On the Infra-Red Spectra of Solutions of O-Chlorophenol and Phenol

<p><b>a–d Immunoblotting of different proteins in control and study groups of gastric adenocarcinoma.</b> (a) Representative immunoblots of different target proteins in gastric epithelium of the control and study groups (from patients 1 to 4) with GAPDH as a loading control. M, marker. Relative protein abundance of NKA α1 (b), NKA β1 (c), and E-cadherin (d) in gastric epithelium of the control and study groups. The asterisks indicate a significant difference between the control and study groups. Values were expressed as the means ± SEM. A.u., arbitrary unit. ***, P < 0.001; ****, P < 0.0001.</p

Early Response of Protein Quality Control in Gills Is Associated with Survival of Hypertonic Shock in Mozambique tilapia

<div><p>The protein quality control (PQC) mechanism is essential for cell function and viability. PQC with proper biological function depends on molecular chaperones and proteases. The hypertonicity-induced protein damage and responses of PQC mechanism in aquatic organisms, however, are poorly understood. In this study, we examine the short-term effects of different hypertonic shocks on the levels of heat shock proteins (HSPs, e.g., HSP70 and HSP90), ubiquitin-conjugated proteins and protein aggregation in gills of the Mozambique tilapia (<i>Oreochromis mossambicus</i>). Following transfer from fresh water (FW) to 20‰ hypertonicity, all examined individuals survived to the end of experiment. Moreover, the levels of branchial HSPs and ubiquitin-conjugated proteins significantly increased at 3 and 24 h post-transfer, respectively. Up-regulation of HSPs and ubiquitin-conjugated proteins was sufficient to prevent the accumulation of aggregated proteins. However, the survival rate of tilapia dramatically declined at 5 h and all fish died within 7 h after direct transfer to 30‰ hypertonicity. We presumed that this result was due to the failed activation of gill PQC system, which resulted in elevating the levels of aggregated proteins at 3 and 4 h. Furthermore, in aggregated protein fractions, the amounts of gill Na⁺/K⁺-ATPase (NKA) remained relatively low when fish were transferred to 20‰ hypertonicity, whereas abundant NKA was found at 4 h post-transfer to 30‰ hypertonicity. This study demonstrated that the response of PQC in gills is earlier than observable changes in localization of ion-secreting transport proteins upon hypertonic challenge. To our knowledge, this is the first study to investigate the regulation of PQC mechanism in fish and characterize its important role in euryhaline teleost survival in response to hypertonic stress.</p></div

Distributions of the KEGG pathways.

<p>Putative proteins were mapped to the reference canonical pathways in the KEGG database. The bar chart shows the number of sequences in different pathway categories.</p

KEGG map of the oxidative phosphorylation pathway and the levels of complex expression.

<p>The error bars reflect differences in the expression of components of each complex. The black and blue lines represent the fold changes in expression of the ETC complex in fish in FW and SW, respectively, of the 18°C group compared to the 28°C group.</p

(A) Glycolysis in fish in FW at 18°C appeared vigorous because of the up-regulation of glucagon receptor, hexokinase (HK), glucokinase (GCK) and anaerobic LDHa, (B) Up-regulation of pyruvate carboxylase (PC), PDH and aerobic LDHb supported aerobic respiration in fish in SW at 18°C.

<p>Metabolites or compounds are indicated by rectangles with blue frames. Rectangles with black frames are mRNA expression of enzymes that participate in glycogen synthesis. Changes in levels of expression are shown as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134959#pone.0134959.g005" target="_blank">Fig 5</a>; purple represents up-regulation of over 5 fold in the ratio of 18°C to 28°C.</p

(A) The TCA cycle and amino acid catabolism in fish in FW were apparently altered for some specific amino acids, and the enzymes in TCA cycle were not up-regulated unless they were involved in the subsequent step of specific enzymes for amino acid catabolism. (B) In fish in SW at 18°C, mRNA expression of enzymes in the TCA cycle was mainly up-regulated.

<p>For NAD⁺ production, a higher ratio of up-regulation was found for mitochondrial nicotinamide nucleotide adenylyltransferase 3 (NMNAT-3) expression. Metabolites or compounds are indicated in blue frames. Black framed rectangles show mRNA expression of enzymes that participate in glycogen synthesis.Changes in levels of expression are shown as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134959#pone.0134959.g005" target="_blank">Fig 5</a>.</p

Relative abundance of succinate dehydrogenase subunit A mRNA in the liver (A), brain (B), gill (C) and kidney (D).

<p>N = 8 for each experiment. The asterisk indicates a significant difference by Student’s t-test (P < 0.05). Values are means ± S.E.M.</p

Classification of the annotated amino-acid sequences.

<p>Amino-acid sequences were grouped into different functional subcategories: (A) biological process (B) cellular component and (C) molecular function.</p

Length distribution of unigenes and the distribution of top-hit species.

<p>(A) Unigenes assembled from Trinity and Oases, (B) unigenes refined from CLC Genomics Workbench, and (C) the distribution of top-hit species.</p

Fish were divided into four groups: freshwater at 28°C (FW28°C), seawater at 28°C (SW28°C), freshwater at 18°C (FW18°C) and seawater at 18°C (SW18°C).

<p>The red frames indicate differential expression comparisons between different temperatures while green ones represent the salinity effects.</p