Long-term effects of graduated compression stockings on cardiorespiratory performance

The use of graduated compression stockings (GCS) in sport has been increasing in the last years due to their potential positive effects for athletes. However, there is little evidence to support whether these types of garments actually improve cardiorespiratory performance. The aim of this study was to examine the cardiorespiratory responses of GCS during running after three weeks of regular use. Twenty recreational runners performed three tests on different days: test 1) – a 5-min maximal effort run in order to determine the participants’ maximal aerobic speed; and tests 2) and 3) – a fatigue running test of 30 minutes at 80% of their maximal aerobic speed with either GCS or PLACEBO stockings at random. Cardiorespiratory parameters (minute ventilation, heart rate, relative oxygen consumption, relative carbon dioxide production, ventilatory equivalents for oxygen and carbon dioxide, and oxygen pulse) were measured. Before each test in the laboratory, the participants trained with the randomly assigned stockings (GCS or PLACEBO) for three weeks. No significant differences between GCS and PLACEBO were found in any of the cardiorespiratory parameters. In conclusion, the present study provides evidence that running with GCS for three weeks does not influence cardiorespiratory parameters in recreational runners.     

Variability in host plant resistance of Sorghum to Striga Hermonthica infestation in West Africa

Fourteen elite sorghum lines were evaluated for their resistance to Striga hermonthica at three locations in Nigeria and Mali. Results showed that many of the lines especially MALISOR 84-1, SAMSORG 41, 97-SB-F5DT-64 (Keninkédié) and the check SRN 39 remained resistant to Striga in all locations with low emerged Striga counts, while SAMSORG 14 had the highest Striga infestation in all locations. Considerable variation in reaction to Striga infestation was observed on Séguètana, 97-SB-F5DT-63 (Wasa), 97-SB-F5DT-65, CMDT 38, CMDT 39 and CMDT 45 which were susceptible to Striga at Samaru, Nigeria but were resistant to Striga at both locations in Mali. Based on low Striga resistance and high grain yield, lines MALISOR 84-1, SAMSORG 41, 97-SB-F5DT-64, 97-SB-F5DT-65, CMDT 39 and SAMSORT 14 have been nominated for wider evaluation across more West African countries.     

Synthesis and in vitro hydroxyapatite binding of peptides conjugated to calcium-binding moieties

To confer bone-binding properties to proteins and other biological agents that lack specific targeting capacity, model peptide-based molecules were synthesized containing poly(aspartic acid), poly(glutamic acid), or a bisphosphonate (pamidronate). These motifs have well-documented affinities to hydroxyapatite, a property desirable for the targeting of molecules to bone for drug delivery and tissue engineering applications. Model peptides of increasing molecular mass (5-33 amino acids) were directly conjugated to eight aspartic acids (Asp8), eight glutamic acids (Glu8), or pamidronate, purified by high-performance liquid chromatography, and characterized by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectroscopy. The modified peptides were incubated with hydroxyapatite in phosphate-buffered saline at physiological conditions over 24 h. This study revealed a significant amount (>90%) of conjugated peptides adsorbed to the hydroxyapatite as compared to unmodified peptides (<5%). It was found that while there were significant differences between the different hydroxyapatite-binding and control groups for all time points, the size of the peptide had no statistical effect on peptide-hydroxyapatite binding. These results demonstrate that bisphosphonate and oligopeptide conjugates hold great promise for the development of new bioactive molecules for bone-specific applications.     

Synthesis and characterization of injectable, thermally and chemically gelable, amphiphilic poly(N-isopropylacrylamide)-based macromers

In this study, we synthesized and characterized a series of macromers based on poly( N-isopropylacrylamide) that undergo thermally induced physical gelation and, following chemical modification, can be chemically cross-linked. Macromers with number average molecular weights typically ranging from 2000-3500 Da were synthesized via free radical polymerization from, in addition to N-isopropylacrylamide, pentaerythritol diacrylate monostearate, a bifunctional monomer containing a long hydrophobic chain, acrylamide, a hydrophilic monomer, and hydroxyethyl acrylate, a hydrophilic monomer used to provide hydroxyl groups for further chemical modification. Results indicated that the hydrophobic-hydrophilic balance achieved by varying the relative concentrations of comonomers used during synthesis was an important parameter in controlling the transition temperature of the macromers in solution and stability of the resultant gels. Storage moduli of the macromers increased over 4 orders of magnitude once gelation occurred above the transition temperature. Furthermore, chemical cross-linking of these macromers resulted in gels with increased stability compared to uncross-linked controls. These results demonstrate the feasibility of synthesizing poly( N-isopropylacrylamide)-based macromers that undergo tandem gelation and establish key criteria relating to the transition temperature and stability of these materials. The data suggest that these materials may be attractive substrates for tissue engineering and cellular delivery applications as the combination of mechanistically independent gelation techniques used in tandem may offer superior materials with regard to gelation kinetics and stability.     

Synthesis and conformational evaluation of a novel gene delivery vector for human mesenchymal stem cells

We have synthesized a novel gene delivery vector by covalently combining branched polyethylenimine (bPEI) and hyaluronic acid (HA) with the aim of improving transfection of bPEI into human mesenchymal stem cells (hMSCs) while maintaining cell viability. Because of the opposite charges on bPEI and HA, the bPEI-HA vector forms a zwitterionic polymer capable of inter- and intramolecular interactions. We have characterized the hydrodynamic radius of bPEI-HA and bPEI-HA/DNA complexes at ambient and physiological temperatures, as well as at a range of salt concentrations using light scattering, and investigated the effect of the size of transfecting complexes on gene delivery. We found that by increasing the salt concentration from 150 to 1000 mM of NaCl, the mean hydrodynamic radius (R(h)) of bPEI-HA increases from 2.0 +/- 1.1 to 366.0 +/- 149.0 nm. However, increasing the salt concentration decreases the mean R(h) of bPEI-HA/DNA complexes from 595.0 +/- 44.6 to 106.0 +/- 19.2 nm at 25 degrees C and from 767.0 +/- 137.2 to 74.0 +/- 23.0 nm at 37 degrees C. hMSCs transfected with smaller complexes showed a significant increase in transfection from 3.8 +/- 1.5% to 19.1 +/- 4.4%. Similarly, bPEI-HA performed significantly better than bPEI in terms of cell viability (86.0 +/- 6.7% with bPEI-HA versus 7.0 +/- 2.8% with bPEI, 24 h post exposure at the highest concentration of 500 mg/mL) and maximum transfection efficiencies (12.0 +/- 4.2% with bPEI/DNA complexes and 33.6 +/- 13.9% with bPEI-HA/DNA complexes). Thus, modifying bPEI by covalent conjugation with HA improves its performance as a gene delivery vector in hMSCs. This presents a promising approach to altering hMSCs for tissue engineering and other applications.     

Two common nonsynonymous paraoxonase 1 (<Emphasis Type="Italic">PON1</Emphasis>) gene polymorphisms and brain astrocytoma and meningioma

Background  

Human serum paraoxonase 1 (PON1) plays a major role in the metabolism of several organophosphorus compounds. The enzyme is encoded by the polymorphic gene PON1, located on chromosome 7q21.3. Aiming to identify genetic variations related to the risk of developing brain tumors, we investigated the putative association between common nonsynonymous PON1 polymorphisms and the risk of developing astrocytoma and meningioma.     

Synthesis of in situ cross-linkable macroporous biodegradable poly(propylene fumarate-co-ethylene glycol) hydrogels

This study describes a synthesis method of biodegradable macroporous hydrogels suitable as in situ cross-linkable biomaterials. Macroporous hydrogels were based on poly(propylene fumarate-co-ethylene glycol) and prepared via coupled free radical and pore formation reactions. Cross-linking was initiated by a pair of redox initiators, ammonium persulfate and L-ascorbic acid. Pores were formed by the reaction between L-ascorbic acid and sodium bicarbonate, a basic component, which evolved carbon dioxide. Sol fraction of the hydrogels was varied from 0.06 +/- 0.01 to 0.64 +/- 0.01. A stereological approach was used to analyze the morphological properties of the macroporous hydrogels by relating the morphological properties of thin sections to the original three-dimensional macroporous hydrogel. Prepared macroporous hydrogels had porosities between 0.43 +/- 0.08 and 0.84 +/- 0.02 and surface area densities between 55 +/- 3 and 108 +/- 7 cm(-1). Sodium bicarbonate concentration had the greatest effect on both the porosity and surface area density. The effect of copolymer formulation on the porosity and surface area density was insignificant. From thin sections of the macroporous hydrogels, the profile size distributions were determined as an estimate of the pore size distribution. Two formulations synthesized with varying L-ascorbic acid concentration of 0.05 and 0.1 M had median profile sizes of 50-100 and 150-200 microm, respectively. This novel synthesis method allows for the in situ cross-linking of biodegradable macroporous hydrogels with morpholological properties suitable for consideration as an injectable tissue engineering scaffold.     

An optimized method for the chemiluminescent detection of alkaline phosphatase levels during osteodifferentiation by bone morphogenetic protein 2

Differentiation of osteoprogenitor cells into osteoblasts is a pivotal step during the normal development and repair of bone. Upregulation of endogenous cellular alkaline phosphatase activity (AP) is a commonly used intracellular marker for the assessment of osteoprogenitor cell differentiation into the osteoblastic phenotype. Current methods for assaying AP involve colorimetric detection of the enzyme's activity using the synthetic substrate p-nitrophenol phosphate. In this paper, we explored an alternative method of detecting AP using the chemiluminescent substrate disodium 3-(4-methoxyspiro[1,2-dioxetane-3,2'-(5'-chloro)tricyclo[3.3.1.1(3,7)]decan]-4-yl) phenyl phosphate (CSPD) for enhanced AP sensitivity and a more simplified assay. Using calf intestinal alkaline phosphatase as a standardizing enzyme, we determined that the chemiluminescent detection system was four orders of magnitude more sensitive than the standard colorimetric method of detection. Moreover, the chemiluminescent assay was faster and markedly simpler to perform. To maximize the utility of this assay system, two osteoprogenitor cell lines were compared for their ability to generate alkaline phosphatases in vitro when exposed to recombinant human bone morphogenetic protein (rhBMP-2). The W20-17 cell line was substantially more sensitive to rhBMP-2 than the C3H10T1/2 cell line, where each cell line produced detectable increases in AP after exposure to rhBMP-2 levels of 5 and 25 ng/ml, respectively. The experimental design for AP responsiveness to rhBMP-2 was further optimized for chemiluminescent detection with the W20-17 cell line by comparing the effects of reporter cell seeding density and the day of assay. In summary, the data presented in this paper demonstrate a faster, simpler, and more sensitive chemiluminescent method to monitor changes in AP levels during osteodifferentiation.     

Injectable nanocomposites of single-walled carbon nanotubes and biodegradable polymers for bone tissue engineering

We have investigated the dispersion of single-walled carbon nanotubes (SWNTs) and functionalized SWNTs (F-SWNTs) in the unsaturated, biodegradable polymer poly(propylene fumarate) (PPF) and examined the rheological properties of un-cross-linked nanocomposite formulations as well as the electrical and mechanical properties of cross-linked nanocomposites. F-SWNTs were produced from individual SWNTs by a diazonium-based method and dispersed better than unmodified SWNTs in both un-cross-linked and cross-linked PPF matrix. Cross-linked nanocomposites with F-SWNTs were superior to those with unmodified SWNTs in terms of their mechanical properties. Specifically, nanocomposites with 0.1 wt % F-SWNTs loading resulted in a 3-fold increase in both compressive modulus and flexural modulus and a 2-fold increase in both compressive offset yield strength and flexural strength when compared to pure PPF networks, whereas the use of 0.1 wt % SWNTs gained less than 37% mechanical reinforcement. These extraordinary mechanical enhancements considered together with Raman scattering and sol fraction measurements indicate strong SWNT-PPF interactions and increased cross-linking densities resulting in effective load transfer. With enhanced mechanical properties and capabilities of in situ injection and cross-linking, these SWNT/polymer nanocomposites hold significant implications for the fabrication of bone tissue engineering scaffolds.     

Shell variation in sympatric freshwater Lymnaea peregra and L. ovata (Gastropoda: Lymnaeidae)

Snails of the genus Lymnaea are morphologically variable and their taxonomy is unclear. In particular, the forms peregra and ovata , distinguished by shell shape, are often considered variants of the same species, L. peregra. We studied a rare situation in a Swiss mountain lake where both forms are sympatric. First, we found that the forms shows complete separation for a number of allozymes. Second, we examined the response of the two snail forms to infection by the trematode Diplostomum phoxini . For the ovata form, there was a transitory, 10% increase in the growth of infected snails compared to uninfected controls. The peregra form showed no gigantism and had higher parasite-induced mortality. Third, we assessed differences between the forms in reproduction under different environmental conditions. Density negatively influenced egg production to the same degree in both forms. However, decreasing water levels, characteristic of part of the lake studied, led to a 30% decrease in egg production for the ovata form, but only a 10% decrease for the peregra form. These differences are discussed in relation to the microdistributions of the snails in the lake and we conclude that the two forms are almost certainly separate species.