Possible use of synthetic aggregation pheromones to control stinkbug Plautia stali in kaki persimmon orchards

• 1 The brown‐winged green stinkbug Plautia stali Scott (Hemiptera: Pentatomidae) is the most serious pest of all noxious stinkbugs in various orchards in Japan. An area‐wide integrated pest management programme using the species‐specific aggregation pheromone is desirable to control P. stali because nonspecific insecticides may kill arthropod natural enemies and induce the resurgence of other endemic pests.
• 2 The traditional mass‐trapping method is not expected to be effective as a result of huge migrations from cypress forests. Therefore, we chose an ‘attract‐and‐kill’ strategy of intensively installing poisonous eggplants with a synthetic aggregation pheromone lure as enclosures for the target orchards.
• 3 We found no overall control effect of the installation of poisonous eggplants, although regional differences were observed, which might originate from topological configurations or the distance from the source population in cypress forests.
• 4 The poisonous eggplants with an aggregation‐pheromone lure, however, changed the spatial distribution of fruit damage, appearing to induce the majority of the damage within a 100‐m range of the poisonous eggplants. Some damage, although at a low level, was found in regions 100–200 m away from the poisonous eggplants.
• 5 We postulate that such low‐damage regions were created because the majority of the bugs dispersed from the centre of the orchards to the areas with poisonous eggplants. The present study, in which the spatial scale of the damage spillover was estimated at approximately 150 m, has important implications for future strategies of attract‐and‐kill and possibly push–pull.

     

Integration-free iPS cells engineered using human artificial chromosome vectors

Human artificial chromosomes (HACs) have unique characteristics as gene-delivery vectors, including episomal transmission and transfer of multiple, large transgenes. Here, we demonstrate the advantages of HAC vectors for reprogramming mouse embryonic fibroblasts (MEFs) into induced pluripotent stem (iPS) cells. Two HAC vectors (iHAC1 and iHAC2) were constructed. Both carried four reprogramming factors, and iHAC2 also encoded a p53-knockdown cassette. iHAC1 partially reprogrammed MEFs, and iHAC2 efficiently reprogrammed MEFs. Global gene expression patterns showed that the iHACs, unlike other vectors, generated relatively uniform iPS cells. Under non-selecting conditions, we established iHAC-free iPS cells by isolating cells that spontaneously lost iHAC2. Analyses of pluripotent markers, teratomas and chimeras confirmed that these iHAC-free iPS cells were pluripotent. Moreover, iHAC-free iPS cells with a re-introduced HAC encoding Herpes Simplex virus thymidine kinase were eliminated by ganciclovir treatment, indicating that the HAC safeguard system functioned in iPS cells. Thus, the HAC vector could generate uniform, integration-free iPS cells with a built-in safeguard system.     

Anti-Aβ drug screening platform using human iPS cell-derived neurons for the treatment of Alzheimer's disease

Background

Alzheimer''s disease (AD) is a neurodegenerative disorder that causes progressive memory and cognitive decline during middle to late adult life. The AD brain is characterized by deposition of amyloid β peptide (Aβ), which is produced from amyloid precursor protein by β- and γ-secretase (presenilin complex)-mediated sequential cleavage. Induced pluripotent stem (iPS) cells potentially provide an opportunity to generate a human cell-based model of AD that would be crucial for drug discovery as well as for investigating mechanisms of the disease.

Methodology/Principal Findings

We differentiated human iPS (hiPS) cells into neuronal cells expressing the forebrain marker, Foxg1, and the neocortical markers, Cux1, Satb2, Ctip2, and Tbr1. The iPS cell-derived neuronal cells also expressed amyloid precursor protein, β-secretase, and γ-secretase components, and were capable of secreting Aβ into the conditioned media. Aβ production was inhibited by β-secretase inhibitor, γ-secretase inhibitor (GSI), and an NSAID; however, there were different susceptibilities to all three drugs between early and late differentiation stages. At the early differentiation stage, GSI treatment caused a fast increase at lower dose (Aβ surge) and drastic decline of Aβ production.

Conclusions/Significance

These results indicate that the hiPS cell-derived neuronal cells express functional β- and γ-secretases involved in Aβ production; however, anti-Aβ drug screening using these hiPS cell-derived neuronal cells requires sufficient neuronal differentiation.     

Intratumoral injection of Propionibacterium acnes suppresses malignant melanoma by enhancing Th1 immune responses

Malignant melanoma (MM) is an aggressive cutaneous malignancy associated with poor prognosis; many putatively therapeutic agents have been administered, but with mostly unsuccessful results. Propionibacterium acnes (P. acnes) is an aerotolerant anaerobic gram-positive bacteria that causes acne and inflammation. After being engulfed and processed by phagocytes, P. acnes induces a strong Th1-type cytokine immune response by producing cytokines such as IL-12, IFN-γ and TNF-α. The characteristic Th2-mediated allergic response can be counteracted by Th1 cytokines induced by P. acnes injection. This inflammatory response induced by P. acnes has been suggested to have antitumor activity, but its effect on MM has not been fully evaluated.We analyzed the anti-tumor activity of P. acnes vaccination in a mouse model of MM. Intratumoral administration of P. acnes successfully protected the host against melanoma progression in vivo by inducing both cutaneous and systemic Th1 type cytokine expression, including TNF-α and IFN-γ, which are associated with subcutaneous granuloma formation. P. acnes-treated tumor lesions were infiltrated with TNF-α and IFN-γ positive T cells. In the spleen, TNF-α as well as IFN-γ producing CD8(+)T cells were increased, and interestingly, the number of monocytes was also increased following P. acnes administration. These observations suggest that P. acnes vaccination induces both systemic and local antitumor responses. In conclusion, this study shows that P. acnes vaccination may be a potent therapeutic alternative in MM.     

Analysis of GABA-induced inhibition of spontaneous firing in chick accessory lobe neurons

It has been hypothesized that chick accessory lobes (ALs) contain functional neurons and act as a sensory organ of equilibrium. It was reported that neurons located in an outer layer of ALs showed γ-aminobutyric acid (GABA)- and glutamic acid decarboxylase (GAD)-like immunoreactivity more strongly than centrally located neurons, which were surrounded by the GAD-immunoreactive terminals. We investigated effects of GABA on the electrical activity of AL neurons. About 50% of embryonic AL neurons exhibited spontaneous firing. In the on-cell recording, GABA, muscimol, and GABA in combination with CGP35348 inhibited this firing. In whole-cell voltage clamp recordings, GABA and muscimol evoked a transient current. The mean reversal potential of GABA-evoked currents was close to the theoretical reversal potential of Cl⁻. These results indicate that GABA exerts the inhibitory effect on the firing through the activation of GABA_A receptors. In addition, the intracellular concentration of Cl⁻ was estimated to be about 16 mM in measurements with the gramicidin-perforated configuration, indicating the physiological reversal potential of the GABA current was about −60 mV. In conclusion, AL neurons have an intrinsic mechanism to evoke the spontaneous firing, which can be arrested by the inhibitory mechanism through the activation of the GABA_A receptors.     

The effect of enzymatically polymerised polyphenols on CD4 binding and cytokine production in murine splenocytes

High-molecular weight polymerised polyphenols have been shown to exhibit anti-influenza virus, anti-HIV, and anti-cancer activities. The purpose of this study was to evaluate the immunomodulating activities of enzymatically polymerised polyphenols, and to clarify the underlying mechanisms of their effects. The cytokine-inducing activity of the enzymatically polymerised polyphenols derived from caffeic acid (CA), ferulic acid (FA), and p-coumaric acid (CoA) was investigated using murine splenocytes. Polymerised polyphenols, but not non-polymerised polyphenols, induced cytokine synthesis in murine splenocytes. Polymerised polyphenols induced several cytokines in murine splenocytes, with interferon-γ (IFN-γ) and granulocyte-macrophage colony-stimulating factor (GM-CSF) being the most prominent. The underlying mechanisms of the effects of the polymerised polyphenols were then studied using neutralising antibodies and fluorescent-activated cell sorting (FACS) analysis. Our results show that polymerised polyphenols increased IFN-γ and GM-CSF production in splenocytes. In addition, the anti-CD4 neutralised monoclonal antibody (mAb) inhibited polymerised polyphenol-induced IFN-γ and GM-CSF secretion. Moreover, polymerised polyphenols bound directly to a recombinant CD4 protein, and FACS analysis confirmed that interaction occurs between polymerised polyphenols and CD4 molecules expressed on the cell surface. In this study, we clearly demonstrated that enzymatic polymerisation confers immunoactivating potential to phenylpropanoic acids, and CD4 plays a key role in their cytokine-inducing activity.     

Evaluation of a rapid immunochromatographic ODK-0901 test for detection of pneumococcal antigen in middle ear fluids and nasopharyngeal secretions

Since the incidence of penicillin-resistant Streptococcus pneumoniae has been increasing at an astonishing rate throughout the world, the need for accurate and rapid identification of pneumococci has become increasingly important to determine the appropriate antimicrobial treatment. We have evaluated an immunochromatographic test (ODK-0901) that detects pneumococcal antigens using 264 middle ear fluids (MEFs) and 268 nasopharyngeal secretions (NPSs). A sample was defined to contain S. pneumoniae when optochin and bile sensitive alpha hemolytic streptococcal colonies were isolated by culture. The sensitivity and specificity of the ODK-0901 test were 81.4% and 80.5%, respectively, for MEFs from patients with acute otitis media (AOM). In addition, the sensitivity and specificity were 75.2% and 88.8%, respectively, for NPSs from patients with acute rhinosinusitis. The ODK-0901 test may provide a rapid and highly sensitive evaluation of the presence of S. pneumoniae and thus may be a promising method of identifying pneumococci in MEFs and NPSs.