Concentration-dependent, temperature-dependent non-Newtonian viscosity of lung surfactant dispersions

The bulk shear viscosities of aqueous dispersions of lavaged calf lung surfactant (LS) and its chloroform:methanol extract (CLSE) were measured as a function of concentration, shear rate and temperature. At 10-mg phospholipid per milliliter, dispersions of LS and vortexed CLSE in 0.15 M NaCl (saline) had low viscosities near 1 cp over a range of shear rates from 225 to 1125 s(-1). Lung surfactant viscosity increased with phospholipid concentration and became strongly non-Newtonian with higher values at low shear rates. At 37 degrees C and 40 mg/ml, LS and vortexed CLSE in saline had viscosities of 38 and 34 cp (77 s(-1)) and 12 and 7 cp (770 s(-1)), respectively. Viscosity values for LS and CLSE were dependent on temperature and, at fixed shear, were lower at 23 degrees C than at 37 or 10 degrees C. Hysteresis was also present in viscosity measurements depending on whether shear rate was successively increased or decreased during study. Addition of 5 mM Ca(2+) at 37 degrees C markedly reduced CLSE viscosity at all shear rates and decreased LS viscosity at low shear rates. Dispersion by sonication rather than vortexing increased the viscosity of CLSE at fixed shear, while synthetic phospholipids dispersed by either method had low, relatively Newtonian viscosities. The complex viscous behavior of dispersions of LS and CLSE in saline results from their heterogeneous aggregated microstructure of phospholipids and apoproteins. Viscosity is influenced not only by the aggregate surface area under shear, but also by phospholipid-apoprotein interactions and aggregate structure/deformability. Similar complexities likely affect the viscosities of biologically-derived exogenous surfactant preparations administered to patients in clinical surfactant therapy.     

Enhancement of the viscosity of mucin by serum albumin.

The interaction of serum albumin with a model epithelial mucin from pig stomach was explored by rotary viscometry. During 30 min of incubation of human serum albumin(20mg/ml) and pig gastric mucin (8mg/ml) in iso-osmotic buffers at 37 degrees C, the solution became markedly viscous. Viscosity enhancement was proportional to albumin concentration (2-40mg/ml), was most pronounced under conditions of low shear rate (less than 45S-1), and was considerably greater than the additive or multiplicative viscosity values calculated from albumin or mucin solutions measured separately. The viscous mucin-albumin complex was destroyed by high shear rates (greater than 90S-1), but slowly re-formed under zero shear conditions. Elevation of pH (7 to 9), ionic strength (0.1 to 1.0), and addition of disodium EDTA (5mM) did not cause marked or specific alterations in the viscosity of the mixture, suggesting that electrostatic interactions probably do not stabilize mucin-albumin complexes. Urea (7M) and heating (35 to 55 degrees C) caused a major increase in the viscosity of mucin and mucin-albumin mixtures, suggesting that rupture of hydrogen bonds, unfolding and partial denaturation of mucin promotes greater intertangling (possibly hydrophobic interactions) between mucin and albumin molecules. The implications of mucin-albumin interaction in diseases associated with mucus obstruction are briefly discussed.     

Action of shear on enzymes: studies with catalase and urease

Intermittent shear was applied to approximately 1 mg/ml solutions of bovine liver catalase in a coaxial cylindrical viscometer at temperatures from 20 to 60°C and shear rates up to 683 sec^?1. The viscometer was sealed to prevent evaporation. Up to 40°C there were no activity losses during 3 hr total shearing. Above 40°C shearing reduced losses due to thermal inactivation, possibly by interfering with precipitation. At 3440 sec^?1 and 40°C fine precipitates formed but little activity was lost. No activity losses were found with experimental conditions under which Taylor vortexing occurred, nor when shear stresses were increased up 57 times by adding glycerol to raise the, viscosity. There were no significant losses in a capillary rheometer at shear rates up to 10⁶ sec^?1. When low concentration (6 μg/ml) catalase solutions were sheared there was little loss in sealed systems, but there were losses in “open” systems even in low-temperature nonshear experiments. Although there were no losses with 1 mg/ml solutions, 6 μg/ml catalase solutions from an alternative source did lose activity in sealed systems but much less than expected from previously published work. Approximately 1 mg/ml jack bean urease solutions were sheared in the sealed system at 23°C and 683 sec^?1 for 3 hr. No losses were found. No evidence of temporary or permanent inactivation was found with 28°g/ml solutions sheared in the presence of urea. Shear forces alone were not found to be as effective in causing enzyme inactivation as is generally believed and alternative mechanisms for damage are discussed.     

犬脾切除对阻断冠脉后血液流变学变化的影响

本实验旨在观察切除脾脏对犬心肌缺血时血液流变学变化的影响。结果表明,切除脾脏后,阻断冠脉血流120min内血细胞比容与高切变率下全血粘度升高程度明显减轻;心肌缺血40min和80min时低切变率下全血粘度升高程度减轻;但对心肌缺血120min时低切变率下全血粘度变化及血浆纤维蛋白原浓度、血浆粘度变化均无明显影响。     

Viscosity and elasticity during collagen assembly in vitro: relevance to matrix-driven translocation

In order to better understand the gelation process associated with collagen assembly, and the mechanism of the in vitro morphogenetic phenomenon of "matrix-driven translocation" [S.A. Newman et al. (1985) Science, 228, 885-889], the viscosity and elastic modulus of assembling collagen matrices in the presence and absence of polystyrene latex beads was investigated. Viscosity measurements at very low shear rates (0.016-0.0549 s(-1)) were performed over a range of temperatures (6.9-11.5 degrees C) in a Couette viscometer. A magnetic levitation sphere rheometer was used to measure the shear elastic modulus of the assembling matrices during the late phase of the gelation process. Gelation was detected by the rapid increase in viscosity that occurred after a lag time tL that varied between O and approximately 500 s. After a rise in viscosity that occurred over an additional approximately 500 s, the collagen matrix was characterized by an elastic modulus of the order of several Pa. The lag time of the assembly process was relatively insensitive to differences in shear rate within the variability of the sample preparation, but was inversely proportional to the time the sample spent on ice before being raised to the test temperature, for test temperatures > 9 degrees C. This suggests that structures important for fibrillogenesis are capable of forming at 0 degrees C. The time dependence of the gelation process is well-described by an exponential law with a rate constant K approximately 0.1 s(-1). Significantly, K was consistently larger in collagen preparations that contained cell-sized polystyrene beads. From these results, along with prior information on effective surface tension differences of bead-containing and bead-lacking collagen matrices, we conclude that changes in matrix organization contributing to matrix-driven translocation are initiated during the lag phase of fibrillogenesis when the viscosity is < or = 0.1 Poise. The phenomenon may make use of small differentials in viscosity and/or elasticity, resulting from the interaction of the beads with the assembling matrix. These properties are well described by standard models of concentrated solutions.     

Molecular weight distribution and solution properties of silk fibroins with different dissolution conditions

Four regenerated silk fibroin (SF) samples were prepared under different dissolution conditions and their molecular weight (MW) distributions and solution properties in water and formic acid were examined. SFL, produced by dissolving in LiBr aqueous solution for 6h, showed the highest MW level. In the three SFC samples, produced by dissolving SF in CaCl(2)/H(2)O/EtOH solution for dissolution times ranging from 3 to 180 min, the MW of the SFs decreased with increasing dissolution time and a new band appeared at low MW. Interestingly, SFL presented as a relatively transparent aqueous solution with 10-30 nm particle size, whereas the three SFC samples exhibited a turbid solution with 100-300 nm particle size. SF formic acid solutions showed a higher viscosity than SF aqueous solutions and exhibited almost Newtonian fluid behavior, whereas SF aqueous solutions displayed abrupt shear thinning in the low shear rate region (0.1-3 s(-1)).     

Disruption of leaves and initial extraction of wildtype CPMV virus as a basis for producing vaccines from plants.

Wildtype cowpea mosaic virus (CPMV) was extracted from fresh and frozen plant material by methods suitable for large-scale application. Deep freezing, crushing, and thawing in water or buffers gave 0.6+/-0.2 mg g(-1) of virus after up to 24 h. Release from sliced fresh leaves was lower at 0.14+/-0.03 mg g(-1). Homogenisation of frozen leaves for 1 min increased yield to a maximum, on average of 3.5 mg g(-1) but varying between batches from 2.2 to 4.8 mg g(-1) virus Long term storage at -80 degrees C increased subsequent yield by 2 mg g(-1) per year on average; the maximum was 10.4+/-1.9 mg g(-1) (665 days storage). Within a batch, similar yields were obtained between individual fresh plants, and from frozen versus fresh leaves. After homogenisation for 1 min, 90% of debris particles were smaller than 12 microm, half under 5 microm and 10% less than 1 microm. Homogenate (4% dry weight) was rheologically complex, exhibiting shear thinning with hysteresis at low shear rates which bears on subsequent processing. At shear rates above 200 s(-1), its apparent viscosity was 0.02 N s m(-2).     

Quantitative analysis of the effects of hyaluronan and aggrecan concentration and hyaluronan size on the elasticity of hyaluronan-aggrecan solutions

We determined elasticity (G') and viscosity (G') of various aggrecan-hyaluronan solutions using a controlled-stress rheometer with high (10 Hz) to low (0.1 Hz) frequencies. Aggrecan solution (50 mg/ml) alone showed little elasticity at any frequency, but the addition of 3300 kDa hyaluronan at 0.001-0.1 mg/ml markedly increased the elasticity, but not the viscosity, at all frequencies. Increasing hyaluronan concentration at >0.1 mg/ml did not further increase the elasticity of the aggrecan solution, and the elasticity of the aggrecan-hyaluronan complex solution reached a plateau at a 500:1 (w/w) ratio. In studies with increasing concentrations of aggrecan and a constant concentration (0.5 mg/ml) of 3300 kDa hyaluronan, aggrecan induced elasticity only at >20 mg/ml, indicating the presence of a critical concentration for elasticity. In the presence of 50 mg/ml aggrecan, 1000 kDa hyaluronan had far less effect on the elasticity of the aggrecan solution than did 3300 kDa hyaluronan. These findings suggest that only approximately 50% reduction in aggrecan concentration (<20 mg/ml), or reduced hyaluronan size (<1000 kDa)--compared with their physiological levels in young cartilage--can abolish the elastic network of the aggrecan-hyaluronan complex.     

Measuring the force production of the hormogonia of Mastigocladus laminosus

The cyanobacterium Mastigocladus laminosus forms hormogonia, which glide slowly away from the parent colony by extruding slime out of nozzles. Using video microscopy, we observed hormogonia embedded in and moving through 1-4% agar solutions with an average velocity of 0.5 microm/s. Agar is non-Newtonian and is subject to shear-thinning so that its viscosity greatly increases at low shear rates. We measured the viscosity of these agar solutions at the very low shear rates appropriate for gliding hormogonia and found it to vary from 1 to 52 million centipoise. Then, by applying a Newtonian drag coefficient for a 100-microm-long, cigar-shaped hormogonium, we found that it produced a force of several million pN. A typical hormogonium has 10-100 thousand 9-nm-wide slime extrusion nozzles. Wolgemuth et al. have proposed hydration-driven swelling of the polyelectrolyte slime ejected from these nozzles as the force production mechanism, and our experiment found a large nozzle force that was consistent with this hypothesis. Average single nozzle force depended on viscosity, being large when the viscosity was high: 71 pN in 3% and 126 pN in 4% agar.     

Viscoelastic properties of proteoglycan subunits and aggregates in varying solution concentrations

Using a cone-on-plate mechanical spectrometer, we have measured the linear and non-linear rheological properties of cartilage proteoglycan solutions at concentrations similar to those found in situ. Solutions of bovine nasal cartilage proteoglycan subunits (22S) and aggregates (79S) were studied at concentrations ranging from 10 to 50 mg ml-1. We determined: (1) the complex viscoelastic shear modulus G (omega) under small amplitude (0.02 radians) oscillatory excitation at frequencies (omega) ranging from 1.0 to 20.0 Hz, (2) the non-linear shear rate (gamma) dependent apparent viscosity napp (gamma) in continuous shear, and (3) the non-linear shear rate dependent primary normal stress difference sigma 1 (gamma) in continuous shear. Both the apparent viscosity and normal stress difference were measured over four decades of shear rates ranging from 0.25 to 250 s-1. Analysis of the experimental results were performed using a variety of materially objective non-linear viscoelastic constitutive laws. We found that the non-linear, four-coefficient Oldroyd rate-type model was most effective for describing the measured flow characteristics of proteoglycan subunit and aggregate solutions. Values of the relaxation time lambda 1, retardation time lambda 2, zero shear viscosity no, and nonlinear viscosity parameter muo were computed for the aggregate and subunit solutions at all of the solute concentrations used. The four independent material coefficients showed marked dependence on the two different molecular conformations, i.e. aggregate or subunit, of proteoglycans in solution.     

Systemic and renal hemodynamics after moderate hemodilution with HbOCs in anesthetized rabbits

Hb-based O(2)-carrying solutions (HbOCs) have been developed as red blood cell substitutes for use in patients undergoing hemodilution. Variously modified Hb with diverse solution properties have been shown to produce variable hemodynamic responses. We examined, through pulsed-Doppler velocimetry, the systemic and renal hemodynamic effects of dextran-benzene-tetracarboxylate-conjugated (Hb-Dex-BTC), bis(3,5-dibromosalicyl)fumarate cross-linked (alphaalpha-Hb), and o-raffinose-polymerized (o-raffinose-Hb) Hb perfused in rabbits after moderate hemodilution (30% hematocrit), and we compared the effects of these Hb solutions with the effects elicited by plasma volume expanders. In addition, vascular hindrance (resistance/blood viscosity at 128.5 s(-1)) was calculated to determine whether a moderate decrease in the viscosity of blood mixed with HbOCs may impair vasoconstriction as a result of autoregulation after infusion of cell-free Hb. No changes were observed in renal hemodynamics after hemodilution with reference or Hb solutions. Increase in blood pressure and vascular resistance was found with Hb-Dex-BTC and alphaalpha-Hb (for 180 min) and, to a lesser extent, with o-raffinose-Hb (for 120 min). Furthermore, Hb-Dex-BTC (high viscosity) and o-raffinose-Hb (medium viscosity) induced comparable increases in vascular hindrance (from 0.091 to 0. 159 and from 0.092 to 0.162 cm(-1), respectively) but far less than that produced by alphaalpha-Hb (low viscosity, from 0.092 to 0.200 cm(-1)). These results suggest that maintaining the viscosity of blood by infusing solutions with high viscosity makes it possible to limit vasoconstriction due to autoregulation mechanisms and mainly caused by hemodilution per se.     

Use of a gel-forming dipeptide derivative as a carrier for antigen presentation

A dipeptide of the formula Fmoc-Leu-Asp and some other related dipeptides were synthesized in solution by standard methods. When such peptides are dissolved in water at concentrations below 1% at 100 °C and cooled below 60 °C they form turbid solutions and eventaully visocelastic gels at lower temperatures. Such gels are thermoreversible and can also be disrupted by mechanical agitation. At a concentration of 2 mg/ml the peptide Fmoc-Leu-Asp forms an aqueous gel at 60 °C with a shear modulus of 80 Pa measured at a frequency of 1 rad/s. Peptide solutions undergo an abrupt increase in light scattering between 1 and 1.5 mg/ml at both 23 and 60 °C. By analogy with previous observations of other systems, this increse appears to be due to the formation of filamentous micelles and the aggregation of filamaents into a three-dimensional network. When low molecular weight adamantanamine derivatives, which are inherently non-antigenci antiviral drugs, were incorporated into the Fmoc-Leu-Asp gel and injected into rabbits, high titre specific antibodies were efficiently produced without the need for additional adjuvant. Both the physical properties of the gel and its effect on the antigenicity of low molecular weight compounds suggest a number of practical applications.     

Rotational diffusion of TEMPONE in the cytoplasm of Chinese hamster lung cells.

The correlation time for rotational diffusion (tau R) of 2,2,6,6-tetramethyl-4-piperidone-N-oxide (TEMPONE) in Chinese hamster lung (V79) cells has been measured. For these cells in an isosmotic solution at 20 degrees C, tau R = 4.18 X 10(-11) s, approximately 3.6 times greater than tau R = 1.17 X 10(-11) s in water. The relationship between tau R and viscosity was investigated in a number of glycerol-water (0-50%) and sucrose-water (20-40%) solutions and a constant Stokes-Einstein volume of 44 A3 was found for TEMPONE in solutions of less than 20% glycerol and sucrose. This gives an average shear viscosity (for rotation of a small molecule) of 0.038 poise for the cytoplasm. When nonsecular terms were used in the calculation of tau R, the activation energies for rotation of TEMPONE in the above solutions correlated well with the activation energies for shear viscosity. The viscosity increases as the cell is shrunk in hypertonic solutions. It also increases with decreasing temperature with an activation energy of 3.7 kcal/mol, about the same as the activation energy for the viscosity of pure water. The rotational correlation times were carefully calculated considering inhomogeneous line broadening, non-Lorentzian line shapes, the need for accurate tensor values and nonsecular terms.     

Chondroitin sulfate reduces the friction coefficient of articular cartilage

The objective of this study was to investigate the effect of chondroitin sulfate (CS)-C on the frictional response of bovine articular cartilage. The main hypothesis is that CS decreases the friction coefficient of articular cartilage. Corollary hypotheses are that viscosity and osmotic pressure are not the mechanisms that mediate the reduction in the friction coefficient by CS. In Experiment 1, bovine articular cartilage samples (n=29) were tested in either phosphate buffered saline (PBS) or in PBS containing 100mg/ml of CS following 48h incubation in PBS or in PBS+100mg/ml CS (control specimens were not subjected to any incubation). In Experiment 2, samples (n=23) were tested in four different solutions: PBS, PBS+100mg/ml CS, and PBS+polyethylene glycol (PEG) (133 or 170mg/ml). In Experiment 3, samples (n=18) were tested in three solutions of CS (0, 10 and 100mg/ml). Frictional tests (cartilage-on-glass) were performed under constant stress (0.5MPa) for 3600s and the time-dependent friction coefficient was measured. Samples incubated or tested in a 100mg/ml CS solution exhibited a significantly lower equilibrium friction coefficient than the respective PBS control. PEG solutions delayed the rise in the friction coefficient relative to the PBS control, but did not reduce the equilibrium value. Testing in PBS+10mg/ml of CS did not cause any significant decrease in the friction coefficient. In conclusion, CS at a concentration of 100mg/ml significantly reduces the friction coefficient of bovine articular cartilage and this mechanism is neither mediated by viscosity nor osmolarity. These results suggest that direct injection of CS into the joint may provide beneficial tribological effects.     

Shear flow induced changes in apolipoprotein C-II conformation and amyloid fibril formation

The misfolding and self-assembly of proteins into amyloid fibrils that occur in several debilitating diseases are affected by a variety of environmental factors, including mechanical factors associated with shear flow. We examined the effects of shear flow on amyloid fibril formation by human apolipoprotein C-II (apoC-II). Shear fields (150, 300, and 500 s(-1)) accelerated the rate of apoC-II fibril formation (1 mg/mL) approximately 5-10-fold. Fibrils produced at shear rates of 150 and 300 s(-1) were similar to the twisted ribbon fibrils formed in the absence of shear, while at 500 s(-1), tangled ropelike structures were observed. The mechanism of the shear-induced acceleration of amyloid fibril formation was investigated at low apoC-II concentrations (50 μg/mL) where fibril formation does not occur. Circular dichroism and tryptophan fluorescence indicated that shear induced an irreversible change in apoC-II secondary structure. Fluorescence resonance energy transfer experiments using the single tryptophan residue in apoC-II as the donor and covalently attached acceptors showed that shear flow increased the distance between the donor and acceptor molecules. Shear-induced higher-order oligomeric species were identified by sedimentation velocity experiments using fluorescence detection, while fibril seeding experiments showed that species formed during shear flow are on the fibril formation pathway. These studies suggest that physiological shear flow conditions and conditions experienced during protein manufacturing can exert significant effects on protein conformation, leading to protein misfolding, aggregation, and amyloid fibril formation.     

Physicochemical characterization of konjac glucomannan

Four commercial konjac glucomannan (KGM) samples and a glucomannan derived from yeast were characterized by aqueous gel permeation chromatography coupled with multi angle laser light scattering (GPC-MALLS). Disaggregation of aqueous glucomannan solutions through controlled use of a microwave bomb facilitated reproducible molar mass distribution determination alleviating the need for derivatization of the polymer or the use of aggressive solvents. Further characterization was undertaken by use of capillary viscometry and photon correlation spectroscopy (PCS). The weight average molecular masses (M(w)) determined were in the region of 9.0 +/- 1.0 x 10(5) g mol(-1) for KGM samples and 1.3 +/- 0.4 x 10(5) g mol(-1) for the yeast glucomannan. The values determined for KGM in aqueous solution are in agreement with those reported for KGM in aqueous cadoxen. The degradation of samples observed upon autoclaving has been quantified by GPC-MALLS and intrinsic viscosity determination, allowing comparison with reported Mark-Houwink parameters. Shear flow experiments were undertaken for a range of KGM solutions of concentration 0.05 to 2.0% using a combination of controlled stress and controlled strain rheometers. The concentration dependence of the zero shear specific viscosity was determined by analysis of the data using the Ellis model. The dependence of the zero shear specific viscosity on the coil overlap parameter was defined and interpretation discussed in terms of the Martin and Tuinier equations.     

Some chemical and physical properties of extracellular polysaccharides produced by Butyrivibrio fibrisolvens strains

Most strains of Butyrivibrio fibrisolvens are known to produce extracellular polysaccharides (EPs). However, the rheological and functional properties of these EPs have not been determined. Initially, 26 strains of Butyrivibrio were screened for EP yield and apparent viscosities of cell-free supernatants. Yields ranged from less than 1.0 to 16.3 mg per 100 mg of glucose added to the culture. Viscosities ranged from 0.71 to 5.44 mPa.s. Five strains (CF2d, CF3, CF3a, CE51, and H10b) were chosen for further screening. The apparent viscosity of the EP from each of these strains decreased by only 50 to 60% when the shear rate was increased from 20 to 1,000 s-1. Strain CE51 produced the EP having the highest solution viscosity. A detailed comparison of shear dependency of the EP from strain CF3 with xanthan gum showed that this EP was less shear sensitive than xanthan gum and, at a shear rate of 1,000 s-1, more viscous. EPs from strains CF3 and H10b were soluble over a wide range of pH (1 to 13) in 80% (vol/vol) ethanol-water or in 1% (wt/vol) salt solutions. The pH of 1% EP solutions was between 4.5 and 5.5. Addition of acid increased solution viscosities, whereas addition of base decreased viscosity. EPs from strains CF3, CE51, and H10b displayed qualitatively similar infrared spectra. Calcium and sodium were the most abundant minerals in the three EPs. The amounts of magnesium, calcium, and iron varied considerably among the EPs, but the potassium contents remained relatively constant.     

Some chemical and physical properties of extracellular polysaccharides produced by Butyrivibrio fibrisolvens strains.

Most strains of Butyrivibrio fibrisolvens are known to produce extracellular polysaccharides (EPs). However, the rheological and functional properties of these EPs have not been determined. Initially, 26 strains of Butyrivibrio were screened for EP yield and apparent viscosities of cell-free supernatants. Yields ranged from less than 1.0 to 16.3 mg per 100 mg of glucose added to the culture. Viscosities ranged from 0.71 to 5.44 mPa.s. Five strains (CF2d, CF3, CF3a, CE51, and H10b) were chosen for further screening. The apparent viscosity of the EP from each of these strains decreased by only 50 to 60% when the shear rate was increased from 20 to 1,000 s-1. Strain CE51 produced the EP having the highest solution viscosity. A detailed comparison of shear dependency of the EP from strain CF3 with xanthan gum showed that this EP was less shear sensitive than xanthan gum and, at a shear rate of 1,000 s-1, more viscous. EPs from strains CF3 and H10b were soluble over a wide range of pH (1 to 13) in 80% (vol/vol) ethanol-water or in 1% (wt/vol) salt solutions. The pH of 1% EP solutions was between 4.5 and 5.5. Addition of acid increased solution viscosities, whereas addition of base decreased viscosity. EPs from strains CF3, CE51, and H10b displayed qualitatively similar infrared spectra. Calcium and sodium were the most abundant minerals in the three EPs. The amounts of magnesium, calcium, and iron varied considerably among the EPs, but the potassium contents remained relatively constant.     

Promotion of 5-aminolevulinic acid on photosynthesis of melon (Cucumis melo) seedlings under low light and chilling stress conditions

When melon seedlings (Cucumis melo L. Ximiya No. 1) were cultured in a growth chamber with about 150 micro mol m(-2) s(-1) photon flux density, the leaf photosynthetic ability reduced dramatically as leaf position decreased from the top. The application of 5-aminolevulinic acid (ALA) solutions significantly increased the net photosynthetic rate (P(n)) as well as apparent quantum yield (AQY), carboxylation efficiency (CE) and stomata conductance (G(s)). After irrigation with 10 ml of ALA solution (10 mg l(-1) or 100 mg l(-1)) per container filled with approximately 250 g clean sand for 3 days, the leaf P(n) was about 40-200% higher than that of controls, and AQY, CE and G(s) increased 21-271%, 55-210% and 60-335%, respectively. Furthermore, ALA treatments increased leaf chlorophyll content and soluble sugar levels, as well as the rate of dark respiration, but decreased the rate of respiration under light. On the other hand, after melon seedlings that had been cultured in the chamber suffered chilling at 8 degrees C for 4 h and then recovered at 25-30 degrees C for 2 and 20 h, the P(n) of the water-irrigated plants was only 12-18% and 37-47%, respectively, compared with the initial P(n) before chilling treatment. If the seedlings underwent the same treatment but with ALA (10 mg l(-1)), the respective P(n) was 22-38% and 76-101%, compared with that of the control before chilling stress. If chilling was prolonged for 6 h, the ALA-pre-treated plants only showed a few symptoms in the leaf margins whereas all water-irrigated plants died, which suggested that ALA presumably promoted chilling tolerance of the plants under low light.     

On shear properties of trabecular bone under torsional loading: effects of bone marrow and strain rate

To further improve our understanding of trabecular bone mechanical behavior in torsion, our objective was to determine the effects of strain rate, apparent density, and presence of bone marrow on trabecular bone shear material properties. Torsion tests of cylindrical trabecular bone specimens from sheep lumbar vertebrae with and without bone marrow were conducted. The bones with marrow were divided into two groups and tested at shear strain rates of 0.002 and 0.05s(-1) measured at the specimen perimeter. The bones without marrow were divided into three groups and tested at shear strain rates of 0.002, 0.015, and 0.05s(-1). Comparing the results of bones with and without marrow tested at low (0.002s(-1)) and high (0.05s(-1)) strain rates, presence of bone marrow did not have any significant effect on trabecular bone shear modulus and strength. In specimens without marrow, power relationships were used to define shear strength and modulus as dependent variables in terms of strain rate and apparent density as independent variables. The shear strength was proportional to the apparent density raised to the 1.02 power and to the strain rate raised to the 0.13 power. The shear modulus was proportional to the apparent density raised to the 1.08 power and to the strain rate raised to the 0.07 power. This study provides further insight into the mechanism of bone failure in trauma as well as failure at the interface between bone and implants as it relates to prediction of trabecular bone shear properties.