Search

In recent years, biotechnology has had a significant impact on the aquaculture industry, particularly in the field of breeding. Molecular selection breeding has emerged as a novel approach to breeding. Reducing the cost of genetic information for individuals with desirable traits after breeding has become an important research direction. Cryopreservation technology allows bypassing time and space constraints in genetic breeding, simplifying broodstock management. This study presents a detailed cryopreservation method for black seabream sperm, evaluating extender type, glucose concentration, cryoprotectant type and concentration, sperm-dilution ratio, and cooling protocols. Sperm motility parameters were analyzed using computer-assisted sperm analysis (CASA) before and after two days of freezing. This involved using an RS solution with a glucose concentration of 15 g/L and adding a 5% final concentration of EG as the sperm cryoprotectant. After mixing the sperm and solution at a ratio of 1:2, we subjected it to 5 min fumigation at 5 cm above the liquid nitrogen surface before plunging it into the nitrogen. Sperm motility reached 85.46 ± 7.32% after two days. Various enzymatic activities showed changes over 20 days post-cryopreservation. This improved cryopreservation protocol for black seabream sperm is beneficial for genetic breeding and reproduction and provides reference for studying the cryodamage mechanisms of black seabream sperm., Competing Interests: Declaration of competing interest None., (Copyright © 2023. Published by Elsevier Inc.)

Interferon regulatory factor 7 (IRF7) regulates type I interferon (IFN) genes via combining to the ISRE region in the immune response against bacteria. Streptococcus iniae is one of the dominant pathogenic bacteria of yellowfin seabream, Acanthopagrus latus. However, the regulatory mechanisms of A. latus IRF7 (AlIRF7) mediated by the type I IFN signalling pathway against S. iniae was ambiguously. In the present study, IRF7, and two IFNa3s (IFNa3 and IFNa3-like) were authenticated from A. latus. The total length of AlIRF7 cDNA is 2142 bp, containing a 1314 bp open reading frame (ORF) encoding an inferred 437 amino acids (aa). Three typical regions, a serine-rich domain (SRD), a DNA-binding domain (DBD), and an IRF association domain (IAD), are conserved in AlIRF7. Furthermore, AlIRF7 is fundamentally expressed in various kinds of organs, with high levels in the spleen and liver. Additionally, S. iniae challenge promoted AlIRF7 expression in the spleen, liver, kidney, and brain. AlIRF7 is confirmed to be located at the nucleus and cytoplasm by overexpression of AlIRF7. Moreover, truncation mutation analyses shows that the regions, -821 bp to +192 bp and -928 bp to +196 bp, were known as core promoters from AlIFNa3 and AlIFNa3-like, respectively. The point mutation analyses and electrophoretic mobile shift assay (EMSA) verified that AlIFNa3 and AlIFNa3-like transcriptions are depended on the M2/5 and M2/3/4 binding sites with AlIRF7 regulation, respectively. Additionally, an overexpression experiment showed that AlIRF7 can dramatically decrease the mRNA levels of two AlIFNa3s and interferon signalling molecules. These results suggest that two IFNa3s may mediate the regulation of AlIRF7 in the immune responses of A. latus against S. iniae infection., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Microplastic (MPs) pollution is a global marine environmental problem. The effects of MPs on the gut microbiota of aquatic organisms have received considerable attention. For example, microbes colonizing MPs in pond cultures alter the structure and function of the intestinal microbes of shrimp and fish. It was hypothesized that bacteria on MPs in natural mariculture areas also interact with the intestinal flora of golden pompano (Trachinotus ovatus) because biofilms can form on the surface of MPs during long-term floating in seawater. To our knowledge, this study is the first to investigate MPs pollution in T. ovatus aquaculture. DNA sequencing and bioinformatics analysis confirmed the effect of microbial colonization of MPs on the intestinal flora of T. ovatus. The MPs detected in the gut wet weight (w.w.) of golden pompano (546 ± 52 items/g) were mainly pellets and fragments of blue or green, whereas the sediment MPs dry weight (d.w.) (4765 ± 116 items/kg) were mainly black fibers. The MPs richness in the sediment gradually increased from the open-sea aquaculture area to the estuarine aquaculture area and was positively correlated with the MPs richness in the intestinal tract of golden pompano. MPs 20-200 μm were the most common in the gut and sediment. The intake of MPs increased the abundance of Proteobacteria and decreased that of Firmicutes in the intestinal flora. The functional compositions of MP-colonizing microbes and gut microbiota were similar, suggesting that the two communities influence each other. Network analysis further confirmed this and revealed that Vibrio plays a key role in the intestinal flora and surface microorganisms of MPs. Overall, the intake of MPs by aquatic animals not only affects the intestinal flora and intestinal microbial function, but also poses potential risks to aquaculture., Competing Interests: Declaration of competing interest No potential conflict of interest was reported by the authors., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

This study aimed to investigate the intoxication mechanism of golden pompano (Trachinotus ovatus) exposed to high ammonia levels and the effects on the immune and antioxidant mechanisms of gills. Juvenile golden pompano was exposed to ammonia (total ammonia: 26.9 mg/L) to induce 96 h of ammonia stress, and a 96 h recovery experiment was performed after poisoning. Then, we evaluated hematological parameters, the histological structure and the expression of related genes. In this experiment, continuous exposure to high levels of ammonia led to a significant increase in plasma alkaline phosphatase (ALP), acid phosphatase (ACP) and lactate dehydrogenase (LDH) levels (P < 0.05), and the levels of triiodothyronine (T3) and tetraiodothyronine (T4) were significantly reduced (P < 0.05). Moreover, the expression of antioxidant genes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and inflammatory cytokines such as tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) increased (P < 0.05). These results indicate that ammonia activates the active osmotic regulatory mechanism of fish gills and participates in defense and immune responses. However, with prolonged exposure to ammonia, the balance of the defense system is disrupted, leading to oxidative damage and inflammation of the gill tissue. This research not only helps elucidate the intoxication mechanism of golden pompano by ammonia at the molecular level but also provides a theoretical basis for further research on detoxification mechanisms., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Aims: Long noncoding RNA (LncRNA) urothelial cancer associated 1 (UCA1) was dysregulated in colorectal cancers (CRC) and promoted tumor progression of CRC. The aims of this study are to further investigate the underlying mechanism., Main Methods: Short hairpin RNAs (shRNAs) were applied for gene knockdown. microRNA mimic and pcDNA-UCA1 plasmids were transfected for miR-495 and UCA1 overexpression, respectively. MTT was applied to determine cell viability and sensitivity of 5-fluorouracil (FU). Transwell assays were performed to evaluate cell migration/invasion. Angiogenesis was evaluated by tube formation. Western blotting and quantitative PCR were utilized for protein and mRNA detection, respectively. The interaction of UCA1, miR-495 and SP1/SP3 were explored by dual-luciferase assay. RNA pulldown was adopted to determine the UCA1/miR-495 interaction., Key Findings: UCA1 was significantly upregulated in CRC tissues. UCA1 enhanced cell proliferation, migration/invasion, angiogenesis, epithelial-mesenchymal transition, and resistance to 5-FU in CRC cell lines. MiR-495 was inversely correlated to the expression of UCA1. The results indicated that UCA1 sponged miR-495, leading to the disinhibition of SP1/SP3 expression. SP1/SP3 induced the expression of DNA methyltransferases and, in turn, contributed to UCA1 mediated tumor-promoting actions. Reduction of SP1/SP3 exerted anti-cancer effects, which can be reversed by forced expression of UCA1., Significance: UCA1-miR-495-SP1/SP3 axis is dysregulated in CRC and contributed to malignant phenotypes of CRC. UCA1-SP1/SP3 may form a positive feedback loop in CRC., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Fatty acyl desaturase 2 (fads2) is a rate-limiting enzyme in long chain polyunsaturated fatty acids (LC-PUFAs) biosynthesis. In mammals, the lipid metabolism is modulated by a transcription factor, peroxisome proliferator-activated receptor alpha β (pparαβ); however, the detailed mechanism via pparαβ regulates fads2 remains unclear in fish. In the present study, we identified the sequence features of Trachinotus ovatus fatty acyl desaturase 2a (Tofads2a) and fatty acyl desaturase 2b (Tofads2b), which both encoded 442 amino acid polypeptides containing cytochrome-b5-like domains and three representative histidine-rich domains. The Phylogenetic and genome organization analyses revealed characteristic phylogeny: the majority of fads2s exhibited a highly conserved exon/intron architecture. Tissue expression patterns by quantitative real-time PCR (qRT-PCR) showed that the two Tofads2s were prominently expressed in the brain. A nutritional study indicated that the transcription of the two Tofads2s was significantly implicated by treatment with a 1: 1 ratio of fish oil: soybean oil (FO:SO) in the liver and brain. Furthermore, functional characterization in yeast demonstrated that both Tofads2a and Tofads2b possessed Δ4/Δ5/Δ8 desaturation activity. Furthermore, promoter activity assays showed that the expressions of the two Tofads2s were actively regulated by pparαβ. Moreover, mutation analyses showed that the M1 and M4/M5 binding sites of pparαβ were functionally vital for binding to Tofads2a and Tofads2b promoters, respectively. Transcriptional activities of the two Tofads2s promoters were significantly reduced after targeted mutation of M1 or M4/M5. Electrophoretic mobile shift assays (EMSAs) verified that pparαβ interacted with the M1 binding site in Tofads2a promoter to accommodate Tofads2a transcription. Briefly, pparαβ plays an important role in Tofads2 expression and may promote the LC-PUFAs biosynthesis by regulating the expression of two Tofads2s., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Myoblast determination protein (MyoD), a muscle-specific basic helix-loop-helix (bHLH) transcription factor, plays a pivotal role in regulating skeletal muscle growth and development. However, the regulation mechanism of MyoD has not been determined in marine fishes. In the present study, we isolated the MyoD1 (AlMyoD1) and MyoD2 (AlMyoD2) genomic sequences and analyzed the expression patterns in different tissues of yellowfin seabream (Acanthopagrus latus). The open reading frame (ORF) sequences of AlMyoD1 and AlMyoD2 encoded 297 and 271 amino acids possessing three common characteristic domains, respectively, containing a myogenic basic domain, a bHLH domain, and a ser-rich region (helix III). Phylogenetic and genome structure analyses exhibited classic phylogeny and highly conserved exon/intron architecture. Furthermore, the AlMyoD1 and AlMyoD2 transcription levels were higher in white muscle than in the other tissues. In order to further study AlMyoD function in muscle, promoter sequence analysis found that several E-box binding sites were present. Additionally, binding sites of Almyomixer involved in mammal myoblast fusion, which expression was also the highest in white muscle, were found in the promoter of AlMyoD. Pomoter activity assays further confirmed that both AlMyoD1 and AlMyoD2 can dramatically activate Almyomixer expression, and the AlMyoD1 M2 and AlMyoD2 M5 E-box binding sites were functionally important for Almyomixer transcription based on mutation analysis and electrophoretic mobile shift assays (EMSA). In summary, two MyoDs play a core role in Almyomixer regulation and may promote myofibre formation during muscle development and growth by regulating Almyomixer expression., Competing Interests: Declaration of competing interest The authors declare no competing financial interests., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Several non-invasive diagnostic scores for non-alcoholic fatty liver (NAFL) have been developed, but the clinical application is limited because of their complexity., Aim: To develop and validate an easy-to-calculate scoring system to identify ultrasound-diagnosed NAFL., Methods: 48,489 patients from 2 centers were included in this study. Multivariable logistic regression models were employed for model development. Ultrasonography was applied to diagnose NAFL. The selected variables were assigned an integer score proportional to the estimated coefficient from the logistic regression analysis, namely NAFL Screening Score (NSS). The ability of the NSS to identify NAFL was assessed by analyzing the area under the receiver operating characteristic curve (AUROC) and was tested in an independent validation cohort. Additionally, the performance of NSS was compared with existing models., Results: NSS was developed as a basic score comprising of age, body mass index (BMI), triglyceride (TG), ALT/AST, fasting plasma glucose (FPG) and uric acid (UA) in both sexes. NSS showed a relatively good discriminative power (AUROC=0.825 for males, 0.861 for females in the validation cohort) in comparison with other models. The optimal cut-off point was 32 for males and 29 for females., Conclusion: We developed and validated NSS, an easy-to-use score sheet identify ultrasound-diagnosed NAFL. NSS may be clinically useful for initial diagnosing NAFL., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Although several risk factors for non-alcoholic fatty liver (NAFL) have been reported, there are few clinical scores that predict its incidence in the long term. We developed and validate a scoring model for individual prediction of 4-y risk for NAFL., Methods: Four-year follow-up data of 8226 initially NAFL-free subjects enrolled for an annual physical examination from Wenzhou Medical Center were analyzed. These subjects are randomly split into the training and the validation cohort. Univariate and multivariable logistic regression models were employed for model development. The selected variables were assigned an integer or half-integer risk score proportional to the estimated coefficient from the logistic model. Risk scores were tested in a validation cohort. We also compared the predictive performance of with that of the NAFLD Index by computing the area under the receiver operating characteristic curve (AUROC)., Results: The NAFL Risk Score was developed as 0 to 18 points comprising of BMI, TG×GGT, ALT/AST, LDL-C/HDL-C and UA in both sexes. Comparison of the observed with the estimated incidence of NAFL at both cohorts showed satisfactory precision. In addition, the NAFL Risk Score showed relatively good discriminative power (AUROC=0.739 for males, 0.823 for females) compared with the NAFLD Index (AUROC=0.661 for males, 0.729 for females) in these Chinese subjects., Conclusions: We developed and validated the NAFL Risk Score, a new scoring model to predict 4-y risk for NAFL. The NAFL Risk Score may be clinically simple and useful for assessing individual risk for NAFL., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Effects of thermomechanical treatment on the microstructure and superelasticity of Ti-7.5Nb-4Mo-2Sn biomedical alloy were investigated by using XRD measurement, optical microscope (OM), transmission electron microscope (TEM) and tensile tests. The titanium alloy samples were prepared by annealing at a temperature in the range of 600 to 1000°C after severe cold rolling; and the samples that were annealed at 800°C were further aged at 600 and 700°C. The volume fraction of α phases decreased while that of ω phases increase with increasing annealing temperature. The α→β transformation temperature of the alloy was determined to be between 700 and 800°C. The alloy that was annealed at 700°C exhibited a high level of superelasticity with relatively high first yield stress (σSIM) at room temperature because it contained a fine α phase. A certain amount of ω phases also resulted in an increase in σSIM, leading to an improvement in the superelasticity of the alloys that were annealed at 900 and 1000°C. Aging treatment led to the precipitations of α and ω phases in the alloy after annealing at 800°C; and the volume fraction of α phases decreased while that of ω phases increased with increasing aging temperature. Excellent superelasticity with high recovered strain (εrecoverable) and strain recovery rate (η) were obtained in the aged alloy due to the reinforcement of α and ω phases induced by aging treatment. The alloy annealed at 700°C for 0.5h exhibited the best superelasticity in all the thermomechanically treated alloys due to the strengthening from the subgrain refining and the precipitating of fine α phases., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Small-diameter vascular grafts can easily lead to thrombosis and intimal hyperplasia. So the rate of the long-term patency is not satisfactory. Scaffolds that can support the growth of cells and remain stable and passable in vivo are in great demand. Vascular scaffolds were fabricated by electrospinning RGD-recombinant spider silk protein (pNSR32), polycaprolactone (PCL) and chitosan (CS). In addition, the cytocompatibility, the stability and the patency of scaffolds in vivo were studied. The results demonstrated that Sprague-Dawley rat aortic endothelial cells (SDRAECs) could highly enhanced adhesion and proliferation, together with increasing stress of fiber formation, and intensive biological function (the expression of PECAM-1 and vWF, the secretion of NO), in the pNSR32/PCL/CS scaffolds, compared with in the pNSR32/PCL and PCL scaffolds. The cell cycle studies of SDRAECs also revealed that the scaffolds promoted the cells to enter the stage of divisional proliferation. Furthermore, pNSR32/PCL/CS scaffolds could support the growth of cells under physiologic conditions and are able to maintain the structural integrity and patency for at least 8 weeks in a SD rat abdominal aortic defect model., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The superelasticity of a β Ti alloy, Ti-7.5Nb-4Mo-2Sn (in atom percent) was evaluated by using loading and unloading cyclic tensile tests under different thermomechanical conditions, and the effects of the plastic deformation, temperature, strain rate and cyclic loading on the superelasticity of the alloy were studied. It is found that, with the applied strain increasing, the stress inducing the reverse martensitic transformation σα″-β and the strain recovery rate η decreases. The increase of deformation temperature promotes σβ-α″, σα″-β and Δσ, and the temperature dependency of the stresses obeys the Clausius-Clapeyron relation. σβ-α″, σα″-β and Δσ are independent on the strain rate when it is lower than 8.35×10(-4)s(-1). However, when the strain rate is higher than 8.35×10(-4)s(-1), σβ-α″ and Δσ increase, but σα″-β decreased with increasing the strain rate. By cyclic loading and unloading to the maximum strain of 6% at 25°C under the strain rate of 1.67×10(-4)s(-1), the alloy exhibits a improved superelasticity after seventh cycles due to the training effect., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Ti-7.5Nb-4Mo-xSn (x=0-4at%) alloys were developed as the biomedical materials. The effect of the Sn content on the microstructure and superelasticity of the alloys was investigated. It is found that Sn is a strong stabilizer of the β phase, which is effective in suppressing the formation of α″ and ω phases in the alloys. Moreover, the Sn addition has a significant impact on the mechanical properties of the alloys. With the increase of Sn addition, the yield stress of the alloys increase, but their elastic modulus, the fracture strength and the ductility decrease, and the deformation mode of the alloys changes from (322) twining to α″ transformation and then to slip. The Ti-7.5Nb-4Mo-1Sn and Ti-7.5Nb-4Mo-3Sn alloys exhibit a good superelasticity with a high σ(SIM) due to the relatively high athermal ω phases containing or the solution hardening at room temperature. Under the maximum strain of 5%, Ti-7.5Nb-4Mo-3Sn (at%) alloy exhibits higher super elastic stability than that of Ti-7.5Nb-4Mo-1Sn alloy., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

A polyphasic approach combining culture-based methods with molecular methods is useful to expand knowledge on microbial diversity in contaminated soil. Microbial diversity was examined in soil samples from a former industrial site in the European Alps (mainly used for aluminum production and heavily contaminated with petroleum hydrocarbons) by culture-dependent and culture-independent methods. The physiologically active eubacterial community, as revealed by fluorescence-in-situ-hybridization (FISH), accounted for 6.7% of the total (DAPI-stained) bacterial community. 4.4% and 2.0% of the DAPI-stained cells could be attributed to culturable, heterotrophic bacteria able to grow at 20°C and 10°C, respectively. The majority of culturable bacterial isolates (34/48) belonged to the Proteobacteria (with a predominance of Alphaproteobacteria and Gammaproteobacteria), while the remaining isolates were affiliated with the Actinobacteria, Cytophaga-Flavobacterium-Bacteroides and Firmicutes. A high fraction of the culturable, heterotrophic bacterial population was able to utilize hydrocarbons. Actinobacteria were the most versatile and efficient degraders of diesel oil, n-alkanes, phenol and PAHs. The bacterial 16S rRNA gene clone library contained 390 clones that grouped into 68 phylotypes related to the Proteobacteria, Bacteroidetes, Actinobacteria and Spirochaetes. The archaeal 16S rRNA gene library contained 202 clones and 15 phylotypes belonging to the phylum Euryarchaeota; sequences were closely related to those of methanogenic archaea of the orders Methanomicrobiales, Methanosarcinales, Methanobacteriales and Thermoplasmatales. A number of bacterial and archaeal phylotypes in the clone libraries shared high similarities with strains previously described to be involved in hydrocarbon biodegradation. Knowledge of the bacterial and archaeal diversity in the studied soil is important in order to get a better insight into the microbial structure of contaminated environments and to better exploit the bioremediation potential by identifying potential hydrocarbon degraders and consequently developing appropriate bioremediation strategies., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A general mathematical model of cell invasion is developed and validated with an experimental system. The model incorporates two basic cell functions: non-directed (diffusive) motility and proliferation to a carrying capacity limit. The model is used here to investigate cell proliferation and motility differences along the axis of an invasion wave. Mathematical simulations yield surprising and counterintuitive predictions. In this general scenario, cells at the invasive front are proliferative and migrate into previously unoccupied tissues while those behind the front are essentially nonproliferative and do not directly migrate into unoccupied tissues. These differences are not innate to the cells, but are a function of proximity to uninvaded tissue. Therefore, proliferation at the invading front is the critical mechanism driving apparently directed invasion. An appropriate system to experimentally validate these predictions is the directional invasion and colonization of the gut by vagal neural crest cells that establish the enteric nervous system. An assay using gut organ culture with chick-quail grafting is used for this purpose. The experimental results are entirely concordant with the mathematical predictions. We conclude that proliferation at the wavefront is a key mechanism driving the invasive process. This has important implications not just for the neural crest, but for other invasion systems such as epidermal wound healing, carcinoma invasion and other developmental cell migrations.

This study proposed a novel headspace gas chromatographic (HS-GC) method for determination of adhesive contaminants (microstickies) in recycled whitewater, a fiber containing process stream, in the paper mill. It is based on the adsorption behavior of toluene (as a tracer) on the hydrophobic surface of microstickies, which affects the apparent vapor-liquid equilibration partitioning of toluene. It was found that the equilibrium concentration of toluene in the vapor phase is inversely proportional to the apparent effective surface area of microstickies that remain in the corresponding solution. Thus, the amount of microsticky materials in the recycled whitewater can be quantified by HS-GC via indirect measurement of the toluene content in the vapor phase of the sample without any pretreatment. The presented method is simple, rapid and automated.

Background: Despite three decades of debate, no conclusion has been reached concerning the effectiveness of postoperative radiotherapy for resected esophageal carcinoma. From 1986 through 1997, a prospective randomized study was carried out with 495 patients in an attempt to define the value of this therapeutic modality., Methods: A total of 495 patients with esophageal cancer who had undergone radical resection were randomized by the envelope method into a surgery-alone group (S) of 275 patients and a surgery plus radiotherapy group (S + R) of 220 patients. Radiation treatment was started 3 to 4 weeks after the operation. The portals encompassed the entire mediastinum and bilateral supraclavicular areas. A midplane dose of 50 to 60 Gy in 25 to 30 fractions was delivered over 5 to 6 weeks., Results: The overall 5-year survival rate was 31.7% for the S group and 41.3% (p = 0.4474) for the S + R group. The 5-year survival rates of patients who were lymph node positive were 14.7% and 29.2% (p = 0.0698), respectively. Five-year survival rates of stage III patients were 13.1% and 35.1% (p = 0.0027), respectively., Conclusions: Postoperative prophylactic radiotherapy improved the 5-year survival rate in esophageal cancer patients with positive lymph node metastases and in patients with stage III disease compared with similar patients who did not receive radiation therapy. These results were almost significant for patients with positive lymph node metastases and highly significant for patients with stage III disease.