Geographically associated endophytic fungi contribute to the tropane alkaloids accumulation of Anisodus tanguticus

Anisodus tanguticus is a valuable plant for extracting tropane alkaloids. However, the mechanisms by which plant microbiome mediate the accumulation of tropane alkaloids in Anisodus tanguticus are still not well understood. In this study, we collected 55 wild Anisodus tanguticus populations on the Tibetan Plateau and the tropane alkaloids content, and root-related bacteria and fungi diversity were analyzed using HPLC and 16 s rDNA and ITS sequencing. The results showed that tropane alkaloids content has obvious geographical distribution characteristics. Anisodine content had a significant positive correlation with latitude, while anisodamine and atropine content had a significant negative correlation with latitude. Variation partition analysis (VPA) showed that root endophytes play a significant role in promoting tropane alkaloid production in Anisodus tanguticus roots. The root endophytes alone explained 14% of the variation, which was the largest contributor. Soil properties variables could independently explain 5% of the variation, and climate variables could explain 1% of the variation. Of these, endophytic fungi alone accounted for 11%, while bacteria explained only 5%. Random forests and Mantel test showed that different regionally enriched endophytic fungi have a greater impact on the accumulation of tropane alkaloids than the whole endophytic fungi. Richness and relative abundance of enriched endophytic fungi in Hengduan-Qilian Mountains (HQ) group has a significant positive correlation with anisodine content, while richness and relative abundance of enriched endophytic fungi in Himalayas-Hengduan Mountains (HH) group has a significant positive correlation with anisodamine and atropine content. And, these enriched endophytic fungi have high network connectivity and distributed in separate network modules. This study further confirmed that endophytes were closely related to tropane alkaloids accumulation in Anisodus tanguticus and contribute to promote sustainable development, cultivation, and precision medicine of Anisodus tanguticus

Use of Praziquantel as an Adjuvant Enhances Protection and Tc-17 Responses to Killed H5N1 Virus Vaccine in Mice

BACKGROUND: H5N1 is a highly pathogenic influenza A virus, which can cause severe illness or even death in humans. Although the widely used killed vaccines are able to provide some protection against infection via neutralizing antibodies, cytotoxic T-lymphocyte responses that are thought to eradicate viral infections are lacking. METHODOLOGY/PRINCIPAL FINDINGS: Aiming to promote cytotoxic responses against H5N1 infection, we extended our previous finding that praziquantel (PZQ) can act as an adjuvant to induce IL-17-producing CD8(+) T cells (Tc17). We found that a single immunization of 57BL/6 mice with killed viral vaccine plus PZQ induced antigen-specific Tc17 cells, some of which also secreted IFN-γ. The induced Tc17 had cytolytic activities. Induction of these cells was impaired in CD8 knockout (KO) or IFN-γ KO mice, and was even lower in IL-17 KO mice. Importantly, the inoculation of killed vaccine with PZQ significantly reduced virus loads in the lung tissues and prolonged survival. Protection against H5N1 virus infection was obtained by adoptively transferring PZQ-primed wild type CD8(+) T cells and this was more effective than transfer of activated IFN-γ KO or IL-17 KO CD8(+) T cells. CONCLUSIONS/SIGNIFICANCE: Our results demonstrated that adding PZQ to killed H5N1 vaccine could promote broad Tc17-mediated cytotoxic T lymphocyte activity, resulting in improved control of highly pathogenic avian influenza virus infection

Praziquantel Facilitates IFN-γ-Producing CD8+ T Cells (Tc1) and IL-17-Producing CD8+ T Cells (Tc17) Responses to DNA Vaccination in Mice

BACKGROUND: CD8(+) cytotoxic T lymphocytes (CTLs) are crucial for eliminating hepatitis B virus (HBV) infected cells. DNA vaccination, a novel therapeutic strategy for chronic virus infection, has been shown to induce CTL responses. However, accumulated data have shown that CTLs could not be effectively induced by HBV DNA vaccination. METHODOLOGY/PRINCIPAL FINDINGS: Here, we report that praziquantel (PZQ), an anti-schistoma drug, could act as an adjuvant to overcome the lack of potent CTL responses by HBV DNA vaccination in mice. PZQ in combination with HBV DNA vaccination augmented the induction of CD8(+) T cell-dependent and HBV-specific delayed hypersensitivity responses (DTH) in C57BL/6 mice. Furthermore, the induced CD8(+) T cells consisted of both Tc1 and Tc17 subtypes. By using IFN-γ knockout (KO) mice and IL-17 KO mice, both cytokines were found to be involved in the DTH. The relevance of these findings to HBV immunization was established in HBsAg transgenic mice, in which PZQ also augmented the induction of HBV-specific Tc1 and Tc17 cells and resulted in reduction of HBsAg positive hepatocytes. Adoptive transfer experiments further showed that PZQ-primed CD8(+) T cells from wild type mice, but not the counterpart from IFN-γ KO or IL-17 KO mice, resulted in elimination of HBsAg positive hepatocytes. CONCLUSIONS/SIGNIFICANCE: Our results suggest that PZQ is an effective adjuvant to facilitate Tc1 and Tc17 responses to HBV DNA vaccination, inducing broad CD8(+) T cell-based immunotherapy that breaks tolerance to HBsAg

A Review on Application of Biochar in the Removal of Pharmaceutical Pollutants through Adsorption and Persulfate-Based AOPs

Increasing quantities of pharmaceutical pollutants have been found in aquatic ecosystems. The treatment of pharmaceutical pollutants has been a major task that people have been committed to in recent years. The removal of pharmaceutical pollutants can be achieved by adsorption and advanced oxidation processes (AOPs). Compared with other carbon materials, biochar has a strong adsorption capacity and persulfate activation ability, and more importantly, biochar is cheap and easy to obtain; thus, it has higher economic benefits. This study firstly reviews the application of biochar in the removal of drugs (tetracycline (TC), sulfamethoxazole (SMX), acetaminophen (ACT), cephalexin (CPX), levofloxacin (LEV), etc.) through adsorption and persulfate-based AOPs. In addition, we summarize the adsorption mechanism of biochar for various pharmaceutical pollutants and the main attack sites on different pharmaceutical pollutants in persulfate-based AOPs catalyzed by biochar. Finally, the challenges and prospects of biochar with respect to the removal of pharmaceutical pollutants are put forward

Degradation of Tetracycline in Water by Fe-Modified Sterculia Foetida Biochar Activated Peroxodisulfate

Tetracycline (TC) is a broad-spectrum antibiotic commonly, made use of in aquaculture and animal husbandry. After entering water bodies, it will represent a major threat to human health. In this study, sterculia foetida biochar (SFC) was readied by the combined hydrothermal pyrolysis (co-HTP) method with sterculia foetida as raw materials. Fen-SFC (Fe2-SFC, Fe3-SFC, and Fe4-SFC) was obtained by doping SFH with different concentrations of FeCl3. Finally, activation of peroxodisulfate (PDS) was achieved, using Fe3-SFC to degrade TC. The degradation of TC obeyed pseudo-second-order kinetics, and the constant of the reaction rate was 0.491 L mg−1 min−1. Radical trapping experiments, EPR test and electrochemical tests evidenced that the high catalytic performance of the Fe3-SFC/PDS system was ascribed to free radical pathway (•OH and SO4•−) and non-radical pathway (1O2 and electron transfer), in which the latter plays a dominant role. This research not only demonstrates a new kind of biochar as an effective catalyst for PS activation, but also offers an avenue for the value-added reuse of sterculia foetida

Degradation of Tetracycline in Water by Fe-Modified Sterculia Foetida Biochar Activated Peroxodisulfate

Tetracycline (TC) is a broad-spectrum antibiotic commonly, made use of in aquaculture and animal husbandry. After entering water bodies, it will represent a major threat to human health. In this study, sterculia foetida biochar (SFC) was readied by the combined hydrothermal pyrolysis (co-HTP) method with sterculia foetida as raw materials. Fen-SFC (Fe2-SFC, Fe3-SFC, and Fe4-SFC) was obtained by doping SFH with different concentrations of FeCl3. Finally, activation of peroxodisulfate (PDS) was achieved, using Fe3-SFC to degrade TC. The degradation of TC obeyed pseudo-second-order kinetics, and the constant of the reaction rate was 0.491 L mg−1 min−1. Radical trapping experiments, EPR test and electrochemical tests evidenced that the high catalytic performance of the Fe3-SFC/PDS system was ascribed to free radical pathway (•OH and SO4•−) and non-radical pathway (1O2 and electron transfer), in which the latter plays a dominant role. This research not only demonstrates a new kind of biochar as an effective catalyst for PS activation, but also offers an avenue for the value-added reuse of sterculia foetida

Manipulating plant RNA-silencing pathways to improve the gene editing efficiency of CRISPR/Cas9 system

Background: The CRISPR/Cas9 system, composed of a single-guide RNA for target recognition and a Cas9 protein for DNA cleavage, has the potential to revolutionize agriculture as well as medicine. Even though extensive work has been done to improve the gene editing activity of CRISPR/Cas9, little is known about the regulation of this bacterial system in eukaryotic host cells, especially at the post-transcriptional level. Results: Here, we evaluate the expression levels of the two CRISPR/Cas9 components and the gene editing efficiency in a set of Arabidopsis mutants involved in RNA silencing. We find that mutants defective in the post-transcriptional gene-silencing pathway display significantly higher Cas9 and sgRNA transcript levels, resulting in higher mutagenesis frequencies than wild-type controls. Accordingly, silencing of AGO1 by introduction of an AGO1-RNAi cassette into the CRISPR/Cas9 vector provides an increase in gene editing efficiency. Co-expression of the viral suppressor p19 from the tomato bushy stunt virus to suppress the plant RNA-silencing pathway shows a strong correlation between the severity of the phenotypic effects caused by p19 and the gene editing efficiency of the CRISPR/Cas9 system for two different target genes, AP1 and TT4. Conclusions: This system has useful practical applications in facilitating the detection of CRISPR/Cas9-induced mutations in T1 plants as well as the identification of transgene-free T2 plants by simple visual observation of the symptom severity caused by p19. Our study shows that CRISPR/Cas9 gene editing efficiency can be improved by reducing RNA silencing in plants

The Application and Challenge of Binder Jet 3D Printing Technology in Pharmaceutical Manufacturing

Three-dimensional (3D) printing is an additive manufacturing technique that creates objects under computer control. Owing to the rapid advancement of science and technology, 3D printing technology has been widely utilized in processing and manufacturing but rarely used in the pharmaceutical field. The first commercial form of Spritam® immediate-release tablet was approved by FDA in 2015, which promoted the advancement of 3D printing technology in pharmaceutical development. Three-dimensional printing technology is able to meet individual treatment demands with customized size, shape, and release rate, which overcomes the difficulties of traditional pharmaceutical technology. This paper intends to discuss the critical process parameters of binder jet 3D printing technology, list its application in pharmaceutical manufacturing in recent years, summarize the still-open questions, and demonstrate its great potential in the pharmaceutical industry