Large-Scale Microcarrier Culture of Chinese Perch Brain Cell for Viral Vaccine Production in a Stirred Bioreactor

Mandarin fish (Siniperca chuatsi) is one of the important cultured fish species in China. Infectious spleen and kidney necrosis virus (ISKNV) and Siniperca Chuatsi rhabdovirus (SCRV) have hindered the development of mandarin fish farming industry. Vaccination is the most effective method for control of viral diseases, however viral vaccine production requires the large-scale culture of cells. Herein, a suspension culture system of Chinese perch brain cell (CPB) was developed on Cytodex 1 microcarrier in a stirred bioreactor. Firstly, CPB cells were cultured using Cytodex 1 microcarrier in 125 mL stirring flasks. With the optimum operational parameters, CPB cells grew well, distributed uniformly, and could fully cover the microcarriers. Then, CPB cells were digested with trypsin and expanded step-by-step with different expansion ratios from the 125 mL stirring bottle to a 500 mL stirring bottle, and finally to a 3-L bioreactor. Results showed that with an expansion ratio of 1:3, we achieved a high cell density level (2.25 × 106 cells/mL) with an efficient use of the microcarriers, which also confirmed the data obtained from the 125 mL stirring flask. Moreover, obvious cytopathic effects (CPE) were observed in the suspended CPB cells post-infection with ISKNV and SCRV. This study provided a large-scale culture system of CPB cells for virus vaccine production.

Book review: Nietzsche’s Culture War: The Unity of the Untimely Meditations

Shilo Brooks’ study on Nietzsche’s Untimely Meditations (UM) (1873-76) is one of the few scholarly works that examines all four of these early essays in combination. Even taken separately, there are fewer independent studies of the UM compared with The Birth of Tragedy (BT) (1872) and the middle works (1878-82), except for “The Use and Disadvantage of History for Life” (HL) (1874), which has garnered the most critical attention. Brooks suggests a compelling reason to investigate all four works together. His organizing principle is reflected in his title: the four essays were individual constituents of a large-scale “culture war”—a philosophical Kulturkampf (p. 12)—that the young Nietzsche waged against Bismarck and the political, social, and cultural conditions in his newly established Reich. Brooks’ decision to treat all four pieces in a single monograph makes eminent sense, and the fact that Nietzsche assembled the essays under the title UM suggests he saw them as part of a common endeavor—or at least as reflective of a certain prevailing mindset at the time: “[W]hen viewed from the perspective of his later works, the critique of German culture featured in the Untimely Meditations and the plan Nietzsche sketches to revitalize it provide a holistic if early blueprint for his later attempt at a revaluation of values” (p. 15).

High Value Phycotoxins From the Dinoflagellate Prorocentrum

Marine dinoflagellates produce chemically diverse compounds, with a wide range of biological activity (antimicrobial, anticancer, treatment of neurodegenerative disease along with use as biomedical research tools). Chemical diversity is highlighted by their production of molecules such as the saxitoxin family of alkaloids (C10H17N7O4 – 299 g/mol) to the amphipathic maitotoxin (C164H256O68S2Na2 – 3,422 g/mol), representing one of the largest and most complex secondary metabolites characterized. Dinoflagellates, are most well-known for the production of red tides which are frequently toxic, including okadaic acid and related dinophysistoxins, which are tumor promoters. The mode of action for these phycotoxins, is by specific inhibition of protein phosphatases, enzymes essential in regulation of many cellular processes. Hence, these compounds are being used for vital cell regulation studies. However, the availability of useful amounts of these compounds has restricted research. Chemical synthesis of some compounds such as okadaic acid has been investigated, but the complexity of the molecule resulted in many lengthy steps and achieved only a poor yield. The use of naturally occurring phytoplankton has been investigated as a potential source of these compounds, but it has been shown to be unreliable and impractical. The most practical option is large scale culture with down-stream processing/purification which requires specialist facilities and expertise. This review, describes the biotechnological potential of these organisms and the challenges to achieve useful yields of high quality phycotoxins using Prorocentrum spp. as an example to produce okadaic acid.

Large-scale culture as a complementary and practical method for discovering natural products with novel skeletons

Large-scale culture is a complementary and practical method for genome mining and OSMAC approaches to discover natural products of unprecedented skeletal classes from fungi.

Self-Organizing 3D Human Choroid Plexus-Ventricle-Cortical Organoids

ABSTRACTBoth the choroid plexus (CP) and the cortex are derived from the rostral neural tube during early embryonic development. In addition to producing CSF, the CP secretes essential factors that orchestrate cortical development and later neurogenesis. Previous brain modeling efforts with human pluripotent stem cells (hPSCs) generated either cortical or CP tissues in 3D culture. Here, we used hPSC-derived neuroectodermal cells, the building blocks of the anterior body, to simultaneously generate CP that forms ventricles and cortical cells in organoids (CVCOs), which can be maintained as 3D organoid cultures. Large scale culture revealed reproducibility of the protocol independent of cell lines, clones or batches. CVCOs contain mature and functional CP that projects multiple cilia into the ventricle-like fluid filled cysts and is in direct contact with appropriately patterned cortical cells. CVCOs thus recapitulate key features of developing forebrain structures observed in in vivo and constitute a useful for dissecting the role of CP in human forebrain development in health and disease.

Applying a New Feeding Protocol for Enhancing Mass Culture and Nutritional Value of the Rotifer Brachionus plicatilis Müller, 1786

The study tested a new feeding protocol to find out the best program using cheap and available products to enhancement the mass culture and the nutritional value of B. plicatilis. The feeding regime was six artificial treatments formulated from dried yeast (Saccharomyces cervisiae) with sucrose sugar and yeast with molasses in different concentrations. Additionally, a live Cyclotella sp. had been applied as supplementary food to each treatment and as independent treatment (control). B. plicatilis was cultured in small scale to find the best food regime and technique to apply it in the large one. B. plicatilis attained its highest density in the small scale culture (370 Ind.ml1 ) with T1 (30 % yeast, 70 % sugar and Cyclotella sp.) at the 12th day, while the highest population growth rate (PGR) (0.65) was calculated at the 9th day. According to the analysis of variance (ANOVA) and PGR, T1 had been applied in the large scale culture. B. plicatilis samples were composed of 51.6% total protein with 16 amino acids and 33.01% total lipid with 19 identified fatty acids. The study concluded that the treatment (T1) is a suitable diet for enrichment the mass culture and the nutritional value of B. plicatilis.

Does feeding on pollen grains affect the performance of Amblyseius swirskii (Acari: Phytoseiidae) during subsequent generations?

Diet is a critical component of the mass-rearing of biological control agents, but the impacts of diet are not always immediately obvious and can take several generations to manifest, resulting in poor survival, reproduction, and ability to kill prey under natural conditions. Our present study aimed to investigate the performance of a commercially-reared phytoseiid mite, Amblyseius swirskii, after four (G4) and six (G6) consecutive generations on pollen grains of two plant species, as well as its ability to find and kill its natural prey, Tetranychus urticae, after long-term rearing on each diet. We found no significant difference between the two diets in intrinsic and finite rates in G4. However, both diet and generation exerted a significant influence on the fecundity of A. swirskii. By G6, females reared on almond pollen had greater net reproductive and intrinsic rate compared to those reared on maize pollen. Conversely, A. swirskii fed on maize pollen consumed fewer prey than those reared on other diets, especially at higher prey densities. The findings have important implications for developing the mass-rearing program of A. swirskii on non-prey diets. Further research must explore the suitability of almond pollen in the large-scale culture of A. swirskii.

In Defense of Erasmus’ Critics

The article confirms Andrew J. Brown’s thesis that despite carrying colophons with dates in the first decade of the sixteenth century, the four sumptuous manuscripts of Erasmus’ translation of the New Testament produced by the scribe Pieter Meghen could not have been finished until the 1520s and in fact preserve a version of Erasmus’ translation not available until the 1520s. Nonetheless, the article goes on to prove that Erasmus was working on a translation of the New Testament already at the time of the colophons in the Meghen manuscripts. Erasmus’ translation was part of his large-scale culture war against medieval scholasticism that he embarked upon at the end of the fifteenth century and continued until his death in 1536. The world around Erasmus changed radically, however, in those four decades, in significant measure because of his own scholarship and writings, but Erasmus himself changed amazingly little in his basic attitudes. The result was that by the end his critics from the Protestant as much as from the Catholic side were rightly frustrated by his incoherent reaction to the changed situation. Emblematic of his inability to face up to the transformed reality is his annotation to I Timothy1:6, which became an unconscious parody of his incoherent stance on religious doctrine and which is translated in an appendix to the article.

Secondary Metabolites of Aeromonas veronii Strain A134 Isolated from a Microcystis aeruginosa Bloom

Aeromonas veronii strain A134 was isolated from Microcystis aeruginosa colonies collected from Lake Kinneret (Sea of Galilee), Israel. The Aeromonas culture media inhibited the growth of M. aeruginosa (strain MGK). The crude extract of a large-scale culture of A. veronii A134 was separated in a few chromatographic steps to yield three new secondary metabolites, 9-chlorolumichrome (1), veronimide (2) and veronipyrazine (3), along with a known lumichrome and several known diketopiperazines. The structures of the new compounds were established by analyses of the data from 1D and 2D NMR experiments and HRMS data of the compounds, as well as a single-crystal X-ray analysis of synthetic 1. The structure elucidation and proposed biogenesis of the new compounds are described below.

Coralmycin Derivatives with Potent Anti-Gram Negative Activity Produced by the Myxobacteria Corallococcus coralloides M23

Seven new coralmycin derivatives, coralmycins C (1), D (2), E (3), F (4), G (5), H (6), and I (7), along with three known compounds, cystobactamids 891-2 (8), 905-2 (9), and 507 (10), were isolated from a large-scale culture of the myxobacteria Corallococcus coralloides M23. The structures of these compounds, including their relative stereochemistries, were elucidated by interpretation of their spectroscopic and CD data. The structure-activity relationships of their antibacterial and DNA gyrase inhibitory activities indicated that the para-nitrobenzoic acid unit is critical for the inhibition of DNA gyrase and bacterial growth, while the nitro moiety of the para-nitrobenzoic acid unit and the isopropyl chain at C-4 could be important for permeability into certain Gram-negative bacteria, including Pseudomonas aeruginosa and Klebsiella pneumoniae, and the β-methoxyasparagine moiety could affect cellular uptake into all tested bacteria. These results could facilitate the chemical optimization of coralmycins for the treatment of multidrug-resistant Gram-negative bacteria.