Identification of Novel HLA-A*24:02-Restricted Epitope Derived from a Homeobox Protein Expressed in Hematological Malignancies

The homeobox protein, PEPP2 (RHOXF2), has been suggested as a cancer/testis (CT) antigen based on its expression pattern. However, the peptide epitope of PEPP2 that is recognized by cytotoxic T cells (CTLs) is unknown. In this study, we revealed that PEPP2 gene was highly expressed in myeloid leukemia cells and some other hematological malignancies. This gene was also expressed in leukemic stem-like cells. We next identified the first reported epitope peptide (PEPP2^271-279). The CTLs induced by PEPP2^271-279 recognized PEPP2-positive target cells in an HLA-A*24:02-restricted manner. We also found that a demethylating agent, 5-aza-2’-deoxycytidine, could enhance PEPP2 expression in leukemia cells but not in blood mononuclear cells from healthy donors. The cytotoxic activity of anti-PEPP2 CTL against leukemic cells treated with 5-aza-2’-deoxycytidine was higher than that directed against untreated cells. These results suggest a clinical rationale that combined treatment with this novel antigen-specific immunotherapy together with demethylating agents might be effective in therapy-resistant myeloid leukemia patients.     

Knockdown of gene expression by antisense morpholino oligos in preimplantation mouse embryos cultured in vitro

Knockdown of gene expression by antisense morpholino oligos (MOs) is a simple and effective method for analyzing the roles of genes in mammalian cells. Here, we demonstrate the efficient delivery of MOs by Endo-Porter (EP), a special transfection reagent for MOs, into preimplantation mouse embryos cultured in vitro. A fluorescein-labeled control MO was applied for monitoring the incorporation of MOs into developing 2-cell embryos in the presence of varying amounts of EP and bovine serum albumin. In optimized conditions, fluorescence was detected in 2-cell embryos within a 3-h incubation period. In order to analyze the validity of the optimized conditions, an antisense Oct4 MO was applied for knockdown of the synthesis of OCT4 protein in developing embryos from the 2-cell stage. In blastocysts, the antisense Oct4 MO induced a decrease in the amount in OCT4 protein to less than half. An almost complete absence of OCT4-positive cells and nearly complete disappearance of the inner cell mass in the outgrowths of blastocysts were also noted. These phenotypes corresponded with those of Oct4-deficient mouse embryos. Overall, we suggest that the delivery of MOs using EP is useful for the knockdown of gene expression in preimplantation mouse embryos cultured in vitro.     

Phylogeography of Arabis serrata (Brassicaceae) in the Japanese Archipelago based on chloroplast DNA variations

Arabis serrata (Brassicaceae), a perennial plant widely distributed along the Japanese Archipelago, occurs in various habitats: for example, limestone zones, serpentine barrens, volcanic soils, and roadsides. It likely survived by adapting to its surrounding environment, resulting in great morphological and ecological variation. In this study, we performed a phylogeographic analysis to examine past changes in the distribution of A. serrata following climate oscillations during the Pleistocene. To cover most of A. serrata's range, leaves were collected from eight to ten individuals randomly selected from each of 37 populations in the Japanese Archipelago. Two chloroplast noncoding regions of the samples were amplified and sequenced: trnT(GGU)‐psbD and trnH(GUG)‐psbA spacers. Twenty‐five haplotypes were detected and distinguished by 31 substitutions. Four main haplotypes were observed in many populations distributed throughout the Japanese Archipelago. According to the genetic boundaries detected using the Monmonier algorithm, A. serrata is clustered into four groups, each including several populations: Hokkaido Island, northern mainland Honshu, central Japan, and western Japan. The boundaries, however, were not robust because all genetic parameters did not support the differentiation among groups. These results indicate the absence of an obvious geographic structure in the distribution of A. serrata, suggesting that this species has experienced a rapid range expansion in postglacial times.     

Pharmacological Evaluation of SKL-18287, a New Long-Acting Glucagon-Like Peptide-1 Receptor Agonist with Enhanced Aggregation Propensity,in Rodent Models

International Journal of Peptide Research and Therapeutics - Very recently, we have reported a new glucagon-like peptide-1 (GLP-1) receptor agonist, SKL-18287, (EC50: 1.2 and 0.13 nM for...     

Growth of zinnia,Italian ryegrass,and alfalfa and their remediation effects in diesel oil-contaminated soils

Abstract

Our objective in this study was to compare the growth of zinnia, Italian ryegrass, and alfalfa, and their remediation effects in oil-contaminated soils. The soils were prepared by mixing 2, 4, or 8% diesel oil by weight with soil. The plant height and dry weights of shoots and roots were highest for zinnia in the 2 and 4% oil treatments, and highest for Italian ryegrass in the 8% oil treatment. The reduction ratios in soil total petroleum hydrocarbons concentration (TPH) for 3 plants were lower in the 4 and 8% oil treatments than those in the 2% treatment. The reduction ratios for Italian ryegrass and zinnia contaminated with 2, 4, and 8% diesel oil treatments were significantly higher than those for alfalfa and the non-cultivation treatment at 45?days after sowing, and there were no significant differences in reduction ratios between Italian ryegrass and zinnia. The reduction ratio of soil TPH concentration brought about by zinnia was also comparable to that of Italian ryegrass. Therefore, we conclude that zinnia shows growth and remediation effects that are equivalent to those of Italian ryegrass, in soils contaminated with less than 8% oil.     

Diversity and host associations of dipteran insects exploiting fungal fruiting bodies in Hokuriku,central Japan

To clarify the diversity and host associations of dipteran insects exploiting fungal fruiting bodies, we collected fruiting bodies at 18 localities in Hokuriku region, central Japan, from 2012 to 2015 and examined them for the emergence of insects. In total, 14,107 dipteran individuals belonging to 20 families emerged from fungi of 8 orders, 25 families, 49 genera and 129 species. Approximately 79% of dipteran individuals belonged to three families, Phoridae, Muscidae and Drosophilidae. The faunal similarity at the family level was relatively high between central (warm‐temperate) and northern (cool‐temperate) areas of Japan. However, the species composition of Drosophilidae was much different between central and northern Japan. The difference in the species composition was discussed in relation to the climatic conditions and fungal flora. None of the species from Drosophilidae, Phoridae, Muscidae, Mycetophilidae, Lonchaeidae and Chloropidae were specialists (they exploited more than one species of fungi), but they showed differences their fungi preference. Adults of some families, especially Drosophilidae, were frequently collected from fruiting bodies, but those of other families were seldom collected, probably reflecting differences in adult feeding ecology.