Effects of genotype and age on eggshell cuticle coverage and color profile in modern laying hen strains

none3noThe aim of this research was to investigate the effects of laying hen genotype and age on eggshell cuticle deposition. A total of 4,320 brown eggs were obtained from 3 modern hen strains (A, B, and C), currently used worldwide for commercial egg production, at different intervals of age (20–30, 40–50, and 60–70 wk). Four samplings of 120 randomly collected eggs were carried out for each genotype/interval of age. Eggs were individually weighed and cuticle blue staining was used to assess quality and degree of cuticle coverage. On each egg, the eggshell color profile was assessed before and after staining using the CIE L*a*b* system and these values were used to calculate ΔE*ab. A 4-point scale visual score (VS) system was also applied to estimate the degree of cuticle coverage after staining (0 = no coverage, 1 = partial coverage, 2 = total coverage - low degree, 3 = total coverage - high degree). The effects of genotype and age and their interaction on eggshell color attributes were assessed by means of factorial ANOVA, while omnibus Chi-Square and Chi-squared Automatic Interaction Detector algorithm were applied for the analysis of VS data. Overall, both genotype and age affected the eggshell color profile as well as the degree of cuticle coverage. Hen strain A showed better cuticle deposition in comparison with B and particularly C one, being ΔE*ab values significantly higher. The VS evaluation revealed that eggs with impaired cuticle coverage degree increased with the hen age (23, 34, and 37%, respectively for 20–30, 40–50, and 60–70 wk; P < 0.05). However, a significant interaction between genotype and age was observed: transition from early to late hen age resulted in a significantly different pattern of ΔE*ab changes in each genotype. The classification tree analysis confirmed that the hen genotype has a greater effect than the age on cuticle deposition. In conclusion, considering the importance of the cuticle in table egg production, these results highlight the crucial role exerted by the genotype on eggshell cuticle coverage.openSirri F.; Zampiga M.; Berardinelli A.Sirri F.; Zampiga M.; Berardinelli A

Variability and interaction of some egg physical and eggshell quality attributes during the entire laying hen cycle

The aim of this study was to investigate the variability and relationships between some egg physical (egg weight, width, length, shape index and surface area) and eggshell parameters (weight and percentage, thickness, breaking strength, L*, a*, and b* values) during the entire laying hen cycle. A total of 8,000 eggs was collected every 5 weeks, from 30 to 81 weeks of hens age (10 samplings of 400 eggs/house), in 2 identical poultry houses equipped with enriched cages. For the statistical analysis, ANOVA, Bivariate Correlation, Principal Component Analysis (PCA) and hierarchical cluster analysis were used. An increase of egg weight, length and eggshell lightness (L*) associated with a reduction of eggshell percentage, breaking strength, and redness (a*) was observed as the hen aged (P10% of eggshell breaking strength and a*. According to the PCA, the highest changes during the laying cycle are related to egg physical parameters (32%) and to eggshell breaking strength, percentage, and thickness (26%). The egg physical parameters appeared to be strongly correlated to each other, whereas a slight correlation between eggshell breaking strength and color attributes were evidenced (-0.231 and 0.289 respectively for L* and a*; P<0.01). Hierarchical cluster analysis, based on principal components of the overall egg attributes is hereby considered, evidenced dissimilarities for eggs laid from peak production up for 39 weeks of hen age from the eggs laid afterwards. The latter group could also be divided into two subgroups, one comprising eggs laid from 44 and 53 weeks of hen age and the other from 58 weeks to the end. In conclusion, the large dataset created in this study allowed to extrapolate some robust information regarding the variability and correlations of the egg physical and eggshell quality attributes throughout the entire laying hen cycle

Inhibition of Bromodomain and Extraterminal Domain (BET) Proteins by JQ1 Unravels a Novel Epigenetic Modulation to Control Lipid Homeostasis

The homeostatic control of lipid metabolism is essential for many fundamental physiological processes. A deep understanding of its regulatory mechanisms is pivotal to unravel prospective physiopathological factors and to identify novel molecular targets that could be employed to design promising therapies in the management of lipid disorders. Here, we investigated the role of bromodomain and extraterminal domain (BET) proteins in the regulation of lipid metabolism. To reach this aim, we used a loss-of-function approach by treating HepG2 cells with JQ1, a powerful and selective BET inhibitor. The main results demonstrated that BET inhibition by JQ1 efficiently decreases intracellular lipid content, determining a significant modulation of proteins involved in lipid biosynthesis, uptake and intracellular trafficking. Importantly, the capability of BET inhibition to slow down cell proliferation is dependent on the modulation of cholesterol metabolism. Taken together, these data highlight a novel epigenetic mechanism involved in the regulation of lipid homeostasis

Status report of a systematic investigation on low-dose ionizing radiation effects in mammalian cells

In the last 15 years a growing interest in the biological effects induced by low doses of ionizing radiation has arisen in the scientific community, due to an increasing number of experimental evidences showing a plethora of non-linear effects occurring after low-dose irradiations. In particular, hyper-radiosensitivity and induced radioresistance (HRS/IRR) have been reported after exposure to low- and high-LET radiation, in human (normal and tumoural) and other mammalian cells in vitro. In this framework, Chinese hamster V79 cells, human primary fibroblasts (HFFF2) and murine embryonic fibroblasts (MEFs) were irradiated with broadbeams of protons in the dose range 0.1–5.0Gy and at 1 Gy/min dose-rate. Cellular response has been evaluated in terms of cell survival, micronuclei induction, chromosomal aberrations and telomere length alterations. For comparison purpose, the same end-points were studied after X/γ-rays irradiation

Evidence that chronic hypoxia causes reversible impairment on male fertility

Aim: To evaluate the effect of chronic hypoxia on human spermatogenic parameters and their recovery time. Methods: Seminological parameters of six male healthy mountain trekkers were evaluated in normoxia at sea level. After 26 days exposure to altitude (ranging from 2 000 m to 5 600 m, Karakorum Expedition) the same parameters were again evaluated after returning to sea level. These parameters were once again evaluated after 1 month and then again after 6 months. Results: Sperm count was found to be lower immediately after returning to sea level (P = 0.0004) and again after a month (P = 0.0008). Normal levels were reached after 6 months. Spermatic motility (%) shows no reduction immediately after returning to sea level (P = 0.0583), whereas after 1 month this reduction was significant (P = 0.0066). After 6 months there was a recovery to pre-hypoxic exposure values. Abnormal or immature spermatozoa (%) increased immediately after returning to sea level (P = 0.0067) and then again after 1 month (P = 0.0004). After 6 months there was a complete recovery to initial values. The total number of motile sperm in the ejaculate was found to be lower immediately after returning to sea level (P = 0.0024) and then again after 1 month (P = 0.0021). After 6 months there was a recovery to pre-hypoxic exposure values. Conclusion: Chronic hypoxia induces a state of oligospermia and the normalization of such seminological parameters at the restoration of previous normoxic conditions after 6 months indicate the influence of oxygen supply in physiological mechanisms of spermatogenesis and male fertility. (Asian J Androl 2008 Jul; 10: 602–606

Hypoxia-Mimetic CoCl2 Agent Enhances Pro-Angiogenic Activities in Ovine Amniotic Epithelial Cells-Derived Conditioned Medium

Amniotic epithelial stem cells (AECs) are largely studied for their pro-regenerative properties. However, it remains undetermined if low oxygen (O2) levels that AECs experience in vivo can be of value in maintaining their biological properties after isolation. To this aim, the present study has been designed to evaluate the effects of a hypoxia-mimetic agent, cobalt chloride (CoCl2), on AECs’ stemness and angiogenic activities. First, a CoCl2 dose-effect was performed to select the concentration able to induce hypoxia, through HIF-1α stabilization, without promoting any cytotoxicity effect assessed through the analysis of cell vitality, proliferation, and apoptotic-related events. Then, the identified CoCl2 dose was evaluated on the expression and angiogenic properties of AECs’ stemness markers (OCT-4, NANOG, SOX-2) by analysing VEGF expression, angiogenic chemokines’ profiles, and AEC-derived conditioned media activity through an in vitro angiogenic xeno-assay. Results demonstrated that AECs are sensitive to the cytotoxicity effects of CoCl2. The unique concentration leading to HIF-1α stabilization and nuclear translocation was 10 µM, preserving cell viability and proliferation up to 48 h. CoCl2 exposure did not modulate stemness markers in AECs while progressively decreasing VEGF expression. On the contrary, CoCl2 treatment promoted a significant short-term release of angiogenic chemokines in culture media (CM). The enrichment in bio-active factors was confirmed by the ability of CoCl2-derived CM to induce HUVEC growth and the cells’ organization in tubule-like structures. These findings demonstrate that an ap-propriate dose of CoCl2 can be adopted as a hypoxia-mimetic agent in AECs. The short-term, chemical-induced hypoxic condition can be targeted to enhance AECs’ pro-angiogenic properties by providing a novel approach for stem cell-free therapy protocols

Pilot Study on Quantitative Cervical Cord and Muscular MRI in Spinal Muscular Atrophy: Promising Biomarkers of Disease Evolution and Treatment?

Introduction: Nusinersen is a recent promising therapy approved for the treatment of spinal muscular atrophy (SMA), a rare disease characterized by the degeneration of alpha motor neurons (αMN) in the spinal cord (SC) leading to progressive muscle atrophy and dysfunction. Muscle and cervical SC quantitative magnetic resonance imaging (qMRI) has never been used to monitor drug treatment in SMA. The aim of this pilot study is to investigate whether qMRI can provide useful biomarkers for monitoring treatment efficacy in SMA. Methods: Three adult SMA 3a patients under treatment with nusinersen underwent longitudinal clinical and qMRI examinations every 4 months from baseline to 21-month follow-up. The qMRI protocol aimed to quantify thigh muscle fat fraction (FF) and water-T2 (w-T2) and to characterize SC volumes and microstructure. Eleven healthy controls underwent the same SC protocol (single time point). We evaluated clinical and imaging outcomes of SMA patients longitudinally and compared SC data between groups transversally. Results: Patient motor function was stable, with only Patient 2 showing moderate improvements. Average muscle FF was already high at baseline (50%) and progressed over time (57%). w-T2 was also slightly higher than previously published data at baseline and slightly decreased over time. Cross-sectional area of the whole SC, gray matter (GM), and ventral horns (VHs) of Patients 1 and 3 were reduced compared to controls and remained stable over time, while GM and VHs areas of Patient 2 slightly increased. We found altered diffusion and magnetization transfer parameters in SC structures of SMA patients compared to controls, thus suggesting changes in tissue microstructure and myelin content. Conclusion: In this pilot study, we found a progression of FF in thigh muscles of SMA 3a patients during nusinersen therapy and a concurrent slight reduction of w-T2 over time. The SC qMRI analysis confirmed previous imaging and histopathological studies suggesting degeneration of αMN of the VHs, resulting in GM atrophy and demyelination. Our longitudinal data suggest that qMRI could represent a feasible technique for capturing microstructural changes induced by SMA in vivo and a candidate methodology for monitoring the effects of treatment, once replicated on a larger cohort

Tendon Immune Regeneration: Insights on the Synergetic Role of Stem and Immune Cells during Tendon Regeneration

Tendon disorders represent a very common pathology in today’s population, and tendinopathies that account 30% of tendon-related injuries, affect yearly millions of people which in turn cause huge socioeconomic and health repercussions worldwide. Inflammation plays a prominent role in the development of tendon pathologies, and advances in understanding the underlying mechanisms during the inflammatory state have provided additional insights into its potential role in tendon dis-orders. Different cell compartments, in combination with secreted immune modulators, have shown to control and modulate the inflammatory response during tendinopathies. Stromal compartment represented by tenocytes has shown to display an important role in orchestrating the inflammatory response during tendon injuries due to the interplay they exhibit with the immune-sensing and infiltrating compartments, which belong to resident and recruited immune cells. The use of stem cells or their derived secretomes within the regenerative medicine field might represent synergic new therapeutical approaches that can be used to tune the reaction of immune cells within the damaged tissues. To this end, promising opportunities are headed to the stimulation of macrophages polarization towards anti-inflammatory phenotype together with the recruitment of stem cells, that possess immunomodulatory properties, able to infiltrate within the damaged tissues and improve tendinopathies resolution. Indeed, the comprehension of the interactions between tenocytes or stem cells with the immune cells might considerably modulate the immune reaction solving hence the inflammatory response and preventing fibrotic tissue formation. The purpose of this review is to compare the roles of distinct cell compartments during tendon homeostasis and injury. Furthermore, the role of immune cells in this field, as well as their interactions with stem cells and tenocytes during tendon regeneration, will be discussed to gain insights into new ways for dealing with tendinopathies