Dynamic spatiotemporal change of net anthropogenic phosphorus inputs and its response of water quality in the Liao river basin.

The Liao river is one of the seven major rivers in China, and the process of phosphorus (P) cycling and change of water quality in this basin are influenced to a considerable extent human activities. In this work, the traditional net anthropogenic phosphorus inputs (NAPI) model was improved by considering the dynamic change of wastewater treatment capacity and P deposition (PDEP) and reclassifying the sources of phosphorus into human P consumption (PHUM), agriculture P consumption (PAGR), livestock P consumption (PANIM) and PDEP to analyze its dynamic spatio-temporal change in the Liao river basin. The results showed that the annual mean NAPI was 785.53 kg P km-2 yr-1 (2001-2020), the maximum value was 940.49 kg P km-2 yr-1 in 2009, and the minimum value was 586.04 kg P km-2 yr-1 in 2001. The temporal variation of NAPI presented an increasing-fluctuation-increasing trend and was basically in line with that of the water quality throughout the three stages, and the spatial distribution of NAPI gradually increased from upstream to downstream. During the two decades, PANIM was the predominant component of NAPI with a share of 64.32%. PHUM, PAGR, and PDEP accounted for 15.97%, 11.54%, and 8.17%, respectively, and the point source NAPI (NAPIP) contributed to 4.95% of NAPI. Further, the INAPI (Improved NAPI) -MR (Multiple Regression) -SWAT (Soil and Water Assessment Tool) model was developed to predict the spatial distribution of P flux under two scenarios. The results showed that the Liao river basin experienced a reduction in P flux to different degrees due to the improvement of the wastewater treatment system, which was more significant in its downstream area. Long-term water quality monitoring is encouraged to develop refined water quality models in the future.

Scene-dependent, feedforward eye gaze metrics can differentiate technical skill levels of trainees in laparoscopic surgery.

INTRODUCTION: In laparoscopic surgery, looking in the target areas is an indicator of proficiency. However, gaze behaviors revealing feedforward control (i.e., looking ahead) and their importance have been under-investigated in surgery. This study aims to establish the sensitivity and relative importance of different scene-dependent gaze and motion metrics for estimating trainee proficiency levels in surgical skills. METHODS: Medical students performed the Fundamentals of Laparoscopic Surgery peg transfer task while recording their gaze on the monitor and tool activities inside the trainer box. Using computer vision and fixation algorithms, five scene-dependent gaze metrics and one tool speed metric were computed for 499 practice trials. Cluster analysis on the six metrics was used to group the trials into different clusters/proficiency levels, and ANOVAs were conducted to test differences between proficiency levels. A Random Forest model was trained to study metric importance at predicting proficiency levels. RESULTS: Three clusters were identified, corresponding to three proficiency levels. The correspondence between the clusters and proficiency levels was confirmed by differences between completion times (F2,488 = 38.94, p < .001). Further, ANOVAs revealed significant differences between the three levels for all six metrics. The Random Forest model predicted proficiency level with 99% out-of-bag accuracy and revealed that scene-dependent gaze metrics reflecting feedforward behaviors were more important for prediction than the ones reflecting feedback behaviors. CONCLUSION: Scene-dependent gaze metrics revealed skill levels of trainees more precisely than between experts and novices as suggested in the literature. Further, feedforward gaze metrics appeared to be more important than feedback ones at predicting proficiency.

Accumulation characteristic of nitrogen in reservoirs during the ice-covered period under superimposed influence of ice and sediments: A case study of Biliuhe reservoir.

Reservoirs located in middle and high latitudes freeze for months in winter, where the accumulation characteristics of pollutants are changed by superimposed influence of salt exclusion from ice on the surface and pollution release from sediments at the bottom. Taking total nitrogen (TN) of Biliuhe reservoir in Northeast China as an example, we developed a model to simulate TN accumulation characteristics influenced by ice and sediments during the freezing period (NACISF), and quantified contributions of TN from ice and sediments. Model parameters of ice and sediments were determined by laboratory freeze-up simulation experiment and sediment release flux simulation experiment, and water quality data were obtained from field investigations. Results showed that the annual average amount of TN input during the ice-covered period from 2015 to 2020 was 220.77 t, the output was 400.11 t, and the accumulated amount was 589.52 t. TN excluded from ice and released from sediments contributed 8.12% and 7.17% of the total TN inputs in winter, respectively. Analysis showed that the TN excluded from ice was positively correlated with ice thickness and initial TN concentration. The maximum ice thickness of Biliuhe reservoir had a 13 year cyclic feature, and the proportion of TN excluded from ice to the total TN inputs in different periods ranged from 10.68% to 17.30% (mean 13.18%). Meanwhile, TN accumulated seasonally as summer > autumn > winter > spring. The TN exclusion effect in 2050 would be weakened when considering the combined effects of climate change and human activities, with a reduction of about 40.85% compared to the current. It is concluded that the NACISF model took into account the influences of both ice and sediments, which provided a detailed understanding of the accumulation characteristics of TN during freezing period, and had important reference significance for water quality management in winter.

Identifying long-range synaptic inputs using genetically encoded labels and volume electron microscopy.

Enzymes that facilitate the local deposition of electron dense reaction products have been widely used as labels in electron microscopy (EM) for the identification of synaptic contacts in neural tissue. Peroxidases, in particular, can efficiently metabolize 3,3'-diaminobenzidine tetrahydrochloride hydrate (DAB) to produce precipitates with high contrast under EM following heavy metal staining, and can be genetically encoded to facilitate the labeling of specific cell-types or organelles. Nevertheless, the peroxidase/DAB method has so far not been reported to work in a multiplexed manner in combination with 3D volume EM techniques (e.g. Serial blockface electron microscopy, SBEM; Focused ion beam electron microscopy, FIBSEM) that are favored for the large-scale ultrastructural assessment of synaptic architecture However, a recently described peroxidase with enhanced enzymatic activity (dAPEX2) can efficienty deposit EM-visible DAB products in thick tissue without detergent treatment opening the possibility for the multiplex labeling of genetically defined cell-types in combination with volume EM methods. Here we demonstrate that multiplexed dAPEX2/DAB tagging is compatible with both FIBSEM and SBEM volume EM approaches and use them to map long-range genetically identified synaptic inputs from the anterior cingulate cortex to the periaqueductal gray in the mouse brain.

Monte Carlo determination of dose coefficients at different developmental stages of zebrafish (Danio rerio) in experimental condition.

The release of liquid effluent of nuclear power into aquatic system increases with the rapid development of nuclear facilities in coastal and inland regions. Aquatic model animals are very important for the study of the radiation hazards to non-human biota in water environment and its extrapolation of dose-effect relationship to human models. However, the study of the radiation dose rate calculation model of the aquatic animal zebrafish is still on the homogeneous isotropic model used for the protection of the environment. A series of zebrafish models (including adults, larvae and embryos, named zebrafish-family: ZF-family) with multiple internal organs are established in this study to investigate the mechanism of radiation damage effect in order to protect non-human species. The internal and external dose coefficients (DCs) of the whole body, heart and gonads of zebrafishes are calculated in water environment with the combination of the real experimental culture condition, using Monte Carlo application package GATE (Geant4 Application for Emission Tomography) and eight nuclides, i.e., 3H, 14C, 90Sr, 60Co, 110mAg, 134Cs, 137Cs, 131I, which are commonly found in the liquid effluent of nuclear power plants, as the source items, The results show that the level of nuclide γ energy determines the external DCs (DCext), and 90Sr plays the most important role in internal DCs (DCint). The comparison between the external DCs of the heart and gonad and that of the whole body shows that DCs (DCext) of heart and gonad for females are 80% and 43% lower than that of whole body, respectively, while for males, the DCs (DCext) of heart is 44% lower than that of the whole body, and DCs (DCext) of gonad is slightly higher than that of the whole body for most nuclides (up to 25%).The dose of internal radiation makes greater contribution than that of external radiation to pure beta emitter (3H, 14C, 90Sr). This internal DCs of ZF-family model with complex internal structure turns out to demonstrate more sensitive DCs change trend and higher calculation values compared with the internal DCs of the simple ellipsoid model. In this model, the photon emitter with strong penetrating power has higher internal DCs, while the low-energy pure beta nuclide does not alter much. In conclusion, it is vital to carry out refined systematic modeling for model organisms, and the determination of DCs of model organs can promote the evaluation of the radiation effects on non-human species.

A non-exhaustive survey revealed possible genetic similarity in mitochondrial adaptive evolution of marine fish species in the northwestern Pacific.

Mitochondrial coding genes involved in the oxidative phosphorylation pathway play vitally important roles in energy production and thermal adaptation. Investigating the underlying molecular mechanism of mitochondrial adaptive evolution is crucial for understanding biodiversity and ecological radiation. In this study, we collated population genetic studies of marine fish species in the northwestern Pacific based on mitochondrial cytochrome b gene sequences, to investigate whether similar patterns could be detected in mitochondrial adaptive evolution. After filtering, nine studies containing eight marine fish species (Ammodytes personatus, Boleophthalmus pectinirostris, Larimichthys polyactis, Mugil cephalus, Pampus argenteus, Platycephalus sp.1, Sebastiscus marmoratus, and Trachidermus fasciatus) belonging to eight different families were retained. Multiple codon-based approaches were used to identify potential sites under selection in each species. By comparison, our results showed that the posterior part of the mitochondrial cytochrome b gene (particularly codon 372 and its neighboring sites) seemed to be involved in the adaptive evolution process, suggesting potential genetic similarity among distantly related species. We also summarized four types of adaptive patterns in the reviewed species, and suggest that the level of genetic differentiation and mitochondrial adaptive evolution might be correlated. Further studies are needed to confirm such relationship by detecting RNA-level evidence and investigating more species and samples.

Dissolution and oral bioavailability enhancement of praziquantel by solid dispersions.

The aim of the present investigation was to enhance the solubility, dissolution, and oral bioavailability of praziquantel (PZQ), a poorly water-soluble BCS II drug (Biopharmaceutical Classification System), using a solid dispersion (SD) technique involving hydrophilic copolymers. The SD formulations were prepared by a solvent evaporation method with PZQ and PEG 4000 (polyethylene glycol 4000), PEG 6000, or P 188 polymers at various weight ratios or a combination of PEG 4000/P 188. The optimized SD formulation, which had the highest solubility in distilled water, was further characterized by its surface morphology, crystallinity, and dissolution in 0.1 M HCl with 0.2% w/v of sodium dodecyl sulfate (SDS). X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) revealed the amorphous form of PZQ in the SDs. Moreover, at an oral dosage of 5 mg/kg PZQ, the SDs had higher Cmax values and areas under the curve (AUCs) compared to those of commercial PZQ tablets. Preparation of PZQ-loaded SDs using PEG 4000/P 188 is a promising strategy to improve the oral bioavailability of PZQ.

Expression Profile Analysis of miR-221 and miR-222 in Different Tissues and Head Kidney Cells of Cynoglossus semilaevis, Following Pathogen Infection.

Half-smooth tongue sole (Cynoglossus semilaevis) is an important marine commercial fish species in China, which suffers from widespread disease outbreaks. Recently, in this regard, our group identified immune-related microRNAs (miRNAs) of C. semilaevis following Vibrio anguillarum infection. Furthermore, miRNA microarray was utilized to characterize the immune roles of important miRNA candidates in response to bacterial infection. Therefore, in the present study, we characterized miR-221 and miR-222 and profiled their expression after challenge. Here, miR-221 and miR-222 precursors were predicted to have a typical hairpin structure. Both miRNAs were expressed in a broad range of tissues in C. semilaevis, while miR-221 and miR-222 were significantly differentially expressed in the immune tissues of C. semilaevis among three small RNA libraries [control group (CG), bacteria-challenged fish without obvious symptoms of infection (NOSG), and bacteria-challenged fish with obvious symptoms of infection (HOSG)]. In order to further characterize and understand the immune response of miR-221 and miR-222, therefore, we profiled miR-221 and miR-222 expression in selected immune tissues after challenge with V. anguillarum. Both miR-221 and miR-222 were upregulated in the liver and spleen, while different expression patterns were observed in the head kidney. In addition, in half-smooth tongue sole head kidney cell line after challenge with lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (poly I:C), peptidoglycan (PGN), and red-spotted grouper nervous necrosis virus (RGNNV), both miR-221 and miR-222 showed significant difference in expression response to pathogen. Meanwhile, the target gene of miR-221 and miR-222 was predicted, which indicated that tumor necrosis factor receptor-associated factor 6 (TRAF6) and interleukin-1 beta (IL-1ß) were the target genes of miR-221 and miR-222, respectively. Collectively, these findings indicated that miR-221 and miR-222 have putative roles in innate immune response during C. semilaevis exposure to pathogens. Our findings could expand the knowledge of immune function of C. semilaevis miRNA and guide future studies on C. semilaevis immunity.

Establishment and characterization of an astroglial cell line derived from the brain of half-smooth tongue sole (Cynoglossus semilaevis).

An astroglial cell line was established from the brain of half smooth tongue sole (Cynoglossus semilaevis) and was designated as CSAC. CSAC shows the morphological homogeneity of epithelial cells. The cell identity was tested by the presence of glial fibrillary acidic protein (GFAP), which was revealed by RT-PCR and immunofluorescence. The cell line was optimally maintained at 24 °C in minimum essential medium supplemented with HEPES, antibiotics, 20% fetal bovine serum, 2-Mercaptoethanol (2-Me) and basic fibroblast growth factor. Chromosome analysis revealed that the CSAC cells maintained a normal diploid chromosome number (2n=42). The fluorescent signals were observed in CSAC after the cells were transfected with green fluorescent protein (GFP) reporter plasmids. The CSAC cell line may serve as a valuable tool for studies on the potential functions of fish astroglial cells.

Expression profiling analysis of the microRNA response of Cynoglossus semilaevis to Vibrio anguillarum and other stimuli.

To investigate the roles of microRNAs (miRNA) of Cynoglossus semilaevis in response to Vibrio anguillarum that were previously identified using high-throughput sequencing, microarray analyses was performed on three small RNA libraries (CG, NOSG, and HOSG) prepared from C. semilaevis immune tissues. In total, of 1279 designed probes, 739 (57.78 %) were detectable. The expression levels of these miRNAs were analyzed using pairwise comparisons among the three libraries, and a total of 99 miRNAs were observed to be significantly differentially expressed. The expression patterns of 10 differentially expressed miRNAs were validated by real-time quantitative PCR (RT-qPCR). In addition, expression of miR-142-5p, miR-223, and miR-181a in response to V. anguillarum at numerous time-points in four tissues, as well as the responses to lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (poly I:C), peptidoglycan (PGN), and red-spotted grouper nervous necrosis virus (RGNNV) in head kidney cells, were studied by qRT-PCR. Taken together, all of the expression profiles showed significant differences compared to the control group; both similarities and differences in the expression responses to the same pathogen were observed. Collectively, these findings highlighted the putative roles for miRNAs in the context of the innate immune response of C. semilaevis exposing to pathogens and that further studies are needed to understand the molecular mechanisms of miRNA regulation in C. semilaevis host-pathogen interactions.

sghC1q, a novel C1q family member from half-smooth tongue sole (Cynoglossus semilaevis): identification, expression and analysis of antibacterial and antiviral activities.

The C1q family includes many proteins that contain a globular (gC1q) domain, and this family is widely conserved from bacteria to mammals. The family is divided into three subgroups: C1q, C1q-like and ghC1q. In this study, a novel C1q family member, sghC1q, was cloned and identified from Cynoglossus semilaevis (named CssghC1q). The full-length CssghC1q cDNA spans 905 bp, including an open reading frame (ORF) of 768 bp, a 5'-untranslated region (UTR) of 25 bp and a 3'-UTR of 112 bp. The ORF encodes a putative protein of 255 amino acids (aa) with a deduced molecular weight of 28 kDa. The predicted protein contains a signal peptide (aa 1-19), a coiled-coil region (aa 61-102) and a globular C1q (gC1q) domain (aa 117-255). Protein sequence alignment indicated that the C-terminus of CssghC1q is highly conserved across several species. Phylogenetic analysis indicated that CssghC1q is most closely related to Maylandia zebra C1q-like-2-like. The CssghC1q genomic sequence spanned 1562 bp, with three exons and two introns. CssghC1q is constitutively expressed in all evaluated tissues, with the highest expression in the liver and the weakest in the heart. After a challenge with Vibrio anguillarum, CssghC1q transcript levels exhibited distinct time-dependent response patterns in the blood, head kidney, skin, spleen, intestine and liver. Recombinant CssghC1q protein exhibited antimicrobial activities against Gram-negative bacteria, Gram-positive bacteria and viruses. The minimum inhibitory concentration (MIC) values against Vibrio harveyi, Vibrio anguillarum, Pseudomonas aeruginosa and Staphylococcus aureus were 0.043 mg/mL, 0.087 mg/mL, 0.174 mg/mL and 0.025 mg/mL, respectively. A low concentration (0.06 mg/mL) of CssghC1q showed significant antiviral activity in vitro against nervous necrosis virus (NNV). These results suggest that CssghC1q plays a vital role in immune defense against bacteria and viruses.

Molecular cloning and multifunctional characterization of GRIM-19 (gene associated with retinoid-interferon-induced mortality 19) homologue from turbot (Scophthalmus maximus).

GRIM-19 (gene associated with retinoid-interferon-induced mortality 19), a novel cell death regulatory gene, plays important roles in cell apoptosis, embryogenesis, mitochondrial respiratory chain and immune response. To date, little information is known about fish GRIM-19 characteristics except orange-spotted grouper (Epinephelus coioides). Here a new GRIM-19 gene is identified and characterized from turbot (Scophthalmus maximus), an economic marine fish in China and Europe. Briefly, turbot GRIM-19 is a 595-bp gene encoding a 144 amino acids protein, which shares the closest relationship with Atlantic halibut (Hippoglossus hippoglossus). The expression of turbot grim-19 in liver, spleen and kidney is up-regulated by the infection of Vibrio anguillarum and LCDV (lymphocystis disease virus). Subsequently, a recombinant protein of turbot GRIM-19 is acquired and the anti-bacterial function is proved by liquid culture inhibition experiment. The subcellular location indicates that turbot GRIM-19 is co-localized with STAT3 in the cytoplasm, which is mainly determined by GRIM-19 41-84 amino acids and STAT3 1-321 amino acids. Finally, the involvements of turbot GRIM-19 in cell apoptosis and NF-κB pathway are investigated. All these data help to understand GRIM-19 function in fish, as well as provide the application possibility of GRIM-19 in fish disease resistance breeding.

[Histological observations of Medaka(Oryzias latipes)gonad sexual differentiation and development].

Over the past few decades, Medaka (Oryzias latipes) has become a model animal in teleost species research due both to its short reproductive cycle and efficient proliferate capacity. Unfortunately, however, systematic data of its sexual differentiation and development have yet to be obtained. In the present study, we observed Medaka gonad development from the earliest recognizable stages through differentiation to maturation under a light microscope, after paraffin sectioning and hematoxylin-eosin staining. The results showed that among juveniles aged 5 to 10 days, the gonad located on the right side of the dorsal abdominal cavity and the germ cells was significantly larger than the surrounding somatic cells. Ten days after hatching, sexual differences between females and males became obvious, and germline cysts derived from oogonium division were present in the gonad. At 50 days after hatching, ovarian cavities and mature sperm were observed. Moreover, during the study we also observed the coexistence of two sexual characteristics during a male's development. Together, these observations fill current gaps in developmental and genetic biology that will allow a more efficient use of Medaka as an experimental model.

[Analysis of milk and dairy products by near-infrared spectroscopy: a review].

As a highly efficient, convenient and nondestructive detection method, near-infrared spectroscopy (NIRS) is appropriate for the analysis of milk and dairy products. The present review discusses the principle of the near-infrared spectroscopy analysis technology and the evaluation standard criteria of milk and dairy products, and also reviews the hardware device technology and the data processing technique for the analysis of milk and dairy products in order to promote the application of near-infrared spectroscopy analysis technology in milk and dairy products quality control in China.