Integrated extraction, structural characterization, and activity assessment of squid pen protein hydrolysates and ß-chitin with different protease hydrolysis.

Squid pen (SP) is a valuable source of protein and ß-chitin. However, current research has primarily focused on extracting ß-chitin from SP. This study innovatively extracted both SP protein hydrolysates (SPPHs) and SP ß-chitin (SPC) simultaneously using protease hydrolysis. The effects of different proteases on their structural characteristics and bioactivity were evaluated. The results showed that SP alcalase ß-chitin (SPAC) had the highest degree of deproteinization (DP, 98.19 %) and SP alcalase hydrolysates (SPAH) had a degree of hydrolysis (DH) of 24.47 %. The analysis of amino acid composition suggested that aromatic amino acids accounted for 17.44 % in SPAH. Structural characterization revealed that SP flavourzyme hydrolysates (SPFH) had the sparsest structure. SPC exhibited an excellent crystallinity index (CI, over 60 %) and degree of acetylation (DA, over 70 %). During simulated gastrointestinal digestion (SGD), the hydroxyl radical scavenging activity, ABTS radical scavenging activity, Fe2+ chelating activity, and reducing power of the SPPHs remained stable or increased significantly. Additionally, SPFC exhibited substantial inhibitory effects on Staphylococcus aureus and Escherichia coli (S. aureus and E. coli), with inhibition circle diameters measuring 2.4 cm and 2.1 cm. These findings supported the potential use of SPPHs as natural antioxidant alternatives and suggested that SPC could serve as a potential antibacterial supplement.

Preparation and Modification of Collagen/Sodium Alginate-Based Biomedical Materials and Their Characteristics.

(1) Background: Collagen and sodium alginate are commonly used in the field of biomedical materials due to their excellent biocompatibility. This study focuses on the preparation, modification, and characterization of collagen/sodium alginate (C/SA)-based biomedical materials. (2) Methods: The characteristics, including surface chemistry, mechanical properties, hygroscopicity, and porosity, were analyzed. The hemostatic activity in vitro was measured using a blood clotting assay and dynamic blood clotting assay. (3) Results: The results from microstructure and porosity measurement revealed that all of the sponges exhibited a porosity of more than 95 percent. The sponge cross-linked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) showed better tensile strength and lower elongation at break. The sponges cross-linked with EDC/NHS and oxidized sodium alginate (OSA) exhibited the highest hygroscopicity in comparison with the uncross-linked sponge. (4) Conclusions: Our study demonstrated that the C/SA-based material we prepared exhibited a high level of porosity, enabling efficient absorption of tissue exudate and blood. Additionally, the materials revealed excellent hemocompatibility, making them suitable for use as a hemostatic dressing in the field of biomedical materials.

Mulberry-derived miR168a downregulates BmMthl1 to promote physical development and fecundity in silkworms.

Plant-derived miRNAs and their interactions with host organisms are considered important factors in regulating host physiological processes. In this study, we investigated the interaction between the silkworm, an oligophagous insect, and its primary food source, mulberry, to determine whether mulberry-derived miRNAs can penetrate silkworm cells and regulate their functions. Our results demonstrated that miR168a from mulberry leaves enters the silkworm hemolymph and binds to the silkworm Argonaute1 BmAGO1, which is transported via vesicles secreted by silkworm cells to exert its regulatory functions. In vivo and in vitro functional studies revealed that miR168a targets the mRNA of silkworm G protein-coupled receptor, BmMthl1, thereby inhibiting its expression and activating the JNK-FoxO pathway. This activation reduces oxidative stress responses, prolongs the lifespan of silkworms, and improves their reproductive capacity. These findings highlight the challenges of replacing mulberry leaves with alternative protein sources and provide a foundation for developing silkworm germplasms suitable for factory rearing.

On the thermal response of multiscale nanodomains formed in trans-anethol/ethanol/water surfactant-free microemulsion.

HYPOTHESIS: Surfactant-free microemulsion (SFME), an emerging phenomenology that occurs in the monophasic zone of a broad category of ternary mixtures 'hydrophobe/hydrotrope/water', has attracted extensive interests due to their unique physicochemical properties. The potential of this kind of ternary fluid for solubilization and drug delivery make them promising candidates in many industrial scenarios. EXPERIMENTS: Here the thermodynamic behavior of these multiscale nanodomains formed in the ternary trans-anethol/ethanol/water system over a wide range of temperatures is explored. The macroscopic physical properties of the ternary solutions are characterized, with revealing the temperature dependence of refractive index and dynamic viscosity. FINDINGS: With increasing temperature, the ternary system shows extended areas in the monophasic zone. We demonstrate that the phase behavior and the multiscale nanodomains formed in the monophasic zone can be precisely and reversibly tuned by altering the temperature. Increasing temperature can destroy the stability of the multiscale nanodomains in equilibrium, with an exponential decay in the scattering light intensity. Nevertheless, molecular-scale aggregates and mesoscopic droplets exhibit significantly different response behaviors to temperature stimuli. The temperature-sensitive nature of the ternary SFME system provides a crucial step forward exploring and industrializing its stability.

The Structural Characteristics and Bioactivity Stability of Cucumaria frondosa Intestines and Ovum Hydrolysates Obtained by Different Proteases.

The study aimed to investigate the effects of alcalase, papain, flavourzyme, and neutrase on the structural characteristics and bioactivity stability of Cucumaria frondosa intestines and ovum hydrolysates (CFHs). The findings revealed that flavourzyme exhibited the highest hydrolysis rate (51.88% ± 1.87%). At pH 2.0, the solubility of hydrolysate was the lowest across all treatments, while the solubility at other pH levels was over 60%. The primary structures of hydrolysates of different proteases were similar, whereas the surface hydrophobicity of hydrolysates was influenced by the types of proteases used. The hydrolysates produced by different proteases were also analyzed for their absorption peaks and antioxidant activity. The hydrolysates of flavourzyme had ß-fold absorption peaks (1637 cm-1), while the neutrase and papain hydrolysates had N-H bending vibrations. The tertiary structure of CFHs was unfolded by different proteases, exposing the aromatic amino acids and red-shifting of the λ-peak of the hydrolysate. The alcalase hydrolysates showed better antioxidant activity in vitro and better surface hydrophobicity than the other hydrolysates. The flavourzyme hydrolysates displayed excellent antioxidant stability and pancreatic lipase inhibitory activity during gastrointestinal digestion, indicating their potential use as antioxidants in the food and pharmaceutical industries.

On evaporation dynamics of an acoustically levitated multicomponent droplet: Evaporation-triggered phase transition and freezing.

HYPOTHESIS: Multi-component droplet evaporation has received significant attention in recent years due to the broad range of applications such as material science, environmental monitoring, and pharmaceuticals. The selective evaporation induced by the different physicochemical properties of components is expected to influence the concentration distributions and the separation of mixtures, thereby leading to rich interfacial phenomena and phase interactions. EXPERIMENTS: A ternary mixture system containing hexadecane, ethanol, and diethyl ether is investigated in this study. The diethyl ether exhibits both surfactant-like and co-solvent properties. Systematic experiments were performed using acoustic levitation technique to achieve a contact-less evaporation condition. The evaporation dynamics and temperature information are acquired in the experiments, using high-speed photography and infrared thermography technologies. FINDINGS: Three distinct stages, namely, 'Ouzo state', 'Janus state', and 'Encapsulating state', are identified for the evaporating ternary droplet in acoustic levitation. A self-sustaining periodic freezing & melting evaporation mode is reported. A theoretical model is developed to characterize the multi-stage evaporating behaviors. We demonstrate the capability to tune the evaporating behaviors by varying the initial droplet composition. This work provides a deeper understanding of the interfacial dynamics and phase transitions involved in multi-component droplets and proposes novel strategies for the design and control of droplet-based systems.

Improving the Sustainability of Processing By-Products: Extraction and Recent Biological Activities of Collagen Peptides.

Society and consumers are increasingly concerned about food safety and the sustainability of food production systems. A significant amount of by-products and discards are generated during the processing of aquatic animals, which still needs to be fully utilized by the food industry. The management and sustainable use of these resources are essential to avoiding environmental pollution and resource waste. These by-products are rich in biologically active proteins, which can be converted into peptides by enzymatic hydrolysis or fermentation treatment. Therefore, exploring the extraction of collagen peptides from these by-products using an enzymatic hydrolysis technology has attracted a wide range of attention from numerous researchers. Collagen peptides have been found to possess multiple biological activities, including antioxidant, anticancer, antitumor, hypotensive, hypoglycemic, and anti-inflammatory properties. These properties can enhance the physiological functions of organisms and make collagen peptides useful as ingredients in food, pharmaceuticals, or cosmetics. This paper reviews the general methods for extracting collagen peptides from various processing by-products of aquatic animals, including fish skin, scales, bones, and offal. It also summarizes the functional activities of collagen peptides as well as their applications.

Coupling Effects of Ionic Surfactants and Electrolytes on the Stability of Bulk Nanobubbles.

As interest in the extensive application of bulk nanobubbles increases, it is becoming progressively important to understand the key factors affecting their anomalous stability. The scientific intrigue over nanobubbles originates from the discrepancy between the Epstein-Plesset prediction and experimental observations. Herein, the coupling effects of ionic surfactants and electrolytes on the stability of bulk nanobubbles is studied. Experimental results show that ionic surfactants not only reduce the surface tension but also promote the accumulation of net charges, which facilitate the nucleation and stabilization of bulk nanobubbles. The addition of an electrolyte in a surfactant solution further results in a decrease in the zeta potential and the number concentration of nanobubbles due to the ion shielding effect, essentially colloidal stability. An adsorption model for the coexistence of ionic surfactants and electrolytes in solution, that specifically considers the effect of the adsorption layer thickness within the framework of the modified Poisson-Boltzmann equation, is developed. A quantitative agreement between the predicted and experimental surface tension is found in a wide range of bulk concentrations. The spatial distribution of the surface potential, surfactant ions and counterions in the vicinity of the interface of bulk nanobubbles are described. Our study intrinsically paves a route to investigate the stability of bulk nanobubbles.

Spontaneously formed multiscale nano-domains in monophasic region of ternary solution.

HYPOTHESIS: Ternary mixtures, containing one hydrotrope and two immiscible fluids, both being soluble in the hydrotrope at any proportion, exhibit unexpected solubilization power and unusual mesoscopic properties, which are a subject of long-standing controversies for decades. EXPERIMENTS: This work investigates the entire monophasic region of ternary trans-anethol/ethanol/water system, where multiscale nanostructurings with correlation lengths exceeding dimensions of individual molecules are identified by dynamic light scattering. The physical properties of the ternary mixture are characterized, with revealing the compositional dependence of refractive index and dynamic viscosity. FINDINGS: In surfactant-free microemulsion (SFME) regime, the single phase consists of two distinct nanoscopic domains in equilibrium, one trans-anethol-rich aggregate at the molecular scale (∼1 nm) and one mesoscopic droplet at the mesoscale (∼100 nm). However, only a tiny fraction of the hydrophobic component trans-anethol (<∼0.025%) are initially incorporated in mesoscale structures, and the vast majority are molecularly dissolved or in the form of aggregates. The concentration of mesoscopic droplets increases sharply when shifting toward the miscibility gap, but their size exhibits a weak compositional dependence. The nanoscopic domains exhibit long-term physical stability, and the Ostwald ripening is still the primary mechanism governing such mesoscopic droplet aging, but with a rather low ripening rate.

Thermally Assisted Heterogeneous Cavitation through Gas Supersaturation.

We demonstrate that besides gaseous pockets also a gas supersaturated spot on a substrate can be a nucleus for cavitation. The supersaturation is achieved by either a formerly dissolved bubble or by heating locally the surface below the boiling temperature. The experiments are conducted in a thin film of water; one side of the water film is in contact with a gold coated substrate that is heated by a continuous laser through plasmonic heating. For nucleation of a bubble, the pressure at the heated spot is reduced by a transient rarefaction wave. The experimental findings suggest that the local gas supersaturation is responsible to nucleate cavitation and thus connects the phase transitions of cavitation and boiling. Additionally, the pressure waves in the liquid gap are studied numerically.

Reduced Circulating Levels of miR-491-5p and miR-485-3p Are Associated with the Occurrence of Vertebral Fractures in Postmenopausal Women with Osteoporosis.

Objective: Postmenopausal women experiences osteoporotic structural damage and bone fragility resulting from reduced bone formation and increased bone resorption. Osteoporosis frequently affects the vertebral column and causes compression fractures. This study aims to characterize roles of miRNAs in osteoporosis and subsequent incidence risk of vertebral fractures for postmenopausal women. Methods. Differentially expressed miRNAs between osteoporotic patients with vertebral fractures and osteoporotic patients without fracture were identified. This retrospective study included 78 osteoporotic patients with vertebral fractures and 82 osteoporotic patients without vertebral fractures. The plasma levels of bone metabolic markers, 25-hydroxyvitamin D (25-(OH)VitD), propeptide of type I procollagen (PINP), and ß-Carboxyl terminal peptide (ß-CTx), were detected using the patented electro-chemiluminescence (ECLIA) method. The expression levels of miR-491-5p and miR-485-3p were determined by qRT-PCR. Pearson correlation analysis was carried out to assess the relationship between miR-491-5p, miR-485-3p, and bone metabolic markers. Receiver operating characteristic (ROC) curves and the area under the ROC curve (AUC) were used to evaluate the performance of miR-491-5p and miR-485-3p in diagnosing the occurrence of vertebral fractures in osteoporotic patients.Results: The plasma levels of PINP and ß-CTx were elevated but the plasma level of 25-(OH)VitD was declined in osteoporotic patients with vertebral fractures when comparable to those without (< 0.05). The plasma expression levels of miR-491-5p and miR-485-3p were declined osteoporotic patients with vertebral fractures when comparable to those without (< 0.001). Pearson correlation analysis revealed that the relative expression level of miR-491-5p was negatively correlated with the level of 25-(OH)VitD (r = -0.518, < 0.001) but positively correlated with the levels of PINP (r = 0.547, < 0.001) and ß-CTx (r = 0.380, < 0.001). We also observed a negative correlation between the relative expression level of miR-485-3p and 25-(OH)VitD (r = -0.388, < 0.001), a positive correlation between miR-485-3p and PINP (r = 0.422,< 0.001). ROC curves for prediction of vertebral fracture following osteoporosis in postmenopausal women by miR-491-5p expression yielded 0.866 AUC and by miR-485-3p expression produced 0.848 AUC. Conclusion: The data suggest that downregulated expressions of miR-491-5p and miR-485-3p may be involved in the occurrence of vertebral fractures in postmenopausal women with osteoporosis.

Fluoride contamination, spatial variation, and health risk assessment of groundwater using GIS: a high-density survey sampling in Weifang City, North China.

The present study, with the aid of GIS, utilizes high-density groundwater (GW) sampling data (1398 samples) to analyze the spatial variation characteristics of GW fluoride in Weifang City (WFC), and evaluate the health risks associated with drinking water routes. The concentration of fluoride in the GW of WFC is observed to be between 0.08 and 9.16 mg/L, with a mean value of 0.62 mg/L. The fluoride concentration of a total of 192 GW samples exceeded the limit of China's GW quality standards (1 mg/L), accounting for 14.74%. The GW fluoride concentration in most areas of WFC is less than 1 mg/L. However, the relatively high-value zones are mostly concentrated in the upper reaches of Wen River, the east of Shouguang, the southeast of Anqiu, the east of Qingzhou, the east of Fangzi, and the southeast and northwest of Gaomi. The hydrochemical types of GW in WFC are mostly HCO3-Ca·Mg and SO4·Cl-Ca·Mg, while GW samples with hydrochemical types HCO3-Na and SO4·Cl-Na are characterized by high fluoride content. The hydrochemical characteristics of GW in WFC are mostly dominated by rock weathering. In addition, the northern coastal plain is evidently influenced by seawater intrusion. The concentration of fluoride in GW is affected by the dissolution of fluorine-containing minerals, cation exchange, and alkaline environmental factors. The effect of GW by seawater intrusion and very high content of Na+ will decrease the fluoride content of the GW through cation exchange. Health risk assessment demonstrated that the mean values of non-carcinogenic hazard quotient (HQ) for infants, children, teenagers, and adults were 0.52, 0.35, 0.31, and 0.30, respectively. In addition, the distribution characteristics of GW fluoride in high health risk areas (HQ > 1) in WFC are further consistent with the spatial variation of GW fluoride content. Overall, the health risk distribution area of GW fluoride in WFC is decreasing in the following order: infants > children > teenagers > adults.

Ion adsorption stabilizes bulk nanobubbles.

The mechanism leading to the extraordinary stability of bulk nanobubbles in aqueous solutions remains an outstanding problem in soft matter, modern surface science, and physical chemistry science. In this work, the stability of bulk nanobubbles in electrolyte solutions under different pH levels and ionic strengths is studied. Nanobubbles are generated via the technique of ultrasonic cavitation, and characterized for size, number concentration and zeta potential under ambient conditions. Experimental results show that nanobubbles can survive in both acidic and basic solutions with pH values far away from the isoelectric point. We attribute the enhanced stability with increasing acidity or alkalinity of the aqueous solutions to the effective accumulation of net charges, regardless of their sign. The kinetic stability of the nanobubbles in various aqueous solutions is evaluated within the classic DLVO framework. Further, by combining a modified Poisson-Boltzmann equation with a modified Langmuir adsorption model, we describe a simple model that captures the influence of ion species and bulk concentration and reproduce the dependence of the nanobubble's surface potential on pH. We also discuss the apparent contradiction between quantitative calculation by ion stabilization model and experimental results. This essentially requires insight into the structure and dynamics of interfacial water on the atomic-scale.

A collagen-based bi-layered composite dressing for accelerated wound healing.

AIM OF THE STUDY: The aim of the study was to fabricate collagen-based composite dressings, evaluate the efficiency for wound healing and reveal the mechanism of promoting wound healing. MATERIALS AND METHODS: An innovative bi-layered composite wound dressing was developed using two marine biomacromolecules (collagen and chitosan). Full-thickness skin defect model was performed to evaluate the wound healing activity in vivo. The levels of inflammatory cytokines including tumor necrosis factor alpha (TNF-α), interleukin (IL-1, IL-6, IL-8) and growth factors like transforming growth factor beta (TGF-ß), vascular epidermal growth factor (VEGF) and basic fibroblast growth factor (bFGF) were quantified by ELISA assays. The total amount of collagen was quantified by hydroxyproline content. The proliferation and viability of fibroblast cells cultured on collagen sponges were determined by CCK-8 assay. RESULTS: The results of wound closure and histopathological analysis indicated that non-crosslinked collagen-based bi-layered composite dressing stimulated wound healing, accelerated re-epithelialization and accomplished wound healing within a time span of 28 days. The results of levels of inflammatory cytokines and growth factors showed that collagen-based composite dressings could reduce the inflammatory response and upregulate growth factors levels to accelerate the wound healing. The results of hydroxyproline content and CCK-8 assay indicated that collagen-based composite dressings could also promote collagen synthesis and fibroblasts viability and proliferation. CONCLUSION: The non-crosslinked collagen-based bi-layered composite dressing could be applied for an efficient and ideal wound dressing. Therefore, the findings provided the essential theoretical basis for the potential of collagen-based composite dressing applied in wound healing fields.

Preparation and evaluation of an innovative antibacterial bi-layered composite dressing for skin wound healing.

AIM OF THE STUDY: The aim of the current study was to develop collagen-based bi-layered composite dressings with antibacterial property and evaluate the efficiency for wound healing. MATERIALS AND METHODS: A bi-layered composite wound dressing was fabricated using two marine biomacromolecules (collagen and chitosan or carboxymethyl chitosan). Non-crosslinked and N-Ethyl-N'-(3-dimethylaminopropyl) carbodiimide/N-Hydroxy succinimide (EDC/NHS) cross-linked collagen sponges fabricated by vacuum freeze-drying technology was used as the inner layer. The medical spun-laced nonwoven coated with chitosan and carboxymethyl chitosan was used as the outer layer. The antibacterial activities against E. coli and S. aureus were evaluated by the inhibition zone assay. Deep second-degree scald model was performed to evaluate the efficiency of bi-layered composite dressings for wound healing. RESULTS: In view of comprehensive evaluation of appearance and in vitro antibacterial activity, medical spun-laced nonwoven coated with 3% of chitosan solution was chosen to be used as the optimized preparation conditions to produce the outer layer of composite dressing, which acted as a barrier against microorganisms and provided mechanical support. Furthermore, the results of wound closure and histopathological analysis indicated that EDC/NHS cross-linked collagen-based bi-layered composite dressing was superior to non-crosslinked and commercial products, which stimulated the wound healing process and accomplished deep second-degree scalded skin healing within a time span of 28 days. CONCLUSION: The EDC/NHS cross-linked collagen-based bi-layered composite dressing had immense potential to be applied for an ideal wound dressing for more efficient and faster wound healing. Therefore, the findings provided the essential theoretical basis for great potential of collagen-based composite dressing used in wound healing applications.

How bulk nanobubbles are stable over a wide range of temperatures.

HYPOTHESIS: Bulk nanobubbles are nanoscopic gaseous domains in an aqueous solution. Their surprising long-term stability remains controversial due to the widespread assumption that spherical bubbles cannot achieve stable equilibrium. To uncover the intrinsic mechanisms underlying stabilization, the thermodynamic behavior of nanobubbles in water over a wide range of temperatures is explored. EXPERIMENTS: Bulk nanobubbles with a typical radius of 50 - 200 nm are generated using acoustic cavitation. Increasing temperature significantly narrows the bubble-size distribution and their mean radius shrinks to a minimum of approx. 50 nm at 45 °C. For higher temperatures a slight increase is observed. The thermal induced shrinkage is reversible: upon cooling they return to the original state. FINDINGS: The observation can be explained with a charge-stabilization mechanism. The intricate balance of competing interactions between water self-ionization and mobility of ions on the surface gives rise to this non-monotonic dependency. Nanobubbles consequently undergo charge loss at lower temperatures and charge conservation at higher temperatures, corresponding to their shrinkage and slight expansion. With theoretical calculations, we further quantity the equilibrium properties of nanobubbles and their zeta potential under various initial conditions. The temperature-sensitive nature of bulk nanobubbles offers a vital step forward exploring and industrializing their stability.

Genome-wide analysis of long noncoding RNA and mRNA profiles in PRRSV-infected porcine alveolar macrophages.

Porcine reproductive and respiratory syndrome (PRRS), which is caused by PRRS virus (PRRSV), is one of the most globally devastating swine diseases. It is essential to develop new strategy to control PRRS via an understanding of mechanisms that PRRSV utilizes to interfere with the host's innate immunity. In this study, we deeply sequenced and analyzed long noncoding RNA (lncRNA) and mRNA expression profiles of the porcine alveolar macrophages (PAMs) after PRRSV infection. 126 lncRNAs and 753 mRNAs were differentially expressed between PRRSV-infected and control PAMs. The co-expressed genes of down-regulated lncRNAs were significantly enriched within NF-kappa B and toll-like receptor signaling pathways. Co-expression network analysis indicated that part of the dysregulated lncRNAs associated with the interferon-induced genes. These dysregulated lncRNAs may play an important role in the host's innate immune responses to PRRSV infection. However, further research is required to characterize the function of these lncRNAs.

Carbazole alkaloids with antiangiogenic activities from Clausena sanki.

Two new carbazole alkaloids 1 and 2, and eleven known congeners 3-13 were isolated and identified from Clausena sanki for the first time. Their structures were elucidated on the basis of extensive UV, IR, MS, NMR spectroscopic data and comparison with literatures. The compounds 1-13 were evaluated by MTT assay to determine whether they decreased VEGF-mediated cell proliferation in HUVECs with Axitinib as positive control. Among them, compounds 1, 2, 6, 8, and 13 (µM) exhibited moderate antiangiogenic activities, which inhibited VEGF-induced HUVEC proliferation in vitro with IC50 values of 12.1 (C.I. 8.2-15.2), 58.1 (C.I. 56.3-63.4), 13.7 (C.I. 9.2-15.4), 16.0 (C.I. 9.5-16.4), and 63.2 (C.I. 57.8-65.7) µM, respectively. Moreover, the antiangiogenic activities of compounds 1-13 were evidenced in vivo in the zebrafish embryo model. As a result, compounds 1, 2, 6, 8, and 13 showed effectively suppress angiogenesis. These research results may guide the search for new natural products with antiangiogenic attributes.

Genome-wide identification of allele-specific expression in response to Streptococcus suis 2 infection in two differentially susceptible pig breeds.

Although allele expression imbalance has been recognized in many species, and strongly linked to diseases, no whole transcriptome allele imbalance has been detected in pigs during pathogen infections. The pathogen Streptococcus suis 2 (SS2) causes serious zoonotic disease. Different pig breeds show differential susceptibility/resistance to pathogen infection, but the biological insight is little known. Here we analyzed allele-specific expression (ASE) using the spleen transcriptome of four pigs belonging to two phenotypically different breeds after SS2 infection. The comparative analysis of allele specific SNPs between control and infected animals revealed 882 and 1096 statistically significant differentially expressed allele SNPs (criteria: ratio ≧ 2 or ≦ 0.5) in Landrace and Enshi black pig, respectively. Twenty nine allelically imbalanced SNPs were further verified by Sanger sequencing, and later six SNPs were quantified by pyrosequencing assay. The pyrosequencing results are in agreement with the RNA-seq results, except two SNPs. Looking at the role of ASE in predisposition to diseases, the discovery of causative variants by ASE analysis might help the pig industry in long term to design breeding programs for improving SS2 resistance.

Environmental Enrichment during Gestation Improves Behavior Consequences and Synaptic Plasticity in Hippocampus of Prenatal-Stressed Offspring Rats.

Prenatal stress can result in various behavior deficits in offspring. Here we tested the effects of environmental enrichment during gestation used as a preventive strategy on the behavior deficits of prenatal-stressed offspring rats as well as the underlying structure basis. We compared the effect size of environmental enrichment during gestation on prenatal-stressed offspring to that of environmental enrichment after weaning. Our results showed that environmental enrichment during gestation partially prevented anxiety and the damage in learning and memory in prenatal-stressed offspring as evaluated by elevated plus-maze test and Morris water maze test. At the same time, environmental enrichment during gestation inhibited the decrease in spine density of CA1 and dentate gyrus neurons and preserved the expression of synaptophysin and glucocorticoid receptors (GRs) in the hippocampus of prenatal-stressed offspring. There was no significant difference in offspring behavior between 7-day environmental enrichment during gestation and 14-day offspring environmental enrichment after weaning. These data suggest that environmental enrichment during gestation effectively prevented the behavior deficits and the abnormal synapse structures in prenatal-stressed offspring, and that it can be used as an efficient preventive strategy against prenatal stresses.