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Abstract Superoxide dismutase is a key enzyme that scavenges superoxide anion and plays vital roles in plant antioxidant system. This study identified six SOD genes from the deciduous conifer Larix kaempferi , which is widely distributed across the cooler regions of the northern hemisphere. These six SOD genes were classified into three types: Cu/Zn-SOD (LkSOD1, 2, 3 and 4), Fe-SOD (LkSOD5) and Mn-SODs (LkSOD6). Three Cu/Zn-SOD proteins (LkSOD1, 3 and 4) were cytosolic-localized, while the other three proteins (LkSOD2, 5 and 6) were chloroplast-localized. Larix SOD proteins displayed catalytic activities toward superoxide anion, and retained >55% of its maximum enzymatic activity between 10 °C and 40 °C. Over expressions of three Larix SOD genes (LkSOD2 , 4 and 6) in Arabidopsis thaliana , respectively, showed increased germination rates and root lengths during salt stress. LkSOD5 gene could rescue pale green and dwarf phenotype of Arabidopsis atfsd2-2 mutant. Taken together, this study provided comprehensive insight into the gymnosperm SOD gene family. Highlights • Six SOD genes were identified from Larix kaempferi. • Larix SOD proteins displayed different subcellular location. • LkSOD5 gene could rescue the pale green and dwarf phenotype of atfsd2-2. • Over expressing of three Larix SOD genes in Arabidopsis could increase salt tolerance. [ABSTRACT FROM AUTHOR]

Sirtuin 3 (SIRT3) is a major deacetylase in the mitochondria and sensitive to diverse nutrient signals. However, how dietary fatty acids regulate SIRT3 expression remains poorly understood. Here, sirt3 gene from large yellow croaker was characterized and can be activated by the oxidized nicotinamide adenine dinucleotide (NAD+) precursor, nicotinamide mononucleotide (NMN). Moreover, the expression of SIRT3 is regulated differently by dietary fatty acids. In vivo , soybean oil increased the mRNA expression of sirt3 in head kidney. In vitro , stearic acid (SA) decreased the expression of SIRT3 in macrophages, while DHA and EPA increased the mRNA expression of sirt3. Meanwhile, all long-chain polyunsaturated fatty acids (LC-PUFAs) enhanced the protein levels of SIRT3, which was consistent with the trend of fatty acids affecting NAD+ levels. Furthermore, inhibition of NAD+ de novo synthesis blocked DHA-induced increases in SIRT3 expression, and NMN supplement reversed SA-induced decreases in SIRT3 expression. In conclusion, these findings suggest that dietary fatty acids may regulate SIRT3 expression by affecting intracellular NAD+ synthesis. Regarding the important role of SIRT3 in regulating mitochondrial function, appropriate activation of SIRT3 may be an important way to alleviate metabolic disorders in aquaculture. • SIRT3 in large yellow croaker was a conserved sirtuin family member and could be activated by NMN. • The effects of fatty acids on SIRT3 regulation is affected by carbon chain saturation rather than carbon chain length. • Dietary fatty acids affected the expression of SIRT3 by regulating intracellular NAD+ synthesis. [ABSTRACT FROM AUTHOR]

Background: The single nucleotide polymorphism (SNP) rs1344706 in ZNF804A (2q32.1) has been associated with schizophrenia in a genome-wide association study (GWAS). A recent candidate gene study, which replicated the positive association with rs1344706, identified another positive SNP (rs7597593) in ZNF804A associated with schizophrenia. Methods: We performed an association study of rs7597593 in four GWAS cohorts of European ancestry. Postmortem human brain expression data of normal Caucasian individuals (n = 89) was also analyzed for examining the effect of rs7597593 on ZNF804A messenger RNA expression, using logistic regression and linear regression. Results: We found that rs7597593 was significantly associated with schizophrenia in the combined GWAS datasets (n = 5023, odds ratio [OR]combined = 1.15, p = .0011). Analysis of stratification by sex showed that the association was driven by the female subjects (OR = 1.29, p = .0002) and was not significant in male subjects (OR = 1.08, p = .148) in the combined sample of four cohorts. A sex by genotype interaction was near significant in both the Genetic Association Information Network sample (p = .0532) and the combined sample of four cohorts (p combined = .0531). Gene expression analysis showed no main effects but a significant female-specific association (p female = .047, p male = .335) and sex by genotype interaction (p = .0166) for rs7597593. Conclusions: Our data suggest a clinical and molecular modulation by sex of the association of ZNF804A SNP rs7597593 and risk of schizophrenia. [Copyright &y& Elsevier]

Nanoplastics are common pollutants in aquatic environments and have attracted widespread research attention. However, few studies focus on the effects of nanoplastic exposure on energy metabolism in crustaceans. Accordingly, we exposed juvenile oriental river prawns (Macrobrachium nipponense) to different concentrations of 75-nm polystyrene nanoplastics (0, 5, 10, 20, and 40 mg/L) for 7, 14, 21, or 28 days. Thereafter, the effects of nanoplastic exposure on metabolite content, energy metabolism-related enzyme activity, and gene expression were evaluated. Our results showed that (1) with increasing nanoplastic concentration and exposure time, the survival rate decreased, while weight gain rate and molting number increased and then decreased; glycogen, triglyceride, and total cholesterol content all declined while lactic acid content increased with higher exposure to nanoplastic concentrations; (2) the activities of acetyl-CoA carboxylase (ACC), hexokinase (HK), carnitine palmitoyl transferase-1, pyruvate kinase (PK), lipase, and fatty acid synthase tended to decrease, while the activity of lactate dehydrogenase (LDH) increased. In particular, the activity of 6-phosphofructokinase exposed to 5 mg/L nanoplastics increased significantly (P < 0.05). (3) Expression of the metabolism-related genes 6-phosphate glucokinase (G-6-Pase), HK , PK , ACC , Acetyl-CoA-binding protein (ACBP), CPT-1 , and fatty-acid-binding protein 10 (FABP 10) increased and then decreased, while expression of the LDH gene showed an upward trend. These results indicate that nanoplastics affect growth, enzyme activity, and the gene expression of energy metabolism in M. nipponense , and that high concentrations of nanoplastics have a negative impact on energy metabolism. Image 1 • Polystyrene nanoplastic exposure could reduce survival and growth in shrimp. • Polystyrene nanoplastic exposure induced perturbation of glucose and lipid metabolism in shrimp. • Time- and dose-related effects of nanoplastic on oriental river prawns were examined. Our results identify potential nanoplastic toxicity on energy metabolism in aquatic organisms. [ABSTRACT FROM AUTHOR]

Two isoforms of Glutathione S-Transferase (GST) genes, belonging to mu (Dp-GSTm1 and Dp-GSTm2) and sigma (Dp-GSTs1 and Dp-GSTs2) classes, were cloned and characterised in the freshwater Daphnia pulex. No signal peptide was found in any of the four GST proteins, indicating that they were cytosolic GST. A highly conserved glutathione (GSH) binding site (G-site) occurred in the N-terminal sequence, and a substrate binding site (H-site), interacting non-specifically with the second hydrophobic substrate, was present in the C-terminal. A Tyr residue, for the stabilization of GSH, was found to be conserved in the analysed sequences. The secondary and tertiary structures indicated that these genes possess the typical cytosolic GST structure, including a conserved N-terminal domain with a βαβαββα motif. The μ loop (NVGPAPDYDR and NFIGAEWDR in Dp-GSTm1 and Dp-GSTm2, respectively) was identified between the βαβ (β1α1β2) and αββα motifs (α2β3β4α3) in the N-terminal domain. The expressions of Dp-GSTs1 , Dp-GSTs2 , and Dp-GSTm1 were higher in other age groups compared to the newly-born neonates (1 d); however, the expression of Dp-GSTm2 first increased and then decreased with age. Gene expression was significantly reduced by high concentration (1 and 2 mg/L) of 75 nm polystyrene nanoplastic. However, nanoplastic exposure at the predicted environmental concentration (1 μg/L) had a low effect. Exposure of mothers to nanoplastic (1 μg/L) elevated the Dp-GSTs2 level in their neonates. These results improve our understanding on the response of different types of Daphnid GST to environmental contaminants, especially nanoplastic. • Four GST genes of D. pulex exhibited all the features typical to the GST family. • Glutathione S-Transferase played a positive role against nanoplastic exposure. • Exposure of mothers to nanoplastic elevated the Dp-GSTs2 level in their neonates. [ABSTRACT FROM AUTHOR]

LncRNA TUC338 has recently been characterized as an oncogene in several types of cancer. Our study aimed to characterize the functionality of TUC338 in prostate carcinoma. It was observed that TUC338 was upregulated in tumor tissues comparing to adjacent healthy tissues of prostate carcinoma patients. Plasma levels of TUC338 were also higher in prostate carcinoma patients than in healthy controls. A 5-year follow-up study showed that high plasma level of TUC338 was correlated with poor survival. miR-466 was downregulated in tumor tissues compared with adjacent healthy tissues of prostate carcinoma patients. TUC338 and miR-466 were inversely correlated in tumor tissues. miR-466 overexpression failed to affect TUC338 expression, while TUC338 overexpression led to downregulated miR-466 expression. TUC338 overexpression failed to significantly affect cancer cell proliferation, but promoted cancer cell migration and invasion. MiR-466 overexpression resulted in reduced rates of cancer cell migration and invasion, and also attenuated the effect of TUC338 overexpression. Therefore, TUC338 may serve as an oncogenic lncRNA in prostate carcinoma by downregulating miR-466. • Our study aimed to characterize the functionality of TUC338 in prostate carcinoma. • TUC338 may regulate miR-466 through unknown pathways. • TUC338 is upregulated and may serve as an oncogene in prostate carcinoma patients by downregulating miR-466. [ABSTRACT FROM AUTHOR]

Myoglobin (Mb) is a monomeric hemoprotein which plays an important role in oxygen storage and transport in cardiac and skeletal muscle under hypoxia. The red tail toad-headed lizard Phrynocephalus erythrurus (Lacertilia: Agamidae), which inhabits at an elevation of 4500–5300 m on the Qinghai–Tibetan plateau, is known to be the highest living lizard in the world. To investigate the characters of myoglobin of this unique species, another low altitude lizard Phrynocephalus przewalskii (Lacertilia: Agamidae) was selected as a reference. The open reading frame (ORF) of myoglobin in two lizards was 465 bp which encodes a polypeptide of 154 amino acids with different theoretical molecular weight and isoelectric point. The amino acid substitutions of myoglobin between two species were found at Thr13Ile, Lys87Thr and His118Asn. Homology modeling results indicated that P. erythrurus myoglobin has a greater heme pocket, which may be more favorable to oxygen binding and unloading. On the other hand, the mRNA levels of myoglobin in both cardiac and skeletal muscle in P. erythrurus were significantly larger than those in low altitude P. przewalskii . At protein level, myoglobin concentration in skeletal muscle in P. erythrurus was notably increased, but no significant difference was observed in cardiac muscle. [ABSTRACT FROM AUTHOR]

Abstract: Fab antibody display on filamentous phage is widely applied to de novo antibody discovery and engineering. Here we describe a phagemid system for the efficient display and affinity selection of Fabs through linkage to the minor coat protein pIX. Display was successful by fusion of either Fd or Lc through a short linker to the amino terminus of pIX and co-expression of the counter Lc or Fd as a secreted, soluble fragment. Assembly of functional Fab was confirmed by demonstration of antigen-specific binding using antibodies of known specificity. Phage displaying a Fab specific for RSV-F protein with Fd linked to pIX showed efficient, antigen-specific enrichment when mixed with phage displaying a different specificity. The functionality of this system for antibody engineering was evaluated in an optimization study. A RSV-F protein specific antibody with an affinity of about 2nM was randomized at 4 positions in light chain CDR1. Three rounds of selection with decreasing antigen concentration yielded Fabs with an affinity improvement up to 70-fold and showed a general correlation between enrichment frequency and affinity. We conclude that the pIX coat protein complements other display systems in filamentous phage as an efficient vehicle for low copy display and selection of Fab proteins. [ABSTRACT FROM AUTHOR]

Schizophrenia polygenic risk is plausibly manifested by complex transcriptional dysregulation in the brain, involving networks of co-expressed and functionally related genes. 1. Samples: DLPFC gene expression (RNA-Seq) abundance was quantified (RPKM) in postmortem human brains from the LIBD Postmortem Human Brain Repository (90 controls- CTRL, 76 schizophrenia- SCZ, Caucasians, age: 16–80, RIN≥7). Common Mind Consortium (CMC) RNA-Seq postmortem DLPFC (125 CTRL, 109 SCZ, Caucasians, age, RIN as above) was used for replication of LIBD co-expression networks. 2. Expression measures/ quality control: only genes with median RPKM≥0.1 (N=23132 genes; LIBD samples) were used. Expression data was normalized by log2 transformation and adjusted for RNA quality measures (RIN, PMI, total gene assignment, proportion of mitochondrial RNA) by empirical Bayesian models. 3. Data analysis: WGCNA was applied to the adjusted expression data to construct co-expression networks, separately for LIBD- CTRL and LIBD- SCZ. Modules of co-expressed genes were detected by dynamic tree cutting method and summarized as "module eigengenes" for further analyses. 4. Modules of LIBD (CTRL; SCZ) were tested for enrichment in PGC2 genes within the 108 loci associated with the schizophrenia risk in the latest GWAS. 5. Composite module preservation statistics- median rank and Zsummary was used for assessing the preservation of LIBD SCZ modules in LIBD CTRL network and for replication of LIBD co-expression modules in the CMC independent data sets. Gene enrichment analyses for testing modules' enrichment in Gene Ontology (GO) biological processes (BP) was performed with AmiGO/ PANTHER. 17 co-expression modules were identified in each LIBD group. Module preservation analysis showed a good preservation of co-expression modules between LIBD CTRL and SCZ networks, with all but one SCZ module showing evidence for strong preservation (Zsummary≥10). PGC2 genes were overrepresented in two CTRL modules (p=0.001973, p=0.028297) enriched for GO-BP such as translation, ATP synthesis, oxidative phosphorylation, homophilic cell adhesion via plasma membrane adhesion molecules, chemical synaptic transmission, axonogenesis, and in one SCZ module (p=0.000519) enriched for RNA processing, nervous system development and modulation of synaptic transmission. 58 PGC2 genes overlapped with the SCZ module (e.g., DRD2, FURIN, TSNARE1), whereas 48 + 17 PGC2 genes were overrepresented in the two CTRL modules (e.g., CACNA1C, CHRM4, CACNA1I, TCF20). Of note, only 9 overrepresented PGC2 genes were shared by CTRL and SCZ: AMBRA1, ANKRD63, HSPA9, LRP1, PRR12, PTPRF, RERE, TOM1L2, ZDHHC5. Replication of co-expression LIBD networks in CMC data set showed evidence of weak to strong preservation (Zsummary>2, respectively ≥10) for 16/ 17 modules for each LIBD group. Our results illustrate the complexity of SCZ risk distribution within the DLPFC co-expression networks in postmortem brains of affected and unaffected individuals. Importantly, not the same PGC2 genes are members of co-expression modules in CTRL and SCZ, nor do they aggregate in similar modules. These observations may be explained by differential transcriptional regulation due to pleiotropy, epistasis, epigenetic dysregulation, or even hidden RNA quality artifacts. Future studies are necessary to elucidate the participation of SCZ risk genes in brain co-expression networks. [ABSTRACT FROM AUTHOR]

• Effects of polystyrene nanoplastic were measured at the molecular and biochemical levels. • Nanoplastic induced the overproduction of ROS. • The ERK gene in Daphnia pulex was cloned. • Oxidative stress is a possible mechanism for the nanoplastic effect. Recently, research on the biological effects of nanoplastics has grown exponentially. However, studies on the effects of nanoplastics on freshwater organisms and the mechanisms of the biological effects of nanoplastics are limited. In this study, the content of reactive oxygen species (ROS), gene and protein expression in the MAPK-HIF-1/NFkB pathway, and antioxidant gene expressions and enzyme activities were measured in Daphnia pulex exposed to polystyrene nanoplastic. In addition, the full-length extracellular signal-regulated kinases (ERK) gene, which plays an important role in the MAPK pathway, was cloned in D. pulex , and the amino acid sequence, function domain, and phylogenetic tree were analyzed. The results show that nanoplastic caused the overproduction of ROS along with other dose-dependent effects. Low nanoplastic concentrations (0.1 and/or 0.5 mg/L) significantly increased the expressions of genes of the MAPK pathway (ERK ; p38 mitogen-activated protein kinases, p38 ; c-Jun amino-terminal kinases, JNK ; and protein kinase B, AKT), HIF-1 pathway (prolyl hydroxylasedomain, PHD ; vascular endothelial growth factor, VEGF; glucose transporter, GLUT ; pyruvate kinase M, PKM ; hypoxia-inducible factor 1, HIF1), and CuZn superoxide dismutase (SOD) along with the activity of glutathione- S -transferase. As the nanoplastic concentration increased, these indicators were significantly suppressed. The protein expression ratio of ERK, JNK, AKT, HIF1α, and NFkBp65 (nuclear transcription factor-kB p65) as well as the phosphorylation of ERK and NFkBp65 were increased in a dose-dependent manner. The activities of other antioxidant enzymes (catalase, total SOD, and CuZn SOD) were significantly decreased upon exposure to nanoplastic. Combined with our previous work, these results suggest that polystyrene nanoplastic causes the overproduction of ROS and activates the downstream pathway, resulting in inhibited growth, development, and reproduction. The present study fosters a better understanding of the biological effects of nanoplastics on zooplankton. [ABSTRACT FROM AUTHOR]