Variability in tracheal mucociliary transport is not controlled by beating cilia in lambs in vivo during ventilation with humidified and nonhumidified air.

Mucociliary transport in the respiratory epithelium depends on beating of cilia to move a mucus layer containing trapped inhaled particles toward the mouth. Little is known about the relationship between cilia beat frequency (CBF) and mucus transport velocity (MTV) in vivo under normal physiological conditions and when inspired air is dry or not fully humidified. This study was designed to use video-microscopy to simultaneously measure CBF and MTV in the tracheal epithelium through an implanted optical window in mechanically ventilated lambs. The inspired air in 6 animals was heated to body temperature and fully saturated with water for 4 hours as a baseline. In another series of experiments, 5 lambs were ventilated with air at different temperatures and humidities and the mucosal surface temperature was monitored with infrared macro-imaging. In the baseline experiments, during ventilation with fully humidified air at body temperature, CBF remained constant, mean 13.9 ± 1.6 Hz but MTV varied considerably between 0.1 and 26.1 mm/min with mean 11.0 ± 3.9 mm/min, resulting in a maximum mucus displacement of 34.2 µm/cilia beat. Fully humidified air at body temperature prevented fluctuations in the surface temperature during breathing indicating a thermodynamic balance in the airways. When lambs were ventilated with dryer air, the mucosal surface temperature and MTV dropped without a significant change in CBF. When inspired air was dry, mainly latent heat (92%) was transferred to air in the trachea, reducing the surface temperature by 5 °C. Reduced humidity of the inspired air lowered the surface temperature and reduced MTV in the epithelium during ventilation.

Effect of collagen packing and moisture content on leather stiffness.

Applications for skin derived collagen materials, such as leather and acellular dermal matrices, usually require both strength and flexibility. In general, both the tensile modulus (which has an impact on flexibility) and strength are known to increase with fiber alignment, in the tensile direction, for practically all collagen-based tissues. The structural basis for flexibility in leather was investigated and the moisture content was varied. Small angle X-ray scattering was used to determine collagen fibril orientation, elongation and lateral intermolecular spacing in leather conditioned by different controlled humidity environments. Flexibility was measured by a three point bending test. Leather was prepared by tanning under biaxial loading to create leather with increased fibril alignment and thus strength, but this treatment also increased the stiffness. As collagen aligns, it not only strengthens the material but it also stiffens because tensile loading is then applied along the covalent chain of the collagen molecules, rather than at an angle to it. Here it has been shown that with higher moisture content greater flexibility of the material develops as water absorption inside collagen fibrils produces a larger lateral spacing between collagen molecules. It is suggested that water provides a lubricating effect in collagen fibrils, enabling greater freedom of movement and therefore greater flexibility. When collagen molecules align in the strain direction during tanning, leather stiffens not only by the fiber alignment itself but also because collagen molecules pack closer together, reducing the ability of the molecules to move relative to each other.

Data on collagen structures in leather with varying moisture contents from small angle X-ray scattering and three point bend testing.

The data presented in this article are related to the research article entitled "Effect of collagen packing and moisture content on leather stiffness" (Kelly et al., 2018). This article describes how moisture content affects collagen packing and leather stiffness. Structural changes were experimentally introduced into ovine leather through biaxial strain during tanning (׳stretch tanning׳). Leather samples produced normally without strain (׳non-stretch tanned׳) and those produced by stretch tanning, were conditioned in a range of relative humidity environments and then analysed by small angle X-ray scattering and three point bend testing. The collagen D-spacing, lateral intermolecular spacing and flexural properties were measured under these varying moisture contents.

Haemodynamic effects of parenteral vs. enteral paracetamol in critically ill patients: a randomised controlled trial.

Paracetamol is a commonly used drug in the intensive care unit. There have been reports in the literature of an association with significant hypotension, a potentially important interaction for labile critically ill patients. Route of administration may influence the incidence of hypotension. This single-centre, prospective, open-label, randomised, parallel-arm, active-control trial was designed to determine the incidence of hypotension following the administration of paracetamol to critically ill patients. Fifty adult patients receiving paracetamol for analgesia or pyrexia were randomly assigned to receive either the parenteral or enteral formulation of the drug. Paracetamol concentrations were measured at baseline and at multiple time points over 24 h. The maximal plasma paracetamol concentration was significantly different between routes; 156 vs. 73 micromol.l(-1) [p = 0.0005] following the first dose of parenteral or enteral paracetamol, respectively. Sixteen hypotensive events occurred in 12 patients: parenteral n = 12; enteral n = 4. The incident rate ratio for parenteral vs. enteral paracetamol was 2.94 (95% CI 0.97-8.92; p = 0.06). The incidence of hypotension associated with paracetamol administration is higher than previously reported and tends to be more frequent with parenteral paracetamol.

Hypertriglyceridemic waist and newly-diagnosed diabetes among remote-dwelling Indigenous Australians.

AIMS: Hypertriglyceridemic waist (HTgW) is predictive of cardiovascular disease. The HTgW relationship with diabetes is little studied. METHODS: This study analysed data from diabetes and cardiovascular risk factor screening programmes in remote Indigenous Australian settlements. Elevated waist girth (EW) was defined as ≥90 cm for men (n = 1134) or ≥80 cm for women (n = 1313). Hypertriglyceridemia (ETg) was defined as ≥1.7 mmol/L. Diabetes was defined as fasting plasma glucose ≥7.0 mmol/L. Body mass index (BMI) was categorised as <22, 22-24.9 and >25.0 kg/m(2). Logistic regression was used to analyse the odds of newly-diagnosed diabetes for individuals with either HTgW, ETg or EW, relative to individuals with values below cut-offs. RESULTS: The prevalence of HTgW was 33.2% for men and 34.8% for women. Accounting for age-group and gender, newly-diagnosed diabetes was associated (odds ratio (OR) (95% confidence interval)) with HTgW: 9.6 (6.6, 13.8). The relationship remained strong after accounting for the covariates BMI and smoking (OR = 4.9 (2.7, 8.8)). In BMI-stratified analyses the strongest odds were observed for the lowest category (<22 kg/m(2): OR = 12.9 (4.0, 41.7)). CONCLUSIONS: HTgW has a high prevalence and is associated with newly-diagnosed diabetes in Indigenous people, particularly those with BMI <22 kg/m(2), whom clinicians might not normally consider for screening.

Associations of morbidity in the underweight.

BACKGROUND/OBJECTIVES: There is little research on the demographic characteristics and morbidity of people categorized as 'underweight' from their body mass index (BMI) although they have often been shown to have greater mortality. This uncertainty makes it difficult to determine whether to include or exclude these individuals when estimating the health and mortality impacts of BMI. This project compares the demographic characteristics and morbidity patterns of the underweight with those of acceptable weight and the overweight. SUBJECTS/METHODS: Data on 10 243 community-living residents from the Health Survey for England (2003) were used. Logistic regression models were constructed to compare demographic, biochemical and anthropometric factors in the underweight (BMI<18.5) with those classified as acceptable weight (BMI 18.5-24.9) or overweight (BMI 25.0-29.9). RESULTS: Univariate analyses found, when compared with other BMI categories, underweight individuals were significantly younger, more likely to smoke, alcohol abstainers, inactive, poorer and were less likely to be ethnically white (all P<0.001). U-shaped relationships between BMI and activities of daily living, respiratory disease, physical activity and mental health variables were seen. In multivariate analysis, the fewest number of significant differences in demographic and morbidity factors were between the underweight and those of acceptable weight. CONCLUSIONS: We recognize that these are cross-sectional data and exclude individuals in institutional settings, but these findings are important. Overall, we could not conclude that the underweight were less healthy than individuals in the other BMI categories. We cannot therefore recommend that the underweight should be excluded from analyses that examine the effects of obesity on mortality.

Effects of the AMPA receptor modulator S 18986 on measures of cognition and oxidative stress in aged rats.

RATIONALE: Development of cognitive-enhancing drugs that delay or halt mild cognitive impairment progression to Alzheimer's disease would be of great benefit. OBJECTIVES: The aim of this study was to examine the ability of (S)-2,3-dihydro-[3,4]-cyclopentano-1,2,4-benzothiadiazine-1,1-dioxide (S 18986), a positive allosteric modulator of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors, to improve behavioral performance and alleviate age-related deficits in oxidative stress status in the prelimbic cortex and hippocampus. MATERIALS AND METHODS: Daily administration of S 18986 (0.1, 0.3, and 1.0 mg/kg) or vehicle was given to separate groups of male rats starting at 12 months of age. Additionally, daily vehicle administration was given to a group of rats starting at 3 months of age. Four months after initiation of drug administration, rats were trained and tested in an operant-delayed alternation task and a reinforcer devaluation task. Upon completion of testing, oxidative stress status was assessed in the prelimbic cortex and hippocampus. RESULTS: S 18986 dose-dependently altered responses in the reinforcer devaluation task such that aged rats came to resemble young rats. There were no age or drug effects in the operant-delayed alternation task. Levels of the lipid peroxidation product 4-hydroxy-nonenal (HNE) were increased, and Cu/Zn-superoxide dismutase (SOD) levels were decreased in prelimbic cortex in aged rats, changes that were reversed by S 18986. Similarly, age-related increases in hippocampal HNE levels were prevented by S 18986. CONCLUSIONS: Positive modulation of AMPA receptor activity may be a therapeutic approach to halt or slow progression of mild cognitive impairment via improvement in oxidative stress status in the hippocampus and prelimbic cortex.

Perinatal delta-9-tetrahydrocannabinol exposure disrupts social and open field behavior in adult male rats.

Marijuana is the most frequently used illegal drug among women of reproductive age, but little is known about the consequences of using marijuana during pregnancy. Delta-9-tetrahydrocannabinol (Delta9-THC), one of the active chemicals in marijuana, has been shown to cross the placental barrier easily. In this study, pregnant Long Evans rats were assigned to one of three treatment groups (Delta9-THC-exposed, vehicle control, and non-treated control) on day 1 of gestation. Drug exposure consisted of 2 mg/kg of natural Delta9-THC, administered twice daily by subcutaneous (s.c.) injection, from gestational day 1 through 22. Pups continued to receive drug exposure via s.c. injection from postnatal day 2 through 10. Male rats from each group were tested starting on postnatal day 90 in a battery of tests which included open field activity, active social interaction, and the forced swim test. There were no significant differences in weight gained by dams or weight of offspring when compared to controls. Delta9-THC-exposed rats showed decreased time in the inner part of the open field and an increase in investigation time in the test of social interaction compared to both control groups. There were no differences among groups in the forced swim test. Perinatal Delta9-THC exposure may result in increased susceptibility to anxious behavior and alter social functioning in adult offspring.

Genotyping of Pseudomonas aeruginosa in cystic fibrosis suggests need for segregation.

BACKGROUND: Emerging resistance of Pseudomonas aeruginosa within cystic fibrosis (CF) populations is attributed to antibiotic pressure and spread of transmissible strains. We describe increasing resistance of P. aeruginosa isolates, resulting in the identification of two multiresistant strains and their impact on morbidity. METHODS: Susceptibility reports of all P. aeruginosa isolates since 1998 in our unit were reviewed. Isolates were submitted for genomic finger-printing by pulsed-field gel electrophoresis. Clinical measures and the consumption of treatment resources were compared between those harbouring resistant organisms and those with sensitive strains. RESULTS: Analysis of 407 reports from 43 patients revealed isolation of multiresistant (MR) organisms increased during 1999. Those harbouring MR strains consumed more resources than non-MR. Strain typing showed a new 'Sheffield' strain in seven patients (100% MR), and the 'Liverpool' strain in 10 patients (40% MR). Individuals in these groups consumed significantly more resources than 23 patients with unique, susceptible strains (4% MR). DISCUSSION: Increasing resistance in isolates of P. aeruginosa may herald the arrival of a transmissible strain in CF Units which though sometimes sensitive, may become multiply resistant and require more intensive treatment. We now segregate those with transmissible strains from each other and from those with unique strains.

Ventral neurons in the anterior nerve cord of amphioxus larvae. I. An inventory of cell types and synaptic patterns.

Serial sections were used to map the ventrally positioned neurons of the anterior nerve cord of a 12.5-day amphioxus larva from the infundibular region to the end of somite 2. Synaptic patterns reveal five categories of descending pathways, four of which are associated with the ventral compartment (VC) motoneurons responsible for escape swimming. 1) Pre-, para-, and postinfundibular (tegmental) neurons with large varicosities and mixed vesicle populations provide both synaptic and paracrine input to various components of the tegmental neuropile and primary motor center. Four categories of these neurons are distinguished on the basis of their vesicles. 2) Multiple anterior sensory pathways converge on the large paired neurons (LPNs) located near the junction of somites 1 and 2. LPN synaptic output is almost exclusively contralateral. This, together with the evidence for cross-innervation between the third pair of LPNs, is consistent with the latter acting as locomotory pacemakers. 3) Axons from several classes of tegmental neurons converge in the paraxial region on each side of the cord where they form distinct tracts, the upper paraxial bundles. The right bundle is larger than the left, which suggests a role during early development when myotome contractions are biased to one side. 4) Fibers in the ventral tracts from ipsilateral projection neurons, sensory neurons, and additional ascending fibers synapse repeatedly with VC motoneurons. This may be how the overall level of excitation of the latter is controlled so as to modulate their response to pacemaker input. The fifth pathway consists of fibers involved in controlling the dorsal compartment (DC) motoneurons responsible for slow swimming, which are largely isolated from inputs to the VC locomotory system. The ventral neurons of the primary motor center form a more or less continuous file on either side of the floor plate, with certain cell types showing a tendency to cluster. There are, however, few obvious patterns of the kind expected if development were controlled by a rigid, lineage-based mechanism. The evolutionary implications of the involvement of a midbrain-level pacemaker in controlling larval swimming in amphioxus is discussed.

Effects of alcohol exposure during development on social behavior in rats.

In addition to the cognitive deficits associated with fetal alcohol syndrome (FAS), clinical and animal studies indicate that alcohol exposure might also have detrimental effects on social behavior. In a rat model of FAS, experimental rats were given alcohol from gestational day (GD) 1 to 22 and from postnatal day (PD) 2 to 10, a period roughly equivalent to all three trimesters in humans. Control groups consisted of rats exposed to the administration procedures but not to alcohol and nontreated rats. At 30 days of age, rats were tested for social behavior in an alley maze that contained its cagemate in the goal box. After varying periods of isolation, the animals' latencies to reach the goal box and their social behaviors once inside the goal box were recorded. Alcohol-exposed animals ran faster than control rats to the occupied goal box regardless of the amount of isolation. The alcohol-exposed animals also exhibited aberrant social interactions with their cagemate once inside the goal box compared to one or both of the control groups. Specifically, the alcohol-exposed animals showed greater amounts of anogenital sniffing, chasing, hopping and darting, and retrieving and lesser amounts of pinning and biting compared to one or both of the control groups. The alcohol-induced change in anogenital sniffing varied over increasing amounts of isolation compared to both control groups, but the alterations in the other behaviors did not. It is argued that the altered social behavior of alcohol-exposed animals is not the result of changes in the animals' motivational state or social learning and may be the result of an increased responsiveness to social stimuli.

Maternal urinary tract infection alters water maze performance in the offspring.

The effects of maternal urinary tract infection (UTI) or endotoxin exposure on fetal outcome in rats were investigated. Prior to conception, dams of the UTI group were water-deprived and anesthetized. The urinary tract was then catheterized and injected with 0.2 of 1 x 10(9) Escherichia coli. The endotoxin group was injected with 0.03 mg/kg lipopolysaccharide on the fourth day of gestation and then every third day thereafter. The control groups were treated in the same manner, with the exception that the infection control was not catheterized or injected with E. coli, and the endotoxin control was not exposed to lipopolysaccharide. A nontreated control group was weighed daily. Beginning on postnatal day (PD) 19, offspring were tested daily in a water maze spatial navigation task. The retention latencies (Sessions 7--10) revealed deficits in the infection and endotoxin groups. In the rat model, these findings suggest that exposure during gestation to a maternal immune challenge may result in adverse fetal outcome.

Kinetic properties of Streptococcus pneumoniae hyaluronate lyase.

Streptococcus pneumoniae hyaluronate lyase is a surface antigen of this bacterial pathogen, which causes significant mortality and morbidity in human populations worldwide. The primary function of this enzyme is the degradation of hyaluronan, a major component of the extracellular matrix of the tissues of practically all vertebrates. The enzyme uses a processive mode of action to degrade hyaluronan to a final product, an unsaturated disaccharide hyaluronan unit. This catalysis proceeds via a five-step proton acceptance and donation mechanism that includes substrate binding, catalysis, release of the disaccharide product, translocation of the remaining hyaluronan substrate, and proton exchange with microenvironment. Based on the analysis of the three-dimensional structure of the native enzyme and its complexes with hexasaccharide substrate and disaccharide product, several residues have been chosen for mutation studies. These mutated residues included the catalytic residues Asn349, His399, Tyr408, and residues responsible for substrate binding and translocation, Arg243 and Asn580. The comparison of the kinetic properties of the wild-type with the mutant enzymes allowed for the characterization of every mutant and the correlation of the kinetic properties of the enzyme with its structure. The comparison of the wild-type hyaluronate lyase with other polysaccharide-degrading enzymes, the hydrolases endonuclease and glucoamylase, shows striking similarity of K(m)s for all of these different enzymes.

Vitamin C inhibits the enzymatic activity of Streptococcus pneumoniae hyaluronate lyase.

Enzyme activity measurement showed that L-ascorbic acid (vitamin C (Vc)) competitively inhibits the hyaluronan degradation by Streptococcus pneumoniae hyaluronate lyase. The complex crystal structure of this enzyme with Vc was determined at 2.0 A resolution. One Vc molecule was found to bind to the active site of the enzyme. The Vc carboxyl group provides the negative charges that lead the molecule into the highly positively charged cleft of the enzyme. The Vc ring system forms hydrophobic interactions with the side chain of Trp-292, which is one of the aromatic patch residues of this enzyme responsible for the selection of the cleavage sites on the substrate chain. The binding of Vc inhibits the substrate binding at hyaluronan 1, 2, and 3 (HA1, HA2, and HA3) catalytic positions. The high concentration of Vc in human tissues probably provides a low level of natural resistance to the pneumococcal invasion. This is the first time that Vc the direct inhibition on the bacterial "spreading factor" was reported, and Vc is also the first chemical that has been shown experimentally to have an inhibitory effect on bacterial hyaluronate lyase. These studies also highlight the possible structural requirement for the design of a stronger inhibitor of bacterial hyaluronate lyase.

Critical periods for the effects of alcohol exposure on learning in rats.

Critical periods for alcohol-induced deficits in spatial navigation and passive avoidance learning were investigated with a rat model of fetal alcohol syndrome. Rats were exposed to alcohol prenatally (Gestational Days 1-10 or 11-22) or postnatally (Postnatal Days 2-10) or throughout all 3 periods. Offspring were tested in either a spatial navigation or an avoidance task as juveniles or adults. As juveniles, the combined exposure group took longer to learn the spatial navigation task compared with all other groups. This effect was not seen in adults. Passive avoidance performance was not affected. These results suggest that long-term exposure to alcohol during development has adverse effects on spatial learning. The lack of differences in the short-term exposure groups implies that there may not be 1 critical period of alcohol exposure, but that the adverse effects of alcohol during development may be cumulative on some behaviors.

Structure and molecular mechanism of a functional form of pneumolysin: a cholesterol-dependent cytolysin from Streptococcus pneumoniae.

One of the key steps in understanding human disease arising from gram-positive bacteria lies in the mechanisms of the cholesterol-dependent cytolysins (CDCs). Pneumolysin (PLY), a CDC from Streptococcus pneumoniae, is of special importance due to the severe impacts of pneumococcal infections on mortality and morbidity worldwide. We have overexpressed, purified, and characterized PLY in its fully functional complex form with the enzyme bound to its receptor activator on target cells, cholesterol. The circular dichroism studies of PLY in solution with an excess of cholesterol show a change in the far UV spectrum consistent with a decrease in the beta-sheet and an increase in the random coil structures of the enzyme. Pore formation in membranes leading to cell lysis is the functional target for this cytolysin. The sedimentation velocity and equilibrium analyses of the cholesterol-bound enzyme show hydrodynamic properties different from those of the cholesterol-free form. The soluble form of the cholesterol-free enzyme exists in solution as a mixture of monomers and dimers, whereas the cholesterol-bound form exists only as a monomer. A mechanism of formation of PLY pores in the lipid bilayer of the target cells is discussed.

The binding site of human adenosine deaminase for CD26/Dipeptidyl peptidase IV: the Arg142Gln mutation impairs binding to cd26 but does not cause immune deficiency.

Human, but not murine, adenosine deaminase (ADA) forms a complex with the cell membrane protein CD26/dipeptidyl peptidase IV. CD26-bound ADA has been postulated to regulate extracellular adenosine levels and to modulate the costimulatory function of CD26 on T lymphocytes. Absence of ADA-CD26 binding has been implicated in causing severe combined immunodeficiency due to ADA deficiency. Using human-mouse ADA hybrids and ADA point mutants, we have localized the amino acids critical for CD26 binding to the helical segment 126-143. Arg142 in human ADA and Gln142 in mouse ADA largely determine the capacity to bind CD26. Recombinant human ADA bearing the R142Q mutation had normal catalytic activity per molecule, but markedly impaired binding to a CD26(+) ADA-deficient human T cell line. Reduced CD26 binding was also found with ADA from red cells and T cells of a healthy individual whose only expressed ADA has the R142Q mutation. Conversely, ADA with the E217K active site mutation, the only ADA expressed by a severely immunodeficient patient, showed normal CD26 binding. These findings argue that ADA binding to CD26 is not essential for immune function in humans.

Critical periods for the effects of alcohol exposure on brain weight, body weight, activity and investigation.

Using an animal model of fetal alcohol syndrome - which equates peak blood alcohol concentrations across different developmental periods - critical periods for the effect of alcohol on brain weight, activity and investigative behavior were examined. The periods of alcohol exposure were from gestational day (GD) 1 through 10, GD 11 through 22, postnatal day (PD) 2 through 10, or all three periods combined. The critical period of alcohol exposure for an increase in activity in juveniles was GD 11 through 22. This pattern was not seen in the same animals in adulthood; instead, increases in both activity and investigation were seen in animals exposed from PD 2 through 10 and not seen in animals exposed during all three periods combined. Brain weight was reduced by alcohol exposure from GD 11 through 22, PD 2 through 10 and all three periods combined. The period from PD 2 through 10 was the only period when the brain weight to body weight ratio was reduced. In conclusion, exposure to alcohol during the periods in the latter half of gestation or early postnatal period seem to have the most deleterious effects on the brain, activity and investigation in the rat. In addition, the effects of alcohol exposure over both the prenatal and postnatal period cannot be easily predicted from the effects of shorter periods of exposure.