Perceived attitudes of family and peers toward same-sex marriage as a distal sexual minority stressor for gay and bisexual men in Taiwan.

BACKGROUND: To investigate whether perceived attitudes of family and peers toward same-sex marriage (SSM) is a type of distal sexual minority stressor, as defined in minority stress theory, this cross-sectional study examined the associations of perceived attitudes of family and peers toward SSM with perceived sexual stigma from family and peers, internalized homonegativity, and mental health problems (e.g., depression, loneliness, anxiety) among gay and bisexual men in Taiwan. METHODS: We recruited 400 gay and bisexual men and assessed their perceived attitudes of family and peers toward SSM; perceived sexual stigma from family and peers; internalized homonegativity; and severity of depression, loneliness, and anxiety. RESULTS: Perceived attitudes of family and peers toward SSM (1) significantly correlated with various aspects of perceived sexual stigma from family and peers and (2) were significantly associated with internalized homonegativity, depression, loneliness, and anxiety. CONCLUSIONS: Perceived attitudes of family and peers toward SSM matched the characteristics of a distal sexual minority stressor, and as a new type of distal sexual minority stressor for lesbian, gay, and bisexual individuals, these perceived attitudes and related stress warrant greater attention from mental health professionals for the development of intervention programs.

Pexophagy is critical for fungal development, stress response, and virulence in Alternaria alternata.

Alternaria alternata can resist high levels of reactive oxygen species (ROS). The protective roles of autophagy or autophagy-mediated degradation of peroxisomes (termed pexophagy) against oxidative stress remain unclear. The present study, using transmission electron microscopy and fluorescence microscopy coupled with a GFP-AaAtg8 proteolysis assay and an mCherry tagging assay with peroxisomal targeting tripeptides, demonstrated that hydrogen peroxide (H2 O2 ) and nitrogen depletion induced autophagy and pexophagy. Experimental evidence showed that H2 O2 triggered autophagy and the translocation of peroxisomes into the vacuoles. Mutational inactivation of the AaAtg8 gene in A. alternata led to autophagy impairment, resulting in the accumulation of peroxisomes, increased ROS sensitivity, and decreased virulence. Compared to the wild type, ΔAaAtg8 failed to detoxify ROS effectively, leading to ROS accumulation. Deleting AaAtg8 down-regulated the expression of genes encoding an NADPH oxidase and a Yap1 transcription factor, both involved in ROS resistance. Deleting AaAtg8 affected the development of conidia and appressorium-like structures. Deleting AaAtg8 also compromised the integrity of the cell wall. Reintroduction of a functional copy of AaAtg8 in the mutant completely restored all defective phenotypes. Although ΔAaAtg8 produced wild-type toxin levels in axenic culture, the mutant induced a lower level of H2 O2 and smaller necrotic lesions on citrus leaves. In addition to H2 O2 , nitrogen starvation triggered peroxisome turnover. We concluded that ΔAaAtg8 failed to degrade peroxisomes effectively, leading to the accumulation of peroxisomes and the reduction of the stress response. Autophagy-mediated peroxisome turnover could increase cell adaptability and survival under oxidative stress and starvation conditions.

A zinc finger suppressor involved in stress resistance, cell wall integrity, conidiogenesis, and autophagy in the necrotrophic fungal pathogen Alternaria alternata.

The tangerine pathotype of Alternaria alternata can withstand high-level reactive oxygen species (ROS). By analyzing loss- and gain-of-function mutants, this study demonstrated that a Cys2His2 zinc finger-containing transcription regulator, A. alternata Stress Response Regulator 1 (AaSRR1), plays a negative role in resistance to peroxides and singlet-oxygen-generating compounds. AaSRR1 plays no role in cellular susceptibility or resistance to superoxide-producing compounds. AaSRR1 also negatively regulates conidiogenesis, maintenance of cell wall and membrane integrities, and chitin biosynthesis. Some wild-type hyphae displayed necrosis after exposure to 30 mM H2O2, whereas AaSRR1 deficient mutant (ΔAaSRR1) hyphae had visible granules and vacuoles. sGFP-AaATG8 proteolysis assays revealed that H2O2 and starvation could trigger autophagy formation in both wild type and ΔAaSRR1. Autophagy occurred at higher rates in ΔAaSRR1 than wild type under both conditions, particularly after H2O2 treatments, indicating that autophagy might contribute to ROS resistance. Upon exposure to H2O2 or under starvation, AaSRR1 was translocated into the nucleus, even though the expression of AaSRR1 was decreased. AaSRR1 is required for vegetative growth but is dispensable for fungal virulence as assayed on detached calamondin leaves. AaSRR1 suppressed the expression of the gene encoding a HOG1 mitogen-activated protein (MAP) kinase implicated in ROS resistance. Mutation of AaSRR1 increased catalase activity but decreased superoxide dismutase activity, leading to fewer ROS accumulation in the cytosol. Nevertheless, our results indicated that AaSRR1 is a transcription suppressor for ROS resistance. This study also revealed tradeoffs between stress responses and hyphal growth in A. alternata.

Baculovirus surface display of the HA protein of H5N2 avian influenza virus and its immunogenicity against a lethal challenge with H5N1 virus in chickens.

In the present study, we have generated several H5N2 HA recombinant baculoviruses for production of a HA subunit vaccine against the lethal H5N2 avian influenza virus (AIV). The effective display of functional HA on the cell membrane and baculoviral envelope was examined. Our results reveal that chickens immunized with the chimeric AIV HA protein fused with the baculovirus gp64 cytoplasmic domain (CTD) induced higher HI titer. To further increase the expression level of the H5N2 AIV HA protein, the HA gene of H5N2 AIV was amplified and cloned into three novel baculovirus surface display vectors BacDual DisplayEGFP-2HA, BacDual DisplayEGFP-3HA, BacDual DisplayEGFP-4HA which contains multiple expression cassettes for higher level display of HA proteins on the cell membrane and baculovirus envelope. To determine the optimum conditions for producing HA protein, various MOI, infection times, and shaker times for virus transfection were tested. Our results reveal that the conditions of an MOI of 5, 3 day post infection, and 15 min of shaker time have higher efficiency for HA protein production. Our results reveal that the baculovirus surface display vector pBacDual DisplayEGFP-4HA increases significantly the expression level of the H5N2 AIV HA protein. Chickens that received two doses of BacDual DisplayEGFP-4HA cell lysates formulated with Montanide ISA70 adjuvant elicited efficient immunogenicity and had an average HI titer of 7 log2 at 2 weeks post-vaccination. Challenge studies revealed that vaccinated chickens with HI titers 5 log2 were completely protected against the lethal H5N1 AIV challenge. Furthermore, HI titers could be maintained at 5 log2 for 20 weeks for laying hens. This study suggests that the HA protein expression from the baculovirus surface display system could be a safe and efficacious subunit vaccine for chickens.

Enzymatic hydrolysates obtained from Trametes versicolor polysaccharopeptides protect human skin keratinocyte against AAPH-induced oxidative stress and inflammatory.

BACKGROUND: Polysaccharopeptides (PSPs) extracted from Trametes versicolor show antitumor, anti-inflammatory, and immunomodulation effects. According to our previous report, the enzymatic hydrolysates obtained from T versicolor PSPs by 80 U/mL ß-1,3-D-glucanase (PSPs-EH80) did not change the functional groups of PSPs but enhanced their antioxidative activities. However, the mechanism elevating the antioxidant and anti-inflammatory effect of PSPs-EH80 is not clear. AIMS: This research focused on the protective mechanism(s) of PSPs-EH80 against free radical and 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced oxidative damage in human keratinocyte (HaCaT) cells. METHODS: We evaluated the anti-inflammatory potential of PSPs-EH80 by assessing its free radical-induced oxidative damage. Using the HaCaT cell as the experimental system, we tested the protective effects of PSPs-EH80 on a model of AAPH-induced cellular oxidative damage through the assessment of cell survival rate. Heme oxygenase 1 (HO-1), nuclear factor erythroid 2-related factor 2 (Nrf2), cyclooxygenase-2 (COX-2), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase were determined using MTT assays and Western blotting. RESULTS: We demonstrated that PSPs-EH80 significantly enhanced keratinocyte viability, and augmented the antioxidant HO-1 expressions through upregulation of the Nrf2, compared with PSPs. Furthermore, PSPs-EH80 significantly reduced AAPH-induced COX-2 expressions through downregulation of the ERK, p38, and NF-κB signaling pathways. CONCLUSION: The PSPs-EH80 exhibits a stronger antioxidant and anti-inflammatory capacity than PSPs. Therefore, PSPs-EH80 could be effective for attenuating free radical-induced oxidative damage in human skin and can be applied widely in the fields of cosmetics and medicine.

Chemical analysis, moisture-preserving, and antioxidant activities of polysaccharides from Pholiota nameko by fractional precipitation.

This study explored Pholiota nameko (P. nameko) polysaccharide fractions, PNP-40, PNP-60, and PNP-80, purified by gradient concentrations of ethanol (40%, 60%, and 80% (v/v)). The physicochemical properties, functional group composition, moisture-preserving, and antioxidant ability were determined. The results indicate that the polysaccharide contents of PNP-40, PNP-60, and PNP-80 are 45.12%, 78.04%, and 72.22%, respectively, while the ß-glucan, protein, and uronic acid contents are 20.20%, 12.20%, and 10.15%, respectively; 11.24%, 14.53%, and 26.94%; and 5.99%, 7.73%, and 3.78%. Furthermore, PNP-60 has better moisture absorption, while PNP-80 has better antioxidant ability and H2O2-injury resistance activity. Monosaccharide composition analysis shows that P. nameko belongs to heteropolysaccharides, which consists of galactose, glucose, and mannose with different types and ratios, and the molecular weight are distributed at 4.40-333.49kDa. It was found that different polysaccharide fractions have the potential to be a moisturizer and an antioxidant, and their active ingredients could be used in the development of cosmetic ingredients.

Facile Functionalization of Polymer Surfaces in Aqueous and Polar Organic Solvents via 3-Mercaptopropylsilatrane.

Surface modification of a polymer substrate with a mercapto functionality is crucial in many applications such as flexible circuitry and point-of-care biosensors. We present here a novel bifunctional molecular adhesive, 3-mercaptopropylsilatrane (MPS), as an interface between polymer and metal surfaces. Under ambient conditions, surface modification of polymer surfaces with a mercapto functionality can be achieved with low concentration (0.46 mM) of MPS in aqueous solvent (50% ethanol) in a short time (<30 min). Three popular polymers for optical sensors, polycarbonate, polyethylene terephthalate, and poly(methyl methacrylate), were employed as substrates, and MPS films formed on these substrates were examined and compared with that on a glass substrate. The films were characterized by UV-vis spectroscopy, water contact angle, X-ray photoelectron spectroscopy, and atomic force microscopy. MPS was also used as a bifunctional linker for the construction of a gold nanoparticle (AuNP) sub-monolayer on these polymer surfaces. Under optimized preparation conditions, the absorbance and full width at half-maximum of the plasmon band are comparable to those of a AuNP-modified glass substrate. Hence, MPS may have a potential to be a key component in polymer substrate-based localized surface plasmon resonance sensors. A self-catalytic surface reaction mechanism is also proposed to account for the results. As compared to a glass surface with a high number of silanol groups, the successful formation of an MPS film on polymer surfaces with relatively few reactive sites is probably due to the lateral polymerization of MPS starting from a condensed MPS molecule on a reactive site of a polymer surface.

Real-time 3D particle manipulation visualized using volume holographic gratings.

Holographic optical tweezers (HOTs) extend optical trapping into three dimensions. Volume imaging then becomes a concern as trapped objects are easily moved out of focus of the imaging objective lens. Here we demonstrate a three-dimensional real-time interactive optical trapping, manipulating, and imaging system based on HOTs incorporated with volume holographic microscope. Intensity information about the trapped objects at multiple depths can be captured in a single measurement. This method is compatible with most imaging modes such as bright-field and fluorescence.

Mapping polarization induced surface band bending on the Rashba semiconductor BiTeI.

Surfaces of semiconductors with strong spin-orbit coupling are of great interest for use in spintronic devices exploiting the Rashba effect. BiTeI features large Rashba-type spin splitting in both valence and conduction bands. Either can be shifted towards the Fermi level by surface band bending induced by the two possible polar terminations, making Rashba spin-split electron or hole bands electronically accessible. Here we demonstrate the first real-space microscopic identification of each termination with a multi-technique experimental approach. Using spatially resolved tunnelling spectroscopy across the lateral boundary between the two terminations, a previously speculated on p-n junction-like discontinuity in electronic structure at the lateral boundary is confirmed experimentally. These findings realize an important step towards the exploitation of the unique behaviour of the Rashba semiconductor BiTeI for new device concepts in spintronics.

Baculovirus as a vaccine vector.

Baculovirus is extensively utilized as an excellent tool for production of recombinant protein in insect cells. Baculovirus infects insects in nature and is non-pathogenic to humans. In addition to insect cells, baculovirus is capable of transducing a broad range of animal cells. Due to its biosafety, large cloning capacity, low cytotoxicity, and non-replication nature in the transduced cells as well as the ease of manipulation and production, baculovirus has been utilized as RNA interference mediators, gene delivery vectors, and vaccine vectors for a wide variety of applications. This article focuses on the utilization of baculoviruses as vaccine vectors to prepare antigen or subunit vaccines.