Determination of the Nucleotide Sequence of Bombyx mori Cytoplasmic Polyhedrosis Virus Segment 9 and Its Expression in BmN4 Cells

Cloning and sequencing of segment 9 of Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) strains H and I were performed. The segment consisted of 1,186 bp harboring 5′ and 3′ noncoding regions and an open reading frame from positions 75 to 1037, encoding a protein with 320 amino acids, termed NS5. Comparison of the nucleotide sequences of NS5 for the two strains indicated 37 point differences resulting in only six amino acid replacements. Homology search showed that NS5 has localized similarities to human poliovirus RNA-dependent RNA polymerase and human rotavirus NS26. By Western blot analysis, NS5 was found in BmCPV-infected midgut cells, but not in polyhedra or virus virions, and was mainly detectable in the nucleus in BmCPV-infected BmN4 cells. Immunoblot analysis with anti-NS5 and antipolyhedrin antibodies displayed marked differences in the period of expression of NS5 and polyhedrin: the polyhedrin molecule was first detected 2 or 3 days after infection with BmCPV, whereas the expression of NS5 was initiated within a few hours. In addition, the level of polyhedrin increased as the infection developed, whereas the amount of NS5 remained essentially constant. When segment 9 was expressed with a baculovirus expression system, the resulting NS5 protein possessed the ability to bind to the double-stranded RNA genome. These results suggest that NS5 is expressed in early stages of infection and contributes to regulation of genomic RNA function.     

Restriction fragment length variations and chromosome mapping of two mouse metallothionein genes,Mt-1 andMt-2

Restriction endonuclease fragment length variations (RFLVs) were found through the use of cDNA probes for metallothionein genes 1 (Mt-1) and 2 (Mt-2) in the mouse. RFLVs were detected in restriction patterns generated by BglII and XbaI in the Mt-1 gene and by PvuII in the Mt-2 gene. All laboratory strains carry the Mt-1a and Mt-2a alleles. Among strains of wild origin, some Western European subspecies (Mus mus domesticus and M. m. brevirostris) also carry the Mt-1a and Mt-2a alleles. In contrast, a European subspecies (M. m. musculus) and the great majority of subspecies from East Asian countries (M. m. molossinus, Chinese mice of wild origin, and M. m. yamashinai) carry the Mt-1b and Mt-2b alleles. A domesticus strain from Bulgaria and two castaneus strains from Thailand and Philippines carry the intermediate combination of Mt-1b and Mt-2a alleles. Using the RFLVs, we mapped the Mt-1 and Mt-2 genes on chromosome 8, and they appear to be very closely linked since no recombination was observed between them in any of the mice examined. Data from three-point cross tests showed that the recombination frequencies are 4.31% between Os and Mt, 15.52% between Mt and Prt-2, and 19.83% between Os and Prt-2. The gene order of Os-Mt-1,Mt-2-Prt-2 has been confirmed.     

Sortilin is upregulated during osteoblastic differentiation of mesenchymal stem cells and promotes extracellular matrix mineralization

Osteoblasts and adipocytes are derived from a common precursor in bone marrow, the mesenchymal stem cell (MSC). Factors driving human MSCs (hMSCs) to differentiate down the two lineages play important roles in determining bone density because it has been shown that bone volume loss associated with osteoporosis and aging is accompanied by reduced osteoblastic bone formation and increased marrow adipose tissue. The genes upregulated in hMSCs during osteogenic differentiation were screened using cDNA microarrays and were semi-quantitated by real-time RT-PCR. One of the genes identified was sortilin, which was upregulated one day after osteogenic induction and remained upregulated for a week. The overexpression of sortilin in hMSCs using an adenovirus vector resulted in the acceleration of mineralization during osteogenic differentiation without affecting alkaline phosphatase activity. Lipoprotein lipase (LPL), produced by adipocytes, is bound by sortilin, which may mediate its endocytosis. By adding LPL to osteogenic induction medium, osteoblastic mineralization was inhibited in a dose-dependent manner. Interestingly, sortilin overexpression abolished the LPL-mediated suppression of osteogenic differentiation. hMSCs exist in marrow where LPL-producing adipose cells are abundant and where osteogenesis is negatively regulated by LPL. Sortilin has a counter effect of promoting osteogenesis by acting as a scavenger of LPL.     

Review of the clingfish genus <Emphasis Type="Italic">Kopua</Emphasis> (Gobiesocidae: Trachelochisminae) in Japan,with description of a new species

A taxonomic review of the clingfish genus Kopua (Gobiesocidae: Trachelochisminae) in Japan recognizes three species: K. japonica Moore, Hutchins and Okamoto 2012, K. vermiculata Shinohara and Katayama 2015 and K. yoko sp. nov. Kopua japonica and K. vermiculata are redescribed with revised diagnoses on the basis of 20 specimens (10.4–30.4 mm standard length; SL) and the holotype, respectively. Kopua japonica is similar to K. vermiculata in head sensory pore characters (normally single nasal and postocular canal pores). However, the former differs distinctly from the latter as follows: 6–8 (modally 7) gill rakers (vs. 4 or 6); 31–33 (33) vertebrae (vs. 35); anus slightly closer to posterior margin of disc than to anal-fin origin (vs. much closer to posterior margin of disc); snout length 5.3–8.7 (mean 7.0) % SL (vs. 9.2 % SL); disc length 21.2–24.0 (22.8) % SL (vs. 18.8 % SL); pre-dorsal- and anal-fin lengths 72.9–78.4 (75.2) and 78.1–82.8 (80.1) % SL, respectively (vs. 67.5 and 73.6 % SL); and two stripes on cheek (vs. a triangular blotch). Kopua yoko sp. nov., based on 14 specimens (17.7–28.8 mm SL) from the Pacific coast of southern Japan, Sea of Japan and the East China Sea, is characterized by the following combination of characters: 6 or 7 (modally 6) dorsal-fin rays; 4–6 (5) anal-fin rays; 21 or 22 (21) pectoral-fin rays; 4–6 (5 or 6) gill rakers; 31–33 (31) vertebrae; a single (rarely two) nasal canal pores; two lacrimal and preopercular canal pores; snout length 6.5–7.9 (mean 7.1) % SL; gill opening depth 5.8–7.1 (6.5) % SL; least interorbital width 2.0–3.7 (2.6) % SL; disc length 20.3–25.0 (23.1) % SL, disc region D without flattened papillae; caudal-peduncle depth 8.1–10.2 (9.2) % SL; anus slightly closer to posterior margin of disc than to anal-fin origin; pre-dorsal- and anal-fin lengths 71.6–77.1 (73.9) and 77.0–83.7 (80.4) % SL, respectively; post-dorsal-caudal length 12.6–15.0 (13.8) % SL; arch-shaped blotches on lateral aspect of body; and two reddish-orange stripes on cheek. Morphological changes with growth in K. japonica and K. yoko sp. nov. are also described.     

Synthesis and structure-activity relationships of 3-substituted 1,4-dihydro-4-oxo-1-(2-thiazolyl)-1,8-naphthyridines as novel antitumor agents

In order to obtain clinically useful antitumor agent, we have designed and synthesized various 3-substituted 1,4-dihydro-4-oxo-1-(2-thiazolyl)-1,8-naphthyridines, and evaluated their cytotoxic activity. The series of novel 3-substituted derivatives synthesized in this study showed good antitumor activity against murine P388 leukemia. Particularly, the 3-formyl 1,8-naphthyridine displayed an antitumor activity equal to that of the 3-carboxy 1,8-naphthyridine against murine and human tumor cell lines as well as in vivo test for mouse leukemia. These results demonstrate that the carboxy group at the C-3 position of 1,8-naphthyridine ring is not essential for antitumor activity. In addition, the trend of cytotoxic activity for the 3-substituted 1,8-naphthyridines was different from that of antibacterial activity.     

Antibody-drug conjugates: recent advances in conjugation and linker chemistries

The antibody-drug conjugate (ADC), a humanized or human monoclonal antibody conjugated with highly cytotoxic small molecules (payloads) through chemical linkers, is a novel therapeutic format and has great potential to make a paradigm shift in cancer chemotherapy. Thisnewantibody-based molecular platform enables selective delivery of a potent cytotoxic payload to target cancer cells, resulting in improved efficacy, reduced systemic toxicity, and preferable pharmacokinetics (PK)/ pharmacodynamics (PD) and biodistribution compared to traditional chemotherapy. Boosted by the successes of FDA-approved Adcetris® and Kadcyla®, this drug class has been rapidly growing along with about 60 ADCs currently in clinical trials. In this article, we briefly review molecular aspects of each component (the antibody, payload, and linker) of ADCs, and then mainly discuss traditional and new technologies of the conjugation and linker chemistries for successful construction of clinically effective ADCs. Current efforts in the conjugation and linker chemistries will provide greater insights into molecular design and strategies for clinically effective ADCs from medicinal chemistry and pharmacology standpoints. The development of site-specific conjugation methodologies for constructing homogeneousADCs is an especially promising path to improving ADC design, which will open the way for novel cancer therapeutics.     

New broad-spectrum parenteral cephalosporins exhibiting potent activity against both methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. Part 3: 7beta-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido] cephalosporins bearing 4-[3-(aminoalkyl)-ureido]-1-pyridinium at C-3'

Among the prepared C-3' substituted-pyridinium cephalosporins, a series of 7beta-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido] cephalosporins bearing 4-[3-(aminoalkyl)-ureido]-1-pyridinium at C-3' showed highly potent antibacterial activity against MRSA and Pseudomonas aeruginosa.     

西藏珠峰－卓奥友峰普士拉地区高山稀疏植被的群落特征及小地形的影响

在西藏定日县珠穆朗玛峰-卓奥友峰附近的普士拉地区的-山坡上(海拔5176—5390m),设置13个样方。进行群落调查、小地形测定和土壤剖面调查,分析该地区高寒植被的群落特征及其与小地形的关系。13个样方(面积25—200m^2不等)中,共记载物种80个(含变种和亚种),分别隶属于47个属。出现频度较高的种有高山嵩草(Kobresia pygmaea)、矮兔耳草(Lagottes humilis)、楔叶委陵菜(Potentilla funeata)、华马先蒿(Pedicularis oederi var.sinensis)、高山委陵菜(Potentilla polyjchista)、密生雪灵芝(Arenaria densissima)等;含物种数较多的属有：虎耳草属(Saxifraga)、风毛菊属(Saussurea)、嵩草属(Kobresia)、委陵菜属(Potentilla)、龙胆属(Gentiana)、葶苈属(Draba)等。随着海拔降低,物种和属的丰富度呈现出上升的趋势在该地区,植被盖度能较好地指示生境条件。等级聚类的结果也支持了这一观点：随着植被盖度的增加,转换物种丰富度(TSR)和转换属丰富度(TGR)增加,而物种多样性指数(Shannon-Wiener指数)减小。利用物种矩阵以及海拔、坡度、坡向、土壤深度、植被盖度等环境变量进行CCA排序,结果分出4类生境类型,它们较好地反映了群落特征与地形的关系。     

Correlation between protective antibody response and patent infection with Hymenolepis nana in mice

Correlation between protective antibody response and patent infection with Hymenolepis nana in mice. International Journal for Parasitology16: 197–203. Mice inoculated with mouse-derived cysticercoids of Hymenolepis nana, as well as with eggs, produced IgG and IgE antibodies that were detected by double diffusion (DD) and passive cutaneous anaphylaxis (PCA), respectively. When mice inoculated with eggs (day 0) were challenged with eggs (day 66), all were resistant to the challenge (assessed by the failure of cysticercoid recovery in the intestinal tissue) and produced protective antibodies evidenced by passive transfer, as well as IgG and IgE isotypes. When mice inoculated with eggs (day 0) were treated with a highly efficacious cestocide, praziquantel on day 6 at the beginning of the lumen phase, all were also resistant to the egg challenge on day 66, however, IgE, IgG, and protective antibodies were not detected. When mice treated with praziquantel before patent infection were repeatedly challenged with high doses of eggs, some of them produced IgG and IgE antibodies. From these results, it is suggested that (1) the production of protective antibody is a secondary response after patency (which may be ascribed to eggs released from mature worms), and (2) mice initially given eggs are highly resistant to egg challenge showing that an effector mechanism of acquired resistance to egg challenge may be expressed without high titres of protective antibody, at least in the serum.