Enhanced Immunogenicity of Mitochondrial-Localized Proteins in Cancer Cells.

Epitopes derived from mutated cancer proteins elicit strong antitumor T-cell responses that correlate with clinical efficacy in a proportion of patients. However, it remains unclear whether the subcellular localization of mutated proteins influences the efficiency of T-cell priming. To address this question, we compared the immunogenicity of NY-ESO-1 and OVA localized either in the cytosol or in mitochondria. We showed that tumors expressing mitochondrial-localized NY-ESO-1 and OVA proteins elicit significantdly higher frequencies of antigen-specific CD8+ T cells in vivo. We also demonstrated that this stronger immune response is dependent on the mitochondrial location of the antigenic proteins, which contributes to their higher steady-state amount, compared with cytosolic localized proteins. Consistent with these findings, we showed that injection of mitochondria purified from B16 melanoma cells can protect mice from a challenge with B16 cells, but not with irrelevant tumors. Finally, we extended these findings to cancer patients by demonstrating the presence of T-cell responses specific for mutated mitochondrial-localized proteins. These findings highlight the utility of prioritizing epitopes derived from mitochondrial-localized mutated proteins as targets for cancer vaccination strategies.S

The impact of tides on the capillary transition zone

The capillary transition zone, also known as the capillary fringe, is a zone where water saturations decrease with height above the water table/oil–water contact as a result of capillary action. In some oil reservoirs, this zone may contain a significant proportion of the oil in place. In groundwater assessments, the capillary fringe can profoundly affect contaminant transport. In this study, we investigated the influence of a tidally induced, semi-diurnal, change in water table depth on the water saturation distribution in the capillary fringe/transition zone. The investigation used a mixture of laboratory experiments, in which the change in saturation with depth was monitored over a period of 90 days, and numerical simulation. We show that tidal changes in water table depth can significantly alter the vertical water saturation profile from what would be predicted using capillary–gravity equilibrium and the drainage or imbibition capillary pressure curves

Parasitism by the Tachinid Parasitoid Exorista japonica Leads to Suppression of Basal Metabolism and Activation of Immune Response in the Host Bombyx mori

The dipteran tachinid parasitoids are important biocontrol agents, and they must survive the harsh environment and rely on the resources of the host insect to complete their larval stage. We have previously demonstrated that the parasitism by the tachinid parasitoid Exoristajaponica, a pest of the silkworm, causes pupation defects in Bombyx mori. However, the underlying mechanism is not fully understood. Here, we performed transcriptome analysis of the fat body of B. mori parasitized by E. japonica. We identified 1361 differentially expressed genes, with 394 genes up-regulated and 967 genes down-regulated. The up-regulated genes were mainly associated with immune response, endocrine system and signal transduction, whereas the genes related to basal metabolism, including energy metabolism, transport and catabolism, lipid metabolism, amino acid metabolism and carbohydrate metabolism were down-regulated, indicating that the host appeared to be in poor nutritional status but active in immune response. Moreover, by time-course gene expression analysis we found that genes related to amino acid synthesis, protein degradation and lipid metabolism in B. mori at later parasitization stages were inhibited. Antimicrobial peptides including Cecropin A, Gloverin and Moricin, and an immulectin, CTL11, were induced. These results indicate that the tachinid parasitoid perturbs the basal metabolism and induces the energetically costly immunity of the host, and thus leading to incomplete larval–pupal ecdysis of the host. This study provided insights into how tachinid parasitoids modify host basal metabolism and immune response for the benefit of developing parasitoid larvae

Homeostatic Regulation of the Duox-ROS Defense System: Revelations Based on the Diversity of Gut Bacteria in Silkworms (<i>Bombyx mori</i>)

The Duox-ROS defense system plays an important role in insect intestinal immunity. To investigate the role of intestinal microbiota in Duox-ROS regulation herein, 16S rRNA sequencing technology was utilized to compare the characteristics of bacterial populations in the midgut of silkworm after different time-periods of treatment with three feeding methods: 1–4 instars artificial diet (AD), 1–4 instars mulberry leaf (ML) and 1–3 instars artificial diet + 4 instar mulberry leaf (TM). The results revealed simple intestinal microbiota in the AD group whilst microbiota were abundant and variable in the ML and TM silkworms. By analyzing the relationship among intestinal pH, reactive oxygen species (ROS) content and microorganism composition, it was identified that an acidic intestinal environment inhibited the growth of intestinal microbiota of silkworms, observed concurrently with low ROS content and a high activity of antioxidant enzymes (SOD, TPX, CAT). Gene expression associated with the Duox-ROS defense system was detected using RT-qPCR and identified to be low in the AD group and significantly higher in the TM group of silkworms. This study provides a new reference for the future improvement of the artificial diet feeding of silkworm and a systematic indicator for the further study of the relationship between changes in the intestinal environment and intestinal microbiota balance caused by dietary alterations

Betaine supplementation alleviates corticosterone-induced hepatic cholesterol accumulation through epigenetic modulation of HMGCR and CYP7A1 genes in laying hens

ABSTRACT: Excessive corticosterone (CORT) exposure could cause hepatic cholesterol accumulation in chickens and maternal betaine supplementation could decrease hepatic cholesterol deposition through epigenetic modifications in offspring chickens. Nevertheless, it remains uncertain whether providing betaine to laying hens could protect CORT-induced hepatic cholesterol accumulation via epigenetic mechanisms. This study aimed to examine the effects of dietary betaine on plasma and hepatic cholesterol contents, expression of cholesterol metabolic genes, as well as DNA methylation on their promoters in the liver of laying hens exposed to CORT. A total of 72 laying hens at 130 d of age were randomly divided into 3 groups: control (CON), CORT, and CORT+betaine (CORT+BET) groups. The experiment lasted for 35 d. Chickens in CON and CORT groups were fed a basal diet, whereas the CORT+BET group chickens were fed the basal diet supplemented with 0.1% betaine for 35 d. On d 28 of the experiment, chickens in CORT and CORT+BET groups received daily subcutaneous injections of CORT (4.0 mg/kg body weight), whereas the CON group chickens were injected with an equal volume of solvent for 7 d. The results showed that CORT administration led to a significant increase (P < 0.05) in the contents of cholesterol in plasma and liver, associated with activation (P < 0.05) of sterol regulatory element binding transcription factor 2 (SREBP2), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), lecithin-cholesterol acyltransferase (LCAT) and low-density lipoprotein receptor (LDLR) genes expression, and inhibition of cholesterol-7-alpha hydroxylase (CYP7A1) and sterol 27-hydroxylase (CYP27A1) genes expression in the liver compared to the CON. In contrast, CORT-induced up-regulation of HMGCR mRNA and protein abundances and downregulation of CYP7A1 mRNA and protein abundances were completely normalized (P < 0.05) by betaine supplementation. Besides, CORT injection led to significant hypomethylation (P < 0.05) on HMGCR promoter and hypermethylation (P < 0.05) on CYP7A1 promoter. Moreover, dietary betaine rescued (P < 0.05) CORT-induced changes in methylation status of HMGCR and CYP7A1 genes promoters. These results indicate that dietary betaine addition protects laying hens from CORT-induced hepatic cholesterol accumulation via epigenetic modulation of HMGCR and CYP7A1 genes

Mechanism of enhanced Bombyx mori nucleopolyhedrovirus-resistance by titanium dioxide nanoparticles in silkworm.

The infection of Bombyx mori nucleopolyhedrovirus (BmNPV) in silkworms is often lethal. It is difficult to prevent, and its lethality is correlated with both viral particle characteristics and silkworm strains. Low doses of titanium dioxide nanoparticles (TiO2 NPs) can promote silkworm growth and improve its resistance to organophosphate pesticides. In this study, TiO2 NPs' effect on BmNPV resistance was investigated by analyzing the characteristics of BmNPV proliferation and transcriptional differences in silkworm midgut and the transcriptional changes of immunity related genes after feeding with TiO2 NPs. We found that low doses of TiO2 NPs improved the resistance of silkworm against BmNPV by 14.88-fold, with the mortalities of the experimental group and control group being 0.56% and 8.33% at 144 h, respectively. The proliferation of BmNPV in the midgut was significantly increased 72 h after infection in both experimental and control groups; the control group reached the peak at 120 h, while the experimental group took 24 more hours to reach the maximal value that was 12.63 times lower than the control, indicating that TiO2 NPs can inhibit BmNPV proliferation in the midgut. Consistently, the expression of the BmNPV-resistant gene Bmlipase-1 had the same increase pattern as the proliferation changes. Immune signaling pathway analysis revealed that TiO2 NPs inhibited the proliferation of silkworm BmNPV to reduce the activation levels of janus kinase/signal transducer and activator of transcription (JAK/STAT) and phosphatidylinositol 3-kinase (PI3K)-Akt signaling pathway, while promoting the expression of Bmakt to improve the immunity. Overall, our results demonstrate that TiO2 NPs increase silkworm resistance against BmNPV by inhibiting virus proliferation and improving immunity in silkworms

Molecular Mechanisms of Reduced Nerve Toxicity by Titanium Dioxide Nanoparticles in the Phoxim-Exposed Brain of <i>Bombyx mori</i>

<div><p><i>Bombyx mori</i> (<i>B. mori</i>), silkworm, is one of the most important economic insects in the world, while phoxim, an organophosphorus (OP) pesticide, impact its economic benefits seriously. Phoxim exposure can damage the brain, fatbody, midgut and haemolymph of <i>B. mori</i>. However the metabolism of proteins and carbohydrates in phoxim-exposed <i>B. mori</i> can be improved by Titanium dioxide nanoparticles (TiO₂ NPs). In this study, we explored whether TiO₂ NPs treatment can reduce the phoxim-induced brain damage of the 5th larval instar of <i>B. mori</i>. We observed that TiO₂ NPs pretreatments significantly reduced the mortality of phoxim-exposed larva and relieved severe brain damage and oxidative stress under phoxim exposure in the brain. The treatments also relieved the phoxim-induced increases in the contents of acetylcholine (Ach), glutamate (Glu) and nitric oxide (NO) and the phoxim-induced decreases in the contents of norepinephrine (NE), Dopamine (DA), and 5-hydroxytryptamine (5-HT), and reduced the inhibition of acetylcholinesterase (AChE), Na⁺/K⁺-ATPase, Ca²⁺-ATPase, and Ca²⁺/Mg²⁺-ATPase activities and the activation of total nitric oxide synthase (TNOS) in the brain. Furthermore, digital gene expression profile (DGE) analysis and real time quantitative PCR (qRT-PCR) assay revealed that TiO₂ NPs pretreatment inhibited the up-regulated expression of <i>ace1, cytochrome c</i>, <i>caspase-9</i>, <i>caspase-3</i>, <i>Bm109</i> and down-regulated expression of <i>BmIap</i> caused by phoxim; these genes are involved in nerve conduction, oxidative stress and apoptosis. TiO₂ NPs pretreatment also inhibited the down-regulated expression of <i>H⁺ transporting ATP synthase</i> and <i>vacuolar ATP synthase</i> under phoxim exposure, which are involved in ion transport and energy metabolism. These results indicate that TiO₂ NPs pretreatment reduced the phoxim-induced nerve toxicity in the brain of <i>B. mori</i>.</p></div

Ultrastructure of the brain tissue in fifth-instar larvae after phoxim exposure 48 h.

<p>(a) Control; (b) TiO₂ NPs; (c) Phoxim; (d) TiO₂ NPs + Phoxim. Green arrows indicate karyopyknosis and chromatin marginalization, blue arrows show mitochondria swelling and became deformed, crest broken.</p

Comparison between fold-difference with qRT-PCR results and DGE assay in each group.

<p>*p<0.05, **p<0.01, and ***p<0.001.</p><p>Values represent means ± SEM (<i>n</i> = 5).</p

Effects of TiO₂ NPs on nerve conduction in the brain of phoxim-exposed fifth-instar larvae.

<p>*<i>p</i><0.05, **<i>p</i><0.01, and ***<i>p</i><0.001. Values represent means ± SEM (<i>N</i> = 5). (a) Neurotransmitter contents, (b) Enzyme activity.</p