Angiogenesis is an important process in cell development, especially in cancer. Vascular endothelial growth factor (VEGF) signaling is an important regulator of angiogenesis. Several therapies that ...act against VEGF signal transduction have been developed, including YN968D1, which is a potent inhibitor of the VEGF signaling pathway. This study investigated the antitumor activity of YN968D1 (apatinib mesylate) in vitro and in vivo. YN968D1 potently suppressed the kinase activities of VEGFR‐2, c‐kit and c‐src, and inhibited cellular phosphorylation of VEGFR‐2, c‐kit and PDGFRβ. YN968D1 effectively inhibited proliferation, migration and tube formation of human umbilical vein endothelial cells induced by FBS, and blocked the budding of rat aortic ring. In vivo, YN968D1 alone and in combination with chemotherapeutic agents effectively inhibited the growth of several established human tumor xenograft models with little toxicity. A phase I study of YN968D1 has shown encouraging antitumor activity and a manageable toxicity profile. These findings suggest that YN968D1 has promise as an antitumor drug and might have clinical benefits. (Cancer Sci 2011; 102: 1374–1380)
Food waste (FW) accounts for the largest category of municipal solid wastes in the United States and the proper disposal of FW is a unique challenge with urgent environmental concerns. Solid-state ...anaerobic digestion (SSAD) is a viable option to treat FW but still hindered by poor mass transfer and severe inhibition caused by the accumulation of acidic intermediates. To improve the treatment capacity and process stability using high-solid FW as the sole feedstock via AD, a bench-scale two-stage system comprising of a first-stage solid-state digester (Stage 1; 2 L) and a second-stage liquid digester (Stage 2; 1.4 L) was established and carried out in a semi-batch mode for 99 days. When one-stage SSAD started to encounter acid accumulation and methanogenesis inhibition at a FW loading of 300 g (15% of working volume), the two-stage system secured robust and stable biogas production when the FW loading continued rising to 400 g (20% of working volume), and an increase of 2.4 kJ/gVS (equivalent to 18.2% in one-stage system) in energy yield was achieved. Stage 1 served as a hydrolysis/acidogenesis reactor with a total acids level of 28.5 g/L and a pH of 5.65. A large portion of lactic acid in total acids indicated shifts of metabolic pathways. The transfer of acid-rich leachate from Stage 1 to Stage 2 ensured a high methane yield of 393.0 mL/gVS (90.2% of the biological methane potential), while the liquid effluent recirculation provided buffering to Stage 1. Further increase of FW loading triggered a system instability. The enhanced buffering capacity and increased free ammonia molecules, synergistically induced certain inhibitory effects on FW acidogenesis in Stage 1. To conclude, the two-stage system outpaced the one-stage SSAD with higher FW loading and energy yield.
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•First-stage solid-state digester and second-stage liquid digester were connected.•Stage 1 served hydrolysis and acidogenesis while Stage 2 served methanogenesis.•Shift of acetic and butyric pathways to lactic pathway was shown in Stage 1.•Food waste loading to two-stage increased by 33.3% as compared to one-stage.•Two-stage outweighed one-stage in specific methane yield by 18.2%.
Yeast diversity and dynamics associated with the spontaneous fermentation of Vidal blanc icewine was investigated in icewine from the Huanren region of China. This knowledge is crucial to control ...fermentation and to screen for indigenous strains capable of producing icewines with regional characteristics. Laboratory-scale spontaneous fermentation was performed to monitor wine yeasts involved in the fermentation. Samples were collected from different stages of the fermentation and analyzed using culture-dependent (Wallerstein laboratory nutrient agar) and -independent methods (high-throughput sequencing, HTS). Two hundred eight yeast strains were isolated and identified by sequencing the internal transcribed spacer (ITS) region and the 26S rDNA D1/D2 domain genes; non-Saccharomyces yeasts were predominant in the initial stages, while Saccharomyces yeasts dominated in the final stages of the fermentation. Analysis of the HTS data of the ITS1 region gene revealed a more complex mycobiota. The wine yeast species found by HTS were more abundant than those found by cultivation; non-Saccharomyces yeasts were dominant initially, while Metschnikowia and Saccharomyces were predominant in the end. These results were not completely in agreement with the results obtained by using the cultivation-based method. Moreover, HTS revealed the presence of Mrakiella and Mrakia, which have never been reported in icewine before.
•Spontaneous fermentation of Vidal blanc icewine was monitored.•Cultivation and HTS revealed yeast diversity and evolution during fermentation.•New colonies morphotypes on WL nutrient agar were identified and described.•First study to describe psychrophilic basidiomycetes in icewine fermentation
Klebsiella pneumoniae is an important multidrug-resistant pathogen carrying prophages. Here we explore the contribution of prophages to bacterial evolution and fitness in silico. This study showed ...prophages contribute to remarkable genome plasticity of K. pneumoniae. The strains of CG258 possess several conserved prophages including the couple of P2-P4 prophages. CRISPR-Cas system has limited impact on the presence of prophages. The strong MLST-depended distribution of CRISPR-Cas and prophages and the high proportion of strains with self-targeting spacers may be the causes. Four core ARGs (blaSHV, fosA and oqxAB) were detected on almost all the chromosomes, but the acquired ARGs were only found in CG258 and CRISPR-positive strains. The factors influencing the chromosomal integration of ARGs in CG258 and CRISPR-positive strains may be different. In CG258, prophages may involve the chromosomal integration of ARGs. For CRISPR-positive strains, the immunity of CRISPR-Cas systems against invading ARG-bearing mobile genetic elements may accelerate the process.
•Prophages contribute to remarkable genome plasticity of K. pneumoniae.•The strains of CG258 possess several conserved prophages including the couple of P2-P4 prophages.•CRISPR-Cas system has limited impact on the presence of prophages.•The acquired ARGs were only found in CG258 and CRISPR-positive strains.•In CG258, prophages may involve the chromosomal integration of ARGs.
•The virulent phage vB_AbaS_D0 of Acinetobacter baumannii was isolated, characterized and the genome was sequenced.•Bacteria did not seem able to develop resistance to phage vB_AbaS_D0.•The ...anti-bacterial effect of the phage cocktail (vB_AbaS_D0 and vB_AbaP_D2) was more effective than use of individual phages.•Phage cocktail and vB_AbaS_D0 induced lower rates of phage resistant mutants in vivo.
The control and treatment of multidrug resistant pathogens infections has become a grand challenge for clinicians worldwide. Virulent phage has long been considered as an effective bactericidal agent, which may be a potentially alternative to antibiotics. However, the rapid development of phage resistance seriously hinders the wide and continuous application of virulent phages. In this study, Acinetobacter baumannii phage vB_AbaS_D0 was isolated, characterized and used to control the phage resistance development in bacterial strains. Transmission electron microscopy analysis of vB_AbaS_D0 indicated it belonged to the Siphoviridae family with an icosahedral head. Its whole genome was 43, 051 bp in size, with a GC content of 45.48% and 55 putative open reading frames. The data showed that vB_AbaS_D0 was a virulent phage. Although vB_AbaS_D0 had a very weak bactericidal activity, a wide range of Acinetobacter baumannii strains were sensitive to it. The results suggested that the cocktail of vB_AbaS_D0 and another Acinetobacter baumannii phage vB_AbaP_D2 could improve the therapeutic efficacy in vivo and in vitro. The resistance mutation frequency of A. baumannii cells infected with D0 or phage cocktail was significantly lower than cells treated with D2 (P < 0.01). Phage therapy in the murine bacteremia model results showed that the percentage of phage resistant mutant occurrence in the phage D0 or cocktail treatment group was significantly lower than in phage D2 treatment group (P < 0.01).
Significance
In transcription factors’ search for target genes, one-dimensional diffusion of the protein along DNA is essential. Experimentally, it remains challenging to resolve the individual ...diffusional steps of protein on DNA. Here, we report mainly all-atom equilibrium simulations of a WRKY domain protein in association with and diffusion along DNA. We demonstrate a complete stepping cycle of the protein for one base pair on DNA within microseconds, along with stochastic motions. Processive protein diffusions on DNA have been further sampled in a coarse-grained model. We have also found preferential DNA-strand association of the domain protein, which becomes most prominent at specific DNA binding, and it can be common for small-domain proteins to balance movements on the DNA with the sequence recognition.
Transcription factor (TF) target search on genome is highly essential for gene expression and regulation. High-resolution determination of TF diffusion along DNA remains technically challenging. Here, we constructed a TF model system using the plant WRKY domain protein in complex with DNA from crystallography and demonstrated microsecond diffusion dynamics of WRKY on DNA by employing all-atom molecular-dynamics (MD) simulations. Notably, we found that WRKY preferentially binds to one strand of DNA with significant energetic bias compared with the other, or nonpreferred strand. The preferential DNA-strand binding becomes most prominent in the static process, from nonspecific to specific DNA binding, but less distinct during diffusive movements of the domain protein on the DNA. Remarkably, without employing acceleration forces or bias, we captured a complete one-base-pair stepping cycle of the protein tracking along major groove of DNA with a homogeneous poly-adenosine sequence, as individual hydrogen bonds break and reform at the protein–DNA binding interface. Further DNA-groove tracking motions of the protein forward or backward, with occasional sliding as well as strand crossing to minor groove of DNA, were also captured. The processive diffusion of WRKY along DNA has been further sampled via coarse-grained MD simulations. The study thus provides structural dynamics details on diffusion of a small TF domain protein, suggests how the protein approaches a specific recognition site on DNA, and supports further high-precision experimental detection. The stochastic movements revealed in the TF diffusion also provide general clues about how other protein walkers step and slide along DNA.
Due to increasing safety concerns regarding human consumption of fish products, an increasing number of medicinal chemicals are prohibited from use in aquaculture. As a result, Chinese herbal ...medicines are being increasingly used, coining the use of the term ＂green medicine.＂ Research shows that Chinese herbal medicines have many beneficial effects on fish, including growth promotion, enhancement of disease resistance, and improvement in meat quality. Many effective ingredients have been discovered in Chinese herbal medicines, which function to promote feed intake, improve meat flavor, and increase digestive enzyme activity. They also regulate and participate in processes that improve the specific and non-specific immunity of fish; however, the composition of Chinese herbal medicines is very complex and it is often difficult to identify the effective ingredients. This article reviews the latest research and application progress in Chinese herbal medicines regarding growth and feed utilization, immunity and disease resistance, and the meat quality of cultured fish. It also discusses research on the chemical constituents of classical Chinese medicinal herbs and problems with the application of Chinese herbal medicines in fish culture. This article concludes by proposing that future studies on Chinese herbal medicines should focus on how to cheaply refine and extract the effective ingredients in classical Chinese medicinal herbs, as well as how to use them efficiently in aquaculture.
Botanical oils are staple consumer goods globally, but as a by-product of oil crops, meal is of low utilization value and prone to causing environmental problems. The development of proteins in meal ...into bioactive peptides, such as
peptide, through biotechnology can not only solve environmental problems, but also create more valuable nutritional additives. In the present work, the hydrolysis process of
meal protein suitable for industrial application was optimized with the response surface methodology (RSM) on the basis of single-factor experiments. Alcalase was firstly selected as the best-performing among four proteases. Then, based on Alcalase, the optimal hydrolysis conditions were as follows: enzyme concentration of 7%, hydrolysis temperature of 61.4 °C, liquid-solid ratio of 22.33:1 (mL/g) and hydrolysis time of 4 h. Under these conditions, the degree of hydrolysis (DH) of
meal protein was 26.23 ± 0.83% and the DPPH scavenging capacity of hydrolysate was 94.15 ± 1.12%. The soluble peptide or protein concentration of
meal protein hydrolysate rose up to 5.24 ± 0.05 mg/mL, the ideal yield of which was estimated to be 17.9%. SDS-PAGE indicated that a large proportion of new bands in hydrolysate with small molecular weights appeared, which was different from the original
meal protein. The present data contributed to further, more specific research on the separation, purification and identification of antioxidant peptide from the hydrolysate of
meal protein. The results showed that the hydrolysis of
meal protein could yield peptides with high antioxidant activity and potential applications as natural antioxidants in the food industry.
Zearalenone (ZEA) is present worldwide as a serious contaminant of food and feed and causes male reproductive toxicity. The implication of paraptosis, which is a nonclassical paradigm of cell death, ...is unclear in ZEA-induced male reproductive disorders. In this study, the toxic effects of ZEA on the blood-testis barrier (BTB) and the related mechanisms of paraptosis were detected in goats. ZEA exposure, in vivo, caused a significant decrease in spermatozoon quality, the destruction of seminiferous tubules, and damage to the BTB integrity. Furthermore, ZEA exposure to Sertoli cells (SCs) in vitro showed similar dysfunction in structure and barrier function. Importantly, the formation of massive cytoplasmic vacuoles in ZEA-treated SCs corresponded to the highly swollen and dilative endoplasmic reticulum (ER), and paraptosis inhibition significantly alleviated ZEA-induced SC death and vacuolization, which indicated the important contribution of paraptosis in ZEA-induced BTB damage. Meanwhile, the expression of ER stress marker proteins was increased after ZEA treatment but decreased under the inhibition of paraptosis. The vacuole formation and SC death, induced by ZEA, were remarkably blocked by ER stress inhibition. In conclusion, these results facilitate the exploration of the mechanisms of the SC paraptosis involved in ZEA-induced BTB damage in goats.
Trastuzumab emtansine (T‐DM1), an antibody–drug conjugate (ADC) consisting of human epidermal growth factor receptor 2 (HER2)‐targeted mAb trastuzumab linked to antimicrotubule agent mertansine ...(DM1), has been approved for the treatment of HER2‐positive metastatic breast cancer. Acquired resistance has been a major obstacle to T‐DM1 treatment, and mechanisms remain incompletely understood. In the present study, we established a T‐DM1‐resistant N87‐KR cell line from HER2‐positive N87 gastric cancer cells to investigate mechanisms of acquired resistance and develop strategies for overcoming it. Although the kinetics of binding, internalization, and externalization of T‐DM1 were the same in N87‐KR cells and N87 cells, N87‐KR was strongly resistant to T‐DM1, but remained sensitive to both trastuzumab and DM1. T‐DM1 failed to inhibit microtubule polymerization in N87‐KR cells. Consistently, lysine‐MCC‐DM1, the active T‐DM1 metabolite that inhibits microtubule polymerization, accumulated much less in N87‐KR cells than in N87 cells. Furthermore, lysosome acidification, achieved by vacuolar H+‐ATPase (V‐ATPase), was much diminished in N87‐KR cells. Notably, treatment of sensitive N87 cells with the V‐ATPase selective inhibitor bafilomycin A1 induced T‐DM1 resistance, suggesting that aberrant V‐ATPase activity decreases T‐DM1 metabolism, leading to T‐DM1 resistance in N87‐KR cells. Interestingly, HER2‐targeted ADCs containing a protease‐cleavable linker, such as hertuzumab‐vc‐monomethyl auristatin E, were capable of efficiently overcoming this resistance. Our results show for the first time that a decrease in T‐DM1 metabolites induced by aberrant V‐ATPase activity contributes to T‐DM1 resistance, which could be overcome by HER2‐targeted ADCs containing different linkers, including a protease‐cleavable linker. Accordingly, we propose that V‐ATPase activity in lysosomes is a novel biomarker for predicting T‐DM1 resistance.
To date, acquired resistances arise to be a major obstacle to T‐DM1 treatment and mechanisms remain incompletely understood. Our results demonstrate for the first time that T‐DM1 metabolites reduction induced by V‐ATPase aberation contributes to T‐DM1 resistance, which could be overcome by HER2‐targeted ADC with different linkers including protease‐cleavable linker, and we propose V‐ATPase activity in lysosomes as a novel biomarker to predict T‐DM1 resistance.
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