Very high density of CHO cells in perfusion by ATF or TFF in WAVE bioreactor™. Part I. Effect of the cell density on the process

High cell density perfusion process of antibody producing CHO cells was developed in disposable WAVE Bioreactor? using external hollow fiber filter as cell separation device. Both “classical” tangential flow filtration (TFF) and alternating tangential flow system (ATF) equipment were used and compared. Consistency of both TFF‐ and ATF‐based cultures was shown at 20–35 × 10⁶ cells/mL density stabilized by cell bleeds. To minimize the nutrients deprivation and by‐product accumulation, a perfusion rate correlated to the cell density was applied. The cells were maintained by cell bleeds at density 0.9–1.3 × 10⁸ cells/mL in growing state and at high viability for more than 2 weeks. Finally, with the present settings, maximal cell densities of 2.14 × 10⁸ cells/mL, achieved for the first time in a wave‐induced bioreactor, and 1.32 × 10⁸ cells/mL were reached using TFF and ATF systems, respectively. Using TFF, the cell density was limited by the membrane capacity for the encountered high viscosity and by the pCO₂ level. Using ATF, the cell density was limited by the vacuum capacity failing to pull the highly viscous fluid. Thus, the TFF system allowed reaching higher cell densities. The TFF inlet pressure was highly correlated to the viscosity leading to the development of a model of this pressure, which is a useful tool for hollow fiber design of TFF and ATF. At very high cell density, the viscosity introduced physical limitations. This led us to recommend cell densities under 1.46 × 10⁸ cell/mL based on the analysis of the theoretical distance between the cells for the present cell line. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:754–767, 2013     

单眼远视性弱视儿童P-VEP检查的临床研究

目的:分析单眼远视性弱视儿童图形视觉诱发电位(P-VEP)检查情况,探讨外周发病机制,为临床诊疗提供依据。方法:选取2013年1月到2015年10月我院收治的单眼远视性弱视儿童75例(75只眼),另选取同期正常儿童32例(64只眼)为对照组,根据病情将弱视儿童分为轻度(A组)和对侧健眼组(B组),中度(C组)和对侧健眼组(D组),重度(E组)和对侧健眼组(F组),应用P-VEP检查各组P100波及振幅。结果:A组、C组、E组P100波潜伏期较B组、D组、F组和对照组延长(P0.05),振幅较B组、D组、F组和对照组降低(P0.05),A组、C组和、E组P100波潜伏期和振幅比较具有统计学意义(P0.05),B组、D组、F组P100波潜伏期与对照组无统计学意义(P0.05),B组、D组、F组振幅显著低于对照组(P0.05),B组、D组、F组P100波潜伏期和振幅比较无统计学意义(P0.05)。结论:单眼远视性弱视儿童弱视眼会出现P100波潜伏期延长,振幅降低,对侧健康眼会出现振幅降低。     

Early network activity propagates bidirectionally between hippocampus and cortex

Spontaneous activity in the developing brain helps refine neuronal connections before the arrival of sensory‐driven neuronal activity. In mouse neocortex during the first postnatal week, waves of spontaneous activity originating from pacemaker regions in the septal nucleus and piriform cortex propagate through the neocortex. Using high‐speed Ca²⁺ imaging to resolve the spatiotemporal dynamics of wave propagation in parasagittal mouse brain slices, we show that the hippocampus can act as an additional source of neocortical waves. Some waves that originate in the hippocampus remain restricted to that structure, while others pause at the hippocampus‐neocortex boundary and then propagate into the neocortex. Blocking GABAergic neurotransmission decreases the likelihood of wave propagation into neocortex, whereas blocking glutamatergic neurotransmission eliminates spontaneous and evoked hippocampal waves. A subset of hippocampal and cortical waves trigger Ca²⁺ waves in astrocytic networks after a brief delay. Hippocampal waves accompanied by Ca²⁺ elevation in astrocytes are more likely to propagate into the neocortex. Finally, we show that two structures in our preparation that initiate waves—the hippocampus and the piriform cortex—can be electrically stimulated to initiate propagating waves at lower thresholds than the neocortex, indicating that the intrinsic circuit properties of those regions are responsible for their pacemaker function. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 661–672, 2016     

Imaging Calcium in Drosophila at Egg Activation

Egg activation is a universal process that includes a series of events to allow the fertilized egg to complete meiosis and initiate embryonic development. One aspect of egg activation, conserved across all organisms examined, is a change in the intracellular concentration of calcium (Ca²⁺) often termed a ''Ca²⁺ wave''. While the speed and number of oscillations of the Ca²⁺ wave varies between species, the change in intracellular Ca²⁺ is key in bringing about essential events for embryonic development. These changes include resumption of the cell cycle, mRNA regulation, cortical granule exocytosis, and rearrangement of the cytoskeleton.In the mature Drosophila egg, activation occurs in the female oviduct prior to fertilization, initiating a series of Ca²⁺-dependent events. Here we present a protocol for imaging the Ca²⁺ wave in Drosophila. This approach provides a manipulable model system to interrogate the mechanism of the Ca²⁺ wave and the downstream changes associated with it.     

Simulating effects of climate change under direct and diapause development in a butterfly

Temperature is one of the most important ecological factors affecting species survival and distributions. Therefore, global climate change, involving increases in mean surface temperature and the occurrence of extreme weather events, may pose a substantial challenge to biodiversity. Whereas tropical ectotherms are believed to be very sensitive to climate change, temperate‐zone species may actually benefit from higher temperatures. However, as in temperate zones large parts of the year are unsuitable for growth and reproduction, seasonal time constraints may complicate matters. Against this background we here investigate the impact of simulated climate change, involving increased mean temperatures and heat waves, across developmental pathways of the butterfly Lycaena tityrus (Poda) (Lepidoptera: Lycaenidae). Increased temperatures speeded up development but decreased pupal mass as expected. However, we found no evidence for detrimental effects of increased temperatures or even simulated heat waves. Furthermore, patterns did not differ between indirectly and directly developing individuals, which are assumed to be more time constrained. Our findings support the notion that not all species will be detrimentally affected by climate change, and suggest that species attributes may be more important than potential time constraints imposed by different developmental pathways.     

参松养心胶囊对心房颤动患者P波离散度及血清炎性因子水平的影响*

目的:探讨参松养心胶囊对心房颤动(AF)患者P波离散度(Pd)及血清炎性因子水平的影响,为临床用药提供依据。方法:将西安交通大学第一附属医院2016年3月-2018年1月收治的AF患者纳入本研究,共148例。所有患者根据不同的治疗方式分为对照组和观察组。对照组(n=74)接受常规治疗,观察组(n=74)在对照组基础上加用参松养心胶囊,两组均治疗8周。比较两组患者治疗后的临床疗效,观察并比较两组患者治疗前及治疗后的Pd、最大P波时限(Pmax)变化情况,检测两组患者治疗前及治疗后的血清炎性因子水平,记录不良反应发生情况。结果:观察组总有效率为89.19%(66/74),高于对照组的70.27%(52/74)(P0.05)。两组患者治疗后的Pmax、Pd均明显降低,且观察组患者治疗后的Pmax、Pd均明显低于对照组(P0.05)。两组患者治疗后的血清炎性因子水平均明显降低,且与对照组相比,观察组患者治疗后的血清肿瘤坏死因子-α(TNF-α)、高敏C反应蛋白(hs-CRP)、白介素-6(IL-6)水平均降低(P0.05)。两组不良反应发生情况比较无统计学差异(P0.05)。结论:参松养心胶囊治疗AF患者安全有效,能明显减轻炎性反应,改善患者心功能。     

Rational Structure Optimized Hybrid Nanogenerator for Highly Efficient Water Wave Energy Harvesting

Water wave energy is a promising renewable energy source that may alleviate the rising concerns over current resource depletion, but it is rarely exploited due to the lack of efficient energy harvesting technologies. In this work, a hybrid system with a triboelectric nanogenerator (TENG) and an electromagnetic generator (EMG) based on an optimized inner topological structure is reported to effectively harvest water wave energy. The TENG with etched polytetrafluoroethylene films and Cu electrodes utilizing the contact‐freestanding mode is designed into a cubic structure, in which the EMG is well hybridized. An integration of TENG and EMG achieves mutual compensation of their own merits, enabling the hybrid system to deliver satisfactory output over a broad range of operation frequency. The output performance of TENG with varied inner topological structures is experimentally and theoretically compared, and a concept is proposed to further clarify the energy conversion efficiency, which should be considered in designing energy harvesting devices. The influences of oscillation frequency, amplitude, and dielectric materials on the output performance of the hybrid system are comprehensively studied on different platforms. Furthermore, the optimum operation frequency ranges for TENG and EMG are concluded. The proposed hybrid nanogenerator renders an effective approach toward large‐scale blue energy harvesting over a broad frequency range.