Deficiency of Leishmania Phosphoglycans Influences the Magnitude but Does Not Affect the Quality of Secondary (Memory) Anti-Leishmania Immunity

Despite inducing very low IFN-γ response and highly attenuated in vivo, infection of mice with phosphoglycan (PG) deficient Leishmania major (lpg2-) induces protection against virulent L. major challenge. Here, we show that mice infected with lpg2- L. major generate Leishmania-specific memory T cells. However, in vitro and in vivo proliferation, IL-10 and IFN-γ production by lpg2- induced memory cells were impaired in comparison to those induced by wild type (WT) parasites. Interestingly, TNF recall response was comparable to WT infected mice. Despite the impaired proliferation and IFN-γ response, lpg2- infected mice were protected against virulent L. major challenge and their T cells mediated efficient infection-induced immunity. In vivo depletion and neutralization studies with mAbs demonstrated that lpg2- L. major-induced resistance was strongly dependent on IFN-γ, but independent of TNF and CD8⁺ T cells. Collectively, these data show that the effectiveness of secondary anti-Leishmania immunity depends on the quality (and not the magnitude) of IFN-γ response. These observations provide further support for consideration of lpg2- L. major as a live-attenuated candidate for leishmanization in humans since it protects strongly against virulent challenge, without inducing pathology in infected animals.     

β-Lactoglobulin/polysaccharide interactions during in vitro gastric and pancreatic hydrolysis assessed in dialysis bags of different molecular weight cut-offs

The effects of gum arabic, low methylated (LM) pectin or xylan at levels of 0 and 50 wt.% on beta-lactoglobulin (β-lg) digestibility were studied as well as the interactions between the two macromolecules during in vitro hydrolysis. The proteolysis was performed in a system involving a two-step hydrolysis: either pepsin alone, or pepsin followed by a trypsin/chymotrypsin (T/C) mixture in dialysis bags with molecular weight cut-offs (MWCO) 1000 or 8000 Da. Digestibility was estimated by the N release and by a SDS-PAGE electrophoresis of retentates from the two dialysis bags after hydrolysis. Turbidimetric measurements monitored the structural evolution of mixtures during the two-step hydrolysis. Results showed that β-lg was almost resistant to peptic digestion and that polysaccharides increased the N release despite a reduction of pepsin activity. This is due to the formation of electrostatic complexes between polysaccharides and β-lg, which reduced β-lg aggregation, increasing its solubility. The polysaccharides reduced significantly the β-lg T/C digestibility as determined using a dialysis bag with a MWCO 1000 Da, without a modification of their enzymatic activities. No significant effect of polysaccharides on the β-lg digestibility was detected using the dialysis bag with a MWCO 8000 Da. The electrophoresis pattern did not show differences in the profile of retentates in relation with the dialysis bag used. This suggests that non-specific interactions could occur during the second step of hydrolysis between polysaccharides and amino acids or peptides smaller than 8000 Da.     

Phylogenetic relationship of Clauseneae (Rutaceae) inferred from plastid and nuclear DNA data and taxonomic implication for some major taxa

Using sequences of the nuclear ribosomal ITS region as well as the chloroplast DNA trnL‐trnF and atpB‐rbcL regions, this study aims to provide further insight into the phylogenetic relationships within Clauseneae and its relationship to Citreae (Rutaceae). Using maximum likelihood (ML) and Bayesian inference (BI), we reconstructed the phylogeny of Clauseneae based on trnL‐F, atpB‐rbcL and ITS sequences. Our data matrix contained 91 accessions, representing 72 species and varieties from 31 genera and two outgroups, including new and extensive sampling of species and varieties representing four genera in the tribe Clauseneae. In the subfamily Aurantioideae, six major clades were resolved with strong support: 1) Micromelum clade: Micromelum; 2) Glycosmis clade: Glycosmis; 3) Bergera clade: Murraya sect. Bergera; 4) Clausena clade: Clausena; 5) Murraya clade: Murraya sect. Murraya + Merrillia; 6) Citreae clade: Citreae. Micromelum, Glycosmis, Clausena and Merrillia were confirmed as monophyletic. In contrast, Murraya s.l. was reconstructed as polyphyletic. Murraya sect. Bergera clustered with Clausena while Murraya sect. Murraya and Merrillia together formed a clade that is sister to the tribe Citreae. All members from Citreae were clustered into a natural group. The genus Micromelum was found to be primitive in this subfamily and more close to Glycosmis. Based on the phylogeny and morphological characters, we discuss the taxonomy of some members of Clauseneae and conclude that the current tribal and generic classification need further revision.     

Defects and rescue of the minor salivary glands in Eda pathway mutants

Despite their importance to oral health, the mechanisms of minor salivary gland (SG) development are largely unexplored. Here we present in vivo and in vitro analyses of developing minor SGs in wild type and mutant mice. Eda, Shh and Fgf signalling pathway genes are expressed in these glands from an early stage of development. Developing minor SGs are absent in Eda pathway mutant embryos, and these mice exhibit a dysplastic circumvallate papilla with disrupted Shh expression. Supplementation of Eda pathway mutant minor SG explants with recombinant EDA rescues minor SG induction. Supplementation with Fgf8 or Shh, previously reported targets of Eda signalling, leads to induction of gland like structures in a few cases, but these fail to develop into minor SGs.     

CRNDE enhances neuropathic pain via modulating miR-136/IL6R axis in CCI rat models

Neuropathic pain has been reported as a type of chronic pain due to the primary dysfunction of the somatosensory nervous system. It is the most serious types of chronic pain, which can lead to a significant public health burden. But, the understanding of the cellular and molecular pathogenesis of neuropathic pain is barely complete. Long noncoding RNAs (lncRNAs) have recently been regarded as modulators of neuronal functions. Growing studies have indicated lncRNAs can exert crucial roles in the development of neuropathic pain. Therefore, our present study focused on the potential role of the lncRNA Colorectal Neoplasia Differentially Expressed (CRNDE) in neuropathic pain progression. Firstly, a chronic constrictive injury (CCI) rat model was built. CRNDE was obviously increased in CCI rats. Interestingly, overexpression of CRNDE enhanced neuropathic pain behaviors. Neuroinflammation was induced by CRNDE and as demonstrated, interleukin-10 (IL-10), IL-1, IL-6, and tumor necrosis factor-α (TNF-α) protein levels in CCI rats were activated by LV-CRNDE. For another, miR-136 was obviously reduced in CCI rats. Previously, it is indicated that miR-136 participates in the spinal cord injury via an inflammation in a rat model. Here, firstly, we verified miR-136 could serve as CRNDE target. Loss of miR-136 triggered neuropathic pain remarkably via the neuroinflammation activation. Additionally, IL6R was indicated as a target of miR-136 and miR-136 regulated its expression. Subsequently, we confirmed that CRNDE could induce interleukin 6 receptor (IL6R) expression positively. Overall, it was implied that CRNDE promoted neuropathic pain progression via modulating miR-136/IL6R axis in CCI rat models.     

Temporal and Vertical Distributions of Bacterioplankton at the Gray's Reef National Marine Sanctuary

Large spatial scales and long-term shifts of bacterial community composition (BCC) in the open ocean can often be reliably predicted based on the dynamics of physical-chemical variables. The power of abiotic factors in shaping BCC on shorter time scales in shallow estuarine mixing zones is less clear. We examined the diurnal variation in BCC at different water depths in the spring and fall of 2011 at a station in the Gray''s Reef National Marine Sanctuary (GRNMS). This site is located in the transition zone between the estuarine plume and continental shelf waters of the South Atlantic Bight. A total of 234,516 pyrotag sequences of bacterial 16S rRNA genes were recovered; they were taxonomically affiliated with >200 families of 23 bacterial phyla. Nonmetric multidimensional scaling analysis revealed significant differences in BCC between spring and fall samples, likely due to seasonality in the concentrations of dissolved organic carbon and nitrate plus nitrite. Within each diurnal sampling, BCC differed significantly by depth only in the spring and differed significantly between day and night only in the fall. The former variation largely tracked changes in light availability, while the latter was most correlated with concentrations of polyamines and chlorophyll a. Our results suggest that at the GRNMS, a coastal mixing zone, diurnal variation in BCC is attributable to the mixing of local and imported bacterioplankton rather than to bacterial growth in response to environmental changes. Our results also indicate that, like members of the Roseobacter clade, SAR11 bacteria may play an important role in processing dissolved organic material in coastal oceans.     

Effect of photoperiod manipulation on the growth performance of juvenile lenok,Brachymystax lenok (Pallas, 1773)

The effect of four different light regimes on growth was studied in lenok, Brachymystax lenok. Fish with average weights of 5.5 g were subjected to four different photoperiods (0L:24D, 6L:18D, 12L:12D and 24L:0D) for 35 days. The specific growth rate (SGR) of lenok in 24‐h darkness had a significantly higher SGR than those in the continuous light regime (P < 0.05); however, there was no significant difference among fish exposed to 6L:18D, 12L:12D and 24L:0D photoperiods. There was a tendency for higher food intake over the light period extension from 0L to 24L, and feed intake was significantly higher in the continuous light group than in 24‐h darkness (P < 0.05). No significant difference in feed conversion efficiency (FCE) was observed between fish exposed to 0L:24D and 6L:18D photoperiods, however, the FCE in both photoperiods was significantly higher than that in the other two groups. The final survival rate of juveniles varied from 79.67 to 95.33%, with significant differences among experimental groups. Fish tested in continuous illumination spent much more energy on respiration and excretion while depositing less energy for growth than in the other photoperiods. In contrast, fish in 24‐h darkness deposited more energy for growth and spent less energy on respiration and excretion. Results show that photoperiod manipulation can affect growth, and that a continuous dark regime could improve growth in lenok at this stage of development.