Integrated Transcriptome and Proteome Analyses Reveal Protein Metabolism in Lactobacillus helveticus CICC22171

Lactobacillus helveticus is a homofermentative lactic acid bacterium. It is widely used in the fabrication of Swiss cheese and other dairy products. The aim of this study was to elucidate the mechanism by which L. helveticus utilizes protein. Lactobacillus helveticus CICC22171 were cultured in two different media with various nitrogen sources. The control contained 20 basic amino acids, while the experimental medium contained casein. De novo transcriptome and isobaric tags for relative and absolute quantification (iTRAQ) proteome analyses were applied to determine how L. helveticus utilizes protein. The casein underwent extracellular hydrolysis via ATP-binding cassette (ABC) transporter upregulation and Mn2+-associated cell envelope proteinase (CEP) downregulation. Sigma factors and EF-Tu were upregulated and Mg2+ was reduced in bacteria to accommodate DNA transcription and protein translation in preparation for proteolysis. Hydrolase activity was upregulated to digest intracellular polypeptides and control endopeptidase genes. In these bacteria, casein utilization affected glycolysis, trehalose phosphotransferase system (PTS), and key factors associated with aerobic respiration and reduced glucose consumption.

Integrins Specifically Associated with the Activated Phagocytic Receptor Complexes from Human U937 Macrophages and the Integrins of Human Blood

Receptor complexes are a key locus in the treatment of disease and pain but remain difficult to isolate. The Fc receptor binds to IgG presented on micro particles and triggers phagocytosis that is required for the engulfment of bacteria and non-self, pathogen molecules. Circulating LDL may be converted to oxLDL that is recognized by scavenger or other innate receptors that trigger the production of free radicals and phagocytic engulfment. The Fc receptor complex was captured using its cognate ligand IgG bound to 2 micron polystyrene beads from live cells by live cell affinity receptor chromatography (LARC). Similarly, oxLDL was used to coat micro beads to isolate the oxLDL-scavenger receptor complexes. The ligand coated beads were presented to live human macrophage U937 cells and sampled over the course of cell binding and engulfment in saline experimental medium. The receptor complexes were isolated by disruption with a French press and sucrose gradient centrifugation. The receptor complexes were separated using a salt and acetonitrile gradient prior to digestion with trypsin. The peptides were analyzed by LC-ESI MS/MS using a linear ion trap (Thermo) with the SEQUEST algorithm. In addition ligand affinity chromatography was performed using IgG, oxLDL or anti CD36 microbeads incubated with crude extracts. Controls included uncoated, or ligand coated, beads incubated in crude extract and/or used experimental medium. Certain integrins were shown to be specifically associated with IgG versus oxLDL and/or control treatments. The functional requirement for these integrins was tested using silencing RNA and quantitative assays of phagocytosis using laser scanning confocal microscopy. For the first time we show a small but statistically significant role for integrins in IgG-Fcγ receptor mediated phagocytosis.

Integrins Specifically Associated with the Activated Phagocytic Receptor Complexes from Human U937 Macrophages and the Integrins of Human Blood

Receptor complexes are a key locus in the treatment of disease and pain but remain difficult to isolate. The Fc receptor binds to IgG presented on micro particles and triggers phagocytosis that is required for the engulfment of bacteria and non-self, pathogen molecules. Circulating LDL may be converted to oxLDL that is recognized by scavenger or other innate receptors that trigger the production of free radicals and phagocytic engulfment. The Fc receptor complex was captured using its cognate ligand IgG bound to 2 micron polystyrene beads from live cells by live cell affinity receptor chromatography (LARC). Similarly, oxLDL was used to coat micro beads to isolate the oxLDL-scavenger receptor complexes. The ligand coated beads were presented to live human macrophage U937 cells and sampled over the course of cell binding and engulfment in saline experimental medium. The receptor complexes were isolated by disruption with a French press and sucrose gradient centrifugation. The receptor complexes were separated using a salt and acetonitrile gradient prior to digestion with trypsin. The peptides were analyzed by LC-ESI MS/MS using a linear ion trap (Thermo) with the SEQUEST algorithm. In addition ligand affinity chromatography was performed using IgG, oxLDL or anti CD36 microbeads incubated with crude extracts. Controls included uncoated, or ligand coated, beads incubated in crude extract and/or used experimental medium. Certain integrins were shown to be specifically associated with IgG versus oxLDL and/or control treatments. The functional requirement for these integrins was tested using silencing RNA and quantitative assays of phagocytosis using laser scanning confocal microscopy. For the first time we show a small but statistically significant role for integrins in IgG-Fcγ receptor mediated phagocytosis.

The Biodegradation of Dispersed Oil Does Not Induce Toxicity

ABSTRACT The acute toxicity of dispersed oil is well understood, and oil dilutes to levels below those of acute concern within hours of dispersion whether oil is dispersed by waves or by lower energy turbulence in the presence of chemical dispersants. Once dispersed, the hydrocarbon components of the spilled oil are degraded promptly by the native microbes in seawater, typically with an apparent half-life of 7–30 days even under Arctic conditions. Nevertheless, concern has been raised that this biodegradation might increase the oil's acute toxicity by generating and releasing toxic by-products. We show here, using Americamysis bahia as the test species, that this does not occur when dispersed oil is present at environmentally-relevant concentrations (initially 3 ppm oil dispersed with Corexit 9500 at a dispersant to oil ratio of 1:20). The guidelines for this toxicity test mandate a temperature of 26 ± 1C, rather warmer than the temperature of collection of the seawater from the New Jersey shore that we used as our experimental medium, so it is not surprising that biodegradation was especially rapid with a half-life for the loss of detectable hydrocarbons of approximately 4 days. We conducted sequential 4-day acute toxicity tests for 20 days, by which time the indigenous microorganisms had removed almost 80% of the detectable hydrocarbons in the lightly weathered crude oil. We saw no mortality in any of the five sequential tests.

From the Instinctual to the Cosmic: Jung’s Exploration of the Colour in The Red Book, 1915-1929/30

Jung wrote extensively about colour symbolism in his patients’ dreams, paintings, and active imagination, beginning with his first mandala study in 1929, and continuing during the 1930s as he learned more about alchemy and Eastern esoteric texts. Students of Jung and Jungian analysts are already well acquainted with this material. The publication of The Red Book (2009), and Jung’s visual works in The Art of C.G. Jung (2019), present new opportunities to study how Jung explored colour between 1915 and 1929. This paper will trace Jung’s colour journey, concentrating on imagery that illustrates the instinctual and cosmic energies of the new god, the self and individuation. Jung’s evolving colour symbolism demonstrates The Red Book’s crucial role as an experimental medium, and confirms that Jung had developed a well-established colour hermeneutic by the 1920s. KEY WORDS colour, instinct, cosmos, new god, self, individuation, mandala, Goethe.

From the Instinctual to the Cosmic: Jung’s Exploration of the Colour in The Red Book, 1915-1929/30

Jung wrote extensively about colour symbolism in his patients’ dreams, paintings, and active imagination, beginning with his first mandala study in 1929, and continuing during the 1930s as he learned more about alchemy and Eastern esoteric texts. Students of Jung and Jungian analysts are already well acquainted with this material. The publication of The Red Book (2009), and Jung’s visual works in The Art of C.G. Jung (2019), present new opportunities to study how Jung explored colour between 1915 and 1929. This paper will trace Jung’s colour journey, concentrating on imagery that illustrates the instinctual and cosmic energies of the new god, the self and individuation. Jung’s evolving colour symbolism demonstrates The Red Book’s crucial role as an experimental medium, and confirms that Jung had developed a well-established colour hermeneutic by the 1920s. KEY WORDS colour, instinct, cosmos, new god, self, individuation, mandala, Goethe.

Impact of Fat Graft Thickness and Harvesting Technique on Adipocyte Viability in a New Porcine Experimental Model: An Immunohistochemical Analysis

Background Autologous fat grafting (AFG) has been employed in surgical practice as a filling method. However, controversies remain on the specifics of this technique. So far, few relevant experimental large animal studies have objectively assessed factors related to AFG integration. Objectives This study utilized an experimental, medium-sized animal model to compare the feasibility of AFG collected employing 2 different techniques with instruments of distinct thicknesses. Methods Twenty minipigs (Sus scropha domesticus) were subjected to AFG harvesting via en bloc resection utilizing 3- (Group I) and 5-mm-diameter (Group II) round punch blades (PBs) and liposuction (LS) with 3- (Group III) and 5-mm-diameter cannulas (Group IV). Both samples were grafted intramuscularly (biceps femoralis). Hematoxylin and eosin staining was employed to identify intact adipocytes, fat necrosis, fibrosis, inflammation, and oil cysts. Immunohistochemical staining (perilipin-A, tumor necrosis factor alfa, and cluster of differentiation number 31) was utilized to quantify the feasibility of adipocytes, tissue necrosis, and neoangiogenesis, respectively. Results Hematoxylin and eosin analysis showed that fat necrosis and histiocyte presence were significantly lower in the AFG harvested utilizing a PB than in LS. For perilipin-A, a statistical difference was observed between subgroups I and III (P = 0.001) and I and IV (P = 0.004). Instrument diameter had no effect on graft integration in comparisons between groups II and III (P = 0.059) and II and IV (P = 0.132). Conclusions In this experimental study, fat collected utilizing a PB demonstrated higher adipocyte viability than fat collected with LS. The diameter of the collection instruments, whether PB or LS, had no effect on graft integration.