Stimulation of avermectin B1a biosynthesis in Streptomyces avermilitis by feeding glucose and propionate

The biosynthetic ability of avermectin B1a in Streptomyces avermilitis was improved with propionate addition and glucose feeding. The results showed that B1a production was increased by 12.8–13.8% through supplement of 0.8% propionate at 24 h of cultivation. A stronger stimulation on B1a biosynthesis in S. avermilitis was observed by 3.0% glucose feeding at 5 d of cultivation. The B1a biosynthesis could be further enhanced by repeated glucose fed-batch process in a 10-l bench-top fermentor. A maximal B1a concentration of 780.0 mg/l was obtained by repeated 1.0% glucose addition on 4 d, 5 d and 6 d, which was 2.1-fold higher than that in a control fermentation. Corresponding with this, an additional 6.8% increase of B1a proportion was observed in comparison with the control process. This stimulation on avermectin B1a production was obvious even in a 2000-l fermentor under suitable control of aeration.     

菜缢管蚜、棉铃虫对杀虫混剂及其单剂的抗性遗传力分析

采用阈性状分析法,估算了菜缢管蚜Lipaphis erysimi(Kaltenbach)和棉铃虫Helicoverpa armigera(Hubuer)对菊马混剂及其单剂、棉铃虫对灭铃威混剂及其单剂的抗性现实遗传力,并对抗性风险和混剂延缓抗性的作用进行了评估。结果表明,菜缢管蚜和棉铃虫对混剂的现实抗性遗传力均明显小于各单剂,依次为：拟除虫菊酯>氨基甲酸酯>有机磷>混剂。混剂的使用寿命大于组成其各个单剂的使用寿命之和,抗性风险依次为：拟除虫菊脂>氨基甲酸酯>有机磷>混剂。菊马混剂对菜缢管蚜和棉铃虫,灭铃威混剂对棉铃虫均有延缓抗性发展的作用。灭铃威对棉铃虫抗性发展的延缓作用最显著。     

Numerical approaches for the rapid analysis of prophylactic efficacy against HIV with arbitrary drug-dosing schemes

Pre-exposure prophylaxis (PrEP) is an important pillar to prevent HIV transmission. Because of experimental and clinical shortcomings, mathematical models that integrate pharmacological, viral- and host factors are frequently used to quantify clinical efficacy of PrEP. Stochastic simulations of these models provides sample statistics from which the clinical efficacy is approximated. However, many stochastic simulations are needed to reduce the associated sampling error. To remedy the shortcomings of stochastic simulation, we developed a numerical method that allows predicting the efficacy of arbitrary prophylactic regimen directly from a viral dynamics model, without sampling. We apply the method to various hypothetical dolutegravir (DTG) prophylaxis scenarios. The approach is verified against state-of-the-art stochastic simulation. While the method is more accurate than stochastic simulation, it is superior in terms of computational performance. For example, a continuous 6-month prophylactic profile is computed within a few seconds on a laptop computer. The method’s computational performance, therefore, substantially expands the horizon of feasible analysis in the context of PrEP, and possibly other applications.     

Large numbers of interleukins‐22‐ and ‐17A‐producing T helper cells in cholangiocarcinoma related to liver fluke infection

Cholangiocarcinoma (CCA) associated with liver fluke infection involves inflammatory and immune processes; however, whether these involve the proinflammatory cytokine IL‐17A and proliferative cytokine IL‐22 remains unclear. Here, numbers of IL‐22‐ and IL‐17A‐producing Th cells and cytokine concentrations in 30 patients with CCA and long‐term liver fluke infection, 40 patients with liver‐fluke infection but not CCA, and 16 healthy controls were compared. Analyses were performed using immunohistochemistry, flow cytometry, ELISA and RT‐PCR. Immunohistochemical staining showed weaker expression of IL‐22 and IL‐17A in patients with CCA with than in those without liver fluke infection (P < 0.01). Flow cytometry revealed significantly greater median proportions of IL‐22‐producing T helper cells in patients with CCA (2.2%) than in those without it (0.69%) or controls (0.4%, P < 0.001). Similar results were obtained for IL‐17A‐producing T helper cells. ELISA revealed plasma concentrations of IL‐22 were 1.3‐fold higher in patients with CCA than in those without it and 4.6‐fold higher than in controls (P < 0.001). Plasma concentrations of IL‐17A were 2.5‐fold higher in patients with CCA than in those without it, and 21‐fold higher than in controls (P < 0.001). Amounts of IL‐22 and IL‐17A mRNAs in blood were significantly higher in patients with CCA than in the other two groups. Proportions of CD4⁺CD45RO⁺ T cells producing IL‐22 correlated with proportions producing IL‐17A (r = 0.759; P < 0.001), and plasma concentrations of IL‐22 correlated with those of IL‐17A (r = 0.726; P < 0.001). These results suggest that both IL‐17A and IL‐22 affect development of CCA related to liver fluke infection.

     

Structural basis of Vta1 function in the multivesicular body sorting pathway

The MVB pathway plays essential roles in several eukaryotic cellular processes. Proper function of the MVB pathway requires reversible membrane association of the ESCRTs, a process catalyzed by Vps4 ATPase. Vta1 regulates the Vps4 activity, but its mechanism of action was poorly understood. We report the high-resolution crystal structures of the Did2- and Vps60-binding N-terminal domain and the Vps4-binding C-terminal domain of S. cerevisiae Vta1. The C-terminal domain also mediates Vta1 dimerization and both subunits are required for its function as a Vps4 regulator. Emerging from our analysis is a mechanism of regulation by Vta1 in which the C-terminal domain stabilizes the ATP-dependent double ring assembly of Vps4. In addition, the MIT motif-containing N-terminal domain, projected by a long disordered linker, allows contact between the Vps4 disassembly machinery and the accessory ESCRT-III proteins. This provides an additional level of regulation and coordination for ESCRT-III assembly and disassembly.     

Long-term production of soluble human Fas ligand through immobilization of <Emphasis Type="Italic">Dictyostelium discoideum</Emphasis> in a fibrous bed bioreactor

The production of recombinant glycoproteins in Dictyostelium discoideum by conventional cell culture methods was limited by low cell density as well as low growth rate. In this work, cotton towel with a good adsorption capability for D. discoideum cells was used as the immobilization matrix in an external fibrous bed bioreactor (FBB) system. With batch cultures in the FBB, the concentration of immobilized cells in the cotton fiber carrier increased to 1.37 × 10⁸ cells per milliliter after 110-h cultivation, which was about tenfold higher than the maximal cell density in the conventional free-cell culture. Correspondingly, a high concentration of soluble human Fas ligand (hFasL; 173.7 μg l⁻¹) was achieved with a high productivity (23 μg l⁻¹ h⁻¹). The FBB system also maintained a high density of viable cells for hFasL production during repeated-batch cultures, achieving a productivity of 9∼10 μg l⁻¹ h⁻¹ in all three batches studied during 15 days. The repeated-batch culture using immobilized cells of D. discoideum in the FBB system thus provides a good method for long-term and high-level production of hFasL.