The effect of hydrostatic pressure on proteoglycan production in articular cartilage in vitro: a meta-analysis.

OBJECTIVE: In previous research the use of hydrostatic pressure (HP) has been applied to enhance the formation of engineered cartilage, through the up-regulation of proteoglycan synthesis by mechanotransduction. However, the HP stimulation approach has been shown to vary between studies with a wide disparity in results, including anabolic, catabolic and non-responsive outcomes. To this end, a meta-analysis of HP publications using 3D cultured chondrocytes was performed to elucidate the key experiment factors involved in achieving a mechanotransducive response. DESIGN: The effects of different HP regimes on proteoglycan production were investigated based on the following factors: static vs dynamic application, pressure magnitude, and experiment duration. Meta-analysis was performed on raw data taken from 11 publications which employed either aggrecan gene expression analysis or dimethyl methylene blue colorimetric assay. The measure of effect was calculated based on mean difference using a random effects model. RESULTS: Analysis revealed that a significant anabolic response was most likely achieved when the following factors were employed; a static HP application, a magnitude within the mid-high physiological range of cartilage (≤5-10 MPa) and a study duration of ≥2 weeks. CONCLUSIONS: Thus, we propose that the selection of HP experiment factors can have a significant influence on engineered cartilage development, and that the results of this meta-analysis can be used as a basis for the planning of future HP experiments.

Towards an assessment for organizational participatory research health partnerships: A systematic mixed studies review with framework synthesis.

Within the health sciences, organizational participatory research (OPR) is defined as a blend of research and action, in which academic researchers partner with health organization members. OPR is based on a sound partnership between all stakeholders to improve organizational practices. However, little research on the evaluation of OPR health partnership exists. This systematic mixed studies review sought to produce a new theoretical model that structures the evaluation of the OPR processes and related outcomes of OPR health partnerships. Six bibliographic databases were searched together with grey literature sources for OPR health partnership evaluation questionnaires. Six questionnaires were included, from which a pool of 95 OPR health partnership evaluation items were derived. The included questionnaires were appraised for the quality of their origin, development and measurement properties. A framework synthesis was performed using an existing OPR framework by organizing questionnaire items in a matrix using a hybrid thematic analysis. This led to our proposed Organizational Participatory Research Evaluation Model (OPREM) that includes three axes, Trust, Collective Learning and Sustainability (with specific dimensions) and 95 items. This model provides information to help stakeholders comprehensively structure the evaluation of their partnerships and subsequent improvement; thus, potentially helping to improve health organization practices.

Ecological content validation of the Information Assessment Method for parents (IAM-parent): A mixed methods study.

This mixed methods study content validated the Information Assessment Method for parents (IAM-parent) that allows users to systematically rate and comment on online parenting information. Quantitative data and results: 22,407 IAM ratings were collected; of the initial 32 items, descriptive statistics showed that 10 had low relevance. Qualitative data and results: IAM-based comments were collected, and 20 IAM users were interviewed (maximum variation sample); the qualitative data analysis assessed the representativeness of IAM items, and identified items with problematic wording. Researchers, the program director, and Web editors integrated quantitative and qualitative results, which led to a shorter and clearer IAM-parent.

Factors associated with recurrence of varicose veins after thermal ablation: results of the recurrent veins after thermal ablation study.

BACKGROUND: The goal of this retrospective cohort study (REVATA) was to determine the site, source, and contributory factors of varicose vein recurrence after radiofrequency (RF) and laser ablation. METHODS: Seven centers enrolled patients into the study over a 1-year period. All patients underwent previous thermal ablation of the great saphenous vein (GSV), small saphenous vein (SSV), or anterior accessory great saphenous vein (AAGSV). From a specific designed study tool, the etiology of recurrence was identified. RESULTS: 2,380 patients were evaluated during this time frame. A total of 164 patients had varicose vein recurrence at a median of 3 years. GSV ablation was the initial treatment in 159 patients (RF: 33, laser: 126, 52 of these patients had either SSV or AAGSV ablation concurrently). Total or partial GSV recanalization occurred in 47 patients. New AAGSV reflux occurred in 40 patients, and new SSV reflux occurred in 24 patients. Perforator pathology was present in 64% of patients. CONCLUSION: Recurrence of varicose veins occurred at a median of 3 years after procedure. The four most important factors associated with recurrent veins included perforating veins, recanalized GSV, new AAGSV reflux, and new SSV reflux in decreasing frequency. Patients who underwent RF treatment had a statistically higher rate of recanalization than those treated with laser.

Percutaneous foam sclerotherapy for venous leg ulcers.

The technique of foam sclerotherapy directed at the distal most vessels, draining the ulcer bed was first described in 2010, with excellent penetration into the underlying venous network possible with this technique. Thirty-five patients have now been treated with this technique as the initial treatment at Midwest Vein Laser, USA. There have been no complications with this technique and rapid healing occurred within 4-8 weeks after the initial treatment in 90% of the patients, and all ulcers were healed at 4 months. Here we present the representative case of a 67-year-old man treated with a modified technique that used a percutaneous approach via reticular or spider veins at the margin of the ulcer bed.

A cell shrinkage artefact in growth plate chondrocytes with common fixative solutions: importance of fixative osmolarity for maintaining morphology.

The remarkable increase in chondrocyte volume is a major determinant in the longitudinal growth of mammalian bones. To permit a detailed morphological study of hypertrophic chondrocytes using standard histological techniques, the preservation of normal chondrocyte morphology is essential. We noticed that during fixation of growth plates with conventional fixative solutions, there was a marked morphological (shrinkage) artifact, and we postulated that this arose from the hyper-osmotic nature of these solutions. To test this, we fixed proximal tibia growth plates of 7-day-old rat bones in either (a) paraformaldehyde (PFA; 4%), (b) glutaraldehyde (GA; 2%) with PFA (2%) with ruthenium hexamine trichloride (RHT; 0.7%), (c) GA (2%) with RHT (0.7%), or (d) GA (1.3%) with RHT (0.5%) and osmolarity adjusted to a 'physiological' level of approximately 280mOsm. Using conventional histological methods, confocal microscopy, and image analysis on fluorescently-labelled fixed and living chondrocytes, we then quantified the extent of cell shrinkage and volume change. Our data showed that the high osmolarity of conventional fixatives caused a shrinkage artefact to chondrocytes. This was particularly evident when whole bones were fixed, but could be markedly reduced if bones were sagittally bisected prior to fixation. The shrinkage artefact could be avoided by adjusting the osmolarity of the fixatives to the osmotic pressure of normal extracellular fluids ( approximately 280mOsm). These results emphasize the importance of fixative osmolarity, in order to accurately preserve the normal volume/morphology of cells within tissues.

New insights into function of the growth plate: clinical observations, chondrocyte enlargement and a possible role for membrane transporters.

The mammalian growth plate is a complex structure which is essential for the elongation of long bones. However, an understanding of how the growth plate functions at the cellular level is lacking. This review, summarises the factors involved in growth-plate regulation, its failure and the consequence of injury. We also describe some of the cellular mechanisms which underpin the increase in volume of the growth-plate chondrocyte which is the major determinant of the rate and extent of bone lengthening. We show how living in situ chondrocytes can be imaged using 2-photon laser scanning microscopy to provide a quantitative analysis of their volume. This approach should give better understanding of the cellular control of bone growth in both healthy and failed growth plates.

Enamel demineralization in primary and permanent teeth.

Although enamel demineralization is important for our understanding of caries formation, no consensus has been reached regarding the possible differences in susceptibility of primary and permanent enamel. We used the constant composition (CC) technique to investigate the acid-induced demineralization of these tissues at a relative undersaturation with respect to hydroxyapatite (HAP) of 0.902, pH = 4.5, and ionic strength = 0.15 mol L(-1). The demineralization rates showed significant differences, primary enamel having the greater susceptibility to dissolution during an initial linear stage: 1.5 +/- 0.5 x 10(-10) mol mm(-2) min(-1) compared with 2.6 +/- 0.5 x 10(-11) mol mm(-2) min(-1) for permanent enamel. During the reactions, we observed nanosized crystallites which attached to the enamel surfaces or escaped into the bulk solution. These nanosized crystallites were kinetically protected against further dissolution, even though the solutions remained undersaturated. It is hypothesized that they may contribute to the remarkable mechanical and dynamic characteristics of enamel.

Chondrocyte death associated with human femoral osteochondral harvest as performed for mosaicplasty.

BACKGROUND: Autologous osteochondral transfer is an option for the treatment of articular defects. However, there are concerns about graft integration and the nature of the tissue forming the cartilage-cartilage bridge. Chondrocyte viability at graft and recipient edges is thought to be an important determinant of the quality of repair. The purpose of the present study was to evaluate early cell viability at the edges of osteochondral grafts from ex vivo human femoral condyles. METHODS: Fresh human tissue was obtained from eleven knees at the time of total knee arthroplasty for the treatment of osteoarthritis. Osteochondral cylinders were harvested with use of a 4.5-mm-diameter mosaicplasty osteotome from regions of the anterolateral aspect of the femoral condyle that were macroscopically nondegenerate and histologically nonfibrillated. Plugs were assessed for marginal cell viability by means of confocal laser scanning microscopy. RESULTS: The diameter of the cartilaginous portion of the osteochondral plugs was a mean (and standard error of the mean) of 4.84 +/- 0.12 mm (as determined on the basis of three plugs). This value was approximately 300 microm greater than the measured internal diameter of the osteotome. There was a substantial margin of superficial zone cell death (mean thickness, 382 +/- 68.2 microm), with >99% cell viability seen more centrally (as determined on the basis of five plugs). Demiplugs were created by splitting the mosaicplasty explants with a fresh number-11 scalpel blade. The margin of superficial zone cell death at the curved edge was significantly greater than that at the site of the scalpel cut (390.3 +/- 18.8 microm compared with 34.8 +/- 3.2 microm; p = 0.0286). Similar findings were observed when the cartilage alone was breached and the bone was left intact, with the margin of superficial zone cell death being significantly greater than that obtained in association with the straight scalpel incision (268 +/- 38.9 microm compared with 41.3 +/- 13.4 microm; p = 0.0286). The margin of superficial zone cell death showed no increase during the time-period between fifteen minutes and two hours after plug harvest. A mathematical approximation of the mosaicplasty region suggested that early cell death of this magnitude affects about one third of the superficial graft area. CONCLUSIONS: The results of the present study suggest that mosaicplasty, while capable of transposing viable hyaline cartilage, is associated with an extensive margin of cell death that is likely to compromise lateral integration and articular reconstruction.

Detection of human blood in the bat tick Carios (Ornithodoros) kelleyi (Acari: Argasidae) in Iowa.

The argasid tick Carios (Ornithodoros) kelleyi Cooley & Kohls is a common ectoparasite of bats and has been found in massive numbers in homes with associated bat colonies in eastern Iowa. This tick feeds nearly exclusively on bats in nature. Several inhabitants of infested homes complained of "bug bites" at night while asleep that may have resulted in erythematous, edematous, urticaric skin lesions and constitutional signs and symptoms. We provide laboratory evidence that a single, engorged C. kelleyi nymph contained host blood from a human female. The clinical implications of our findings are intriguing but unclear.

Effects of occlusal load on cervical lesions.

Toothbrush abrasion has been considered to cause cervical lesions. However, some investigators have proposed that occlusal loading factors direct tensile stresses at the cervical area, resulting in wedge-shaped abfraction defects. The purpose of this study is to investigate the role of axial and non-axial load on the development of cervical lesions. Matched paired premolars, extracted for orthodontic purposes, were used in a custom-fabricated toothbrushing apparatus. A periodontal sulcus width of 0b1 mm with 1 mm gingival recession was simulated with denture base resin. In phase 1, eight matched premolar pairs were subjected to 80 h (1b4 million strokes) of brushing and 300 g of toothbrush force. Toothpaste slurry was applied continuously through the toothbrush. One specimen in each pair was subjected to 250 h and 45 kg of continuous axial load, while the other unloaded tooth served as a negative control. In phase 2, 10 matched premolar pairs were subjected to the same conditions; however, the experimental teeth were subjected to 250 h and 45 kg of intermittent non-axial load, directed at a 45 degrees angle to the buccal cusp. Rubber impressions were made of the cervical lesions, then trimmed, weighed, and compared to determine the amount of tooth material lost. When teeth were loaded axially, there was significantly less tooth material loss (P < 0b02); however, when teeth were loaded non-axially, there was no significant difference (P =0b80) when compared with controls. Optical and scanning electron microscopy did not reveal any significant differences in the morphology between pairs. Our data suggest that the application of occlusal load may not necessarily play a significant role in the progression of cervical tooth wear commonly referred to as abfraction.

Precision application of aldicarb to enhance efficiency of thrips (Thysanoptera: Thripidae) management in cotton.

Field studies were conducted during 1999-2001 in two climatic/edaphic areas of Georgia (Southern Piedmont and East Gulf Coastal Plain) to test the hypothesis that precision placement of aldicarb with cotton seed in hill planting at spatially specific intervals could decrease insecticide use for management of tobacco thrips, Frankliniella fusca (Hinds). Precision-placed aldicarb controlled thrips during cotton seedling stages using per ha amounts of one-half or less than standard in-furrow application rates with no significant differences in yield. Residual analysis of cotton plants showed that plants in precision placement plots had as much or more aldicarb and aldicarb metabolites present as compared with cotton treated with conventional in-furrow treatments. Higher rates of precision-placed aldicarb did cause phytotoxic burning early in the growing season, but no significant impact on yield was observed.

Fibrin-type fibrinoid in placentae from pregnancies associated with maternal smoking: association with villous trophoblast and impact on intervillous porosity.

Smoking during pregnancy perturbs maternal haemostasis via activated coagulation which could include greater coagulation (fibrin-type fibrinoid deposition) in the placental intervillous space. This might affect intervillous haemodynamics and transport of oxygen and nutrients to the fetus. Fibrin deposits could influence the sizes and numbers of intervillous spaces ('pores') and perivillous fibrin could reflect changes in the nature or activity of trophoblast. Here, we test whether or not smoking is associated with differences in the composition of villous trophoblast, the amounts and patterns of fibrin and, hence, the dimensions and numbers of intervillous pores. Random samples of placentae were taken from pregnancies classified according to smoking status (non-smokers, light smokers, heavy smokers). Stereology was used to estimate volumes of intervillous space and fibrin, test for differences in trophoblast composition and patterns of fibrin deposition at the villous surface, and determine the impact of deposits on the mean volumes and theoretical numbers of intervillous pores. No group differences were found in total volumes or surfaces of trophoblast or total volume of intervillous fibrin. However, the total surfaces of syncytial knots declined in smokers and the surfaces of syncytial bridges increased. Particularly in heavy smokers, this was associated with reduced deposits of perivillous fibrin at syncytial knots. In all placentae, the greatest deposits occurred where there was trophoblast denudation. Little fibrin was seen on thin regions of syncytium. Regardless of smoking status, intervillous fibrin reduced intervillous pore size and increased pore number. However, heavy smokers had larger pores. Reductions in syncytial knots are consistent with reports that smoking reduces the incidence of trophoblast apoptosis whilst increases in syncytial bridges are consistent with enhanced branching angiogenesis. Results confirm that perivillous fibrin accumulates preferentially at denudation sites. They also suggest that smoking perturbs the normal pattern of fibrin deposition, that the impact is greater in heavy smokers and that the placental site is privileged or active in terms of fibrinolytic or anti-coagulatory activity. This activity seems to reside in thin regions of syncytium.

The volume and morphology of chondrocytes within non-degenerate and degenerate human articular cartilage.

OBJECTIVE: Cartilage swelling is an early event in osteoarthritis (OA). However, the response of chondrocytes to increased tissue hydration is unknown. This work studied the volume and morphology of living in situ human chondrocytes as a function of cartilage degeneration. METHODS: The tibial plateaus from knee joints of 40 patients were obtained following above-knee amputations or knee arthroplasty, and degree of cartilage degeneration from 0 (non-eroded) to 3 (extensive fibrillations) was assessed using several criteria. In situ chondrocytes were labeled with fluorescent indicators (calcein for living cells, propidium iodide for dead cells) permitting the quantification of volume and visualisation of morphology of cells within the cartilage zones by confocal scanning laser microscopy (CLSM). RESULTS: Chondrocyte volume within superficial and mid-zones, but not of deep zone cells, increased significantly (P<0.05 and P<0.02, respectively; one-way analysis of variance), with degree of cartilage hydration and degeneration. The volume increase ( approximately 90% for mid-zone chondrocytes, grade 3 cartilage) was greater than that which might occur following loss/excision of sub-chondral bone (<15% swelling). The CLSM technique utilised here revealed that approximately 40% of chondrocytes within all cartilage grades exhibited at least one cytoplasmic processes of <8 microm. The presence of these processes did not indicate a cell body of larger volume than cells without processes, and did not contribute to cell volume. CONCLUSIONS: The volume of in situ chondrocytes within the superficial and mid-zones increased with cartilage degeneration. Cell swelling was greater than that expected from the increased hydration in OA, suggesting that an increase in chondrocyte volume might play a role in the changes to matrix metabolism occurring in OA.

Asthma self-management: the perspective of children.

This qualitative study explored the knowledge, perceptions, and autonomy of 7- and 12-year-old children relative to the management of their asthma. A total of 32 children with moderate to severe asthma were interviewed using an open-ended drawing interview and a semi-structured interview. The triangulation of results from these two methods revealed developmental differences. Younger children identified medicines by shape, color, or lay terms, relied on adults to manage their asthma, and did not recognize warning symptoms of an attack. Older children mastered biomedical terminology and used medicines independently, although they sometimes asked for the assistance of an adult. All children perceived benefits and non-monetary costs of asthma medicines. However, they lacked understanding of the categories and role of asthma medicines. This study suggests that long-term control and quick-relief metered dose inhalers should be identifiable by consistent color-coding, and that professionals should tailor asthma education and information to children's stages of cognitive development.

The osmotic sensitivity of isolated and in situ bovine articular chondrocytes.

Articular chondrocytes experience changes to matrix hydration during both physiological (static load) and pathophysiological (osteoarthrosis, OA) conditions. Such changes should alter chondrocytes' volume, which has been shown to modify matrix metabolism. However, the osmometric behaviour of chondrocytes is not well understood. Here, using confocal laser scanning microscopy (CLSM), we have investigated the 'passive' osmotic responses of fluorescent-labelled chondrocytes within, and isolated from, the matrix. The volume-regulatory pathways normally activated by cell shrinkage/swelling, were blocked by bumetanide/REV5901, respectively. Chondrocytes in situ were broadly grouped into superficial (SZ), mid (MZ) and deep (DZ) zones, and there was a significant increase in resting cell volume with depth into the cartilage. Variation in medium osmolarity (range 0-530 mOsm; corresponding to an extracellular osmolarity of approximately 150 to approximately 600 mOsm) caused a rapid and sustained change to in situ MZ chondrocytes' volume. Over the range 180-380 mOsm, the change to in situ or isolated chondrocytes' volume was similar. For MZ chondrocytes. ideal osmometric (Boyle-van't Hoff) behaviour was apparent over the extracellular osmolarity range of approximately 250 to approximately 600 mOsm. Chondrocytes within the SZ appeared to be more sensitive to reduced osmolarity, swelling more for a given reduction in osmolarity, than MZ or DZ chondrocytes. These data show that over wide variations in osmolarity, articular chondrocytes in situ were osmotically sensitive, and for MZ chondrocytes behaved as perfect osmometers with the extracellular matrix (ECM) not restraining cell volume changes. Changes to matrix hydration may therefore alter passive chondrocytes' volume and unless compensated by volume-regulatory pathways, could lead to changes in cell volume, and hence matrix metabolism.

The role of a swelling-activated taurine transport pathway in the regulation of articular chondrocyte volume.

Swelling articular chondrocytes by reducing osmolarity stimulates a taurine transport pathway, which is implicated in regulatory volume decrease (RVD) in various cell types. The present study investigated factors controlling the activity of this pathway in chondrocytes, in particular (1) the effects of the acute (seconds) and chronic (hours) exposure of chondrocytes to anisotonic media, and (2) whether there is a role for metabolites from the arachidonic acid cascade in activating the taurine transport pathway. For in situ and isolated chondrocytes, the point at which swelling-activated [14C]taurine efflux was stimulated (the "set-point") corresponded closely to the osmolarity of the incubation medium (180, 280 or 380 mosmol/l). However, the volume of chondrocytes isolated into these media and measured by confocal microscopy was not different ( congruent with 645 microm3). Activity of the swelling-activated taurine transport pathway was inhibited by REV5901 (an inhibitor of steps of the arachidonic acid cascade; K0.5 8+/-4 microM), NDGA (a general lipoxygenase inhibitor; K0.5 28+/-5 microM), or MK886 (an inhibitor of the 5-lipoxygenase-activating protein; 91% inhibition at 10 microM), but weakly by the more potent 5-lipoxygenase inhibitor REV5901 para (K0.5 350+/-100 microM). Addition of the leukotriene (LT) B4 or D4 receptor antagonists, CP-105,696 and L660,711 respectively, or of the leukotrienes LTB4, LTC4, LTD4 and LTE4 or lipoxins (hepoxylin A3 or B3) had no effect on the activity of the pathway in isotonic or hypotonic media. The role of the pathway in RVD was determined in isolated calcein-loaded chondrocytes using fluorescence imaging. RVD was observed and inhibited by REV5901 (50 microM) and by NDGA (75 microM). The data show that despite chronic exposure of chondrocytes to anisotonic media, the cells maintain a pre-determined volume that is the "set-point" for the activation of the taurine transport pathway following acute hypotonic challenge. This pathway appears to play a role in chondrocyte RVD, but its activation does not involve metabolites of the arachidonic acid cascade.

Electronic and structural variation among copper(II) complexes with substituted phenanthrolines.

A series of copper(II) complexes with substituted phenanthroline ligands has been synthesized and characterized electronically and structurally. The compounds that have been prepared include the monosubstituted ligand complexes of the general formula [Cu(5-R-phen)(2)(CH(3)CN)](BF(4))(2), where R = NO(2), Cl, H, or Me, and the disubstituted ligand complex [Cu(5,6-Me(2)-phen)(2)(CH(3)CN)](BF(4))(2). The complexes [Cu(5-NO(2)-phen)(2)(CH(3)CN](BF(4))(2) (1), [Cu(5-Cl-phen)(2)(CH(3)CN)](BF(4))(2) (2), [Cu(o-phen)(2)(CH(3)CN)](BF(4))(2) (3), and [Cu(5-Me-phen)(2)(CH(3)CN)](BF(4))(2) (4) each crystallize in the space group C2/c with compounds 1, 2, and 4 comprising an isomorphous set. The disubstituted complex [Cu(5,6-Me(2)-phen)(2)(CH(3)CN)](BF(4))(2) (5) crystallizes in the space group P2(1)/c. Each structure is characterized by a distorted trigonal bipyramidal arrangement of ligands around the central copper atom with approximate or exact C(2) symmetry. The progression from electron-withdrawing to electron-donating substituents on the phenanthroline ligands correlates with less accessible reduction potentials for the bis-chelate complexes.

A comparison of the wear resistance and hardness of indirect composite resins.

STATEMENT OF PROBLEM: Various new, second-generation indirect composites have been developed with claimed advantages over existing tooth-colored restorative materials. To date, little independent research has been published on these materials, and the properties specified in the advertising materials are largely derived from in-house or contracted testing. PURPOSE: Four second-generation indirect composites (Artglass, belleGlass, Sculpture, and Targis) were tested for wear resistance and hardness against 2 control materials with well-documented clinical application. Human enamel was also tested for comparison. MATERIAL AND METHODS: Twelve specimens of each material were fabricated according to the manufacturers' directions and subjected to accelerated wear in a 3-body abrasion, toothbrushing apparatus. Vickers hardness was measured for each of the tested materials, and energy dispersive x-ray (EDX) spectroscopy was performed to determine the elemental composition of the composite fillers. The statistical tests used for wear and hardness were the Kruskal-Wallis 1-way ANOVA test with Mann-Whitney tests and 1-way ANOVA with multiple comparisons (Tukey HSD). The Pearson correlation coefficient was used to determine the existence of a relationship between the hardness of the materials and the degree to which they had worn. The level of statistical significance chosen was alpha=.05. RESULTS: The control material Concept was superior to the other composites in wear resistance and hardness and had the lowest surface roughness. Significant relationships were observed between depth of wear and hardness and between depth of wear and average surface roughness. Enamel specimens were harder and more wear resistant than any of the composites. EDX spectroscopy revealed that the elemental composition of the fillers of the 4 new composites was almost identical, as was the composition of the 2 control composites. CONCLUSION: The differences in wear, hardness, and average surface roughness may have been due to differences in the chemistry or method of polymerization of the composites. Further research in this area should be encouraged. It was also apparent that the filler present in the tested composites did not exactly fit the manufacturers' descriptions.