Protein and lipid composition of sperm plasma membrane are modified as these gametes continue to mature during their transit along the spermatic tract. Our previous study revealed that during its journey through the spermatic duct of the black tiger prawn, Penaeus monodon, sperm cholesterol content decreases through the action of lipid-binding proteins within the luminal environment. In this study, the full cDNA sequence of epididymal secretory protein E1 (HE1), or Niemann-Pick C2 (NPC2), was cloned from P. monodon (termed Pmnpc2), and its conserved cholesterol/lipid-binding domain was characterized. The putative tertiary structure of PmNPC2 showed high similarity with the structure of Bos taurus NPC2. Pmnpc2 is expressed in many tissues, including the spermatic tract (i.e., testis, vas deferens, terminal ampoule) and the female thelycum. In situ hybridization revealed the presence of Pmnpc2 transcripts in the vas deferens, terminal ampoule, and thelycum epithelia, suggesting that PmNPC2 could be secreted into the lumen of the spermatic duct. A recombinant hexahistidine-tagged PmNPC2 (rPmNPC2-6His) was able to bind cholesterol and sperm lipid extracts, while co-incubation of sperm from the vas deferens with rPmNPC2-6His resulted in the depletion of cholesterol from these gametes. Together, these results suggest that PmNPC2 participates in sperm cholesterol efflux during the sperm maturation process in P. monodon. Mol. Reprod. Dev. 83: 259-270, 2016. © 2016 Wiley Periodicals, Inc.

Turbostratic graphene is a multilayer graphene, which has exotic electrical properties similar to those of monolayer graphene due to the low interlayer interaction. Additionally, the stacking structure of the turbostratic multilayer graphene can decrease the effect of attachment of charge impurities and surface roughness. This paper explores the growth of high-purity and high-quality turbostratic graphene with different interlayer spacings by calcining ferric chloride and sucrose at 1000 °C for 1 h under an argon atmosphere. X-ray diffraction patterns and Raman results imply that the turbostratic graphene contains two different interlayer spacings: 3.435 and 3.55 Å. The 3.55 Å turbostratic graphene is on top of the 3.435 Å turbostratic graphene, and there is an AB stacking pattern between the topmost graphene layer of 3.435 Å turbostratic graphene and the first graphene layer of the 3.55 Å turbostratic graphene, with an interlayer spacing of 3.35 Å. The two different interlayer spacings of turbostratic graphene arise from different cooling rates between the higher temperature ranges (>700 °C) and lower temperatures (<700 °C).

The growth of turbostratic stacked graphene using waste ferric chloride solution as a feedstock.

Purpose: To evaluate the toxicity of the dried seaweed, Sargassum plagiophyllum, extract (SPE) pretreatment in constipated mice.Methods: The dried seaweed powder was mixed with distilled water and extracted by autoclave at 121°C. Antioxidant activity of the extract was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Human normal colon cells were pretreated with SPE at 0 - 100 μg/mL for 24 h before challenging them with 100 μM hydrogen peroxide (H2O2). Intracellular reactive oxygen species (ROS) were quantified using 2',7'- dichlorodihydrofluorescein diacetate (H2DCFDA). Male ICR mice were pretreated for 14 consecutive days with SPE at 100, 500 and 1,000 mg/kg or lactulose at 500 mg/kg. Body weight and food intake were recorded daily. Constipation was induced with loperamide on days 12, 13 and 14 and fecal pellets evacuated over a 4-hr period. The ileum, liver, kidney, and spleen were collected for histopathological examination.Results: The IC50 for the radical scavenging capacity of SPE was 343.90 ± 4.21 μg/mL compared to 14.14 ± 0.71 μg/mL for ascorbic acid. Pretreatment with SPE was significantly reduced ROS production in human normal colon cells. Oral administration of all doses of SPE and lactulose for 14 consecutive days had no effect on food intake or body weight when compared to the normal control group. Defecation was significantly more frequent in mice pretreated with SPE at 100 mg/kg than in the constipation control group. Histopathological examination of the ileum, liver, kidney and spleen of pretreated constipated mice revealed no toxic effect from either SPE or lactulose. On the other hand, the loss of mucus-producing cells in the ileum of constipated mice was significantly lower in mice pretreated with SPE.Conclusions: These findings support the safety of SPE supplementation and may broaden itsapplication in clinical fields as an alternative drug or supplement for constipation management.

Contaminations of chemicals in foods and drinks are raising public concerns. Among these, styrene, a monomer for plastic production, receives increasing interest due to its ability to leach from the packaging and contaminate in foods and drinks causing many health problems. The present study was designed to investigate the effects of styrene monomer (STR) and its metabolite styrene oxide (STO) on C2C12 myoblast proliferation and differentiation. Based on an MTT assay, both STR and STO showed no cytotoxic effect at 10–100 μM. However, at 50–100 μM STO, but not STR, significantly inhibited cell proliferation. The STO-treated cells were accumulated in S-phase of cell cycles as revealed by flow cytometry. The antioxidant enzyme (catalase and superoxide dismutase) activities and the gene expressing these enzymes of the arrested cells were decreased and ultimately led to nuclear condensation and expression of apoptotic markers such as cleaved caspase-3 and-9, but not cleaved caspase-8. In addition, STO significantly suppressed myogenic differentiation by decreasing both the number and size of differentiated myotubes. Biochemical analysis showed attenuations of total protein synthesis and myosin heavy chain (MHC) protein expression. In conclusion, a metabolite of styrene, STO, leached from plastic packaging of foods and beverages suppressed both myoblast proliferation and differentiation, which would affect skeletal muscle development and regeneration.