Hexarelin Protects Rodent Pancreatic Β-Cells Function from Cytotoxic Effects of Streptozotocin Involving Mitochondrial Signalling Pathways In Vivo and In Vitro

Mitochondrial functions are crucial for pancreatic β-cell survival and glucose-induced insulin secretion. Hexarelin (Hex) is a synthetic small peptide ghrelin analogue, which has been shown to protect cardiomyocytes from the ischemia-reperfusion process. In this study, we used in vitro and in vivo models of streptozotocin (STZ)-induced β-cell damage to study the protective effect of Hex and the associated mechanisms. We found that STZ produced a cytotoxic effect in a dose- and time-dependent manner in MIN6 cells (a mouse β-cell line). Hex (1.0 μM) decreased the STZ-induced damage in β-cells. Rhodamine 123 assay and superoxide DHE production assay revealed that Hex ameliorated STZ-induced mitochondrial damage and excessive superoxide activity in β-cells. In addition, Hex significantly reduced STZ-induced expression of cleaved Caspases-3, Caspases-9 and the ratio of pro-apoptotic protein Bax to anti-apoptotic protein Bcl-2 in MIN6 cells. We further examined the in vivo effect of Hex in a rat model of type 1 diabetes induced by STZ injection. Hex ameliorated STZ-induced decrease in plasma insulin and protected the structure of islets from STZ-induced disruption. Hex also ameliorated STZ-induced expression of cleaved Caspase-9 and the Bax in β-cells. In conclusion, our data indicate that Hex is able to protects β-cell mass from STZ-caused cytotoxic effects involving mitochondrial pathways in vitro and in vivo. Hex may serve as a potential protective agent for the management of diabetes.     

Anti-Inflammatory and Antinociceptive Properties of Flavonoids from the Fruits of Black Mulberry (Morus nigra L.)

We analyzed the anti-inflammatory and antinociceptive activities of total flavonoids (TF) found in black mulberry fruits. The TF content was 20.9 mg/g (dry weight). Two anthocyanins, cyanidin-3-O-glucoside (8.3 mg/g) and cyanidin-3-O-rutinoside (2.9 mg/g), were identified in the fruits by UPLC. The TF of black mulberry fruits had significant reducing power and radical (OH^-,O2.−, DPPH and ABTS) scavenging activities that was demonstrated in a dose-response curve. The TF had inhibitory activities on xylene-induced ear edema and carrageenan-induced paw edema in mice. In addition, TF had antinociceptive activities in the two nociceptive phases of formalin test. We used ELISA to detect the pro-inflammatory cytokines IL-1β, TNF-α, IFN-γ, and NO in the serum of mice. These cytokines were significantly inhibited or scavenged by TF (50 and 100 mg/kg). The results demonstrated that TF of black mulberry possess anti-inflammatory and analgesic effects that might correlate to its antioxidant activities and inhibition of pro-inflammatory cytokines.     

Tim-3 Is Upregulated in NK Cells during Early Pregnancy and Inhibits NK Cytotoxicity toward Trophoblast in Galectin-9 Dependent Pathway

NK cells accumulate at the maternal-fetal interface (MFI) and play essential roles in maintaining immune tolerance during pregnancy. The mechanisms that facilitate NK cells tolerance to fetal tissue are largely unknown. T cell Ig and mucin domain-containing protein 3 (Tim-3) is a newly defined molecule with essential immunological function in many physiological and pathological processes. Recent study showed that Tim-3 was involved in the regulation of immune tolerance at MFI. However, whether Tim-3 regulates NK cells cytotoxicity toward trophoblasts is unclear. Here, we showed Tim-3 was mainly expressed by decidual NK cells (dNK) and Tim-3 level in dNK was higher than peripheral NK cells (pNK). Tim-3⁺ dNK expressed more levels of mature markers CD94 and CD69 than Tim-3^- dNK cells and blocking Tim-3 significantly inhibited dNK IFN-γ and TNF-α secretion. Furthermore, we found TGF-β1 may contribute to such up-regulation of Tim-3 in NK cells. Interestingly, blocking Tim-3 enhanced NK cytotoxicity toward trophoblast cell line HTR-8 but not K562. We found HTR-8 expressed Tim-3 ligand Galectin-9, in contrast K562 did not. Small interfering RNA-mediated silencing of Galectin-9 expression enhanced NK cytotoxicity toward HTR-8. We further showed Tim-3/Galecin-9 inhibited NK cytotoxicity toward trophoblast partially via impairing the degranulation process. In addition, clinical data showed that abnormal Tim-3 level on pNK might be associated with recurrent spontaneous abortion (RSA). Thus, our data demonstrate Tim-3/Galectin-9 pathway maintains local tolerance by suppressing NK cytotoxicity toward trophoblasts which may represent a new immunologic tolerance mechanism at MFI.     

Simultaneous Removal of Mn(II) and Nitrate by the Manganese-Oxidizing Bacterium Acinetobacter sp. SZ28 in Anaerobic Conditions

A novel bacterium, strain SZ28, identified as Acinetobacter sp., showed anaerobic denitrification ability using Mn(II) as the electron donor. Nitrate-nitrogen concentration decreased from nearly 16.52–mg L^?1 to 4.4–mg L^?1, without accumulation of nitrite as an intermediate, with a maximum of 0.063–mg NO₃^?-N L^?1 h^?1, reaching a peak of 0.085–mg NO₃^?-N L^?1 h^?1 in sodium acetate. The nitrate removal rate reached 0.067–mg NO₃^?-N L^?1 h^?1, 0.059–mg NO₃^?-N L^?1 h^?1, and 0.078 mg NO₃^?-N L^?1 h^?1 using Mn(II), S(II), and Fe(II) as electron donors, respectively. The optimum pH was 6.0, with a removal rate of 0.063–mg NO₃^?-N L^?1 h^?1     

Estimation of genetic parameters for disease‐resistance traits in Cynoglossus semilaevis (Günther, 1873)

The objective of this study was to estimate genetic parameters for three traits based on 28 Cynoglossus semilaevis families approximately 6 months of age (at least 5 cm total length), including trait_1 (survival of 26 families, 3434 individuals in total subjected to challenge tests with Edwardsiella tarda), trait_2 (survival of 20 families, 2016 individuals in total subjected to challenge tests with Vibrio anguillarum) and trait_3 (survival of 27 families, 9340 individuals tagged at circa 180 days of age and reared in indoor ponds for circa another 5 months). The result showed that there were large differences in the survival of the families after challenge (11.11–65.31% for E. tarda and 9.18–70.54% for V. anguillarum). Additionally, the survival of families reared in indoor ponds was also different, varying from 21.00% to 73.67%. Heritabilities of the three traits varied from 0.14 to 0.26, as estimated by the linear model (LM) and the threshold model (TM). The trait_1 heritabilities (0.26 and 0.19 estimated by LM and TM) were higher than those of the others (0.20 and 0.23 estimated by LM, 0.14 and 0.19 estimated by TM). The estimates of heritabilities using LM were consistently higher than those of TM in this study. There were significant medium genetic correlations of 0.44 and 0.42 between trait_1 and trait_2 obtained from LM and TM (P < 0.05). However, very low and non‐significant genetic correlations of trait_1 and trait_3 (?0.10 for LM, ?0.05 for TM), as well as those of trait_2 and trait_3 (0.05 for LM, 0.04 for TM) were obtained. Therefore, indirect selection for trait_1 and trait_2 was effective, but almost ineffectual for trait_1 and trait_3 as well as trait_2 and trait_3. Otherwise, there was no significant difference in the predictive abilities of LM and TM. Two families resistant to both Edwardsiella tarda and Vibrio anguillarum were selected plus one family resistant to both Vibrio anguillarum and naturally infected by unknown pathogens through family selection. As there was very low and non‐significant genetic correlation of trait_3 and trait_1 as well as trait_2, superior strains are anticipated with the ability to resist two or more kinds of diseases, through the crossing of families selected for the three traits described above. The results support the hypothesis that genetic variation exists for disease survival, which could be used to design a breeding program for selecting strains of Cynoglossus semilaevis with high disease resistance.