Detection and quantitation of forty eight cytokines, chemokines, growth factors and nine acute phase proteins in healthy human plasma, saliva and urine

Cytokines, chemokines, growth factors (CCGFs) and other low abundance proteins/peptides in human body fluids or in tissues are potential biomarkers. Human body fluids such as plasma, saliva, urine, etc. are being analyzed more frequently than tissues primarily because of ease of sample collection. However, available information on concentrations of a large number of CCGFs in various body fluids of the same healthy individuals and gender-specific CCGFs is limited. In this work concentrations of 48 CCGFs were measured using multiplex bead assays and compared between plasma, saliva and urine collected from 20 male and female healthy volunteers. Forty three CCGFs were detected at least in one sample type of which 37 were in plasma, 41 were in saliva, and 34 were in urine; five CCGFs were not detected in any sample. Concentrations of detected CCGFs differed significantly between sample types but similar between gender groups. Gender-specific CCGFs were also observed. Concentrations of nine acute phase proteins were also measured from plasma, saliva and urine to determine general health conditions of the volunteers. This work will provide an idea of which CCGFs are detectable and their relative concentrations in healthy human plasma, saliva and urine and which CCGFs are gender-specific.     

Rapid throughput analysis of filamentous fungal growth using turbidimetric measurements with the Bioscreen C: a tool for screening antifungal compounds

A rapid method for screening antifungal compounds and performing ecophysiological studies with filamentous fungi has been developed by the use of specific semi-solid media and spectrophotometric/turbidimetric measurements using the Bioscreen C with 2×100 microtitre well plates. The medium composition and preparation, inoculum size, medium volumes, and incubation parameters for measuring initial germination and growth dynamics have been optimised. These have been applied to assess the effectiveness of 18 concentrations of propyl propane thiosulfinate (PTS) against Aspergillus flavus in YES medium under different environmental regimes. Minimum inhibitory concentrations (MIC), and non-inhibitory concentration (NIC) values, and a newly developed MIC(50), have been calculated for the efficacy of these concentrations of PTS under four different environmental conditions (0.995 and 0.95 water activity (a(w)); 20 and 25°C) over 7d periods with automated measurements every 20 min. Data were modelled using the Lambert-Pearson Model, a mathematical modelling approach previously used for bacterial inhibition. Results have been compared with traditional growth rate data and Lethal Dose(50) (LD(50)) values. This approach could have major implications for rapid screening assays for growth and secondary metabolite production by filamentous organisms and has major advantages in terms of time reduction, culture medium volumes required, measurement and the ability to integrate and model key parameters for comparing efficacy.     

Response of preosteoblasts to thermal stress conditioning and osteoinductive growth factors

Conditioning protocols involving mechanical stress independently or with chemical cues such as growth factors (GFs) possess significant potential to enhance bone regeneration. However, utilization of thermal stress conditioning alone or with GFs for bone therapy has been under-investigated. In this study, a preosteoblast cell line (MC3T3-E1) was exposed to treatment with water bath heating (44°C, 4 and 8 min) and osteoinductive GFs (bone morphogenetic protein-2 and transforming growth factor-β1) individually or in combination to investigate whether these stimuli could promote induction of bone-related markers, an angiogenic factor, and heat shock proteins (HSPs). Cells remained viable when heating durations were less than 20 min at 40oC, 16 min at 42oC, and 10 min at 44oC. Increasing heating duration at 44°C, promoted gene expression of HSPs, osteocalcin (OCN), and osteopontin (OPN) at 8 h post-heating (PH). Heating in combination with GFs caused the greatest gene induction of osteoprotegerin (OPG; 6.9- and 1.6-fold induction compared to sham-treated and GF only treated groups, respectively) and vascular endothelial growth factor (VEGF; 16.0- and 1.6-fold compared to sham and GF-only treated groups, respectively) at 8 h PH. Both heating and GFs independently suppressed the matrix metalloproteinase-9 (MMP-9) gene. GF treatment caused a more significant decrease in MMP-9 protein secretion to non-detectable levels compared to heating alone at 72 h PH. Secretion of OCN, OPN, and OPG increased with the addition of GFs but diminished with heating as measured by ELISA at 72 h PH. These results suggest that conditioning protocols utilizing heating and GFs individually or in combination can induce HSPs, bone-related proteins, and VEGF while also causing downregulation of osteoclastic activity, potentially providing a promising bone therapeutic strategy.     

Dietary administration of zingerone to enhance growth, non-specific immune response, and resistance to Vibrio alginolyticus in Pacific white shrimp (Litopenaeus vannamei) juveniles

Zingerone, one of the active components of ginger, is a phenolic alkanone with antioxidant and anti-inflammatory properties. The effects of zingerone supplementation on the growth, immunity, and disease resistance of Pacific white shrimp (Litopenaeus vannamei) juveniles were studied. Four experimental diets, including a control diet (without zingerone enrichment) and 1, 2.5, and 5 mg zingerone (kg diet)⁻¹ were used. After 56 days of culture, shrimp fed diets supplemented with 1, 2.5, and 5 mg zingerone (kg diet)⁻¹ had significantly greater weight gain and feed efficiency than the controls. Furthermore, after 56 days of culture, shrimp fed all doses of the zingerone diet had higher survival rates compared to the controls after 24-72 h of challenge by the pathogen, Vibrio alginolyticus. Significantly increased phenoloxidase levels were found in shrimp fed the zingerone diets at all doses, and respiratory bursts, lysozyme and phagocytic activities of shrimp fed 2.5 and 5 mg zingerone (kg diet)⁻¹ also significantly increased. Neither the total hemocyte count nor superoxide dismutase activity of the experimental and control groups revealed significant differences at any dose. The results indicate that zingerone can be recommended as a supplement to shrimp feed to increase growth, immunity, and disease resistance against the pathogen, V. alginolyticus. Use of zingerone as appetizer and immunostimulant in shrimp is promising.     

短期饥饿和再投喂对岩原鲤仔鱼生存生长以及RNA/DNA和RNA/蛋白质比率的影响(英文)

研究通过对岩原鲤仔鱼在饥饿和再投喂条件下其生存、生长率、RNA/DNA和RNA/蛋白质比率的测定,评估了仔鱼对饥饿的耐受能力和恢复能力。在(19.5±0.5)℃水温下,将岀膜后第16天的岩原鲤仔鱼随机分成6个组:1个持续投饲对照组,实验组分别禁食1、2、3、4、5d后再投喂,实验共进行10d。每天分别从各组取9尾鱼测定体重、体长、RNA、DNA、蛋白质含量。实验结果显示,饥饿处理组仔鱼存活率和以上各项生长指标均随饥饿时间的增加而下降,在恢复投喂后均表现不同程度的补偿生长,其中饥饿1、2、3d的仔鱼在恢复投喂后显示出完全补偿生长,几乎弥补了饥饿所产生的影响,平均终体重与对照组比较无显著差异。饥饿4、5d的仔鱼显示部分补偿生长,恢复投喂只少量减轻了饥饿的影响,平均终体重与对照组相比存在显著差异。饥饿1、2、3d的仔鱼和4、5d的仔鱼在恢复投喂后分别需要1—2d和4d时间才能达到与对照组无显著差异水平。仔鱼生长率变动范围从0.59%到8.00%WW/day,仔鱼RNA/DNA比率、RNA/蛋白质比率与生长率的回归方程为:GR=3.63RNA/DNA 1.74(R2=0.80)和GR=120.14RNA/Protein 2.33(R2=0.31),两种比率均与生长率呈显著线性相关,RNA/DNA比率对生长变化的拟合度更好。结果表明,仔鱼阶段食物缺乏很可能是影响岩原鲤仔鱼存活、生长的主要因素。RNA/DNA更适合作为评定岩原鲤仔鱼营养条件和生长的指标。     

Advanced photobioreactor LED illumination system: Scale‐down approach to study microalgal growth kinetics

A newly designed and constructed LED illumination device for commercial cylindrical bioreactors is presented for application in microalgal cultivations and investigation of growth kinetics. An ideally illuminated volume is achieved by focusing the light toward the center of the reactor and thereby compensating the mutual shading of the cells. The relevant biomass concentration for homogeneous illumination depending on reactor radius was determined by light distribution measurements for Chlamydomonas to 0.2 g/L (equal 0.435 optical density at 750 nm). It is shown that cultivation experiments with the newly designed illumination device operated in batch mode can be successfully applied for determination of growth rates and photo conversion efficiencies. The exact knowledge of physiological reactions of specific strain(s) and the estimation of relevant parameters for scale‐up can be used for construction of economic pilot plant photobioreactors. The determination of light‐dependent kinetics of growth and product formation is the first necessary step to achieve this. A wide variety of different parameters can be examined like the effect of different illumination conditions (light intensity, frequency of day/night cycles, flashing light, light color…) and thereby for each single application specific, relevant, and interesting parameters will be examined.     

IGF-Ⅰ和 MGF 长期基因治疗对小鼠血糖代谢的影响

为研究IGF-Ⅰ和MGF长期基因治疗对哺乳动物血糖代谢的生理影响,首先构建了包含MGF和IGF-Ⅰ全长编码基因的真核表达载体,然后使用活体基因导入仪将其导入小鼠的左侧股四头肌,每2周一次共导入3次,首次导入15周后进行糖耐量测定实验。实验证实IGF-Ⅰ的长期持续表达会导致小鼠的糖代谢能力降低,血糖最高值可达到12.07±1.35mmol/L比对照(10.15±0.87mmol/L)和MGF组(10.58±0.61mmol/L)有显著性的升高(P<0.05),MGF单独存在情况下在糖代谢方面则没有上述作用,但是IGF-Ⅰ和MGF共同导入的小鼠的血糖调节能力更明显减弱,其最高值(16.30±2.69mmol/L)明显高于其它3组(P<0.001)。因此推测与胰岛素进行拮抗作用的主要是IGF-Ⅰ的N段序列,而产生与糖代谢相关功能的则为暴露的IGF-ⅠC段序列;另一种可能是MGF促进了肌肉细胞对外源DNA的吸收和表达因此其可以作为基因导入的佐剂。