Validation of a Cleanroom Compliant Sonication-Based Decellularization Technique: A New Concept in Nerve Allograft Production

Defects of the peripheral nervous system are extremely frequent in trauma and surgeries and have high socioeconomic costs. If the direct suture of a lesion is not possible, i.e., nerve gap > 2 cm, it is necessary to use grafts. While the gold standard is the autograft, it has disadvantages related to its harvesting, with an inevitable functional deficit and further morbidity. An alternative to autografting is represented by the acellular nerve allograft (ANA), which avoids disadvantages of autograft harvesting and fresh allograft rejection. In this research, the authors intend to transfer to human nerves a novel technique, previously implemented in animal models, to decellularize nerves. The new method is based on soaking the nerve tissues in decellularizing solutions while associating ultrasounds and freeze–thaw cycles. It is performed without interrupting the sterility chain, so that the new graft may not require post-production γ-ray irradiation, which is suspected to affect the structural and functional quality of tissues. The new method is rapid, safe, and inexpensive if compared with available commercial ANAs. Histology and immunohistochemistry have been adopted to evaluate the new decellularized nerves. The study shows that the new method can be applied to human nerve samples, obtaining similar, and, sometimes better, results compared with the chosen control method, the Hudson technique.     

Mechanical behavior of cold-water fish gelatin gels crosslinked with 1,4-butanediol diglycidyl ether

In this work, chemical crosslinking with 1,4-butanediol diglycidyl ether (BDDGE) is used as strategy to enhance mechanical performance of fish gelatin (FG) gels in order to meet the properties' range of mammalian gelatin physical gels. Joint analysis of free amino groups, swelling ratio, and total soluble material indicates that crosslinking degree increases with increasing FG concentration and it is favored by a 0.2 BDDGE/FG ratio. Increasing crosslinking degree enhances gel indentation strength and shear modulus (μ) while decreases fracture toughness (G_IC). Measured μ and G_IC values lies within the range exhibited by mammalian gelatin physical gels, but the relationship between these parameters is opposite. This is due to the different fracture mechanisms occurring in chemically crosslinked and physical gels.     

Supervised Multivariate Analysis of Hyper-spectral NIR Images to Evaluate the Starch Index of Apples

Fruit maturity indexes are crucial in harvest time determination and commercial context. The harvest time of apples, matching the desired commercial characteristics, is assessed through starch–iodine test in practice. Fruit halves are dipped into iodine solution and patterns are visually evaluated by experts comparing them to reference charts. Aim of the work was to study the relationships of near infrared (NIR) spectral images (1,000–1,700 nm), starch/starch-free patterns visually assessed and RGB color images. Spectral images of 88 Golden Delicious Klon B apples were sampled at seven different maturity stages. Partial least-squares discriminant analysis (PLSDA) technique was used on hyper-spectral NIR images to classify single pixels using its NIR reflectance spectrum. The response variable (i.e. the classification for each pixel) was identified through the matching of single pixel obtained with the color images, segmented in two classes (starch and starch-free), and the NIR hyper-spectral matrix. Mean hyper-spectral classification obtained through PLSDA modeling on individual apple correctly classified 80.81% of the total pixels, while the unique model, i.e. a single model including all the fruits, resulted in 66.33%. In the latter case, the relationship with the RGB classification showed high values (Pearson correlation coefficient r = 0.95). The present work shows the feasibility of NIR imaging spectroscopy as a tool for apple fruit maturity determination, avoiding expert’s subjective interpretation by traditional starch index assignments.     

Composition of phospholipid fraction in raw chicken meat and pre-cooked chicken patties: influence of feeding fat sources and processing technology

The effect of dietary lipid sources (animal fat, AF vs. vegetable oil, VO) was investigated on the phospholipid (PL) fraction of raw chicken meat and pre-cooked chicken patties. PL classes were determined on extracted lipids by high-performance liquid chromatography (HPLC) connected with an evaporative light-scattering detector (ELSD). The HPLC-ELSD method showed good intraday repeatability (less than 5% for different PL) and limits of detection ranging from 0.006 mg/mL (phosphatidylethanolamine) to 0.045 mg/mL (sphingomyelin). Diet did not consistently influence PL in chicken breast. A significant difference was assessed in meat batter where total PL were at levels of 349.0 and 388.2 mg/100 g of product in batter from broilers fed AF and VO diet, respectively. Cooking oil absorption caused an important decrease in PL (lower than 300 mg/100 g of product in pre-fried patties). Diet was an effective means to control the PL fatty acid profile in chicken, particularly the levels of polyunsaturated fatty acids (PUFA). In minced breast and thigh and meat batter from birds fed a VO-supplemented diet, the relative amount of PUFA was 46, 81 and 28% higher than that determined in the corresponding samples obtained from broilers fed an AF-based diet.     

External Shape Differences between Sympatric Populations of Commercial Clams <Emphasis Type="Italic">Tapes decussatus</Emphasis> and <Emphasis Type="Italic">T. philippinarum</Emphasis>

The actual Italian production of clams is chiefly sustained by the native Tapes decussatus and the fortuitously imported Tapes philippinarum. Both species are commercialized as “Vongola verace”, but the commercial value of T. philippinarum is lower. The discrimination of species by sight is usually difficult and it cannot be done by observation based on shell morphology but only when animals open their valves hence displaying the two siphons. In this study, we propose a new, noninvasive method to discriminate individuals of both species based on the analysis of the external shape of their shells. Accordingly, in sympatric populations at two sites of the Po river outlet, we have chosen individuals (63 for T. decussatus and 57 for T. philippinarum) of comparable commercial size for which a certain genetic discrimination was previously done. Pictures of the left side valve were taken for all specimens. Their profiles were analyzed with the elliptic Fourier analysis. The mean outline for each species was graphically extracted. The coefficients of the harmonic equations were analyzed by multivariate classification (partial least squares discriminant analysis [PLSDA]). Results showed a high percentage of correct classification of individuals of both species (96.6%). Contour analysis reflected the overall shell shape and thus identified morphological aspects that were difficult to recognize and quantify in sight. The high percentage of correct classifications obtained by combining the analysis of elliptic Fourier harmonics with PLSDA demonstrated the feasibility of this method to discriminate species with a high level of resemblance.     

Root Exposure to 5-Aminolevulinic Acid (ALA) Affects Leaf Element Accumulation,Isoprene Emission,Phytohormonal Balance,and Photosynthesis of Salt-Stressed Arundo donax

Arundo donax has been recognized as a promising crop for biomass production on marginal lands due to its superior productivity and stress tolerance. However, salt stress negatively impacts A. donax growth and photosynthesis. In this study, we tested whether the tolerance of A. donax to salinity stress can be enhanced by the addition of 5-aminolevulinic acid (ALA), a known promoter of plant growth and abiotic stress tolerance. Our results indicated that root exposure to ALA increased the ALA levels in leaves along the A. donax plant profile. ALA enhanced Na⁺ accumulation in the roots of salt-stressed plants and, at the same time, lowered Na⁺ concentration in leaves, while a reduced callose amount was found in the root tissue. ALA also improved the photosynthetic performance of salt-stressed apical leaves by stimulating stomatal opening and preventing an increase in the ratio between abscisic acid (ABA) and indol-3-acetic acid (IAA), without affecting leaf methanol emission and plant growth. Supply of ALA to the roots reduced isoprene fluxes from leaves of non-stressed plants, while it sustained isoprene fluxes along the profile of salt-stressed A. donax. Thus, ALA likely interacted with the methylerythritol 4-phosphate (MEP) pathway and modulate the synthesis of either ABA or isoprene under stressful conditions. Overall, our study highlights the effectiveness of ALA supply through soil fertirrigation in preserving the young apical developing leaves from the detrimental effects of salt stress, thus helping of A. donax to cope with salinity and favoring the recovery of the whole plant once the stress is removed.     

Explicit versus implicit method for radiative heat transfer in gray and diffuse enclosures

The paper presents an explicit formulation of the radiative problem in gray enclosures. The method is based on a series expansion that derives the mutual radiation factors by means of summation of the contributions to the radiative thermal exchange given by the multi-reflection process. A mathematical proof that the infinite expansion converges to the well known solution, given by the implicit formulation and based on radiosities balance, is provided. Moreover a parametric analysis of the approximation induced by the explicit algorithm, as a result of the truncation order in the calculation of the series expansion, is carried out. The analysis shows how the precision of every approximation order is mainly related to the surface averaged infrared reflectance and is weakly dependent on the reflectance distribution. For high emissive and low reflecting surfaces, very few summations are required to meet high precision; the same level of precision is achievable also with medium and high reflecting surfaces, by increasing the truncation order of the series. More generally, the explicit formulation provides an alternative approach to problems involving multi-reflecting cavities in different engineering applications, from heat transfer to optics.