20 hr. From the initial rate of rise of Na in the cell on treatment with ouabain the activity of the Na pump can be estimated to be approximately 92 pM/cm2 . sec, a value much smaller than the transepithelial net flux. The results are not consistent with the standard model in which Na-alanine influx stimulates the Na pump and enhances fluid transport by osmotic coupling in the lateral interspace system. A scheme is proposed based upon that for absorption in Necturus gallbladder by which alanine stimulates an active paracellular fluid transfer driven by motile elements of the junction.

Alanine↗
PubMed1997

Fluid recirculation in Necturus intestine and the effect of alanine.

Fluid absorption by Necturus small intestine has been studied using radiolabeled dextrans as molecular probes of the paracellular pathway under voltage-clamped conditions. Fluxes of H3-dextrans of MW up to 20K were followed in both directions between mucosal (M) and serosal (S) baths by fractionating those that passed the epithelium as a function of molecular radius. Consideration of the unstirred layers in the baths and the surface geometry rules out any contribution made by solute polarization. The geometry of the paracellular system was measured by light microscopy, TEM and SEM, and values were used in conjunction with a program that calculates convective-diffusive coupling in the tight junctions, intercellular spaces and subepithelium in series. The results indicate that the net fluxes are due to the convection of fluid through two opposing paracellular fluid circuits with different size selectivity, resulting in net absorption at small radii. Alanine at 20 mM stimulates fluid and salt uptake by a factor of 2. Its effect on the two convective components is to increase the M to S flux and decrease the S to M. The selectivities are not significantly different from those before alanine treatment. The volume absorption predicted from the net probe fluxes is very close to that measured gravimetrically across the epithelium.

Alanine↗
PubMed1997

The influence of flexed-knee gait on the energy cost of walking in children.

There is an understanding that walking with flexed knees contributes to the increased energy cost of walking found in children with neurological conditions. To determine the influence of flexed-knee gait on energy cost of walking in a group of children without neurological abnormality, the gait patterns of 10 normal children were studied using a Vicon system and standard marker set. A telemetric system (Cosmed K2) was then used to measure the oxygen cost of walking of the same children. The tests were repeated restricting the subjects' knee extension bilaterally, using hinged braces, set to 0, 15, 30, and 45 degrees of flexion. Although the braces themselves caused a significant increase in O2 cost (mL/kg/m) (P < 0.05), due to a decrease in walking speed, no further significant increase in oxygen cost was demonstrated regardless of the degree of knee flexion imposed, despite a significant increase in measured hip flexion and ankle dorsiflexion at the 45 degrees of knee flexion setting (P < 0.05). We propose that moderate flexed-knee gait does not of itself cause an increase in the energy cost of walking and that other factors present in the physically disabled child are likely to be implicated.

Adolescent↗
PubMed1996

Energy consumption in children with spina bifida and cerebral palsy: a comparative study.

The authors looked for differences in the energy expenditure patterns of ambulant children with cerebral palsy and spina bifida. Oxygen consumption was measured according to type of cerebral palsy or level of spina bifida lesion, and in healthy children. The rate of oxygen consumption (mL/kg/min) was significantly higher in the children with diplegia than in those with hemiplegia or with spina bifida or the healthy children. Oxygen cost (mL/kg/m) was significantly higher and velocity was significantly slower in all the groups with disability than in the healthy children. The reason children with diplegia consumed more oxygen than other children when walking may be that their abnormal equilibrium reactions impaired their balance and their ability to control their walking speed.

Cerebral Palsy↗