Proximate composition, amino acid and fatty acid composition of fish maws

<div><p>Fish maws are commonly recommended and consumed in Asia over many centuries because it is believed to have some traditional medical properties. This study highlights and provides new information on the proximate composition, amino acid and fatty acid composition of fish maws of <i>Cynoscion acoupa</i>, <i>Congresox talabonoides</i> and <i>Sciades proops</i>. The results indicated that fish maws were excellent protein sources and low in fat content. The proteins in fish maws were rich in functional amino acids (FAAs) and the ratio of FAAs and total amino acids in fish maws ranged from 0.68 to 0.69. Among species, croaker <i>C. acoup</i>a contained the most polyunsaturated fatty acids, arachidonic acid, docosahexaenoic acid and eicosapntemacnioc acid, showing the lowest value of index of atherogenicity and index of thrombogenicity, showing the highest value of hypocholesterolemic/hypercholesterolemic ratio, which is the most desirable.</p></div

Primer sequences for amplification.

<p>Primer sequences for amplification.</p

<i>In situ</i> detection of PmChk1 hybridization.

<p>The hepatopancreas was harvested after dsRNA-Chk1 administration. Brown dots denote positive reactions (arrows). A represents a negative control. B represents the collected hepatopancreas after dsRNA-GFP injection. C represents the collected hepatopancreas after dsRNA-Chk1 injection. Scale bar = 30.</p

Relative expression of <i>PmChk1</i> in different tissues at stage II (chromatin nucleolus).

<p>Relative expression of <i>PmChk1</i> in different tissues according to quantitative real-time PCR using <i>EF-1α</i> as an internal reference. Vertical bars represent the mean ± SD (n = 3 for each group). Different letters above the vertical bars denote significant differences (<i>P</i> < 0.05).</p

The gene information of PmChk1.

<p>(A) Full-length cDNA of <i>PmChk1</i> (schematic). The deduced amino-acid sequence is depicted underneath the nucleotide sequence. Initiation code (ATG) and termination code (TAA) are denoted by boxes. Polyadenylation signal sequence (AATAAA) is emboldened. (B) Multiple alignment of the deduced amino-acid sequences of Chk1 from <i>P</i>. <i>monodon</i> and other species. A sequence logo denoting similarity is shown at the top of alignments, and numbers of amino acids are shown on the right-hand side of alignments. GenBank numbers of Chk1 were: <i>P</i>. <i>monodon</i>: KU958380; <i>Homo sapiens</i>: AAC51736.1; <i>Poeciliopsis prolifica</i>: JAO88875.1; <i>Mus musculus</i>: AAC53334.1; <i>Daphnia pulex</i>: AGN95867.1; <i>Melipona quadrifasciata</i>: KOX69887.1; <i>Cyphomyr mexcostatus</i>: KYN05919.1. The S-TKc domain is indicated with a red box. (C) Neighbor-joining phylogenetic tree of E2F-2s based on amino-acid sequences. Confidence in each node was evaluated by 2000 bootstrap replicates using Mega v5.03. (D) Comparison of the genomic DNA sequence encoding Chk1 in <i>P</i>. <i>monodon</i>. Green-shaded rectangles denote exons, gray horizontal lines denote introns, and the numbers reflect the length (in bp) of exons and introns.</p

Relative expression of <i>PmChk1</i> in the ovaries and hepatopancreas of <i>P</i>. <i>monodon</i> after dsRNA-RBL and dsRNA-p53 treatment.

<p>(A) Relative expression of <i>PmChk1</i> in ovaries. (B) Relative expression of <i>PmChk1</i> in the hepatopancreas. (C) Relative expression of <i>PmChk1</i> in ovaries. (D) Relative expression of <i>PmChk1</i> in the hepatopancreas. Ovary and hepatopancreas tissues harvested from <i>P</i>. <i>monodon</i> injected with dsRNA-RBL were compared with respect to expression of <i>PmChk1</i> mRNA (relative to EF-1α) using the Student’s <i>t</i>-test. Vertical bars represent the mean ± SD (n = 3 for each group). Significant differences from controls are denoted: **<i>P</i> < 0.01, *<i>P</i> < 0.05.</p

Nucleotide sequence of <i>PmChk1</i> demonstrating the 5ʹ upstream genomic sequence.

<p>Numbering of the nucleotide sequence commences from the transcription start site (+1) (arrow) and proceeds as positive numbers in the 3ʹ direction and as negative numbers in the 5ʹ direction. The putative binding sequence motifs for transcription factors and ribosomal promoter site are underlined.</p

Relative expression of <i>PmCDC2</i> and <i>PmCyclin B</i> in the ovaries and hepatopancreas of <i>P</i>. <i>monodon</i> after dsRNA-Chk1 treatment.

<p><b>(A)</b> Relative expression of <i>PmCDC2</i> in ovaries. (B) Relative expression of <i>PmCDC2</i> in the hepatopancreas. (C) Relative expression of <i>PmCyclin B</i> in ovaries. (D) Relative expression level of <i>PmCyclin B</i> in the hepatopancreas. Vertical bars represent the mean ± SD (n = 3 for each group). Significant differences from controls are denoted: **<i>P</i> < 0.01, *<i>P</i> < 0.05.</p

Genomic structure, expression, and functional characterization of checkpoint kinase 1 from <i>Penaeus monodon</i>

<div><p><i>Chk1</i> is a cell-cycle regulator. <i>Chk1</i> has been identified in organisms ranging from yeast to humans, but few researchers have studied <i>Chk1</i> in shrimps. We cloned <i>Chk1</i> from the black tiger shrimp (<i>Penaeus monodon</i>). The full-length cDNA sequence of <i>PmChk1</i> had 3,334 base pairs (bp), with an open reading frame of 1,455 bp. The complete genomic sequence of <i>PmChk1</i> (11,081 bp) contained 10 exons separated by nine introns. qRT-PCR showed that <i>PmChk1</i> was highly expressed in the ovaries and gills of <i>P</i>. <i>monodon</i>. The lowest <i>PmChk1</i> expression was noted in stage III of ovarian development in <i>P</i>. <i>monodon</i>. <i>PmChk1</i> expression decreased significantly after injection of 5-hydroxytryptamine and eyestalk ablation in <i>P</i>. <i>monodon</i> ovaries. RNA interference experiments were undertaken to examine the expression of <i>PmChk1</i>, <i>PmCDC2</i>, and <i>PmCyclin B</i>. <i>PmChk1</i> knockdown in the ovaries and hepatopancreas by dsRNA-Chk1 was successful. The localization and level of <i>PmChk1</i> expression in the hepatopancreas was studied using <i>in situ</i> hybridization, which showed that data were in accordance with those of qRT-PCR. The Gonadosomatic Index of <i>P</i>. <i>monodon</i> after dsRNA-Chk1 injection was significantly higher than that after injection of dsRNA-GFP or phosphate-buffered saline. These data suggest that PmChk1 may have important roles in the ovarian maturation of <i>P</i>. <i>monodon</i>.</p></div