A combined passive and active musculoskeletal model study to estimate L4-L5 load sharing

A number of geometrically-detailed passive finite element (FE) models of the lumbar spine have been developed and validated under in vitro loading conditions. These models are devoid of muscles and thus cannot be directly used to simulate in vivo loading conditions acting on the lumbar joint structures or spinal implants. Gravity loads and muscle forces estimated by a trunk musculoskeletal (MS) model under twelve static activities were applied to a passive FE model of the L4-L5 segment to estimate load sharing among the joint structures (disc, ligaments, and facets) under simulated in vivo loading conditions. An equivalent follower (FL), that generates IDP equal to that generated by muscle forces, was computed in each task. Results indicated that under in vivo loading conditions, the passive FE model predicted intradiscal pressures (IDPs) that closely matched those measured under the simulated tasks (R² = 0.98 and root-mean-squared-error, RMSE = 0.18 MPa). The calculated equivalent FL compared well with the resultant force of all muscle forces and gravity loads acting on the L4-L5 segment (R² = 0.99 and RMSE = 58 N). Therefore, as an alternative approach to represent in vivo loading conditions in passive FE model studies, this FL can be estimated by available in-house or commercial MS models. In clinical applications and design of implants, commonly considered in vitro loading conditions on the passive FE models do not adequately represent the in vivo loading conditions under muscle exertions. Therefore, more realistic in vivo loading conditions should instead be used.     

Response analysis of the lumbar spine during regular daily activities—A finite element analysis

A non-linear poroelastic finite element model of the lumbar spine was developed to investigate spinal response during daily dynamic physiological activities. Swelling was simulated by imposing a boundary pore pressure of 0.25 MPa at all external surfaces. Partial saturation of the disc was introduced to circumvent the negative pressures otherwise computed upon unloading. The loading conditions represented a pre-conditioning full day followed by another day of loading: 8 h rest under a constant compressive load of 350 N, followed by 16 h loading phase under constant or cyclic compressive load varying in between 1000 and 1600 N. In addition, the effect of one or two short resting periods in the latter loading phase was studied.The model yielded fairly good agreement with in-vivo and in-vitro measurements. Taking the partial saturation of the disc into account, no negative pore pressures were generated during unloading and recovery phase. Recovery phase was faster than the loading period with equilibrium reached in only ～3 h. With time and during the day, the axial displacement, fluid loss, axial stress and disc radial strain increased whereas the pore pressure and disc collagen fiber strains decreased. The fluid pressurization and collagen fiber stiffening were noticeable early in the morning, which gave way to greater compression stresses and radial strains in the annulus bulk as time went by. The rest periods dampened foregoing differences between the early morning and late in the afternoon periods. The forgoing diurnal variations have profound effects on lumbar spine biomechanics and risk of injury.     

Implantable MEMS compressive stress sensors: Design,fabrication and calibration with application to the disc annulus

Physiological stresses are fundamental to biomechanical testing, mechanobiological analyses, implant design, and tissue engineering. The purpose of this study was to design, fabricate, and evaluate compressive stress sensors packaged for extended, in vivo implantation in the annulus of the intervertebral disc. A commercial microelectromechanical systems (MEMS) pressure sensor die was selected as the active element for a custom stress sensor. The sensor die was modified and packaged to protect the electrical system from the biochemical and biomechanical environment. Completed sensors were calibrated under hydrostatic pressure and solid contact compression. Calibrations were performed before and after 8 weeks of in vivo implantation in a porcine disc. For the two reported sensors, stress and voltage were linearly correlated over a range of 0–1.8 MPa with less than 5% change in sensitivity. Sensitivity to solid contact stress was within 10% of that from hydrostatic pressure. In contrast to most previous studies, in which disc pressure was measured in the fluidic nucleus pulposus, these sensors may be used to measure in vivo dynamic compressive stresses in the annulus at magnitudes typical of the musculoskeletal system in a large animal over a relatively long post-operative time.     

Effects of eight different ligament property datasets on biomechanics of a lumbar L4-L5 finite element model

Ligaments assist trunk muscles in balancing external moments and providing spinal stability. In absence of the personalized material properties for ligaments, finite element (FE) models use dispersed data from the literature. This study aims to investigate the relative effects of eight different ligament property datasets on FE model responses. Eight L4-L5 models distinct only in ligament properties were constructed and loaded under moment (15 N m) alone or combined with a compressive follower load (FL). Range of motions (RoM) of the disc-alone model matched well in vitro data. Ligament properties significantly affected only sagittal RoMs (∼3.0–7.1° in flexion and ∼3.8–5.8° in extension at 10 N m). Sequential removal of ligaments shifted sagittal RoMs in and out of the corresponding in vitro ranges. When moment was combined with FL, center of rotation matched in vivo data for all models (3.8 ± 0.9 mm and 4.3 ± 1.8 mm posterior to the disc center in flexion and extension, respectively). Under 15 N m sagittal moments, ligament strains were often smaller or within the in vitro range in flexion whereas some posterior ligament forces approached their failure forces in some models. Ligament forces varied substantially within the models and affected the moment-sharing and internal forces on the disc and facet joints. Intradiscal pressure (IDP) had the greatest variation between models in extension. None of the datasets yielded results in agreement with all reported measurements. Results emphasized the important role of ligaments especially under larger moments and the need for their accurate representation in search for valid spinal models.     

Application of S-thanatin,an antimicrobial peptide derived from thanatin,in mouse model of Klebsiella pneumoniae infection

Thanatin was first discovered from the hemipteran insect Podisus maculiventris and showed a promising antimicrobial activity. Multidrug-resistant (MDR) clinical isolates of Klebsiella pneumoniae have developed resistance to current therapies. As an attempt to resolve this problem, the efficacy of thanatin and its analogues against clinical isolates of K. pneumoniae was studied in vitro and in vivo. S-thanatin showed an improved antimicrobial activity with the tested MIC values was 2–8-fold lower than those of other thanatin analogs. Antimicrobial assay indicated a high activity of S-thanatin against K. pneumoniae in vitro with MIC between 4 and 8 μg/ml. Its in vivo activity was evaluated using a K. pneumoniae-infected mice model. Adult male ICR mice were randomly grouped and given an intraperitoneal (i.p.) administration of 2 × 10¹⁰ colony-forming units of K. pneumoniae (CI 120204205). Afterwards, mouse groups were subjected to i.p. administration of saline or S-thanatin (5, 10, or 15 mg/kg). After an inspection of 72 h, the mice were finally sacrificed for analysis of in vivo bacterial growth and plasma endotoxin level. The results showed that S-thanatin administration apparently improved the survival rate and reduced the bacterial CFU from intra-abdominal fluid in mice. The plasma endotoxin level was improved as well. All above implied that S-thanatin, as an alternative, may provide a novel strategy for treating K. pneumoniae infection and other infections due to multidrug-resistant bacteria.     

Concentration-dependent effects of resveratrol and metabolites on the redox status of human erythrocytes in single-dose studies

Dietary trans-resveratrol (RES) is rapidly metabolized into sulfated and glucuronated conjugates in humans. This study focused on the in vitro determination of the antioxidant capacity of RES and its main physiological metabolites and on its relevance in vivo. In vitro, RES, RES-3-O-sulfate (R3S) and 3-O-glucuronide (R3G) showed antioxidant activities at a concentration of 1 mM when compared to Trolox using an assay in which the antioxidant inhibits iron-induced linoleic acid oxidation: 0.87±0.08 mM Trolox equivalents (TE) for RES, 0.52±0.01 mM TE for R3S and 0.36±0.02 mM TE for R3G. At a concentration of 1 μM, compounds promoted linoleic acid peroxidation (RES −0.30±0.09 mM TE, R3S −0.48±0.05 mM TE and R3G −0.57±0.07 mM TE). To elucidate whether these effects were reflected in vivo, total antioxidant capacity, reactive oxygen species (ROS), conjugated fatty acid dienes (CD), superoxide dismutase (SOD) and catalase (CAT) activities were determined in human plasma and erythrocytes over 24 h, after oral intake of either 0.05 g RES as piceid or 5 g RES. Oral administration of RES did not show an impact on total antioxidant capacity, ROS or CD. However, enzymatic activities of ROS scavenging SOD and CAT were significantly lower after high-dose compared to low-dose administration of RES (P<.03 and P<.01). In conclusion, in healthy subjects, neither 0.05 g nor 5 g RES changed blood oxidative state, although our in vitro data point to a prooxidative activity of low concentrations of RES and its metabolites, which could be important in vivo for individuals with compromised antioxidant defense capacity.     

High hydrostatic pressure increased stability and activity of immobilized lipase in hexane

Lipases are important to high value product synthesis, modification, and enhancement. However, they are often unstable above 40 °C. While most current applications of high hydrostatic pressure (HHP) are for inactivating deleterious enzymes, there is evidence that HHP can stabilize and increase activity of some enzymes. This study examines the apparent kinetics of immobilized lipase-catalyzed synthesis of isoamyl acetate at HHP in hexane. HHP reduced thermal inactivation of lipase by up to 152% after 4 h at 80 °C and 400 MPa when compared to incubations at low pressure. No significant differences were found in activation energy (E_a) at different pressures, irrespectively of the pressurization and heating sequence, and were between 35.7 ± 3.5 and 47.8 ± 8.2 kJ mol^?1, depending on the method. In all methods utilized, activity at 63.5 and 80 °C at 400 MPa was greater (from about 20 to 96% increase) than at low pressure. Activity increased by 110% at low pressure versus a 239% increase at 350 MPa when the temperature was increased from 40 to 80 °C. Increasing pressure up to 350 MPa increased lipase activity while pressures greater than 350 MPa maintained or decreased lipase activity. Activation volume (ΔV^≠) appeared negative between ambient pressure and 200 MPa in contrast to a positive ΔV^≠ between 300 and 600 MPa. Apparent ΔV^≠ was 14.3 ± 1.7 or 15.2 ± 2.2 cm³ mol^?1 at 40 or 80 °C, respectively, between 300 and 500 MPa.     

Alginate embedding and subsequent sporulation of in vitro-produced Conidiobolus thromboides hyphae using a pressurised air-extrusion method

Conidiobolus thromboides is an entomophthoralean fungus with potential as a biological control agent of aphids. However, its application in biological control is limited due to its formulation requirements. The objective of this study was to develop and optimise a novel air-extrusion method to embed C. thromboides hyphae at high density in alginate pellets. An orthogonal experimental design was used to investigate selected combinations of parameters known to affect hyphal density within pellets. The diameter of pellets produced, and the calculated density of hyphae within them, ranged from 0.18 ± 0.09 to 3.17 ± 0.06 mm and from 0.02 to 350.56 mg/mm³ respectively. These data were used to predict the optimal parameter combination to deliver the greatest density of hyphae of C. thromboides per pellet: 1% sodium alginate, a 1:2 ratio of hyphae to sodium alginate, an orifice diameter of 0.232 mm and an air pressure of 0.05 MPa. Pellets made under the optimal conditions predicted produced a mean total of 4.3 ± 0.6 × 10⁵ conidia per pellet at 100% relative humidity which was significantly greater than the mean total number of conidia produced from infected aphid cadavers of comparable size (9.35 ± 0.85 × 10⁴) (p < 0.001). In conclusion, air-extrusion embedding appears to be a promising method for formulating in vitro-produced hyphae of C. thromboides for use in biological control.     

Design,synthesis and biological screening of new 4-thiazolidinone derivatives with promising COX-2 selectivity,anti-inflammatory activity and gastric safety profile

Two series of new thiazolidin-4-one derivatives 4a–c and 8a–e were designed and prepared. All the synthesized compounds were evaluated for their in vitro COX-2 selectivity and anti-inflammatory activity in vivo. Compounds 8c and 8d showed the best overall in vitro COX-2 selectivity (selectivity indexes of 4.56 and 5.68 respectively) and in vivo activities (edema inhibition % = 61.8 and 67 after 3 h, respectively) in comparison with the reference drug celecoxib (S.I. = 7.29, edema inhibition % = 60 after 3 h). In addition, 8c and 8d were evaluated for their mean effective anti-inflammatory doses (ED₅₀ = 27.7 and 18.1 μmol/kg respectively, celecoxib ED₅₀ = 28.2 μmol/kg) and ulcerogenic liability (reduction in ulcerogenic potential versus celecoxib = 85%, 92% respectively. Molecular docking studies were performed and the results were in agreement with that obtained from the in vitro COX inhibition assays.     

Stability and activity of Dictyoglomus thermophilum GH11 xylanase and its disulphide mutant at high pressure and temperature

The functional properties of extremophilic Dictyoglomus thermophilum xylanase (XYNB) and the N-terminal disulphide-bridge mutant (XYNB-DS) were studied at high pressure and temperature. The enzymes were quite stable even at the pressure of 500 MPa at 80 °C. The half-life of inactivation in these conditions was over 30 h. The inactivation at 80 °C in atmospheric pressure was only 3-times slower. The increase of pressure up to 500 MPa at 80 °C decreased only slightly the enzyme's stability, whereas in 500 MPa the increase of temperature from 22 to 80 °C decreased significantly more the enzyme's stability. While the high temperature (80–100 °C) decreased the enzyme reaction with short xylooligosaccharides (xylotetraose and xylotriose), the high pressure (100–300 MPa) had an opposite effect. The temperature of 100 °C strongly increased the K_m but did not affect the k_cat to the same extent, thus indicating that the interaction of the substrate with the active site suffers before the catalytic reaction begins to decrease as the temperature rises. Circular dichroism spectroscopy showed the high structural stability of XYNB and XYNB-DS at 93 °C.     

Applying a follower load delivers realistic results for simulating standing

The exact loads acting on the lumbar spine during standing remain hitherto unknown. It is for this reason that different loads are applied in experimental and numerical studies. The aim of this study was to compare intersegmental rotations, intradiscal pressures and facet joint forces for different loading modes simulating standing in order to ascertain, the results for which loading modes are closest to data measured in vivo.A validated osseoligamentous finite element model of the lumbar spine ranging from L1 to the disc L5–S1, was used. Six load application modes were investigated as to how they could simulate standing. This posture was simulated by applying a vertical force of 500 N at the centre of the L1 vertebral endplate with different boundary conditions, by applying a follower load, and by applying upper body weight and muscle forces. The calculated intersegmental rotations and intradiscal pressures were compared to in vivo values.Intersegmental rotations at one level vary by up to 8° for the different loading modes simulating standing. The overall rotation in the lumbar spine varies between 2.2° and 19.5°. With a follower load, the difference to the value measured in vivo is 3.3°. For all other loading cases studied, the difference is greater than 6.6°. Intradiscal pressures vary slightly with the loading mode. Calculated forces in the facet joints vary between 0 and nearly 80 N.Applying a follower load of 500 N is the only loading mode simulating standing for which the calculated values for intervertebral rotations and intradiscal pressures agreed well with in vivo data from literature.     

An in vivo model of a mechanically-induced bone marrow lesion

Bone marrow lesions (BMLs) are radiologic abnormalities in magnetic resonance images of subchondral bone that are correlated with osteoarthritis. Little is known about the physiologic processes within a BML, although BMLs are associated with mechanical stress, bone tissue microdamage and increased bone remodeling. Here we establish a rabbit model to study the pathophysiology of BMLs. We hypothesized that in vivo loads that generate microdamage in cancellous bone would also create BMLs and increase bone remodeling. In vivo cyclic loading (0.2–2.0 MPa in compression for 10,000 cycles at 2 Hz) was applied to epiphyseal cancellous bone in the distal femurs of New Zealand white rabbits (n = 3, right limb loaded, left limb controls experienced surgery but no loading). Magnetic resonance images were collected using short tau inversion recovery (STIR) and T1 weighted sequences at 1 and 2 weeks after surgery/loading and histological analysis of the BML was performed after euthanasia to examine tissue microdamage and remodeling. Loaded limbs displayed BMLs while control limbs showed only a small BML-like signal caused by surgery. Histological analysis of the BML at 2 weeks after loading showed increased tissue microdamage (p = 0.03) and bone resorption (p = 0.01) as compared to controls. The model described here displays the hallmarks of load-induced BMLs, supporting the use of the model to examine changes in bone during the development, progression and treatment of BMLs.     

Metabolic engineering of fatty acyl-ACP reductase-dependent pathway to improve fatty alcohol production in Escherichia coli

Fatty alcohols are important components of surfactants and cosmetic products. The production of fatty alcohols from sustainable resources using microbial fermentation could reduce dependence on fossil fuels and greenhouse gas emission. However, the industrialization of this process has been hampered by the current low yield and productivity of this synthetic pathway. As a result of metabolic engineering strategies, an Escherichia coli mutant containing Synechococcus elongatus fatty acyl-ACP reductase showed improved yield and productivity. Proteomics analysis and in vitro enzymatic assays showed that endogenous E. coli AdhP is a major contributor to the reduction of fatty aldehydes to fatty alcohols. Both in vitro and in vivo results clearly demonstrated that the activity and expression level of fatty acyl-CoA/ACP reductase is the rate-limiting step in the current protocol. In 2.5-L fed-batch fermentation with glycerol as the only carbon source, the most productive E. coli mutant produced 0.75 g/L fatty alcohols (0.02 g fatty alcohol/g glycerol) with a productivity of up to 0.06 g/L/h. This investigation establishes a promising synthetic pathway for industrial microbial production of fatty alcohols.     

Mechanical stress regulates osteogenic differentiation and RANKL/OPG ratio in periodontal ligament stem cells by the Wnt/β-catenin pathway

BackgroundThe balance between osteoblastic and osteoclastic activity is critical in orthodontic tooth movement (OTM). Mesenchymal stem cells (MSCs) play an important role in maintaining bone homeostasis, and periodontal ligament stem cells (PDLSCs) are tissue-specific MSCs in the periodontal ligament. However, whether PDLSCs are required for periodontal tissue remodeling during OTM is not fully understood.MethodsHere, we used PDGFRα and Nestin to trace PDLSCs during OTM in rats. We treat human PDLSCs with 100 kpa static pressure for 1 h or 12 h in vitro, and examined the phenotypic changes and expression of RANKL and OPG in these cells.ResultsIn vivo, we found that positive signals of PDGFRα and Nestin in the PDL gradually increased and then decreased on the pressure side to which pressure was applied. In vitro, the osteogenic differentiation of PDLSCs was significantly increased after force treatment for 1 h relative to 12 h. In contrast, the expression ratio of RANKL/OPG was reduced at 1 h and significantly increased at 12 h. Furthermore, we found that the Wnt/β-catenin pathway was dynamically activated in the PDL and in PDLSCs after mechanical stimulation. Importantly, the canonical Wnt pathway inhibitor DKK1 blocked the osteogenesis effect and rescued the ratio of RANKL/OPG in PDLSCs under force treatment for 1 h.ConclusionsOur findings reveal that PDLSCs participate in OTM and that the Wnt/β-catenin pathway maintains bone homeostasis during tooth movement by regulating the balance between osteoblastic and osteoclastic activity.General significanceWe describe a novel potential mechanism related to tooth movement.     

Decay and termite resistance,water absorption and swelling of thermally compressed wood panels

This study evaluated decay and termite resistance of thermally compressed pine wood panels under pressure at either 5 or 7 MPa and either 120 or 150 °C for 1 h. Wood specimens from the panels were exposed to laboratory decay resistance tests by using the wood degrading fungi, Gloeophyllum trabeum and Trametes versicolor. The thermal compression process caused increases in density and decreases in thickness of the panels; however, laboratory decay resistance tests revealed that thermally compressed wood was not resistant against the wood degrading fungi tested. More interesting results were found in laboratory termite resistance tests by using the Eastern subterranean termites, Reticulitermes flavipes. As pressure and temperature applied to the specimens increased to 7 MPa and 120 °C, mass losses in the specimens gradually decreased in comparison with control specimens. However, the specimens compressed at 7 MPa and 150 °C showed higher mass losses when compared to the specimens compressed at 7 MPa and 120 °C. The lowest water absorption and swelling rates were seen in the specimens exposed to a pressure of 7 MPa at 120 °C. The thermal compression process at 7 MPa and 150 °C resulted in the highest water absorption and swelling in the specimens.     

Phosphorus removal from agricultural runoff by constructed wetland

A free-water surface wetland covering an area of 2800 m² was operated from March 2002 to June 2004 for agricultural runoff treatment in the Dianchi Valley in China. In the wetland were grown Zizania Caduciflora Turez Hand-mazt and Phragmites australis (Cav.) Trin.ex Steud. The instantaneous inflow rate was measured and the integrated flux was recorded by an ultrasonic flow instrument all year round. The average inflow rate, hydraulic loading rate (HLR) and hydraulic retention time (HRT) were kept at 242 m³ d^?1, 12.7 cm d^?1 and 2.0 d, respectively. The annual average total phosphorus (TP) in the inflow was 0.87 mg L^?1, and the corresponding removal efficiency was calculated to be 59.0%. Biannual plant uptake and removal by harvesting and seed transport was the main pathway for TP removal, while the influent TP load was 12.9 g m^?2 year^?1. Hydraulic retention time had a significant positive correlation with the removal of P (r² = 0.88). Water temperature, inflow phosphorus load, inflow and hydraulic load rates were positively correlated with the removal of P. Inflow phosphorus concentrations were negatively correlated with the removal of P. It is shown that the free-water surface wetland was an effective and economical system for agricultural runoff treatment in lake regions.     

Effect of titanium on lipoprotein lipase activity in vivo and in vitro

SummaryLipoprotein lipase (LPL) is a major lipolytic enzyme in the intravascular metabolism of postprandial triglyceride-rich lipoproteins. This enzyme is synthesized and secreted by tissues and transported to the capillary endothelial surface. Decreased activity of this enzyme is suggested to be involved in arterial sequestration of lipoproteins and thus in the progression of atherosclerosis. Titanium salts are widely used in industry, medicine, and pharmacy for tablet coating, pharmaceuticals and cosmetic products. In this study the effect of titanium on post-heparin LPL activity is reported in vivo and in vitro.MethodsGroups of Male Wistar rats were administered (i.p) with an acute dose of 2.5 mg/kg titanium chloride for 10 days and a chronic dose of 0.75 mg/kg for 30 and/or 60 days. Blood samples were then collected for LPL assay. For in vitro study, plasma aliquots were incubated in the presence of up to 50 mM titanium and the enzyme activity was measured.ResultsAnimals exposed to acute dose of titanium showed about 20% reduction in LPL activity, whereas 31% and 36% reductions were observed in animals chronically exposed for 30 and/or 60 days, respectively. Titanium in vitro also led to enzyme inhibition, so that a decrease of 28–53% was seen in the presence of 0.1–50 mM titanium. This inhibition by titanium was potentiated when citrate and/or bicarbonate was present.ConclusionAlthough the mechanism of titanium effect on LPL activity in vivo and in vitro demands more investigations, the inhibitory effect of titanium ion in vivo should be considered seriously in subjects exposed to this metal ion. Changes in LPL activity may affect whole body lipid metabolism, a condition favorable for development and progression of atherosclerosis.