Hexafluoropropylene oxide dimer acid (GenX) exposure induces apoptosis in HepG2 cells.

Hexafluoropropylene oxide dimer acid, also known as GenX, is a poly- and perfluoroalkyl substance (PFAS). PFASs are nonvolatile synthetic substances that can be readily disseminated into the environment during processing and use, making them easy to implement in the soil, drinking water, and air. Compared to other PFASs, GenX has a comparatively short carbon chain length and is expected to have a lower tendency to accumulate in humans; therefore, GenX has recently been used as a substitute to other PFASs. However, the mechanisms underlying GenX action and intoxication in humans remains unclear. In this study, the apoptotic capacity of GenX in human liver cells was investigated. When representative human-derived liver cells (HepG2 cells) were treated with GenX for 12 h, cell viability was reduced, and apoptosis was greatly increased. In addition, GenX increased the generation of intracellular reactive oxygen species (ROS), indicating the induction of oxidative stress in a dose-dependent manner. GenX treatment increased the expression of major apoptosis-related genes relative to the untreated control group. This research indicates that GenX causes apoptosis through ROS mediation in HepG2 cells, which may expand our knowledge of the molecular and toxicological mechanisms of GenX.

Antibacterial Activity of Green-Synthesized Silver Nanoparticles Using Areca catechu Extract against Antibiotic-Resistant Bacteria.

In this work, the antibacterial activity of silver nanoparticles (AgNPs) synthesized using Areca catechu extracts against three species of antibiotic-susceptible and three species of resistant bacteria was investigated. The effects of this plant were more promising when compared with other medicinal plants tested. The hydrothermal extract of Areca catechu was mixed with silver nitrate to synthesize AgNPs. The synthesized particle characteristics were analyzed by UV-Vis spectrophotometry, scanning electron microscopy (SEM), dynamic light scattering (DLS), and Fourier-transform infrared spectroscopy (FT-IR). Minimum inhibitory concentration and minimum bactericidal concentration tests were conducted to confirm antibacterial activity and the results showed that AgNPs synthesized using Areca catechu extracts effectively inhibited the growth of bacterial species. Moreover, the SEM images of the bacterial species treated with AgNPs synthesized with Areca catechu extracts showed that clusters of AgNPs were attached to the surface of the bacterial cell wall, which could induce destruction of the cell membranes. The results suggest that AgNPs synthesized with Areca catechu extracts have the potential to treat antibiotic-resistant bacteria known as the major cause of nosocomial infections.

Biological effects of melatonin on human adipose­derived mesenchymal stem cells.

Mesenchymal stem cells (MSCs) are capable of differentiating into other cell types and exhibit immunomodulatory effects. MSCs are affected by several intrinsic and extrinsic signaling modulators, including growth factors, cytokines, extracellular matrix and hormones. Melatonin, produced by the pineal gland, is a hormone that regulates sleep cycles. Recent studies have shown that melatonin improves the therapeutic effects of stem cells. The present study aimed to investigate whether melatonin enhances the biological activities of human adipose­derived MSCs. The results demonstrated that treatment with melatonin promoted cell proliferation by inducing SRY­box transcription factor 2 gene expression and preventing replicative senescence. In addition, melatonin exerted anti­adipogenic effects on MSCs. PCR analysis revealed that the expression of the CCAAT enhancer binding protein a gene, a key transcription factor in adipogenesis, was decreased following melatonin treatment, resulting in reduced adipogenic differentiation in an in vitro assay. The present study also examined the effect of melatonin on the immunomodulatory response using a co­culture system of human peripheral blood mononuclear cells and MSCs. Activated T cells were strongly inhibited following melatonin exposure compared with those in the control group. Finally, the favorable effects of melatonin on MSCs were confirmed using luzindole, a selective melatonin receptor antagonist. The proliferation­promoting, anti­inflammatory effects of melatonin suggested that melatonin­treated MSCs may be used for effective cell therapy.

Localization patterns of dopamine active transporter synthesizing cells during development of brine shrimp.

There have been many studies on dopamine active transporter (DAT) in humans and laboratory animals; however, there is a lack of information on DAT in brine shrimp. In this study, we demonstrated the neuronal and nonneuronal characteristics of DAT-synthesizing (DAT+ cells) during development of brine shrimp. In neuronal cells, the DAT+ neurons in the central body and lobes of a protocerebrum (PC) controlled the deutocerebrum. The sensory cells of nauplius eyes projected their decussated axons to the PC, and the DAT+ cells at the posterior region were associated with migration and control of the 10 posterior neurons during the early nauplius stage. In nonneuronal cells, the five types of glands, that is, the salt, antennal, mandible, and accessory glands and posterior gland1 and gland2 synthesized DAT protein. In addition, the gut and rectum dilator muscles and renal cells expressed DAT protein. Thus, DAT protein acts in the development of several types of cells during development of brine shrimp.

Effects of PRELI in Oxidative-Stressed HepG2 Cells.

Protein of relevant evolutionary and lymphoid interest (PRELI) is known for preventing apoptosis by mediating intramitochondrial transport of phosphatidic acid. However, the role of PRELI remains unclear. This study has demonstrated functions of PRELI through PRELI-knockdown in hepatocellular carcinoma (HepG2) cells exposed to oxidative stress by hydrogen peroxide. Results show that PRELI has three functions in HepG2 cells with regard to oxidative stress. First, PRELI affects expressional regulation of SOD-1 and caspase-3 genes in HepG2 cells. PRELI knockdown HepG2 cells have shown up-regulation of caspase-3 and down-regulation of SOD-1. Second, PRELI suppresses mitochondrial apoptosis in HepG2 cells. Fluorescence intensity related to mitochondrial apoptosis in PRELI-knockdown HepG2 cells increased more than two-fold compared to normal HepG2 cells. Third, PRELI suppresses senescence of HepG2 cells with oxidative stress. PRELI knockdown HepG2 cells showed higher levels of senescence than normal HepG2 cells. These results suggest that PRELI is a crucial protein in the suppression of apoptosis in HepG2 cells in response to oxidative stress.

DISTINCTIVE LOCALIZATION OF GROUP 3 LATE EMBRYOGENESIS ABUNDANT SYNTHESIZING CELLS DURING BRINE SHRIMP DEVELOPMENT.

Despite numerous studies on late embryogenesis abundant (LEA) proteins, their functions, roles, and localizations during developmental stages in arthropods remain unknown. LEA proteins protect crucial proteins against osmotic stress during the development and growth of various organisms. Thus, in this study, fluorescence in situ hybridization was used to determine the crucial regions protected against osmotic stress as well as the distinctive localization of group 3 (G3) LEA(+) cells during brine shrimp development. Several cell types were found to synthesize G3 LEA RNA, including neurons, muscular cells, APH-1(+) cells, and renal cells. The G3 LEA(+) neuronal cell bodies outside of the mushroom body projected their axonal bundles to the central body, but those inside the mushroom body projected their axonal bundles toward the deutocerebrum without innervating the central body. The cell bodies inside the mushroom body received axons of the G3 LEA(+) sensory cells at the medial ventral cup of the nauplius eye. Several glands were found to synthesize G3 LEA RNA during the nauplius stages of brine shrimp, including the sinus, antennal I and II, salt, and three ectodermal glands. This study provides the first demonstration of the formation of G3 LEA(+) sinus glands at the emergence stages of brine shrimp. These results suggest that G3 LEA protein is synthesized in several cell types. In particular, specific glands play crucial roles during the emergence and nauplius stages of brine shrimp.

Silkworm 30K protein inhibits ecdysone-induced apoptosis by blocking the binding of ultraspiracle to ecdysone receptor-B1 in cultured Bm5 cells.

This study investigates the mechanism through which increased 30K protein inhibits ecdysone-induced apoptosis in the Bm5 silkworm ovarian cell line. Treatment of Bm5 cells with 20-hydroxyecdysone (20E) after transfection with the pIZT/V5-His control vector triggered apoptosis, but 20E treatment did not trigger apoptosis in Bm5 cells transfected with the pIZT/30K/V5-His vector. To confirm its inhibitory effect on apoptosis, 30K protein was first purified from Escherichia coli transformed with a 30K expression vector and used to generate specific antibodies in mice. Anti-30K antiserum was used to confirm synthesis of the 30K protein in pIZT/30K/V5-His-transfected Bm5 cells and to detect 30K protein binding to the ecdysone receptor-B1 (EcR-B1). Anti-30K antiserum was used to immunoprecipitate protein complexes containing 30K from Bm5 cells transfected with pIZT/30K/V5-His vector and treated with 20E. We observed that 30K proteins bound primarily to the EcR-B1 and not to ultraspiracle (USP). Reciprocal immunoprecipitation of EcR-B1-containing complexes from Bm5 cells transfected with control pIZT/V5-His vector and treated with 20E showed that EcR-B1 bound to USP in the absence of 30K but did not bind to USP in pIZT/30K/V5-His-transfected Bm5 cells. These results demonstrate that 30K proteins block USP binding to EcR-B1 through formation of a 30K/EcR-B1 complex, resulting in inhibition of 20E-induced Bm5 cell apoptosis.

Increase of 30K protein in identified motoneurons by hemolymph results in inhibition of programmed cell death in silkworm, Bombyx mori (Lepidoptera, Bombycidae).

This study demonstrates that a 30K protein was gradually synthesized in primary-cultured motoneurons from the accessory planta retractor (APR) of the 6th abdominal ganglion (APR6) in silkworm ventral ganglia through stimulation of hemolymph. An increase in 30K protein synthesis resulted in an inhibition of programmed cell death (PCD) of APR6 motoneurons. The 30K protein was gradually synthesized from the 30Kc6 gene of identified APR6s in day-6 4th instars to day-9 5th instar larvae, but synthesis of the 30K protein ceased in isolated APR6s of day-1 pupa, which normally begin to undergo PCD. When pupal APR6s were treated with larval hemolymph, however, the 30K protein was synthesized suggesting the existence of an anti-PCD factor in the larval hemolymph. An increase of 30K protein within the APR6s was confirmed by antiserum made against the recombinant 30K protein that originated from the APR 30Kc6 gene. Larval APR6, in which PCD was induced with 20-hydroxyecdysone (20E) added to the primary culture, exhibited some PCD characteristics of shrinkage of cell bodies, axonal fragmentation and loss of mitochondrial function. These results provide new insights on the survival or PCD of insect motoneurons through stimulation of hemolymph.