In vivo clonal tracking reveals evidence of haemangioblast and haematomesoblast contribution to yolk sac haematopoiesis.

During embryogenesis, haematopoietic and endothelial lineages emerge closely in time and space. It is thought that the first blood and endothelium derive from a common clonal ancestor, the haemangioblast. However, investigation of candidate haemangioblasts in vitro revealed the capacity for mesenchymal differentiation, a feature more compatible with an earlier mesodermal precursor. To date, no evidence for an in vivo haemangioblast has been discovered. Using single cell RNA-Sequencing and in vivo cellular barcoding, we have unravelled the ancestral relationships that give rise to the haematopoietic lineages of the yolk sac, the endothelium, and the mesenchyme. We show that the mesodermal derivatives of the yolk sac are produced by three distinct precursors with dual-lineage outcomes: the haemangioblast, the mesenchymoangioblast, and a previously undescribed cell type: the haematomesoblast. Between E5.5 and E7.5, this trio of precursors seeds haematopoietic, endothelial, and mesenchymal trajectories.

Can we improve our day-case rate in functional endoscopic sinus surgery?

OBJECTIVE: Day-case functional endoscopic sinus surgery is associated with increased patient satisfaction and reduced costs. This study aimed to identify reasons for overnight admission with a view to improving same-day discharge. METHODS: This was a retrospective observation study over a one-year period. All consecutive patients who underwent elective functional endoscopic sinus surgery were included. RESULTS: A total of 172 patients were included in this study. Functional endoscopic sinus surgery was planned as a day-case procedure in 152 patients (88 per cent), with a planned overnight stay in 20 (12 per cent). The rate of same-day discharge in patients who underwent elective functional endoscopic sinus surgery was 80.2 per cent (n = 138). Reasons for an unplanned overnight admission were: bleeding (n = 8), urinary retention (n = 3), medical co-morbidities (n = 1), post-operative pain (n = 1) and social reasons (n = 1). CONCLUSION: There is scope to further improve the functional endoscopic sinus surgery day-case rate by utilising techniques to minimise post-operative bleeding. This has the potential to improve both patient satisfaction and service efficiency.

Pre-placement screening for tuberculosis in healthcare workers.

BACKGROUND: Healthcare workers (HCWs) are at occupational risk of contracting and transmitting tuberculosis (TB). Despite national guidance, the optimal process for the pre-placement screening of new entrant HCWs for TB in the UK is not certain, nor the appropriateness of using a one-step interferon gamma release assay (IGRA) screening programme. AIMS: To assess the potential for an IGRA-only TB screening programme for new entrant HCWs, and identify cost savings achieved through this process. METHODS: We conducted a retrospective analysis of IGRA and tuberculin skin tests (TST) within our occupational health service over a 3-year period. HCWs with markedly discordant test results (IGRA negative, TST positive) were followed up to determine whether they developed active TB. We also estimated the yearly cost savings if the existing two-step process was replaced with an IGRA-only programme. RESULTS: Totally, 96/1258 (8%) HCWs had positive IGRA results; 788 TSTs were performed for newly screened IGRA-negative HCWs without Bacille Calmette-Guérin scars, among which 597 (76%) tested negative (TST <6 mm). None of the 10 individuals with grossly discordant test results (TST >15 mm) developed active TB during the study period. We calculated savings of £20,453 if the two-step process was replaced with an IGRA-only programme. CONCLUSIONS: The absence of disease progression in individuals with markedly discordant results in this study suggest that an IGRA-only screening programme for new HCWs in the UK is feasible, and may be safe although our follow-up period was insufficient. Our results also suggest that substantial cost savings can be made by using this programme.

The genome of the simian and human malaria parasite Plasmodium knowlesi.

Plasmodium knowlesi is an intracellular malaria parasite whose natural vertebrate host is Macaca fascicularis (the 'kra' monkey); however, it is now increasingly recognized as a significant cause of human malaria, particularly in southeast Asia. Plasmodium knowlesi was the first malaria parasite species in which antigenic variation was demonstrated, and it has a close phylogenetic relationship to Plasmodium vivax, the second most important species of human malaria parasite (reviewed in ref. 4). Despite their relatedness, there are important phenotypic differences between them, such as host blood cell preference, absence of a dormant liver stage or 'hypnozoite' in P. knowlesi, and length of the asexual cycle (reviewed in ref. 4). Here we present an analysis of the P. knowlesi (H strain, Pk1(A+) clone) nuclear genome sequence. This is the first monkey malaria parasite genome to be described, and it provides an opportunity for comparison with the recently completed P. vivax genome and other sequenced Plasmodium genomes. In contrast to other Plasmodium genomes, putative variant antigen families are dispersed throughout the genome and are associated with intrachromosomal telomere repeats. One of these families, the KIRs, contains sequences that collectively match over one-half of the host CD99 extracellular domain, which may represent an unusual form of molecular mimicry.

Porous NiTi for bone implants: a review.

NiTi foams are unique among biocompatible porous metals because of their high recovery strain (due to the shape-memory or superelastic effects) and their low stiffness facilitating integration with bone structures. To optimize NiTi foams for bone implant applications, two key areas are under active study: synthesis of foams with optimal architectures, microstructure and mechanical properties; and tailoring of biological interactions through modifications of pore surfaces. This article reviews recent research on NiTi foams for bone replacement, focusing on three specific topics: (i) surface modifications designed to create bio-inert porous NiTi surfaces with low Ni release and corrosion, as well as bioactive surfaces to enhance and accelerate biological activity; (ii) in vitro and in vivo biocompatibility studies to confirm the long-term safety of porous NiTi implants; and (iii) biological evaluations for specific applications, such as in intervertebral fusion devices and bone tissue scaffolds. Possible future directions for bio-performance and processing studies are discussed that could lead to optimized porous NiTi implants.

Binding of calcium to SP-A, a surfactant-associated protein.

SP-A is a lung-specific pulmonary surfactant-associated protein containing a calcium-dependent carbohydrate recognition domain and collagen-like sequence. The protein is a major component of the extracellular form of surfactant known as tubular myelin. SP-A is thought to influence the surface properties of surfactant lipids and regulate the turnover of extracellular surfactant through interaction with a specific cell-surface receptor. These properties of SP-A are dependent on the presence of calcium. We have estimated calcium binding parameters for SP-A from binding data obtained by equilibrium dialysis and gel permeation chromatography. Our results suggest that each SP-A monomer binds two to three calcium ions in conditions chosen as similar to those found in the alveolar lumen. The binding data are best fit to a model incorporating two calcium binding sites with different affinities. Studies with a fragment of SP-A generated by limited proteolysis suggest the higher affinity site for calcium is located in the noncollagenous carboxy-terminal end of SP-A. This region of SP-A contains a carbohydrate recognition domain homologous to other C-type lectins. The binding of calcium to this region of SP-A causes a conformational change as assessed by a small change in the intrinsic fluorescence spectrum and a marked change in the susceptibility to proteolysis. At physiological calcium concentrations, intact SP-A aggregates in a reversible fashion, a property that may be relevant to the formation of tubular myelin.

Seat height in handrim wheelchair propulsion.

To study the effect of seat height on the cardiorespiratory system and kinematics in handrim wheelchair ambulation, nine non-wheelchair users participated in a wheelchair exercise experiment on a motor-driven treadmill. The subjects conducted five progressive exercise tests. After an initial try-out test, four tests were performed at different standardized seat heights of 100, 120, 140, and 160 degrees elbow extension (subject sitting erect, hands on the rim in top-dead-center = 12.00 hrs; full extension = 180 degrees). Each test consisted of four 3-minute exercise blocks at speeds of respectively 0.55, 0.83, 1.11, and 1.39 m.s-1 (2-5 km.hr-1). Analysis of variance revealed significant effects of seat height (P less than 0.05) on gross mechanical efficiency (ME), oxygen cost, push range, and push duration, and on the ranges of motion in the different arm segments and trunk. Mean ME appeared higher at the lower seat heights of 100 and 120 degrees elbow extension. This is reflected in an enhanced oxygen consumption at seat heights of 140 and 160 degrees elbow extension. Simultaneously, the push range showed a 15 to 20 degree decrease with increasing seat height, which is reflected in a decreased push duration. In the push phase, decreases in retroflexion and abduction/adduction of the upper arm were seen. The trunk shifted further forward, and the motion range in the elbow joint shifted to extension with increasing seat height. No shifts in minimum and maximum angular velocities were seen with increasing seat height. The results showed an interrelationship between wheelchair seat height and both cardiorespiratory and kinematic parameters. With respect to the cardiorespiratory system, the optimization of the wheelchair geometry, based on functional characteristics of the user, appears beneficial.

The major lung surfactant protein, SP 28-36, is a calcium-dependent, carbohydrate-binding protein.

SP 28-36, a major protein of pulmonary surfactant, has striking amino acid sequence homology with soluble mannose-binding proteins isolated from rat liver and contains residues common to the carbohydrate-binding domains of other mammalian lectins. We have used carbohydrate-affinity chromatography to investigate carbohydrate-binding properties of SP 28-36 isolated from canine and human (alveolar proteinosis patients) lung lavage. SP 28-36 binds to immobilized D-mannose, L-fucose, D-galactose, and D-glucose. The protein binds only weakly to N-acetyl-D-galactosamine and N acetyl-D-glucosamine. Binding is Ca2+-dependent. The threshold Ca2+ concentration is 0.6 mM and maximal binding occurs with 1 mM Ca2+. Bound protein is quantitatively recovered by elution with 2 mM EDTA. Ba2+, Sr2+, and Mn2+, but not Mg2+, can substitute for Ca2+. Unlike some other mammalian lectins, SP 28-36 binds to carbohydrate at pH 5.0. Recombinant human SP 28-36 isolated from the media of Chinese hamster ovary cells, transfected with a DNA construct encoding SP 28-36, has similar carbohydrate-binding activity to the native proteins. Mannose affinity chromatography of the culture medium of Chinese hamster ovary cells results in an efficient purification of the secreted recombinant human SP 28-36.

Laboratory and on-ice test comparisons of anaerobic power of ice hockey players.

The suitability of the Wingate Anaerobic Test (WAT40) as a laboratory measure of anaerobic capacity (AnCap) and power (AnPow) of ice hockey players was tested against the Reed Repeat Sprint Skate-RSS (1979) and the Sargeant Anaerobic Skate (SAS40). Twenty-four university and Junior A players (20.2 +/- 1.6 years), assigned by random draw, performed the three tests over a seven day period. Blood lactate taken from an unwarmed finger tip was used to assess work intensity. The AnCap (7.7 +/- 0.2 Watts X kg-1) and AnPow (10.1 +/- 0.2 Watts X kg-1) for WAT40 were significantly lower (p less than 0.05) than for RSS (AnCap 9.3 +/- 0.8 Watts X kg-1; AnPow 11.5 +/- 1.1 Watts X kg-1) and SAS40 (AnCap 9.7 +/- 0.8 Watts X kg-1; AnPow 11.9 +/- 1.8 Watts X kg-1). SAS40 was significantly higher (p less than 0.05) than RSS for both AnCap and AnPow. The RSS (r = 0.96; ME 4.5%) and SAS40 (r = 0.97; ME = 3.6%) showed excellent test-retest reliability and reproducibility for AnCap but were only fair on AnPow (RSS: r = 0.73; ME = 10.7%; SAS40: r = 0.65; ME = 18.4%). While the correlations among the tests (AnCap: SAS40 vs WAT40, r = 0.73; RSS vs WAT40, r = 0.69) were significant (p less than 0.05), the highest predictive capability estimate (r2) was only 53.3%. The correlations for blood lactates (WAT40: 10.8 +/- 1.5 mmol X l-1; SAS40: 10.7 +/- 1.9 mmol X l-1; RSS: 11.5 +/- 1.6 mmol X l-1) were not significant. Based upon the particular protocol used, the laboratory test WAT40 does not demonstrate a high relationship with on-ice measures of AnCap and AnPow in this group of ice hockey players.

Role of calcium ions the structure and function of pulmonary surfactant.

Pulmonary surfactant isolated by centrifugation in buffers containing ions contains at least three different morphologic structures. The presence of one of these, tubular myelin, is dependent on calcium ions, since chelation of the calcium ions causes disruption of this structure. Addition of EDTA also decreases the ability of the surfactant to absorb rapidly to air-water interfaces and lower surface tension. Titration with calcium ions (2.5 or 5 mM) restores rapid surface adsorption and restores the tubular myelin structural forms. Magnesium ions cannot substitute for calcium ions in these processes. The reversibility of structure and function induced by calcium ions and EDTA is also accompanied by reversible isopycnic density shifts probably related to aggregation and disaggregation of the lipid-protein complex with calcium ions and EDTA, respectively.

Inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in hepatoma tissue culture cells by pure cholesterol and several cholesterol derivatives. Evidence supporting two distinct mechanisms.20l.

Pure cholesterol associated in complexes with lipoproteins (whole serum and human low density lipoproteins) or esterified with succinic acid (cholesteryl succinate) and bound to albumin effectively suppresses 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity in hepatoma tissue culture (HTC) cells grown in lipoprotein-poor serum medium during short 4-hour) incubation periods. Simultaneous measurments of the kinetics of uptake of radioactive unesterified cholesterol of whole serum and cholesteryl succinate, their conversion to lipid products, and the decay in enzyme activity, suggest that the cholesterol-induced suppression is mediated by the sterol itself rather than by inhibitory lipid products derived from its metabolism. Several cholesterol derivatives such as cholestenone, 7-ketocholesterol, and 7alpha-and 25-hydroxycholesterol also suppress reductase activiy in HTC cells and are significantly more inhibitory than the pure cholesterol preparations. The decrease in enzyme activity produced by cholesterol and its derivatives is concentration-dependent and specific. [1-14C]Oleate incorporation experiments indicate that cholesterol ester formation in HTC cells is not increased at inhibitory concentrations of the steroids. These data suggest that sterol ester formation is not an obligatory process in the feedback control of HMG-CoA reductase activity. The half-life of the reductase (3 to 4 hours) is not significantly changed by cycloheximide, plus or minus whole serum, and cholesteryl succinate. In contrast, the half-life is strongly reduced when HTC cells are incubated with cycloheximide plus maximal concentrations of 25-hydroxycholesterol, 7-ketocholesterol, or cholestenone, resulting in t1/2 values of 24, 36, and 60 min, respectively. Increasing concentrations of whole serum and cholesteryl succinate have no significant effect on the apparent rate constant of inactivation of the enzyme, whereas its apparent rate of synthesis is decreased 3- and 10-fold, respectively. These results are reversed with oxygenated steroid inhibitors. The rate of synthesis of reductase is essentially unchanged as the concentrations of 25-hydroxycholesterol, 7-ketocholesterol, and cholestenone are increased in the culture medium, whereas the apparent rate constant for degradation is increased 9-, 7-, and 3-fold, respectively. HMG-CoA reductase activity in HTC cells thus appears to be modulated by two different mechanisms in which steroid structure is important. Whole serum and cholesteryl succinate specifically decrease the rate of enzyme synthesis, while 25-hydroxycholesterol, 7-ketocholesterol, and cholestenone increase the rate of inactivation of the reductase.