Protodyne : An immunostimulatory protein component, prepared from gram-positive bacillus subtilis

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Зміна агрохімічних показників сірих лісових ґрунтів лівобережного лісостепу під впливом лісових екосистем

Purpose. The study of changes in the main agrochemical parameters in the upper layers of the gray forest soils of the Left Bank Forest-Steppe of Ukraine that were removed from agricultural processing and forested with pine, in different years. Methods. Theoretical methods included the collection and description of facts, their analysis. Empirical methods involved conducting field research on test plots of the state-owned enterprise “Chuguevo-Babchansky LG” and farm near woodlands. Laboratory and analytical studies were performed using standardized measurement methods. The generalization of the experimental data was performed using application software packages. Results. Studies were carried out on soils under natural forest plantations, soils in intensive agricultural processing and unproductive soils that are derived from agricultural use. In all the studied soils, the main agrochemical parameters were compared among themselves: mobile forms of nitrogen, pH, humus content, content of common forms of nitrogen, phosphorus and potassium. Statistical dependences between agrochemical parameters in the studied variants were established. Conclusions. The results obtained indicate the revitalization and development of humification and accumulation of nutrients in old arable gray forest soils with an increase in the duration of exposure to pine forest. The age stages of the forest determine the specifics of the environmental factors that influence the soil-forming processes. In the soil of a young pine-tree (12 years), there are more cases of an increase in the coefficients of spatial variation. This indicates that the formation of young ecosystems are characterized by low resistance, in these biocenoses the dynamic equilibrium is not reached, which is characteristic of more mature pine forests. Цель. Изучение изменений основных агрохимических показателей в верхних слоях серых лесных почв Левобережной Лесостепи Украины выведенных из сельхозобработки и облесённые сосной обыкновенной в разные годы. Методы. Теоретические методы включали сбор и описание фактов, их анализ. Эмпирические методы предусматривали проведение полевых исследований на пробных площадях ГП «Чугуево-Бабчанский ЛГ» и фермерского хозяйства вблизи лесных массивов. Лабораторно-аналитические исследования проводили с помощью стандартизированных методов выполнения измерений. Обобщение экспериментальных данных выполняли с помощью прикладных программных пакетов. Результаты. Исследования проводились на почвах под естественными лесными насаждениями, почвах в интенсивной сельскохозяйственной обработке и малопродуктивных почвах, которые выведены из сельскохозяйственного использования. Во всех исследуемых почвах сравнивались между собой основные агрохимические показатели: подвижные формы азота, рН, содержание гумуса, содержание общих форм азота, фосфора и калия. Проанализировано статистические зависимости между агрохимическими параметрами в исследуемых вариантах. Выводы. Полученные результаты свидетельствуют о восстановлении и развитии в старопахотных серых лесных почвах процессов гумификации и аккумуляции биогенных элементов при увеличении продолжительности воздействия на них соснового леса. Возрастные стадии леса определяют специфику экологических факторов, влияющих на почвообразующие процессы. В почве молодого сосняка (12 лет) отмечается больше случаев увеличения коэффициентов пространственных вариаций. Это свидетельствует о том, что формирование молодых экосистем характеризуются низкой устойчивостью, в этих биоценозах не достигнуто динамического равновесия, которая свойственна более зрелым соснякам.Мета. Вивчення змін основних агрохімічних показників в верхніх шарах сірих лісових ґрунтів Лівобережного Лісостепу України виведених з сільськогосподарського використання і заліснених сосною звичайною в різні роки. Методи. Теоретичні методи включали збір та опис фактів, їх аналіз. Емпіричні методи передбачали проведення польових досліджень на пробних площах ДП «Чугуєво-Бабчанський ЛГ» та фермерського господарства поблизу лісових масивів. Лабораторно-аналітичні дослідження проводили згідно стандартизованих методів виконання вимірювань. Узагальнення експериментальних даних виконували за допомогою прикладних програмних пакетів. Результати. Дослідження проводились на ґрунтах під природними лісовими насадженнями, ґрунтах в інтенсивному сільськогосподарському обробітку та малопродуктивних ґрунтах, які виведені з с.-г. використання. У всіх досліджуваних ґрунтах порівнювались між собою основні агрохімічні показники: вміст мінерального азоту, рН сольовий, вміст гумусу, вміст загальних форм азоту, фосфору і калію. Проаналізовано статистичні залежності між агрохімічними параметрами в досліджуваних ґрунтах. Висновки. Отримані результати свідчать про відновлення і розвиток в староорних сірих лісових ґрунтах процесів гуміфікації та акумуляції біогенних елементів при збільшенні тривалості впливу на них соснового лісу. Вікові стадії лісу визначають специфіку екологічних факторів, що впливають на ґрунтоутворювальні процеси. В ґрунті наймолодшого сосняку (12 років) відзначається більше випадків зі збільшенням коефіцієнтів просторових варіацій. Це свідчить про те, що формування молодих екосистем характеризуються найнижчою стійкістю, в цих біоценозах не досягнуто динамічної рівноваги, яка властива більш зрілим соснякам

Field Production and Functional Evaluation of Chloroplast-Derived Interferon-α2b

Type I interferons (IFNs) inhibit viral replication and cell growth and enhance the immune response, and therefore have many clinical applications. IFN-α2b ranks third in world market use for a biopharmaceutical, behind only insulin and erythropoietin. The average annual cost of IFN-α2b for the treatment of hepatitis C infection is $26 000, and is therefore unavailable to the majority of patients in developing countries. Therefore, we expressed IFN-α2b in tobacco chloroplasts, and transgenic lines were grown in the field after obtaining United States Department of Agriculture Animal and Plant Health Inspection Service (USDA-APHIS) approval. Stable, site-specific integration of transgenes into chloroplast genomes and homoplasmy through several generations were confirmed. IFN-α2b levels reached up to 20% of total soluble protein, or 3 mg per gram of leaf (fresh weight). Transgenic IFN-α2b had similar in vitrobiological activity to commercially produced PEG-Intron™ when tested for its ability to protect cells against cytopathic viral replication in the vesicular stomatitis virus cytopathic effect (VSV CPE) assay and to inhibit early-stage human immunodeficiency virus (HIV) infection. The antitumour and immunomodulating properties of IFN-α2b were also seen in vivo . Chloroplast-derived IFN-α2b increased the expression of major histocompatibility complex class I (MHC I) on splenocytes and the total number of natural killer (NK) cells. Finally, IFN-α2b purified from chloroplast transgenic lines (cpIFN-α2b) protected mice from a highly metastatic tumour line. This demonstration of high levels of expression of IFN-α2b, transgene containment and biological activity akin to that of commercial preparations of IFN-α2b facilitated the first field production of a plant-derived human blood protein, a critical step towards human clinical trials and commercialization

Detection and Identification of the Atypical Bovine Pestiviruses in Commercial Foetal Bovine Serum Batches

The recently emerging atypical bovine pestiviruses have been detected in commercial foetal bovine serum (FBS) of mainly South American origin so far. It is unclear how widely the viruses are presented in commercial FBS of different geographic origins. To further investigate the possible pestivirus contamination of commercially available FBS batches, 33 batches of FBS were obtained from ten suppliers and analysed in this study for the presence of both the recognised and the atypical bovine pestiviruses. All 33 batches of FBS were positive by real-time RT-PCR assays for at least one species of bovine pestiviruses. According to the certificate of analysis that the suppliers claimed for each batch of FBS, BVDV-1 was detected in all 11 countries and BVDV-2 was detected exclusively in the America Continent. The atypical pestiviruses were detected in 13 batches claimed to originate from five countries. Analysis of partial 5′UTR sequences showed a high similarity among these atypical bovine pestiviruses. This study has demonstrated, for the first time that commercial FBS batches of different geographic origins are contaminated not only with the recognised species BVDV-1 and BVDV-2, but also with the emerging atypical bovine pestiviruses

A novel asymmetric 3D in-vitro assay for the study of tumor cell invasion

<p>Abstract</p> <p>Background</p> <p>The induction of tumor cell invasion is an important step in tumor progression. Due to the cost and slowness of <it>in-vivo </it>invasion assays, there is need for quantitative <it>in-vitro </it>invasion assays that mimic as closely as possible the tumor environment and in which conditions can be rigorously controlled.</p> <p>Methods</p> <p>We have established a novel asymmetric 3D in-vitro invasion assay by embedding a monolayer of tumor cells between two layers of collagen. The cells were then allowed to invade the upper and lower layers of collagen. To visualize invading cells the gels were sectioned perpendicular to the monolayer so that after seeding the monolayer appears as a thin line precisely defining the origin of invasion. The number of invading tumor cells, their proliferation rate, the distance they traverse and the direction of invasion could then be determined quantitatively.</p> <p>Results</p> <p>The assay was used to compare the invasive properties of several tumor cell types and the results compare well with those obtained by previously described assays. Lysyl-oxidase like protein-2 (Loxl2) is a potent inducer of invasiveness. Using our assay we show for the first time that inhibition of endogenous Loxl2 expression in several types of tumor cells strongly inhibits their invasiveness. We also took advantage of the asymmetric nature of the assay in order to show that fibronectin enhances the invasiveness of breast cancer cells more potently than laminin. The asymmetric properties of the assay were also used to demonstrate that soluble factors derived from fibroblasts can preferentially attract invading breast cancer cells.</p> <p>Conclusion</p> <p>Our assay displays several advantages over previous invasion assays as it is allows the quantitative analysis of directional invasive behavior of tumor cells in a 3D environment mimicking the tumor microenvironment. It should be particularly useful for the study of the effects of components of the tumor microenvironment on tumor cell invasiveness.</p

Free exopolysaccharide from Mycoplasma mycoides subsp. mycoides possesses anti-inflammatory properties

In this study we explored the immunomodulatory properties of highly purified free galactan, the soluble exopolysaccharide secreted by Mycoplasma mycoides subsp. mycoides (Mmm). Galactan was shown to bind to TLR2 but not TLR4 using HEK293 reporter cells and to induce the production of the anti-inflammatory cytokine IL-10 in bovine macrophages, whereas low IL-12p40 and no TNF-α, both pro-inflammatory cytokines, were induced in these cells. In addition, pre-treatment of macrophages with galactan substantially reduced lipopolysaccharide (LPS)-induced production of pro-inflammatory cytokines TNF- and IL-12p40 while increasing LPS-induced secretion of immunosuppressive IL-10. Also, galactan did not activate naïve lymphocytes and induced only low production of the Th1 cytokine IFN-γ in Mmm-experienced lymphocytes. Finally, galactan triggered weak recall proliferation of CD4+ T lymphocytes from contagious bovine pleuropneumonia-infected animals despite having a positive effect on the expression of co-stimulatory molecules on macrophages. All together, these results suggest that galactan possesses anti-inflammatory properties and potentially provides Mmm with a mechanism to evade host innate and adaptive cell-mediated immune responses. (Résumé d'auteur