Integration of high-throughput analytics and cell imaging enables direct early productivity and product quality assessment during Chinese Hamster ovary cell line development for a complex multi-subunit vaccine antigen

Mammalian cell line generation typically includes stable pool generation, single cell cloning and several rounds of clone selection based on cell growth, productivity and product quality criteria. Individual clone expansion and phenotype-based ranking is performed initially for hundreds or thousands of mini-scale cultures, representing the major operational challenge during cell line development. Automated cell culture and analytics systems have been developed to enable high complexity clone selection workflows; while ensuring traceability, safety, and quality of cell lines intended for biopharmaceutical applications. Here we show that comprehensive and quantitative assessment of cell growth, productivity, and product quality attributes are feasible at the 200–1,200 cell colony stage, within 14 days of the single cell cloning in static 96-well plate culture. The early cell line characterization performed prior to the clone expansion in suspension culture can be used for a single-step, direct selection of high quality clones. Such clones were comparable, both in terms of productivity and critical quality attributes (CQAs), to the top-ranked clones identified using an established iterative clone screening approach. Using a complex, multi-subunit antigen as a model protein, we observed stable CQA profiles independently of the cell culture format during the clonal expansion as well as in the batch and fed-batch processes. In conclusion, we propose an accelerated clone selection approach that can be readily incorporated into various cell line development workstreams, leading to significant reduction of the project timelines and resource requirements.     

Mujeres mayores de 65 años que realizan ejercicio físico supervisado: diferencias en la capacidad física,la actividad física y la calidad de vida en función de su velocidad de la marcha

IntroductionWalking speed (WS) is an easy, quick and inexpensive measure that could be used to discern between older people with greater and lesser function and thus individualize physical exercise programs.ObjectivesTo analyze the differences in physical capacity, physical activity, and quality of life in people over 65 years of age who attended a physical exercise program according to their WS and age.Methods55 women (mean age: 76.67 ± 6.66 years) were divided into groups based on their WS (low WS: ≤ 1.59 m/s and high WS: > 1.59 m/s) and age (older-younger: ≤ 76 years and older-older: > 76 years). The following parameters were compared: 10 Meters Walk Test (10MWT), Arm Curl Test, Handgrip, Chair Stand Test, 8 Foot Up and Go Test (8FUG), 6 Minute Walk Test (6MWT), and the Minnesota and The Short Form-36 Health Survey (SF-36) questionnaires.ResultsThe level of physical activity was higher than 3000 METs/week in all groups. The high WS group had better results in the Arm Curl Test, 10MWT, 8FUG and 6MWT and in the Physical Role and Vitality dimensions of the SF-36 (P < .05). The older-older group had lower weight, BMI and Handgrip (P < .01).ConclusionsThe best results in physical capacity and quality of life are in those women with higher WS, suggesting that WS could be useful to individualize physical exercise programs.     

Leaf traits and performance vary with plant age and water availability in Artemisia californica

Background and AimsLeaf functional traits are strongly tied to growth strategies and ecological processes across species, but few efforts have linked intraspecific trait variation to performance across ontogenetic and environmental gradients. Plants are believed to shift towards more resource-conservative traits in stressful environments and as they age. However, uncertainty as to how intraspecific trait variation aligns with plant age and performance in the context of environmental variation may limit our ability to use traits to infer ecological processes at larger scales.MethodsWe measured leaf physiological and morphological traits, canopy volume and flowering effort for Artemisia californica (California sagebrush), a dominant shrub species in the coastal sage scrub community, under conditions of 50, 100 and 150 % ambient precipitation for 3 years.Key ResultsPlant age was a stronger driver of variation in traits and performance than water availability. Older plants demonstrated trait values consistent with a more conservative resource-use strategy, and trait values were less sensitive to drought. Several trait correlations were consistent across years and treatments; for example, plants with high photosynthetic rates tended to have high stomatal conductance, leaf nitrogen concentration and light-use efficiency. However, the trade-off between leaf construction and leaf nitrogen evident in older plants was absent for first-year plants. While few traits correlated with plant growth and flowering effort, we observed a positive correlation between leaf mass per area and performance in some groups of older plants.ConclusionsOverall, our results suggest that trait sensitivity to the environment is most visible during earlier stages of development, after which intraspecific trait variation and relationships may stabilize. While plant age plays a major role in intraspecific trait variation and sensitivity (and thus trait-based inferences), the direct influence of environment on growth and fecundity is just as critical to predicting plant performance in a changing environment.     

Intraspecific trait variation influences physiological performance and fitness in the South Africa shrub genus Protea (Proteaceae)

Background and AimsGlobal plant trait datasets commonly identify trait relationships that are interpreted to reflect fundamental trade-offs associated with plant strategies, but often these trait relationships are not identified when evaluating them at smaller taxonomic and spatial scales. In this study we evaluate trait relationships measured on individual plants for five widespread Protea species in South Africa to determine whether broad-scale patterns of structural trait (e.g. leaf area) and physiological trait (e.g. photosynthetic rates) relationships can be detected within natural populations, and if these traits are themselves related to plant fitness.MethodsWe evaluated the variance structure (i.e. the proportional intraspecific trait variation relative to among-species variation) for nine structural traits and six physiological traits measured in wild populations. We used a multivariate path model to evaluate the relationships between structural traits and physiological traits, and the relationship between these traits and plant size and reproductive effort.Key ResultsWhile intraspecific trait variation is relatively low for structural traits, it accounts for between 50 and 100 % of the variation in physiological traits. Furthermore, we identified few trait associations between any one structural trait and physiological trait, but multivariate regressions revealed clear associations between combinations of structural traits and physiological performance (R² = 0.37–0.64), and almost all traits had detectable associations with plant fitness.ConclusionsIntraspecific variation in structural traits leads to predictable differences in individual-level physiological performance in a multivariate framework, even though the relationship of any particular structural trait to physiological performance may be weak or undetectable. Furthermore, intraspecific variation in both structural and physiological traits leads to differences in plant size and fitness. These results demonstrate the importance of considering measurements of multivariate phenotypes on individual plants when evaluating trait relationships and how trait variation influences predictions of ecological and evolutionary outcomes.     

Tolerance to oxidative stress is associated with both oxidative stress response and inherent growth in a fungal wheat pathogen

Reactive oxygen species are toxic byproducts of aerobic respiration that are also important in mediating a diversity of cellular functions. Reactive oxygen species form an important component of plant defenses to inhibit microbial pathogens during pathogen–plant interactions. Tolerance to oxidative stress is likely to make a significant contribution to the viability and pathogenicity of plant pathogens, but the complex network of oxidative stress responses hinders identification of the genes contributing to this trait. Here, we employed a forward genetic approach to investigate the genetic architecture of oxidative stress tolerance in the fungal wheat pathogen Zymoseptoria tritici. We used quantitative trait locus (QTL) mapping of growth and melanization under axenic conditions in two cross-populations to identify genomic regions associated with tolerance to oxidative stress. We found that QTLs associated with growth under oxidative stress as well as inherent growth can affect oxidative stress tolerance, and we identified two uncharacterized genes in a major QTL associated with this trait. Our data suggest that melanization does not affect tolerance to oxidative stress, which differs from what was found for animal pathogens. This study provides a whole-genome perspective on the genetic basis of oxidative stress tolerance in a plant pathogen.     

环丝氨酸联合莫西沙星治疗耐多药肺结核的疗效及对免疫功能和生活质量的影响

摘要 目的：探讨环丝氨酸联合莫西沙星治疗耐多药肺结核（MDR-TB）的疗效及对免疫功能和生活质量的影响。方法：选取2016年7月到2018年10月期间我院收治的MDR-TB患者86例作为研究对象,根据奇偶排序法将患者分为对照组（n=43,基础治疗联合莫西沙星治疗）和研究组（n=43,对照组基础上联合环丝氨酸治疗）,均治疗20个月。对比两组疗效、痰结核菌转阴率、病灶吸收率、空洞缩小率情况、免疫功能、生活质量及不良反应。结果：研究组治疗20个月后的总有效率高于对照组（P<0.05）。研究组治疗12个月后、治疗20个月后痰结核菌转阴率、病灶吸收率、空洞缩小率均高于对照组（P<0.05）。两组治疗20个月后心理功能、躯体功能、社会功能、物质生活评分均升高,且研究组高于对照组（P<0.05）。两组治疗20个月后CD8⁺较治疗前降低,且研究组低于对照组（P<0.05）,CD3⁺、CD4⁺/CD8⁺、CD4⁺较治疗前升高,且研究组高于对照组（P<0.05）。对比两组不良反应发生率无差异（P>0.05）。结论：环丝氨酸联合莫西沙星治疗MDR-TB,能够有效的促进患者机体免疫功能和生活质量的改善,并能够有效的促进临床转归,且用药安全性较高。     

肺癌根治术后并发症危险因素分析及其生活质量变化研究

摘要 目的：探讨肺癌根治术后并发症的危险因素,并分析其生活质量的变化。方法：纳入我院2018年9月～2020年7月收治的行肺癌根治术患者104例,对患者的临床资料进行回顾性分析。根据患者术后3个月的并发症发生情况,分成并发症组（n=32）和非并发症组（n=72）,分析肺癌根治术后并发症发生的影响因素,利用癌症病人生活质量测定量表（QLQ-C30）评估患者术前及术后3个月的生活质量。结果：肺癌根治术后32例出现并发症,其中切口感染6例,肺炎7例,肺不张9例,心律失常6例,脓胸4例。并发症组年龄≥60岁、烟龄≥10年、传统开胸术、慢性阻塞性肺疾病史人数占比高于非并发症组（P＜0.05）。多因素Logistic回归分析显示,年龄≥60岁（OR=2.978,95%CI：1.415-6.267）、烟龄≥10年（OR=3.847,95%CI：1.869-7.918）、传统开胸术（OR=3.065,95%CI：1.544-6.084）、慢性阻塞性肺疾病史（OR=2.848,95%CI：1.481-5.477）是肺癌根治术后患者发生并发症的危险因素（P＜0.05）。非并发症组术后3个月的总体生活质量量表、角色功能、躯体功能、情绪功能、社会功能评分高于术前与并发症组,且恶心呕吐、疲乏、疼痛、便秘、呼吸困难、食欲下降、睡眠障碍、腹泻评分较术前与并发症组明显降低（P＜0.05）。结论：肺癌根治术后并发症的发生主要与患者年龄、烟龄、手术方式、慢性阻塞性肺疾病史有关,且并发症对患者术后生活质量影响较大。