Improved hematopoietic differentiation of human pluripotent stem cells via estrogen receptor signaling pathway

Additional file 2: Table S1. Temporal changes (%) of ER-Îą and hematopoietic phenotypes during hiPSC-derived hematopoietic differentiation

YH29407 with anti-PD-1 ameliorates anti-tumor effects via increased T cell functionality and antigen presenting machinery in the tumor microenvironment

Among cancer cells, indoleamine 2, 3-dioxygenase1 (IDO1) activity has been implicated in improving the proliferation and growth of cancer cells and suppressing immune cell activity. IDO1 is also responsible for the catabolism of tryptophan to kynurenine. Depletion of tryptophan and an increase in kynurenine exert important immunosuppressive functions by activating regulatory T cells and suppressing CD8+ T and natural killer (NK) cells. In this study, we compared the anti-tumor effects of YH29407, the best-in-class IDO1 inhibitor with improved pharmacodynamics and pharmacokinetics, with first and second-generation IDO1 inhibitors (epacadostat and BMS-986205, respectively). YH29407 treatment alone and anti-PD-1 (aPD-1) combination treatment induced significant tumor suppression compared with competing drugs. In particular, combination treatment showed the best anti-tumor effects, with most tumors reduced and complete responses. Our observations suggest that improved anti-tumor effects were caused by an increase in T cell infiltration and activity after YH29407 treatment. Notably, an immune depletion assay confirmed that YH29407 is closely related to CD8+ T cells. RNA-seq results showed that treatment with YH29407 increased the expression of genes involved in T cell function and antigen presentation in tumors expressing ZAP70, LCK, NFATC2, B2M, and MYD88 genes. Our results suggest that an IDO1 inhibitor, YH29407, has enhanced PK/PD compared to previous IDO1 inhibitors by causing a change in the population of CD8+ T cells including infiltrating T cells into the tumor. Ultimately, YH29407 overcame the limitations of the competing drugs and displayed potential as an immunotherapy strategy in combination with aPD-1

Protection of Japanese flounder Paralichthys olivaceus from viral hemorrhagic septicemia (VHS) by Poly(I:C) immunization

In immunization of fish with polyinosinic-polycytidylic acid (poly[I:C], a synthetic double-stranded RNA), injection of Poly(I:C) followed by challenge with a live virus induces a transient, non-specific antiviral state by interferon activity. When exposed to a virus while in this antiviral state, the fish acquire a specific and protective immunity against the corresponding viral disease and survive. In the present study, the effiacy of Poly(I:C) immunization was investigated in japanese flounder Paralichthys olivaceus using viral hemorrhagic septicemia virus (VHSV) as a model; the minimum dose of Poly(I:C) required for inducing protection and the duration of the antiviral state were determined, and a potentially curative effect of Poly(I:C) administration was assessed. The antiviral state was induced by administration of Poly(I:C) doses ranging from 12.5 to 200 µg fish(-1). Minimum dose to induce the antiviral state (relative percentage survival, RPS: 90%) was 12.5 µg fish(-1). No curative effect of Poly(I:C) was observed in fish pre-infected with VHSV. Fish injected with 200 µg Poly(I:C) fish(-1) were highly protected (RPS: 100%) from an artificial challenge with VHSV, and specific antibodies against VHSV were detected. The corresponding high level of antiviral state against VHSV was attained 1 d post Poly(I:C) injection, lasted for 6 d and susequently decreased. Moreover, the surviving fish were highly protected from re-challenge with VHSV (RPS: 100%). Thus, it was considered that an immunity against viral hemorrhagic septicemia was induced in the Japanese flounder by injecting live VHSV following Poly(I:C) administration

ELISA法による抗RSIVブリ抗体の検出

FBS components in viral culture fluid interfere with the detection of specific antibody in immunized fish. In the present study, we developed a method to eliminate the interference of FBS using a model of red sea bream iridovirus (RSIV) vaccine. In an experiment using yellowtail antiserum against BSA, which is a major component of FBS in cell culture media, antibodies against BSA were completely absorbed via incubation of antiserum in 50% FBS at 25 degrees C for 1 h. In antibody detection, ELISA with the sera of yellowtail surviving RSIV-challenge, antibodies against RSIV were detected, but those against FBS disappeared via the pre-treatment of the sera. Therefore, antibodies against RSIV are effectively detected using ELISA with the pre-treatment of fish sera.RSIV培養液を接種したブリからの抗RSIV抗体検出ELISA法について検討した。培養液に含まれるFBSの主要成分であるBSAに対する抗ブリ血清を用いた場合、血清に終濃度50％（v／v）となるようFBSを添加し25℃で1時間処理することで、血清中の抗BSA抗体が完全に吸収された。培養RSIV液で攻撃した生残ブリから得た血清を同様に処理することで、抗体検出ELISAにより抗RSIV抗体が検出できることが明らかになった

Development of a novel real-time RT-PCR method using peptide nucleic acid (PNA) probes for detecting and genotyping of viral haemorrhagic septicaemia virus (VHSV)

Viral haemorrhagic septicaemia (VHS) is one of the most serious viral diseases in salmonid and olive flounder farms. The causative agent of VHS is the VHS virus (VHSV), which has been classified into four genotypes (I–IV), based on sequence analysis of the genes encoding for nucleoprotein, glycoprotein, and non-structural viral protein. Among the various diagnostic methods, real-time reverse transcription PCR method based on TaqMan-probe (RT-qPCR) is a stable, rapid, specific, and highly sensitive method for viral gene detection. However, the currently accepted diagnostic method based on RT-qPCR can only detect viral presence and load, and does not provide information about viral genotype. Peptide nucleic acids (PNAs) are artificially synthesized DNA analogues with an uncharged peptide backbone. PNA probes can effectively detect a target gene by amplification and a specific melting temperature signal. It was reported that PNA probes can effectively distinguish between mismatched sequences based on their different melting temperatures in amplified PCR products. The present study reports a novel real-time RT-PCR method for simultaneous detection and genotyping of VHSV using PNA probes. The newly-developed method showed a sensitivity similar to that of the infectious titre by fish cell cultures inoculated with the virus, except for genotype IVa, where viral inoculation in cell culture showed a 10-fold higher sensitivity than the novel method. The melting point analysis to distinguish the four genotypes was performed on 80 VHSV isolates representing all known genotypes, showing that this novel real-time RT-PCR can distinguish between all VHSV genotypes without the need of further sequencing

Evaluation of the Antimicrobial Effect of Graphene Oxide Fiber on Fish Bacteria for Application in Aquaculture Systems

The growing importance of the domestic aquaculture industry has led not only to its continuous development and expansion but also to an increase in the production of wastewater containing pathogenic microorganisms and antibiotic-resistant bacteria. As the existing water purification facilities have a high initial cost of construction, operation, and maintenance, it is necessary to develop an economical solution. Graphene oxide (GO) is a carbon-based nanomaterial that is easy to manufacture, inexpensive and has excellent antimicrobial properties. In this study, the antimicrobial effect of GO polyester fibers on seven species of fish pathogenic bacteria was analyzed to evaluate their effectiveness in water treatment systems and related products. As a result of incubating GO polyester fibers with seven types of fish pathogenic bacteria for 1, 6, and 12 h, there was no antimicrobial effect in Vibrio harveyi, V. scopthalmi, and Edwardsiella tarda. In contrast, GO fibers showed antimicrobial effects of more than 99% against A. hydrophila, S. parauberis, S. iniae, and P. piscicola, suggesting the potential use of GO fibers in water treatment systems

Modifications of the nucleoprotein of viral haemorrhagic septicaemia virus showed gain of virulence in intraperitoneally infected rainbow trout

International audienceViral haemorrhagic septicaemia virus (VHSV) is the cause of an important listed disease in European rainbow trout (Oncorhynchus mykiss) aquaculture and can be present in a wide range of fish species, including marine fish, which can act as viral reservoir. Recent studies revealed putative genetic virulence markers of VHSV to rainbow trout highlighting the roles of the nucleoprotein, phosphoprotein and non-virion protein. Using reverse genetics, we produced recombinant viruses by introducing parts of or the entire nucleoprotein from a high-virulent isolate VHSV into a low-virulent backbone. Furthermore, we also made recombinant viruses by introducing residue modifications in the nucleoprotein that seem to play a role in virulence. Rainbow trout challenged with these recombinant viruses (rVHSVs) by intraperitoneal injection (IP) developed clinical signs and showed lower survival when compared to the parental rVHSV whereas fish challenged by immersion did not show clinical signs except for the high-virulent control. The mutations did not influence the viral growth in cell culture. The recombinant viruses and parental recombinant were unable to replicate and show cytopathic effect in EPC cells whereas the high-virulent control was well adapted in all the fish cell lines tested. We showed evidence that corroborates with the hypothesis that the nucleoprotein has virulence motifs associated with VHSV virulence in rainbow trout

Vapor Transport Synthesis of Two-Dimensional SnS2 Nanocrystals Using a SnS2 Precursor Obtained from the Sulfurization of SnO2

Manufacturing high-quality, two-dimensional (2D), layered materials with crystal-growth techniques is an important challenge for the advancement of 2D communication technologies. In this study, a simple method was developed for synthesizing 2D nanocrystals based on the model system of SnS2. The method involves the sulfurization of a metal oxide to a metal chalcogenide, which subsequently acts as a source of vapors for the growth of 2D crystals. The effect of the annealing conditions on the thermal sulfurization of SnO2 powder was investigated. The results showed that pure SnS2 powder could be obtained in a N2 atmosphere at 700 °C. SnS2 nanocrystals were successfully synthesized from the as-prepared SnS2 powder by the vapor transport method. The synthesized SnS2 nanocrystals had a 2D layered structure with hexagonal symmetry and exhibited typical n-type semiconducting characteristics, with an optical band gap of 2.05 eV. This novel method, which uses a preferentially prepared source for vapor transport, could provide a simple way to synthesize new types of 2D layered materials. This is because it only requires the volatilization of a source and subsequent condensation to a single crystal for the growth of 2D materials, with no complex chemical reactions occurring during vapor transport. © 2016 American Chemical Society.