Keyhole limpet hemocyanin induces innate immunity via Syk and Erk phosphorylation

Hemocyanin is an extracellular respiratory protein containing copper in hemolymph of invertebrates, such as Mollusk and Arthropod. Keyhole limpet hemocyanin (KLH) is one of hemocyanins and has many years of experience for vaccine developments and immunological studies in mammals including human. However, the association between KLH and the immune systems, especially the innate immune systems, remains poorly understood. The aim of this study is to clarify the direct effects of KLH on the innate immune systems. KLH activated an inflammation-related transcription factor NF-kB as much as lipopolysaccharide (LPS) in a human monocytic leukemia THP-1 reporter cell line. We have found that the KLH-induced NF-kB activation is partially involved in a spleen tyrosine kinase (Syk) pathway. We have also successfully revealed that an extracellular signal-regulated kinase (Erk), a member of mitogen-activated protein kinases, is located in an upstream of NF-kB activation induced by KLH. Furthermore, a Syk phosphorylation inhibitor partially suppressed the Erk activation in KLH-stimulated THP-1. These results suggest that both Syk and Erk associate with the KLH-induced NF-kB activation in the human monocyte

Ultrastructural Evidence for Temperature-Dependent Ca2+ Release from Fish Sarcoplasmic Reticulum During Rigor Mortis

The release or leakage of ca2+ from the sarcoplasmic reticulum (SR) during rigor mortis of fish muscle was investigated by transmission electron microscopy using pyroantimonate and related biochemical changes. Ca2+ -pyroantimonate deposits were observed in the SR immediately after spiking the fish. At the onset of rigor for fish stored at 0°C, no deposits were found in the SR; however, fish stored for the same period at woe which were still in the pre-rigor state, clearly showed Ca2+ deposits in the SR. In association with the Ca2+ translocation, ATP degraded faster at 0 c than at 10 °C, probably due to enhancement of myofibrillar ATPase activity by the increasing Ca2 + concentrations. Therefore, rapid Ca2+ release from the SR at 0°C seemed to trigger the acceleration of fish rigor mort is at this temperature, analogous to the phenomenon called cold shortening

The Yeast Tor Signaling Pathway Is Involved in G2/M Transition via Polo-Kinase

The target of rapamycin (Tor) protein plays central roles in cell growth. Rapamycin inhibits cell growth and promotes cell cycle arrest at G1 (G0). However, little is known about whether Tor is involved in other stages of the cell division cycle. Here we report that the rapamycin-sensitive Tor complex 1 (TORC1) is involved in G2/M transition in S. cerevisiae. Strains carrying a temperature-sensitive allele of KOG1 (kog1-105) encoding an essential component of TORC1, as well as yeast cell treated with rapamycin show mitotic delay with prolonged G2. Overexpression of Cdc5, the yeast polo-like kinase, rescues the growth defect of kog1-105, and in turn, Cdc5 activity is attenuated in kog1-105 cells. The TORC1-Type2A phosphatase pathway mediates nucleocytoplasmic transport of Cdc5, which is prerequisite for its proper localization and function. The C-terminal polo-box domain of Cdc5 has an inhibitory role in nuclear translocation. Taken together, our results indicate a novel function of Tor in the regulation of cell cycle and proliferation

Changes in Intestinal Gene Expression of Zebrafish (Danio rerio) Related to Sterol Uptake and Excretion upon β-Sitosterol Administration

Replacement of fishmeal with plant ingredients will introduce not only plant oil and protein but also phytosterol to the fish diet. Mammals strictly restrict the uptake of phytosterol at intestinal epithelial cells by regulating the gene expressions of sterol uptake and excretion proteins; however, phytosterol is found in the fish muscle and other organs. In order to assess the ability of phytosterol uptake by the intestinal epithelial cells of fish, no-sterol diet, cholesterol-, and β-sitosterol-containing diet was separately administered to zebrafish, and the relative mRNA expressions related to sterol uptake and excretion were evaluated. Gene expression of Niemann-Pick C1-like protein 1 in the sitosterol-fed group was significantly higher than that of the cholesterol-fed group (p < 0.05). The expression of apolipoprotein A-I gene was also higher in the sitosterol-fed group than that in the no-sterol and cholesterol-fed groups. The expressions of ATP-binding cassette, sub-family G, member 5 and 8, were significantly higher in the sitosterol-fed group, compared to the no-sterol group. Regarding the gene expression of ATP-binding cassette sub-family A, member 1, the sitosterol-fed group showed higher expression level compared to the other groups (p < 0.01). These results suggest that fish should be tolerant to phytosterols in contrast to mammals

Utilization of Biological Responses of Fish and Shellfish for Improving Seafood Qualities

Quality control of seafood has been one major purpose of scientific researches on fish and shellfish. In this monograph, we deal with the utilization of biological responses in the preharvest and postharvest stages of aquatic animals, (1) responses of fish muscle energy metabolisms to environmental temperature changes in postmortem stages, (2) responses of fish and shellfish chromatophore to extrinsic stimuli, and (3) responses of fish to the oral administration of phytosterol related compounds. These approaches will not only confer the improvement of seafood qualities but the addition of extra value to seafood for human consumption. Because animals respond biologically to many factors, comprehensive understanding of biological responses of aquatic animals to environmental factors will lead to the further improvement of seafood qualities

Ferulic Acid Promotes Hypertrophic Growth of Fast Skeletal Muscle in Zebrafish Model

As a widely distributed and natural existing antioxidant, ferulic acid and its functions have been extensively studied in recent decades. In the present study, hypertrophic growth of fast skeletal myofibers was observed in adult zebrafish after ferulic acid administration for 30 days, being reflected in increased body weight, body mass index (BMI), and muscle mass, along with an enlarged cross-sectional area of myofibers. qRT-PCR analyses demonstrated the up-regulation of relative mRNA expression levels of myogenic transcriptional factors (MyoD, myogenin and serum response factor (SRF)) and their target genes encoding sarcomeric unit proteins involved in muscular hypertrophy (skeletal alpha-actin, myosin heavy chain, tropomyosin, and troponin I). Western blot analyses detected a higher phosphorylated level of zTOR (zebrafish target of rapamycin), p70S6K, and 4E-BP1, which suggests an enhanced translation efficiency and protein synthesis capacity of fast skeletal muscle myofibers. These changes in transcription and translation finally converge and lead to higher protein contents in myofibers, as confirmed by elevated levels of myosin heavy chain (MyHC), and an increased muscle mass. To the best of our knowledge, these findings have been reported for the first time in vivo and suggest potential applications of ferulic acid as functional food additives and dietary supplements owing to its ability to promote muscle growth