Plant-derived pharmaceuticals for the developing world

Plant-produced vaccines and therapeutic agents offer enormous potential for providing relief to developing countries by reducing the incidence of infant mortality caused by infectious diseases. Vaccines derived from plants have been demonstrated to effectively elicit an immune response. Biopharmaceuticals produced in plants are inexpensive to produce, require fewer expensive purification steps, and can be stored at ambient temperatures for prolonged periods of time. As a result, plant-produced biopharmaceuticals have the potential to be more accessible to the rural poor. This review describes current progress with respect to plant-produced biopharmaceuticals, with a particular emphasis on those that target developing countries. Specific emphasis is given to recent research on the production of plant-produced vaccines toward human immunodeficiency virus, malaria, tuberculosis, hepatitis B virus, Ebola virus, human papillomavirus, rabies virus and common diarrheal diseases. Production platforms used to express vaccines in plants, including nuclear and chloroplast transformation, and the use of viral expression vectors, are described in this review. The review concludes by outlining the next steps for plant-produced vaccines to achieve their goal of providing safe, efficacious and inexpensive vaccines to the developing world.     

我国网络平台本土兰科植物贸易状况

我国兰科植物物种丰富, 很多具有极高的药用和观赏价值。随着网络平台的迅速发展, 兰科植物的线上交易越来越普遍。为了解国内网络平台的本土兰科植物贸易情况, 本文利用PyCharm Community软件爬取网络交易信息和人工浏览收集的方法, 分析了淘宝网、拼多多和中兰网的兰科植物贸易情况。调查发现: 网络贸易的本土兰科植物共计84属339种(含3变种), 包含45个中国特有种, 兰属(Cymbidium)和石斛属(Dendrobium)是最大的贸易对象。其中, 仅以观赏用途作为卖点的兰科植物309种(91.1%), 仅以药用价值作为卖点的兰科植物5种(1.5%), 以双用途作为卖点的兰科植物25种(7.4%)。本次调查共记录到336种(99.1%)野生兰科植物销售信息, 并发现部分销售者刻意包装野生兰科植物的现象。大部分淘宝网和拼多多的兰科植物最大销售量对应的价位为10-49元, 其中拼多多的价格更低且集中, 位于15-35元的占57.5%。商品货源地较集中, 主要包括云南省、广东省、广西壮族自治区、浙江省、福建省和四川省。进一步对本次记录的兰科植物进行濒危等级统计后发现: 受威胁兰科植物188种(55.5%), 其中极危(Critically Endangered, CR) 23种(6.8%), 濒危(Endangered, EN) 85种(25.1%), 易危(Vulnerable, VU) 80种(23.6%)。为加强我国野生兰科植物多样性保护, 强烈建议各网络平台依据最新《国家重点保护野生植物名录》制定兰科植物商品信息发布规则, 重点监督货源地集中的地区, 同时加强对公众的宣传教育, 相关部门应利用网络平台加强兰科资源的信息交流, 共同保护野生兰科植物。     

Foraminifers of the Viséan–Serpukhovian boundary interval in Western Palaeotethys: a review

A detailed revision of foraminiferal zonal schemes in sections throughout Europe and North Africa for the Viséan–Serpukhovian boundary interval suggests that several foraminiferal taxa might have the potential to form reliable markers throughout the Palaeotethys. This would support the currently investigated boundary definition based on the First Appearance Datum of the conodont Lochriea ziegleri. However, correlation of these foraminiferal markers in the Western Palaeotethys region has encountered several problems, partly arising from taxonomic issues, but mainly because of apparent discrepancies between the First Occurrence Data (FOD). Analysis of the available foraminiferal data has revealed that some taxa show marked delays in their FODs, due to the timing of westward dispersal within the Palaeotethys, emanating from a probable source in eastern Russia. As a result of this investigation, two dispersal routes have been identified, a northern branch and a southern branch. In general, the displacements within the southern branch occurred more rapidly than in the northern branch. In addition to different dispersal routes, separation of the main foraminiferal markers in stratigraphical sections from different regions can result from isolation of shallow‐water facies of the inner platform from those of relatively deeper‐water settings in the outer platform, the latter showing more consistent foraminiferal FODs. The differences in palaeobathymetry and associated energy levels have enabled two foraminiferal zonal schemes to be distinguished for the Viséan–Serpukhovian boundary interval in the Western Palaeotethys, one for the inner platform and a second one for the outer platform.     

Observation of a porbeagle shark Lamna nasus aggregation at a North Sea oil platform

A minimum of 20 porbeagles Lamna nasus were observed circling the Alba oil platform (UK North Sea) over several days in July 2014. Although schools of fish often aggregate around oil platforms, less is known about their ability to aggregate pelagic sharks and L. nasus are rarely seen at the surface. This unusual observation provides new insights into L. nasus behaviour and habitat use and the potential role of offshore artificial structures on pelagic predators.     

Electrochromatographic characterization of methacrylate ester-based monolith and capillary electrochromatography separation of flavonoids

A porous polymethacrylate ester-based monolithic column for capillary electrochromatography (CEC) was designed by mean of in situ co-polymerizing lauryl methacrylate (LMA), ethylene dimethacrylate (EDMA) and 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) in a ternary porogenic solvent including cyclohexanol, 1,4-butanediol and water. After investigating the influence factors of the CEC monolithic columns, four flavonoids (i.e., Rutin, Quercetin, Kaempferol, and Quercitrin) were separated and assayed to evaluate this monolithic column with CEC method. Under optimum conditions, the CEC method exhibited high separation efficiency, with rapid separation time of 3–4 min, for the four flavonoid samples using 10 mM phosphate buffer containing 70% acetonitrile (pH 9.0). Importantly, the proposed method could provide a promising approach for rapid separation and detection in biomedicine.     

A simple and sensitive HPLC fluorescence method for determination of tadalafil in mouse plasma

A simple and sensitive high-performance liquid chromatographic (HPLC) method utilizing fluorescence detection was developed for the determination of the phosphodiesterase type 5 inhibitor tadalafil in mouse plasma. This method utilizes a simple sample preparation (protein precipitation) with high recovery of tadalafil (∼98%), which eliminates the need for an internal standard. For constituent separation, the method utilized a monolithic C₁₈ column and a flow rate of 1.0 mL/min with a mobile phase gradient consisting of aqueous trifluoroacetic acid (0.1% TFA in deionized water pH 2.2, v/v) and acetonitrile. The method calibration was linear for tadalafil in mouse plasma from 100 to 2000 ng/mL (r > 0.999) with a detection limit of approximately 40 ng/mL. Component fluorescence detection was achieved using an excitation wavelength of 275 nm with monitoring of the emission wavelength at 335 nm. The intra-day and inter-day precision (relative standard deviation, RSD) values for tadalafil in mouse plasma were less than 14%, and the accuracy (percent error) was within −14% of the nominal concentration. The method was utilized on mouse plasma samples from research evaluating the potential cardioprotective effects of tadalafil on mouse heart tissue exposed to doxorubicin, a chemotherapeutic drug with reported cardiotoxic effects.     

Chromatographic detection of residual cellular DNA on short monolithic columns

Analysis of crude samples from biotechnological processes is often required to demonstrate that residual host cell impurities are reduced or eliminated during purification. Current knowledge suggests that a continuous-cell-line DNA can be considered a cellular contaminant rather than a significant risk factor requiring removal to extremely low levels. Anion-exchange chromatography is one of the most important methods used in the downstream processing and analysis of different biomolecules. In this article, an application using Convective Interaction Media monolithic columns to improve the detection of residual cellular DNA is described.     

Two-dimensional HPLC on-line analysis of phosphopeptides using titania and monolithic columns

Conventional and comprehensive two-dimensional (2D) HPLC systems using the combination of titania and monolithic columns were established for the on-line analysis of phosphopeptides. Compared with immobilized metal affinity chromatography of a general method for the analysis of phosphopeptides, the use of titania columns in the analysis permits the specific isolation of phosphopeptides in a higher yield. Using the current 2D HPLC systems, phosphopeptides were specifically isolated from nonphosphorylated peptides by the first-dimension titania column, followed by the high-speed separation of the phosphopeptides by the second-dimension monolithic column. Proteolytic digests of beta-casein were analyzed within 30 min using the comprehensive 2D HPLC system; all phosphopeptides from beta-casein could be efficiently isolated and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The comprehensive 2D HPLC system coupled with mass spectrometry will be useful for high-throughput and on-line phosphoproteome analyses.     

In situ pH maintenance for mammalian cell cultures in shake flasks and tissue culture flasks

pH in animal cell cultures decreases due to production of metabolites like lactate. pH control via measurement and base addition is not easily possible in small‐scale culture formats like tissue‐culture flasks and shake flasks. A hydrogel‐based system is reported for in situ pH maintenance without pH measurement in such formats, and is demonstrated to maintain pH between 6.8 and 7.2 for a suspension CHO cell line in CD CHO medium and between 7.3 and 7.5 for adherent A549 cells in DMEM:F12 containing 10% FBS. This system for pH maintenance, along with our previous report of hydrogels for controlled nutrient delivery in shake flasks can allow shake flasks to better mimic bioreactor‐based fed batch operation for initial screening during cell line and process development for recombinant protein production in mammalian cells. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012     

Clinical implications of microRNAs in cancer

Abstract

MicroRNAs (miRNAs) are endogenously produced non-coding RNAs that serve as micromanagers by negatively regulating gene expression. MiRNAs are implicated in several biological pathways including development of neoplasia. Because altered miRNA expression is implicated in the pathobiology of various cancers, these molecules serve as potential therapeutic targets. Using miRNA mimics to restore levels of aberrantly down-regulated miRNAs or miRNA inhibitors to inactivate over-expressed miRNAs shows promise as the next generation of therapeutic strategies. Manipulation of miRNAs offers an alternative therapeutic approach for chemo- and radiation-resistant tumors. Similarly, miRNA expression patterns can be used for diagnosis and to predict prognosis and efficacy of therapy. We present here an overview of how miRNAs affect cancers, how they may be used as biomarkers, and the clinical implications of miRNAs in cancer.