A two-layer elasto-visco-plastic rheological model for the material parameter identification of bone tissue

The ability to measure bone tissue material properties plays a major role in diagnosis of diseases and material modeling. Bone’s response to loading is complex and shows a viscous contribution to stiffness, yield and failure. It is also ductile and damaging and exhibits plastic hardening until failure. When performing mechanical tests on bone tissue, these constitutive effects are difficult to quantify, as only their combination is visible in resulting stress–strain data. In this study, a methodology for the identification of stiffness, damping, yield stress and hardening coefficients of bone from a single cyclic tensile test is proposed. The method is based on a two-layer elasto-visco-plastic rheological model that is capable of reproducing the specimens’ pre- and postyield response. The model’s structure enables for capturing the viscously induced increase in stiffness, yield, and ultimate stress and for a direct computation of the loss tangent. Material parameters are obtained in an inverse approach by optimizing the model response to fit the experimental data. The proposed approach is demonstrated by identifying material properties of individual bone trabeculae that were tested under wet conditions. The mechanical tests were conducted according to an already published methodology for tensile experiments on single trabeculae. As a result, long-term and instantaneous Young’s moduli were obtained, which were on average 3.64 GPa and 5.61 GPa, respectively. The found yield stress of 16.89 MPa was lower than previous studies suggest, while the loss tangent of 0.04 is in good agreement. In general, the two-layer model was able to reproduce the cyclic mechanical test data of single trabeculae with an root-mean-square error of 2.91 ± 1.77 MPa. The results show that inverse rheological modeling can be of great advantage when multiple constitutive contributions shall be quantified based on a single mechanical measurement.

     

Evaluation of cetyltrimethylammonium bromide as a potential short-term preservative agent for stripped goat skin

The aim of the present study was to evaluate cetyltrimethylammonium bromide (CTAB) as a short-term preservative agent for stripped goat skin, as an alternative to common salt (sodium chloride), the presently used preservative in the leather industry. Bacteria were isolated from the stripped goat skin at various time intervals and 16S rDNA sequencing results showed the presence of both Gram positive and Gram negative bacteria in stripped goat skin. All the bacterial isolates were inhibited by CTAB with MICs of 0.5–24.5 mg l⁻¹. Application of CTAB against microbial consortium on stripped goat skin showed that skin sample kept in 5% CTAB solution for 10 min did not contain any bacterial growth even after 12 days storage at room temperature. Scanning electron microscopy analysis and physical testing of skin preserved using CTAB did not reveal any bacterial attack on fiber structure of skin. This study shows that CTAB can be applied as a viable alternative to NaCl which generates a huge amount of pollution there by reducing pollution from leather processing.     

In Vitro Percutaneous Permeation and Skin Accumulation of Finasteride Using Vesicular Ethosomal Carriers

In order to develop a novel transdermal drug delivery system that facilitates the skin permeation of finasteride encapsulated in novel lipid-based vesicular carriers (ethosomes)finasteride ethosomes were constructed and the morphological characteristics were studied by transmission electron microscopy. The particle size, zeta potential and the entrapment capacity of ethosome were also determined. In contrast to liposomes ethosomes were of more condensed vesicular structure and they were found to be oppositely charged. Ethosomes were found to be more efficient delivery carriers with high encapsulation capacities. In vitro percutaneous permeation experiments demonstrated that the permeation of finasteride through human cadaver skin was significantly increased when ethosomes were used. The finasteride transdermal fluxes from ethosomes containing formulation (1.34 ± 0.11 μg/cm²/h) were 7.4, 3.2 and 2.6 times higher than that of finasteride from aqueous solution, conventional liposomes and hydroethanolic solution respectively (P < 0.01).Furthermore, ethosomes produced a significant (P < 0.01) finasteride accumulation in the skin, especially in deeper layers, for instance in dermis it reached to 18.2 ± 1.8 μg/cm². In contrast, the accumulation of finasteride in the dermis was only 2.8 ± 1.3 μg/cm² with liposome formulation. The study demonstrated that ethosomes are promising vesicular carriers for enhancing percutaneous absorption of finasteride.     

The layered structure of coronary adventitia under mechanical load

The mechanical loading-deformation relation of elastin and collagen fibril bundles is fundamental to understanding the microstructural properties of tissue. Here, we use multiphoton microscopy to obtain quantitative data of elastin and collagen fiber bundles under in situ loading of coronary adventitia. Simultaneous loading-imaging experiments on unstained fresh coronary adventitia allowed morphometric measurements of collagen and elastin fibril bundles and their individual deformation. Fiber data were analyzed at five different distension loading points (circumferential stretch ratio λ_θ = 1.0, 1.2, 1.4, 1.6, and 1.8) at a physiological axial stretch ratio of λ_axial = 1.3. Four fiber geometrical parameters were used to quantify the fibers: orientation angle, waviness, width, and area fraction. The results show that elastin and collagen fibers in inner adventitia form concentric densely packed fiber sheets, and the fiber orientation angle, width, and area fraction vary transmurally. The extent of fiber deformation depends on the initial orientation angle at no-distension state (λ_θ = 1.0 and λ_axial = 1.3). At higher distension loading, the orientation angle and waviness of fibers decrease linearly, but the width of collagen fiber is relatively constant at λ_θ = 1.0–1.4 and then decrease linearly for λ_θ ≥ 1.4. A decrease of the relative dispersion (SD/mean) of collagen fiber waviness suggests a heterogeneous mechanical response to loads. This study provides fundamental microstructural data for coronary artery biomechanics and we consider it seminal for structural models.     

Vasotocin has the potential to inhibit basolateral Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-pump current across isolated skin of tree frog in vitro<Emphasis Type="Italic">,</Emphasis> via its V<Subscript>2</Subscript>-type receptor/cAMP pathway

Adult frog skin transports Na⁺ from the apical to the basolateral side across the skin. Antidiuretic hormone (ADH) is involved in the regulation of Na⁺ transport in both mammals and amphibians. We investigated the effect of arginine vasotocin (AVT), the ADH of amphibians, on the short-circuit current (SCC) across intact skin and on the basolateral Na⁺/K⁺-pump current across apically nystatin-permeabilized skin of the tree frog, Hyla japonica, in which the V₂-type ADH receptor is expressed in vitro. In intact skin, 1 pM AVT had no effect on the SCC, but 10 nM AVT was sufficient to stimulate the SCC since 10 nM and 1 μM of AVT increased the SCC 3.2- and 3.4-fold, respectively (P > 0.9). However, in permeabilized skin, AVT (1 μM) decreased the Na⁺/K⁺-pump current to 0.79 times vehicle control. Similarly, 500 μM of 8Br-cAMP increased the SCC 3.2-fold, yet 1 mM of 8Br-cAMP decreased the Na⁺/K⁺-pump current to 0.76 times vehicle control. Arachidonic acid (10⁻⁵ M) tended to decrease the Na⁺/K⁺-pump current. To judge from these in vitro experiments, AVT has the potential to inhibit the basolateral Na⁺/K⁺-pump current via the V₂-type receptor/cAMP pathway in the skin of the tree frog.     

Comparison of thermoregulatory responses to heat between Malaysian and Japanese males during leg immersion

The objective of this study was to investigate thermoregulatory responses to heat in tropical (Malaysian) and temperate (Japanese) natives, during 60 min of passive heating. Ten Japanese (mean ages: 20.8 ± 0.9 years) and ten Malaysian males (mean ages: 22.3 ± 1.6 years) with matched morphological characteristics and physical fitness participated in this study. Passive heating was induced through leg immersion in hot water (42°C) for 60 min under conditions of 28°C air temperature and 50% RH. Local sweat rate on the forehead and thigh were significantly lower in Malaysians during leg immersion, but no significant differences in total sweat rate were observed between Malaysians (86.3 ± 11.8 g m⁻² h⁻¹) and Japanese (83.2 ± 6.4  g m⁻² h⁻¹) after leg immersion. In addition, Malaysians displayed a smaller rise in rectal temperature (0.3 ± 0.1°C) than Japanese (0.7 ± 0.1°C) during leg immersion, with a greater increase in hand skin temperature. Skin blood flow was significantly lower on the forehead and forearm in Malaysians during leg immersion. No significant different in mean skin temperature during leg immersion was observed between the two groups. These findings indicated that regional differences in body sweating distribution might exist between Malaysians and Japanese during heat exposure, with more uniform distribution of local sweat rate over the whole body among tropical Malaysians. Altogether, Malaysians appear to display enhanced efficiency of thermal sweating and thermoregulatory responses in dissipating heat loss during heat loading. Thermoregulatory differences between tropical and temperate natives in this study can be interpreted as a result of heat adaptations to physiological function.     

Patterning limnological characteristics of the Chilika lagoon (India) using a self-organizing map

In this study, a self-organizing map (SOM) was utilized to classify habitats in the Chilika lagoon located in India, the largest lagoon ecosystem in Asia (maximum length, 64.3 km; mean width, 20.1 km). The lagoon was marginally eutrophic (nitrate, 0.25 ± 0.22 mg L⁻¹; orthophosphate, 0.26 ± 0.22 mg L⁻¹; n = 1,980, respectively) for six years (1999–2004), and it used to be warm, shallow, turbid and predominantly brackish. The SOM model successfully identified the changing patterns of limnology in the lagoon using the monthly limnological dataset from 30 study sites (July 1999–December 2004). Comparative re-sectoring evaluation of current monitoring sites was accomplished based on the outcome of the modeling. The new site clustering that emerged from the model was similar to conventional ones, and several sites were reorganized. Water physicochemistry was affected by freshwater inflow during monsoon and the new lagoon mouth constructed in September 2000, which resulted in variations in site characteristics in terms of limnology. The results of this study may provide information on the limnological patterns in Chilika lagoon, and they leave room for further study into functional changes in the lagoon ecology with respect to changes in climatic factor, freshwater flow and lagoon morphology.     

The influence of growth conditions on the cell dry weight per unit biovolume of <Emphasis Type="Italic">Klebsormidium flaccidum</Emphasis> (Charophyta), a typical ubiquitous soil alga

The dry weight per unit biovolume of 810 single, living cells of the ubiquitous soil algae Klebsormidium flaccidum from 80 experiments were determined using a Mach–Zehnder double-beam interference microscope. Different substrates such as agarized nutrient solution, different soils, and slag heap material, different pH values, temperatures and light intensities were used and cells from both growth and stationary phase were measured. The total possible range of dry weight with respect to carbon per unit biovolume (C/ubv) values was 93–226 fg μm⁻³. The mean value of all data was 147 fg μm⁻³, which concurs with the average value as taken from literature data of several planktonic algal species and groups within the respective size range (cell volume 300–1,000 μm³). We could show that C/ubv is suitable to quantify environmental stress conditions: C/ubv values ≤140 fg μm⁻³ are characteristic of cells grown under optimum conditions, and values ≥160 fg μm⁻³ reflect quantitatively graded stress situations. We propose integrating the microscope interferometric method using K. flaccidum as a test organism into a soil test system to determine the prevailing environmental conditions.     

Constitutive modeling of an electrospun tubular scaffold used for vascular tissue engineering

In this study, we sought to model the mechanical behavior of an electrospun tubular scaffold previously reported for vascular tissue engineering with hyperelastic constitutive equations. Specifically, the scaffolds were made by wrapping electrospun polycaprolactone membranes that contain aligned fibers around a mandrel in such a way that they have microstructure similar to the native arterial media. The biaxial stress-stretch data of the scaffolds made of moderately or highly aligned fibers with three different off-axis fiber angles \(\alpha \) (30\(^\circ \), 45\(^\circ \), and 60\(^\circ \)) were fit by a phenomenological Fung model and a series of structurally motivated models considering fiber directions and fiber angle distributions. In particular, two forms of fiber strain energy in the structurally motivated model for a linear and a nonlinear fiber stress–strain relation, respectively, were tested. An isotropic neo-Hookean strain energy function was also added to the structurally motivated models to examine its contribution. The two forms of fiber strain energy did not result in significantly different goodness of fit for most groups of the scaffolds. The absence of the neo-Hookean term in the structurally motivated model led to obvious nonlinear stress-stretch fits at a greater axial stretch, especially when fitting data from the scaffolds with a small \(\alpha \). Of the models considered, the Fung model had the overall best fitting results; its applications are limited because of its phenomenological nature. Although a structurally motivated model using the nonlinear fiber stress–strain relation with the neo-Hookean term provided fits comparably as good as the Fung model, the values of its model parameters exhibited large within-group variations. Prescribing the dispersion of fiber orientation in the structurally motivated model, however, reduced the variations without compromising the fits and was thus considered to be the best structurally motivated model for the scaffolds. It appeared that the structurally motivated models could be further improved for fitting the mechanical behavior of the electrospun scaffold; fiber interactions are suggested to be considered in future models.     

Mechanical characterisation of in vivo human skin using a 3D force-sensitive micro-robot and finite element analysis

The complex mechanical properties of skin have been the subject of much study in recent years. Several experimental methods developed to measure the mechanical properties of skin in vivo, such as suction or torsion, are unable to measure skin’s anisotropic characteristics. An experiment characterising the mechanical properties of in vivo human skin using a novel force-sensitive micro-robot is presented. The micro-robot applied in-plane deformations to the anterior forearm and the posterior upper arm. The behaviour of the skin in each area is highly nonlinear, anisotropic, and viscoelastic. The response of the upper arm skin is very dependent on the orientation of the arm. A finite element model consisting of an Ogden strain energy function and quasi-linear viscoelasticity was developed to simulate the experiments. An orthogonal initial stress field, representing the in vivo skin tension, was used as an additional model parameter. The model simulated the experiments accurately with an error-of-fit of 17.5% for the anterior lower forearm area, 6.5% for the anterior upper forearm and 9.3% for the posterior upper arm. The maximum in vivo tension in each area determined by the model was 6.2 Nm⁻¹ in the anterior lower forearm, 11.4 Nm⁻¹ in anterior upper forearm and 5.6 Nm⁻¹ in the posterior upper arm. The results also show that a finite element model with a neo-Hookean strain energy function cannot simulate the experiments with the same accuracy.     

Comparison of various methods of estimating radon dose at underground workplaces in wineries

Levels of radon were surveyed in the air at underground workplaces of eight major Slovenian wineries. Geometric mean and geometric standard deviation values, respectively, obtained with different devices were 81 Bq m⁻³ and 2.3 with alpha scintillation cells, 114 Bq m⁻³ and 2.0 by exposing etched track detectors for 1–5 months, and 183 Bq m⁻³ and 2.6 from 1–4-weeks continuous measurements. The equilibrium factor was 0.25–0.67, and the unattached fraction of radon short-lived decay products was in the range 0.09–0.20. Effective doses were calculated and compared based on radon data obtained with different techniques.     

Estimating material parameters of human skin in vivo

An accurate mathematical representation of the mechanical behaviour of human skin is essential when simulating deformations occurring in the skin during body movements or clinical procedures. In this study constitutive stress–strain relationships based on experimental data from human skin in vivo were obtained. A series of multiaxial loading experiments were performed on the forearms of four age- and gender matched subjects. The tissue geometry, together with recorded displacements and boundary forces, were combined in an analysis using finite element modelling. A non-linear optimization technique was developed to estimate values for the material parameters of a previously published constitutive law, describing the stress–strain relationship as a non-linear anisotropic membrane. Ten sets of material parameters where estimated from the experiments, showing considerable differences in mechanical behaviour both between individual subjects as well as mirrored body locations on a single subject. The accuracy of applications that simulate large deformations of human skin could be improved by using the parameters found from the in vivo experiments as described in this study.     

Spinning of a gigantic bundle of hollow fibrils by a spirally moving higher plant protoplast

A unique fiber spinning was found in protoplasts from white birch (Betula platyphylla) leaves under an acidic medium containing high concentration of Ca²⁺. After expanding from 10 to 100 μm in diameter under the culture condition, the protoplast started secreting a gigantic fiber while moving in a spiral way. Real time video analyses elucidated that the orientation, rate and pattern of the motion were directed due to the inverse force of the fiber spinning. Moreover, observation using several microscopic methods accompanied with histochemical staining and nuclear magnetic resonance (NMR) analysis indicated that the fiber was composed of 400–500 nm wide (1→3)-β-glucan hollow sub-fibrils. This entire phenomenon may be a response against the stress imposed. The observation presented provides an understanding of the unique relationship between fiber spinning and the bottom-up fiber fabrication from nano to micro scales.     

Factors affecting the production of <Emphasis Type="SmallCaps">l</Emphasis>-xylulose by resting cells of recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis>

Factors affecting the production of the rare sugar l-xylulose from xylitol using resting cells were investigated. An E. coli BPT228 strain that recombinantly expresses a gene for xylitol dehydrogenase was used in the experiments. The ratio of xylitol to l-xylulose was three times lower in the cytoplasm than in the medium. The effects of pH, temperature, shaking speed, and initial xylitol concentration on l-xylulose production were investigated in shaking flasks using statistical experimental design methods. The highest production rates were found at high shaking speed and at high temperature (over 44°C). The optimal pH for both productivity and conversion was between 7.5 and 8.0, and the optimal xylitol concentration was in the range 250–350 g l⁻¹. A specific productivity of 1.09 ± 0.10 g g⁻¹ h⁻¹ was achieved in a bioreactor. The response surface model based on the data from the shake flask experiments predicted the operation of the process in a bioreactor with reasonable accuracy.     

Phosphorus uptake and turnover by periphyton in the presence of suspended clays

We investigated the effect of suspended bentonite and kaolinite clays on phosphorus uptake and turnover by lotic periphyton in laboratory microcosms. Clays were characterized for their phosphorus affinity using laboratory batch experiments. Periphyton cultivated on glass microscope slides was subjected to a 0.02 mg L⁻¹ radiolabeled soluble reactive phosphorus solution in which a 200-mg L⁻¹ clay load was suspended. A 1-h uptake experiment was followed by a 10-day turnover experiment. Biomass normalized phosphorus uptake, and turnover rates were described by mean rate constants ranging from 0.14 to 0.17 min⁻¹ for uptake and 0.04–0.07 days⁻¹ for turnover. Mean phosphorus concentrations were compared among treatments using repeated measures analysis of variance (ANOVA). Mean phosphorus concentrations among treatments were compared using one-way ANOVA. No significant differences were found among treatments for either analysis. Under laboratory conditions, these clays appear to have little or no short-term influence upon phosphorus uptake or turnover by periphyton.     

Drug-Cyclodextrin-Vesicles Dual Carrier Approach for Skin Targeting of Anti-acne Agent

In the present study attempt was made for preparation of isotretinoin-hydroxypropyl β cyclodextrin (HP-β-CD) inclusion complex and encapsulate this complex in elastic liposomes to study the effect of dual carrier approach on skin targeting of isotretinoin. The isotretinoin HP-β-CD complex was prepared by freeze-drying method and characterized by IR spectroscopy. The drug and drug-CD complex loaded elastic liposomal formulation were prepared and characterized in vitro, ex-vivo and in vivo for shape, size, entrapment efficiency, no. of vesicles per cubic mm, in vitro skin permeation and deposition study, photodegradation and skin toxicity assay. The transdermal flux for different vesicular formulations was observed between 10.5 ± 0.5 to 13.9 ± 1.6 μg/cm²/h. This is about 15-21 folds higher than that obtained from drug solution (0.7 ± 0.1 μg/cm²/h) and 4-5 folds higher than obtained with drug-CD complex solution (2.7 ± 0.1 μg/cm²/h). The amount of drug deposit was found to increase significantly (p < 0.05) by cyclodextrin complexation (30.1 ± 0.1 μg). The encapsulation of this complex in elastic liposomal formulation further increases its skin deposition (262.2 ± 21 μg). The results of skin irritation study using Draize test also showed the significant reduction in skin irritation potential of isotretinoin elastic liposomal formulation in comparison to free drug. The results of the present study demonstrated that isotretinoin elastic liposomal formulation possesses great potential for skin targeting, prolonging drug release, reduction of photodegradation, reducing skin irritation and improving topical delivery of isotretinoin.     

Increase in silicon concentrations and release from suspended solids and bottom sediments in Lake Kasumigaura, Japan

Increasing trends of dissolved Si measured by a colorimetric method and ICP (DSi_col and DSi_ICP, respectively) and total Si concentrations were detected at the center of Lake Kasumigaura during 1980–2006 (mean DSi_col concentration in the 1980s and 2000s was 1.3 and 4.0 mg l⁻¹, respectively). The observation of such trends is rare; therefore, the elucidation of the causes could be useful to understanding silicon dynamics in inland waters. Based on statistical analysis, we found that the increases in DSi_col and lithogenic Si accounted for most of the total Si increase (44 and 45%, respectively) and that biogenic Si, consisting of diatom frustules, also increased with them. Increases in DSi_ICP concentration were not detected near the mouth of the inflowing rivers, suggesting that the increase was caused by in-lake processes. Because the increase in suspended solids (SS) caused by sediment resuspension had been observed in the lake for the same period, we assumed that the Si release from SS containing diatom frustules contributed to the increase. The results of the laboratory experiments in which surface sediments were stirred in lake waters showed that the change in DSi_col concentration depended mainly on SS concentration, water temperature and the elapsed time of diatom frustules dissolution. An estimation of the released amount of Si from SS using the sediment resuspension model was (1.0–2.7) × 10⁹ g year⁻¹ in the 2000s, which was about 30–90% of the increase in the DSi_col outflow of 3.0 × 10⁹ g year⁻¹ from the 1980s to the 2000s. We also determined the Si release rates from bottom sediments through laboratory experiments. The Si amount released from bottom sediments in the lake in the 2000s was estimated to be 4.3 × 10⁹ g year⁻¹, which was about 2–4 times higher than the estimated Si amount released from SS. These findings suggest that the sediment resuspension might be the cause of the latest DSi increase.     

Microstructural and biomechanical alterations of the human aorta as a function of age and location

While it is known that the aorta stiffens with location and age, little is known about the underlying mechanisms that govern these alterations. The purpose of this study was to investigate the relationship between the anisotropic biomechanical behavior and extracellular matrix microstructure of the human aorta and quantify how each changes with location and age. A total of 207 specimens were harvested from 5 locations (ascending n = 33, arch n = 38, descending n = 54, suprarenal n = 52, and abdominal n = 30) of 31 autopsy donor aortas (aged 3 days to 93 years). Each specimen underwent planar biaxial testing in order to derive quantitative biomechanical endpoints of anisotropic stiffness and compliance. Quantitative measures of fiber alignment and degree of fiber alignment were also generated on the same samples using a small-angle light scattering (SALS) technique. Circumferential and axial stiffening occurred with age and increased from the proximal to distal aorta, and the abdominal region was found to be more stiff than all others (p ≤ 0.006). Specimens from donors aged 61 and above were drastically more stiff than younger specimens (p < 0.001) and demonstrated greater circumferential compliance and axial stiffening (p < 0.001). Fiber direction for all ages and locations was predominantly circumferential (p < 0.001), and the degree of fiber alignment was found to increase with age (p < 0.001). Our results demonstrate that the aorta becomes more biomechanically and structurally anisotropic after age 60; with significant changes occurring preferentially in the abdominal aorta, these changes may play an important role in the predisposition of disease formation (e.g., aneurysm) in this region with age.