Acellularization-Induced Changes in Tensile Properties Are Organ Specific - An In-Vitro Mechanical and Structural Analysis of Porcine Soft Tissues

Introduction

Though xenogeneic acellular scaffolds are frequently used for surgical reconstruction, knowledge of their mechanical properties is lacking. This study compared the mechanical, histological and ultrastructural properties of various native and acellular specimens.

Materials and Methods

Porcine esophagi, ureters and skin were tested mechanically in a native or acellular condition, focusing on the elastic modulus, ultimate tensile stress and maximum strain. The testing protocol for soft tissues was standardized, including the adaption of the tissue’s water content and partial plastination to minimize material slippage as well as templates for normed sample dimensions and precise cross-section measurements. The native and acellular tissues were compared at the microscopic and ultrastructural level with a focus on type I collagens.

Results

Increased elastic modulus and ultimate tensile stress values were quantified in acellular esophagi and ureters compared to the native condition. In contrast, these values were strongly decreased in the skin after acellularization. Acellularization-related decreases in maximum strain were found in all tissues. Type I collagens were well-preserved in these samples; however, clotting and a loss of cross-linking type I collagens was observed ultrastructurally. Elastins and fibronectins were preserved in the esophagi and ureters. A loss of the epidermal layer and decreased fibronectin content was present in the skin.

Discussion

Acellularization induces changes in the tensile properties of soft tissues. Some of these changes appear to be organ specific. Loss of cross-linking type I collagen may indicate increased mechanical strength due to decreasing transverse forces acting upon the scaffolds, whereas fibronectin loss may be related to decreased load-bearing capacity. Potentially, the alterations in tissue mechanics are linked to organ function and to the interplay of cells and the extracellular matrix, which is different in hollow organs when compared to skin.     

Dibutyryl-cAMP affecting fat deposition of finishing pigs by decreasing the inflammatory system related to insulin sensitive or lipolysis

Background

The mechanism of db-cAMP regulating fat deposition and improving lean percentage is unclear and needs to be further studied.

Methods

Eighteen 100-day-old Duroc × Landrance × Large White barrows (49.75?±?0.75 kg) were used for experiment 1, and 15 eighteen 135-day-old barrows (78.34?±?1.22 kg) were used for experiment 2 to investigate the effects of dietary dibutyryl-cAMP (db-cAMP) on fat deposition in finishing pigs. Pigs were fed with a corn-soybean meal-based diet supplemented with 0 or 15 mg/kg db-cAMP, and both experiments lasted 35 days, respectively.

Results

The results showed that db-cAMP decreased the backfat thickness, backfat percentage, and diameter of backfat cells without changing the growth performance or carcass characteristics in both experiments, and this effect was more marked in experiment 1 than in experiment 2; db-cAMP enhanced the activity of the growth hormone–insulin-like growth factor-1 (GH-IGF-1) axis and pro-opiomelanocortin (POMC) system in both experiments, which suppressed the accumulation of backfat deposition; microarray analysis showed that db-cAMP suppressed the inflammatory system within the adipose tissue related to insulin sensitivity, which also reduced fat synthesis.

Conclusions

In summary, the effect of db-cAMP on suppressing fat synthesis and accumulation is better in the earlier phase than in the later phase of finishing pigs, and db-cAMP plays this function by increasing the activity of the GH-IGF-1 axis and POMC system, while decreasing the inflammatory system within the adipose tissue related to insulin sensitive or lipolysis.

     

Effect of Hydrothermal Modifications on Functional,Pasting and Structural Properties of False Banana (<Emphasis Type="Italic">Ensete ventricosum</Emphasis>) Starch

Starch extracted from ensete (Ensete ventricosum, Musaceae) also called false banana, was modified by hydrothermal methods of annealing (ANN) and heat moisture treatment (HMT) processes. The effects of treatments on functional, pasting properties, morphology and diffraction pattern of the starch were studied. Swelling power and solubility changed significantly (p < 0.05) with modification. Water absorption (89.3–152.4 %) and oil absorption (105.0–161.3 %) capacities increased significantly (p < 0.05) with ANN and HMT. Alkaline water retention decreased with ANN but increased significantly (p < 0.05) with HMT. Hydrothermal modifications led to reduction in least gelation capacity of ensete starch. In terms of the pasting properties studied, the hydrothermal modifications imparted improved gel strength, increased paste stability, reduced retrogradation tendency and slowed staling rate on ensete starch. Scanning electron micrographs depicted fairly angular and elliptical shapes with diverse sizes for the starch granules. Clustering of granules, mucilage formation, fissures and surface indentation which were gaining prominence with increasing moisture level and temperature of treatment were the hallmarks of modified samples. Native and modified ensete starches showed similar type-B diffraction pattern with maximum peak range of 19.8–20.0^o (2θ). Findings of this work showed that hydrothermally modified ensete starches possess excellent value-added potentials for utilization in pharmaceuticals and food applications.     

Simplifying Hansen Solubility Parameters for Complex Edible Fats and Oils

Hansen solubility parameters (HSPs), often used to predict the miscibility between two compounds, are an alternative tool in evaluating the ability of the solvent to interact via dispersion, dipole-dipole, and hydrogen bonding interactions. The aim of this paper is to find a simple way to calculate HSPs for complex mixtures of triglycerides (TAGs). HSPs were calculated using two approaches: the first assumes that the contributions to the dispersion, dipole-dipole, and hydrogen bonding interactions may be subdivided into larger functional moieties (i.e., fatty acids and fatty acid methyl esters) that are additive, while the second approach assumes that vegetable oils are comprised of mixtures of simple TAGs in the same mass fractions as the fatty acids. The HSPs obtained using the two approaches are compared to reference values determined using the “Hansen Solubility Parameters in Practice” software (HSPiP) considering the complex TAG profile for each vegetable oil.HSPs for vegetable oils, obtained with the HSPiP software, did not correspond well to the HSPs obtained from the group contribution approach, when using fatty acids, fatty acids + glycerol or fatty acid methyl esters. In contrast, the HSPs calculated for vegetable oils, assuming that all TAGs are simple and in the same mass fractions as the fatty acids, provide similar values to the HSPs obtained from the HSPiP software. Therefore, it is possible to calculate the HSPs for complex oils by simply knowing the fatty acid composition. Knowledge of HSPs may be used to rationalize the ability of certain low molecular weight molecules to develop organogels in vegetable oils as well as the crystallization of triglycerides.     

Population size and land use affect the genetic variation and performance of the endangered plant species <Emphasis Type="Italic">Dianthus seguieri</Emphasis> ssp. <Emphasis Type="Italic">glaber</Emphasis>

Human activity and land use changes in the past decades have led to landscape homogenization and small-scale fragmentation of grassland habitats in most regions of central Europe. As a result, populations of many grassland species are small and strongly fragmented, facing extinction due to genetic depauperation and local maladaptation in remnant habitats. In this study, remaining populations of the strongly endangered grassland species Dianthus seguieri ssp. glaber (“Ragged Pink”) in Bavaria were investigated in order to evaluate the environmental factors influencing its genetic variation and performance. We first evaluated habitat, vegetation and population structure. Species performance was then studied by assessing the number of generative shoots, flowers and fertile capsules; and evaluating seed weight and seed viability. Finally, genetic variation was analyzed using molecular markers (AFLPs). Our analyses revealed that population size and land use abandonment have the strongest impact on genetic variation and species’ performance. Large and extended populations were most variable. 72 % of overall genetic variability of Dianthus seguieri ssp. glaber was found to be within populations, whereas 28 % remained between populations. Increased vegetation height and coverage, and a high proportion of gramineous species resulting from the lack of land use, reduced genetic variation, effective fruit and seed set. Our study shows that both population size and land use abandonment need to be considered to ensure the long term protection of endangered plant species. Maintaining an open habitat structure and adequate soil nutrient conditions through targeted annual mowing regime, over-storey vegetation trimming and green waste removal and the establishment of vegetation buffer strips will allow this species’ persistence and continuous recruitment.     

Genetic evidence of recent migration among isolated-by-sea populations of the freshwater blenny (<Emphasis Type="Italic">Salaria fluviatilis</Emphasis>)

Genetic diversity is the raw material for evolutionary change, so a species’ capacity to maintain its genetic diversity is a major concern in conservation genetics. Although genetic diversity within a population is reduced through time by genetic drift, gene flow among populations can act to recover or add new genetic variants. The goal of this study was to infer potential connectivity among isolated-by-sea populations of the vulnerable freshwater blenny (Salaria fluviatilis) and to determine if gene flow could contribute to maintaining genetic diversity in connected populations. Four genetic clusters (one small at the North, one large at the South for both East and West coasts) were detected with different clustering methods (FLOCK, STUCTURE, UPGMA, AMOVA). The two larger genetic clusters with higher migration-rate estimates among localities had higher genetic diversity and allelic richness and lower relatedness between individuals, compared to isolated localities found in smaller clusters. Our results also suggest that sea currents may facilitate fish movements among neighbouring rivers. Overall, gene flow among isolated-by-sea but close rivers could maintain the evolutionary potential of freshwater blenny populations. This finding should be considered when elaborating a conservation program for this species.     

Integrating ecological and genetic structure to define management units for caribou in Eastern Canada

Genetic diversity is a key parameter to delineate management units, but many organisms also display ecological characteristics that may reflect potential local adaptations. Here, we used ecological and genetic information to delineate management units for a complex system involving several ecotypes of caribou (Rangifer tarandus) from Québec and Labrador, eastern Canada. We genotyped 560 caribou at 16 microsatellite loci and used three Bayesian clustering methods to spatially delineate and characterize genetic structure across the landscape. The different approaches employed did not converge on the same solution, and differed in the number of inferred genetic clusters that best fit the dataset but also in the spatial distribution of genetic variation. We reconciled variability among the methods using a synthetic approach that considers the sum of the partitions obtained by each of them and retrieved six genetically distinct groups that differ in their spatial extent across the range of caribou in the study area. These genetic groups are not consistent with the presently defined ecological designations for this species. Combining both genetic and ecological criteria, we distinguished eight independent management units. Overall, the management units we propose should be the focus of conservation and management actions aimed to maximize genetic and ecological diversity and ensure the persistence of caribou populations inhabiting increasingly disturbed landscapes.     

Barn owls (<Emphasis Type="Italic">Tyto alba</Emphasis>) in western North America: phylogeographic structure,connectivity, and genetic diversity

The barn owl (Tyto alba) is a non-migratory species widely distributed across much of North America in areas with extensive old-field and grassland habitat and without extensive winter snow cover. We investigated the genetic diversity and phylogeographic patterns of barn owl populations in western North America, ranging from British Columbia (BC) to southern California, and one eastern population from Pennsylvania. We also determined the genetic distinctiveness of a population off the coast of southern California, Santa Barbara Island, as management plans to control the local owl population are being considered to decrease predation rate on the now threatened Scripps’s Murrelet (Synthliboramphus scrippsi). Using 8 polymorphic microsatellite markers (N = 126) and ND2 mitochondrial sequences (N = 37), we found little to no genetic structure among all sampled regions, with the exception of Santa Barbara Island. The BC mainland population, despite its northwestern geographically peripheral location and ongoing habitat degradation, is not genetically depauperate. However, individuals from Vancouver Island, likewise a peripheral population in BC, exhibited the lowest genetic diversity of all sampled locations. The low global F_ST value (0.028) estimated from our study suggests that old-field agricultural habitats are well connected in North America. Since the BC population has declined by about 50 % within the last three decades, it is vital to focus on preserving the remaining barn owl habitats in BC to allow successful establishment from neighbouring populations. Additionally, our microsatellite data revealed that the population on Santa Barbara Island showed genetic divergence from its continental counterpart. Mitochondrial data, however, demonstrated that this island population is not a monophyletic lineage containing unique haplotypes, and hence cannot be designated as an Evolutionarily Significant Unit.