New cell lines from Lymantria xylina (Lepidoptera: Lymantriidae): characterization and susceptibility to baculoviruses

Four new cell lines, designated as NTU-LY-1 to -4, respectively, were established from the pupal tissues of Lymantria xylina Swinhoe (Lepidoptera: Lymantriidae). These cell lines have been cultured approximately 80 passages during 2 years in TNM-FH medium supplemented with 8% fetal bovine serum, at a constant temperature of 28 degrees C. Each line consists of three major morphological types: round cells, spindle-shaped cells, and giant cells. The characterization of these cell lines showed that they are different from previously established lines derived from related Lepidopteran species. All new lines were susceptible to the L. xylina multiple nucleopolyhedrovirus (LyxyMNPV) and appeared to have a good potential for studying this virus.     

Decoy receptor 3 suppresses TLR2-mediated B cell activation by targeting NF-κB

Decoy receptor 3 (DcR3) is a soluble protein in the TNFR superfamily. Its known ligands include Fas ligand, homologous to lymphotoxin, showing inducible expression, and competing with HSV glycoprotein D for herpes virus entry mediator, a receptor expressed by T lymphocytes, TNF-like molecule 1A, and heparan sulfate proteoglycans. DcR3 has been reported to modulate the functions of T cells, dendritic cells, and macrophages; however, its role in regulating B cell activation is largely unknown. In this study, we found that the DcR3.Fc fusion protein bound to human and mouse B cells and suppressed the activation of B cells. DcR3.Fc attenuated Staphylococcus aureus, IgM-, Pam(3)CSK(4)-, and LPS-mediated B cell proliferation but did not affect cytokine-induced B cell growth. In the presence of these mitogens, DcR3.Fc did not induce B cell apoptosis, suggesting that DcR3 may inhibit the signal(s) important for B cell activation. Because the combination of Fas.Fc, LT-βR.Fc (homologous to lymphotoxin, showing inducible expression, and competing with HSV glycoprotein D for herpes virus entry mediator, a receptor expressed by T lymphocytes receptor), and DR3.Fc (TNF-like molecule 1A receptor) did not suppress B cell proliferation and because the biological effect of DcR3.Fc on B cells was not blocked by heparin, we hypothesize that a novel ligand(s) of DcR3 mediates its inhibitory activity on B cells. Moreover, we found that TLR2-stimulated NF-κB p65 activation and NF-κB-driven luciferase activity were attenuated by DcR3.Fc. The TLR2-induced cytokine production by B cells was consistently reduced by DcR3. These results imply that DcR3 may regulate B cell activation by suppressing the activation of NF-κB.     

Low-Dose Dextromethorphan,a NADPH Oxidase Inhibitor,Reduces Blood Pressure and Enhances Vascular Protection in Experimental Hypertension

Background

Vascular oxidative stress may be increased with age and aggravate endothelial dysfunction and vascular injury in hypertension. This study aimed to investigate the effects of dextromethorphan (DM), a NADPH oxidase inhibitor, either alone or in combination treatment, on blood pressure (BP) and vascular protection in aged spontaneous hypertensive rats (SHRs).

Methodology/Principal Findings

Eighteen-week-old WKY rats and SHRs were housed for 2 weeks. SHRs were randomly assigned to one of the 12 groups: untreated; DM monotherapy with 1, 5 or 25 mg/kg/day; amlodipine (AM, a calcium channel blocker) monotherapy with 1 or 5 mg/kg/day; and combination therapy of DM 1, 5 or 25 mg/kg/day with AM 1 or 5 mg/kg/day individually for 4 weeks. The in vitro effects of DM were also examined. In SHRs, AM monotherapy dose-dependently reduced arterial systolic BP. DM in various doses significantly and similarly reduced arterial systolic BP. Combination of DM with AM gave additive effects on BP reduction. DM, either alone or in combination with AM, improved aortic endothelial function indicated by ex vivo acetylcholine-induced relaxation. The combination of low-dose DM with AM gave most significant inhibition on aortic wall thickness in SHRs. Plasma total antioxidant status was significantly increased by all the therapies except for the combination of high-dose DM with high-dose AM. Serum nitrite and nitrate level was significantly reduced by AM but not by DM or the combination of DM with AM. Furthermore, in vitro treatment with DM reduced angiotensin II-induced reactive oxygen species and NADPH oxidase activation in human aortic endothelial cells.

Conclusions/Significance

Treatment of DM reduced BP and enhanced vascular protection probably by inhibiting vascular NADPH oxidase in aged hypertensive animals with or without AM treatment. It provides the potential rationale to a novel combination treatment with low-dose DM and AM in clinical hypertension.     

Inhibition Effect of a Custom Peptide on Lung Tumors

Cecropin B is a natural antimicrobial peptide and CB1a is a custom, engineered modification of it. In vitro, CB1a can kill lung cancer cells at concentrations that do not kill normal lung cells. Furthermore, in vitro, CB1a can disrupt cancer cells from adhering together to form tumor-like spheroids. Mice were xenografted with human lung cancer cells; CB1a could significantly inhibit the growth of tumors in this in vivo model. Docetaxel is a drug in present clinical use against lung cancers; it can have serious side effects because its toxicity is not sufficiently limited to cancer cells. In our studies in mice: CB1a is more toxic to cancer cells than docetaxel, but dramatically less toxic to healthy cells.     

A COMPARATIVE STUDY OF THE FOLIAR ANATOMY OF LYCOPODIUM SPECIES

A comparative study of the leaves of 31 species of Lycopodium was made. The pattern of tracheids varies with the species. Usually they exhibit annular, helical, reticulate, modified or transition forms; none of the species have advanced phylogenetically beyond the scalariform tracheid stage. Mucilage canals have been found in four species. Tracheids are enclosed by a few to several layers of oblique ended, elongated parenchyma cells in most species, but an endodermis is absent. Vascular bundles are encircled by fiber cells in three species. The mesophyll of most species consists of identically shaped cells, although palisade-like tissue has been observed in a few species. The epidermal cells vary from elongated to isodiametric and have either undulated or smooth anticlinal walls, which are deeply pitted in some species. The outer epidermal walls are usually thick and heavily cutinized. Stomates are distributed on both surfaces in 18 species, on the abaxial surface in 11 species, and on the adaxial side in only two species. Most stomates are practically isodiametric in surface view, broader than epidermal cells, usually parallel to the vein, and at the same level as the adjacent epidermal cell. A typical guard cell has a prominent outer ledge and a less developed inner ledge of cutin in most species. Six groupings are suggested, based on similarity of leaf structure and the known chromosome numbers.     

Synthesis and anticancer activity of a novel series of 9-O-substituted berberine derivatives: A lipophilic substitute role

To alter its hydrophobicity, a series of compounds bearing 9-O-alkyl- or 9-O-terpenyl- substituted berberine were synthesized and evaluated for anticancer activity against human cancer HepG2 and HT29 cell lines. We found that the lipophilic substitute of 9-O-alkyl- and 9-O-terpenyl berberine derivatives plays a role in inhibiting the human cancer cell growth and its activity could be maximized with the optimized substitute type and chain length. Most strikingly, nonetheless, of the six compounds prepared, sample 8, a farnesyl 9-O-substituted berberine, showed either comparable or better cytotoxic activity against human cancer HepG2 cell line than that of berberine. Compound 8 had also shown a 104-fold antiproliferation activity in compare with berberine against human hepatoma HepG2 cell lines after 48 incubation hours. Further, in Hoechst 33258 and annexin V-FITC/PI staining analyses it induced apoptosis in HepG2 cells at lower concentration than that of berberine for 24 h. Take all; farnesyl 9-O-substituted berberine could be a potential candidate for new anticancer drug development.     

A case for "StopGo": reprogramming translation to augment codon meaning of GGN by promoting unconventional termination (Stop) after addition of glycine and then allowing continued translation (Go)

When a eukaryotic mRNA sequence specifying an amino acid motif known as 2A is directly followed by a proline codon, two nonoverlapping proteins are synthesized. From earlier work, the second protein is known to start with this proline codon and is not created by proteolysis. Here we identify the C-terminal amino acid of an upstream 2A-encoded product from Perina nuda picorna-like virus that is glycine specified by the last codon of the 2A-encoding sequence. This is an example of recoding where 2A promotes unconventional termination after decoding of the glycine codon and continued translation beginning with the 3' adjacent proline codon.     

Antioxidant activities and phytochemical characteristics of extracts from Acacia confusa bark

The present study was carried out to evaluate the antioxidant activities of bark extract of Acacia confusa and some of the isolated constituents from its ethyl acetate (EtOAc) fraction in various in vitro systems to gain mechanistic insights. Results from antioxidant assays together with authentic antioxidant standards revealed that EtOAc fraction showed strong superoxide radical scavenging activity, reducing power, and ferrous ion-chelating ability. Following an in vitro antioxidant activity-guided fractionation procedure, 16 constituents including 12 benzoic acids, three cinnamic acids and one lignans were isolated and identified from the EtOAc fraction. We also evaluated the structure-activity relationships of benzoic and cinnamic acid derivatives. Results obtained indicated that the bark extracts and the derived phytochemicals from A. confusa have a great potential to prevent disease caused by the overproduction of radicals and also it might be used as a potential source of natural antioxidant agent.     

Bacterial community of very wet and acidic subalpine forest and fire-induced grassland soils

The subalpine forest and grassland ecosystems at Tatajia in Yushan National Park, Taiwan, at an elevation of 2,700 m, mean annual precipitation of 4,100 mm, mean annual temperature of 9.5°C, and soil pH near 3.5, represent land types whose bacterial communities have not been previously characterized. To this end, small subunit (SSU) rRNA libraries were prepared from environmental DNA, and 319 clones were sequenced and characterized. Despite differences in vegetation, Acidobacteria, Proteobacteria and Firmicutes were the most abundant phyla in soil communities from the forest and grassland. Although not significantly different, on the basis of Chao1, Shannon and other indices and rarefaction analyses, the diversity of the bacterial community of grassland appeared higher than that of the forest. The composition of the most abundant operational taxonomic units (OTUs) also differed between the grassland and forest communities. Because the grassland was formed by fire 30 years ago from forest, these results indicated a different bacterial community could form within that time. Moreover, most of the OTUs abundant in Tatajia soils had been previously detected in other studies, but in lower numbers. Therefore, the bacterial communities in Tatajia differed in relative abundance but not in types of bacteria present. However, one acidobacterial OTU abundant in Tatajia had previously been found to be abundant in soils from around the world. Thus, this OTU may represent a particularly abundant and cosmopolitan bacterial phylotype.