LXR-dependent gene expression is important for macrophage survival and the innate immune response

The liver X receptors (LXRs) are nuclear receptors with established roles in the regulation of lipid metabolism. We now show that LXR signaling not only regulates macrophage cholesterol metabolism but also impacts antimicrobial responses. Mice lacking LXRs are highly susceptible to infection with the intracellular bacteria Listeria monocytogenes (LM). Bone marrow transplant studies point to altered macrophage function as the major determinant of susceptibility. LXR-null macrophages undergo accelerated apoptosis when challenged with LM and exhibit defective bacterial clearance in vivo. These defects result, at least in part, from loss of regulation of the antiapoptotic factor SPalpha, a direct target for regulation by LXRalpha. Expression of LXRalpha or SPalpha in macrophages inhibits apoptosis in the setting of LM infection. Our results demonstrate that LXR-dependent gene expression plays an unexpected role in innate immunity and suggest that common nuclear receptor pathways mediate macrophage responses to modified lipoproteins and intracellular pathogens.     

Stimulation of the chemiluminescence of human polymorphonuclear leukocytes in various stages of the intraerythrocyte development of Plasmodium falciparum

The synchronized cultures of Plasmodium falciparum were used to stimulate in vitro the chemiluminescence of human polymorphonuclear leukocytes in the presence of immune serum. The schizonts were concentrated by Percoll gradient centrifugation method (density 1.085 and osmolarity 285 mOsmol), and placed in culture, treated 6 hours later by sorbitol. Under incubation at constant temperature and pressure, the rate of synchronization reached 85% for schizonts during 5 replicative cycles. Every asexual stages of Plasmodium falciparum were used separately to stimulate polymorphonuclear leukocytes: merozoites were the most effective, followed by schizonts, trophozoites, and lastly supernatants of cultures containing degradation products of parasites.     

GAP promoter‐based fed‐batch production of highly bioactive core streptavidin by Pichia pastoris

Streptavidin is a homotetrameric protein binding the vitamin biotin and peptide analogues with an extremely high affinity, which leads to a large variety of applications. The biotin‐auxotrophic yeast Pichia pastoris has recently been identified as a suitable host for the expression of the streptavidin gene, allowing both high product concentrations and productivities. However, so far only methanol‐based expression systems have been applied, bringing about increased oxygen demand, strong heat evolution and high requirements for process safety, causing increased cost. Moreover, common methanol‐based processes lead to large proportions of biotin‐blocked binding sites of streptavidin due to biotin‐supplemented media. Targeting these problems, this paper provides strategies for the methanol‐free production of highly bioactive core streptavidin by P. pastoris under control of the constitutive GAP promoter. Complex were superior to synthetic production media regarding the proportion of biotin‐blocked streptavidin. The optimized, easily scalable fed‐batch process led to a tetrameric product concentration of up to 4.16 ± 0.11 µM of biotin‐free streptavidin and a productivity of 57.8 nM h^?1 based on constant glucose feeding and a successive shift of temperature and pH throughout the cultivation, surpassing the concentration in un‐optimized conditions by a factor of 3.4. Parameter estimation indicates that the optimized conditions caused a strongly increased accumulation of product at diminishing specific growth rates (μ ≈ D < 0.01 h^?1), supporting the strategy of feeding. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:855–864, 2016     

Real-time PCR detection and quantification of fish probiotic Phaeobacter strain 27-4 and fish pathogenic Vibrio in microalgae, rotifer, Artemia and first feeding turbot (Psetta maxima) larvae

Aims:  To develop a SYBR Green quantitative real-time PCR protocol enabling detection and quantification of a fish probiotic and two turbot pathogenic Vibrio spp. in microcosms.
Methods and Results:  Phaeobacter 27-4, Vibrio anguillarum 90-11-287 and Vibrio splendidus DMC-1 were quantified as pure and mixed cultures and in presence of microalgae ( Isochrysis galbana ), rotifers ( Brachionus plicatilis ), Artemia nauplii or turbot ( Psetta maxima ) larvae by real-time PCR based on primers directed at genetic loci coding for antagonistic and virulence-related functions respectively. The optimized protocol was used to study bioencapsulation and maintenance of the probiont and pathogens in rotifers and for the detection and quantification of Phaeobacter and V. anguillarum in turbot larvae fed rotifers loaded with the different bacteria in a challenge trial.
Conclusions:  Our real-time PCR protocol is reproducible and specific. The method requires separate standard curve for each host organism and can be used to detect and quantify probiotic Phaeobacter and pathogenic Vibrio bioencapsulated in rotifers and in turbot larvae.
Significance and Impact of the Study:  Our method allows monitoring and quantification of a turbot larvae probiotic bacteria and turbot pathogenic vibrios in in vivo trials and will be useful tools for detecting the bacteria in industrial rearing units.     

Baculiferins A–O,O-sulfated pyrrole alkaloids with anti-HIV-1 activity,from the Chinese marine sponge Iotrochota baculifera

Fifteen new DOPA-derived pyrrole alkaloids, named baculiferins A–O (2–16), were isolated from the Chinese marine sponge Iotrochota baculifera, together with the known alkaloids purpurone (1) and ningalin A (17). Most of the new compounds contain one to three O-sulfate units. Their structures were determined by extensive spectroscopic analysis including ¹H and ¹³C NMR (COSY, HMQC, HMBC) and ESIMS data. A possible pathway for the biosynthetic origin of the isolated alkaloids is proposed, in which DOPA is assumed to be a joint biogenetic precursor. Baculiferins C, E–H, and K–N (4, 6–9, 12–15) were found to be potent inhibitors against the HIV-1 IIIB virus in both, MT4 and MAGI cells. Additional bioassay revealed that baculiferins could dramatically bind to the HIV-1 target proteins Vif, APOBEC3G, and gp41, for which structure–activity relationships are discussed.     

Cultivation of Ulva lactuca with manure for simultaneous bioremediation and biomass production

The potential of liquid manure as sole nutrient source for cultivation of Ulva lactuca was investigated with the perspective of utilizing the produced biomass for feed and/or energy. Algae grown with manure demonstrated equal growth rates to algae grown with standard f/2-medium. The optimum manure concentration, expressed as ammonium concentration, was 25?μM. At these conditions, the biomass produced was potentially suitable for anaerobic digestion, due to a relative high carbon/nitrogen ratio (approximately 19). At higher manure concentrations, tissue concentrations of nitrogen, phosphorus, proteins, and amino acids increased, making the biomass less suitable for anaerobic digestion but potentially interesting as a feed supplement. Cultivating U. lactuca with manure as nutrient source has potential in terms of bioremediation as well as production of bioenergy and protein-feed.     

Identification of Armillaria field isolates using isozymes and mycelial growth characteristics

This research was conducted to develop procedures based on mycelial growth characteristics and patterns of esterase (EST) and polyphenol oxidase (PPO) production by diffuse mycelia for identification of Armillaria field isolates from Quercus-Carya-Pinus forests in the Ozark Mountains (central USA). The 285 isolates collected were first identified by standard diploid-haploid pairing tests as A. gallica, A. mellea, or A. tabescens. A strong PPO band was diagnostic for A. gallica. All A. mellea isolates tested and 91% of the A. tabescens isolates tested were distinguished based on production of EST bands in three standardized R f ranges. A procedure based on mycelial growth and morphology on tannic acid medium (TA) at 24 °C and on malt extract medium (ME) at 33 °C correctly identified 98% of A. gallica isolates and all A. mellea and A. tabescens isolates. On TA, A. gallica grew slowest. On ME, A. mellea grew slowest: mycelial morphology differed among species; A. gallica typically stained the agar and produced an appressed/submerged growth pattern with concentric bands of decreasing hyphal density, A. mellea typically did not stain the agar and produced round mycelia with smooth margins and abundant aerial hyphae, A. tabescens typically stained the agar and grew appressed/submerged with very irregular margins and patchy hyphal density. These are the first published systems evaluating the potential for identifying Armillaria field isolates based on their mycelial growth characteristics and EST and PPO complements. This revised version was published online in June 2006 with corrections to the Cover Date.     

Alkaloids and ethnobotany of Mexican peyote cacti and related species

Some Mexican cacti of the genera Ariocarpus, Mammillaria, Obregonia, Pelecyphora, Solisia and Turbinicarpus have been studied with respect to their alkaloids and ethnobotany. Seven previously known cactus alkaloids were identified. N,N-Dimethyl-4-hydroxy-3-methoxyphenethylamine was found to occur in Ariocarpus agavoides. In Pelecyphora aselliformis N,N-dimethyl-3-hydroxy-4, 5-dimethoxyphenethylamine was identified as the major alkaloid. The possible relationship between the vernacular names of these plants and their alkaloid content is discusse.