Design of a PKCδ-specific small peptide as a theragnostic agent for glioblastoma

Glioblastoma is an aggressive malignant brain tumor that starts in the brain or spine and frequently recurs after anticancer treatment. The development of an accurate diagnostic system combined with effective cancer therapy is essential to improve prognosis of glioma patients. Peptides, produced from phage display, are attractive biomolecules for glioma treatment because of their biostability, nontoxicity, and small size. In this study, we employed phage display methodology to screen for peptides that specifically recognize the target PKCδ as a novel biomarker for glioma. The phage library screening yielded four different peptides displayed on phages with a 20- to 200-pM K_d value for the recombinant PKCδ catalytic domain. Among these four phage peptides, we selected one to synthesize and tagged it with fluorescein isothiocyanate (FITC) based on the sequence of the PKCδ-binding phage clone. The synthetic peptide showed a relative binding affinity for antibody and localization in the U373 glioma cell. The kinase activity of PKCδ was inhibited by FITC-labeled peptide with an IC₅₀ of 1.4 μM in vitro. Consequently, the peptide found in this study might be a promising therapeutic agent against malignant brain tumor.     

Embryology and relationships of Lauraceae (Laurales)

Embryology of Lauraceae, hitherto poorly known, was investigated on the basis of 35 species from 23 genera to contribute to a better understanding of familial and generic relationships. Data from the genera investigated and from the literature show that the genera of Lauraceae are very similar embryologically, but that differences do exist in a few characters among the genera. Based on comparisons with other families of Laurales, Lauraceae consistendy had a pachychalazal ovule or seed with a ramified raphal vascular bundle at chalaza (an apomorphy) in common with Hernandiaceae. However, since several core lauralean families such as Amborellaceae, Monimiaceae, and Gomortegaceae are little known embryologically, these must be studied for critical comparison. Wimin Lauraceae, Cassytha is clearly distinct from the rest of the family in having an ab initio cellular type endosperm (a plesiomorphy, also reported in Umbellularia ) instead of a nuclear type endosperm (an apomorphy) as in the rest of the family, in lacking the nucellar cap and in having the micropyle formed by bom the inner and outer integument, facts supporting the traditional taxonomic placement of Cassytha in its own subfamily and the remainder of the family in the other subfamily. Widiin the rest of the family, the amoeboid tapetum (an apomorphy) distinguishes 15 genera ( Actinodaphne, Cinnamomum, Laurus , etc.) from the six genera with the glandular tapetum. In addition, a mature embryo sac protruding from the nucellus (an apomorphy) distinguishes five genera ( Beibchmiedia, Caryodaphnopsis, Cryptocarya, Endiandra, Potamria) and one species of Ocotea (O. rubra ) from the rest of the family. These results can properly be incorporated in a future suprageneric classification. The distinctness of Ocotea rubra wimin the genus is also discussed.     

A palynotaxonomic study of the genus Filipendula (Rosaceae)

Pollen grains from 15 species (18 taxa) of the genus Filipendula were examined with light and scan-ning electron microscopy. It was revealed that the pollen grains are isopolar, tricolporate, with scabrate or scabrate-microechinate surface. The pollen morphology was compared with the conventional classification sys-tems of the genus by different authors, and supported Shimizu's system (1961), in which the genus was divided into three subgenera. The monotypic subgen. Hypogyna is characterized by pollen lacking fastigium and thickened costae colpi. The other monotypic subgen. Filipendula differs from others by pollen having larger grain, larger pore size, longitudinally elliptic fastigium and thickened costae colpi. The largest subgen. Ulmaria is distinguished by pollen having rounded or latitudinally elliptic fastigium and thickened costae colpi. Sectional classification was not supported by the pollen morphology due to insufficient variability.     

A new species of Monoliropus Mayer, 1903 (Crustacea,Amphipoda, Caprellidae) from Korean waters

A new species of the genus Monoliropus belonging to the family Caprellidae was collected from the Yellow Sea, Korea. The new species differs from Monoliropus agilis Mayer, 1903, Monoliropus kazemii Momtazi & Sari, 2013, and Triprotella amica Arimoto, 1970 as follows: 1) gnathopod 1 subrectangular; 2) pereonites 2–3 with acute triangular processes anterolaterally; 3) mandibular palp, apical article with four simple setae subdistally. The new species is fully illustrated and extensively compared with related species. This is the first record of the genus Monoliropus from Korean waters. A key to Monoliropus species is also given.     

Characterization of acetohydroxyacid synthase I from Escherichia coli K-12 and identification of its inhibitors

The first step in branched-chain amino acid biosynthesis is catalyzed by acetohydroxyacid synthase (EC 2.2.1.6). This reaction involves decarboxylation of pyruvate followed by condensation with either an additional pyruvate molecule or with 2-oxobutyrate. The enzyme requires three cofactors, thiamine diphosphate (ThDP), a divalent ion, and flavin adenine dinucleotide (FAD). Escherichia coli contains three active isoenzymes, and acetohydroxyacid synthase I (AHAS I) large subunit is encoded by the ilvB gene. In this study, the ilvB gene from E. coli K-12 was cloned into expression vector pETDuet-1, and was expressed in E. coli BL21 (DH3). The purified protein was identified on a 12% SDS-PAGE gel as a single band with a mass of 65 kDa. The optimum temperature, buffer, and pH for E. coli K-12 AHAS I were 37 °C, potassium phosphate buffer, and 7.5. Km values for E. coli K-12 AHAS I binding to pyruvate, Mg(+2), ThDP, and FAD were 4.15, 1.26, 0.2 mM, and 0.61 μM respectively. Inhibition of purified AHAS I protein was determined with herbicides and new compounds.     

The growth of K562 human leukemia cells was inhibited by therapeutic neural stem cells in cellular and xenograft mouse models

To confirm the anti-tumor effect of engineered neural stem cells (NSCs) expressing cytosine deaminase (CD) and interferon-β (IFN-β) with prodrug 5-fluorocytosine (FC), K562 chronic myeloid leukemia (CML) cells were co-cultured with the neural stem cell lines HB1.F3.CD and HB1.F3.CD.IFN-β in 5-FC containing media. A significant decrease in the viability of K562 cells was observed by the treatment of the NSC lines, HB1.F3.CD and HB1.F3.CD.IFN-β, compared with the control. A modified trans-well assay showed that engineered human NSCs significantly migrated toward K562 CML cells more than human normal lung cells. In addition, the important chemoattractant factors involved in the specific migration ability of stem cells were found to be expressed in K562 CML cells. In a xenograft mouse model, NSC treatments via subcutaneous and intravenous injections resulted in significant inhibitions of tumor mass growth and extended survival dates of the mice. Taken together, these results suggest that gene therapy using genetically engineered stem cells expressing CD and IFN-β may be effective for treating CML in these mouse models.     

Identification of quantitative trait loci and a candidate locus for freezing tolerance in controlled and outdoor environments in the overwintering crucifer Boechera stricta

Development of chilling and freezing tolerance is complex and can be affected by photoperiod, temperature and photosynthetic performance; however, there has been limited research on the interaction of these three factors. We evaluated 108 recombinant inbred lines of Boechera stricta, derived from a cross between lines originating from Montana and Colorado, under controlled long day (LD), short‐day (SD) and in an outdoor environment (OE). We measured maximum quantum yield of photosystem II, lethal temperature for 50% survival and electrolyte leakage of leaves. Our results revealed significant variation for chilling and freezing tolerance and photosynthetic performance in different environments. Using both single‐ and multi‐trait analyses, three main‐effect quantitative trait loci (QTL) were identified. QTL on linkage group (LG)3 were SD specific, whereas QTL on LG4 were found under both LD and SD. Under all conditions, QTL on LG7 were identified, but were particularly predictive for the outdoor experiment. The co‐localization of photosynthetic performance and freezing tolerance effects supports these traits being co‐regulated. Finally, the major QTL on LG7 is syntenic to the Arabidopsis C‐repeat binding factor locus, known regulators of chilling and freezing responses in Arabidopsis thaliana and other species.