Leukocyte accumulation promoting fibrin deposition is mediated in vivo by P-selectin on adherent platelets.

The glycoprotein P-selectin is a cell adhesion molecule of stimulated platelets and endothelial cells, which mediates the interaction of these cells with neutrophils and monocytes. It is a membrane component of cell storage granules, and is a member of the selectin family which includes E-selectin and L-selectin. P-selectin recognizes both lineage-specific carbohydrate ligands on monocytes and neutrophils, including the Lewis x antigen, sialic acid, and a protein component. In inflammation and thrombosis, P-selectin may mediate the interaction of leukocytes with platelets bound in the region of tissue injury and with stimulated endothelium. To evaluate the role of P-selectin in platelet-leukocyte adhesion in vivo, the accumulation of leukocytes within an experimental thrombus was explored in an arteriovenous shunt model in baboons. A Dacron graft implanted within an arteriovenous shunt is thrombogenic, accumulating platelets and fibrin within its lumen. These bound platelets express P-selectin. Here we show that antibody inhibition of leukocyte binding to P-selectin expressed on platelets immobilized on the graft blocks leukocyte accumulation and inhibits the deposition of fibrin within the thrombus. These results indicate that P-selectin is an important adhesion molecule on platelets, mediating platelet-leukocyte binding in vivo, that the presence of leukocytes in thrombi is mediated by P-selectin, and that these leukocytes promote fibrin deposition.     

Protein kinase D inhibitor CRT0066101 suppresses bladder cancer growth in vitro and xenografts via blockade of the cell cycle at G2/M

The protein kinase D (PKD) family of proteins are important regulators of tumor growth, development, and progression. CRT0066101, an inhibitor of PKD, has antitumor activity in multiple types of carcinomas. However, the effect and mechanism of CRT0066101 in bladder cancer are not understood. In the present study, we show that CRT0066101 suppressed the proliferation and migration of four bladder cancer cell lines in vitro. We also demonstrate that CRT0066101 blocked tumor growth in a mouse flank xenograft model of bladder cancer. To further assess the role of PKD in bladder carcinoma, we examined the three PKD isoforms and found that PKD2 was highly expressed in eight bladder cancer cell lines and in urothelial carcinoma tissues from the TCGA database, and that short hairpin RNA (shRNA)-mediated knockdown of PKD2 dramatically reduced bladder cancer growth and invasion in vitro and in vivo, suggesting that the effect of the compound in bladder cancer is mediated through inhibition of PKD2. This notion was corroborated by demonstrating that the levels of phospho-PKD2 were markedly decreased in CRT0066101-treated bladder tumor explants. Furthermore, our cell cycle analysis by flow cytometry revealed that CRT0066101 treatment or PKD2 silencing arrested bladder cancer cells at the G2/M phase, the arrest being accompanied by decreases in the levels of cyclin B1, CDK1 and phospho-CDK1 (Thr161) and increases in the levels of p27^Kip1 and phospho-CDK1 (Thr14/Tyr15). Moreover, CRT0066101 downregulated the expression of Cdc25C, which dephosphorylates/activates CDK1, but enhanced the activity of the checkpoint kinase Chk1, which inhibits CDK1 by phosphorylating/inactivating Cdc25C. Finally, CRT0066101 was found to elevate the levels of Myt1, Wee1, phospho-Cdc25C (Ser216), Gadd45α, and 14-3-3 proteins, all of which reduce the CDK1-cyclin B1 complex activity. These novel findings suggest that CRT0066101 suppresses bladder cancer growth by inhibiting PKD2 through induction of G2/M cell cycle arrest, leading to the blockade of cell cycle progression.     

Response of murine spermatocytes to the metaphase-arresting effect of several mitotic arrestants

Summary Three mitotic arrestants, colchicine, Colcemid and vinblastine, were evaluated for their effects on producing meiotic arrest in the laboratory mouse. Colchicine is the most effective and its effect is both dose- and time-dependent. Treatment of mice with 40 mg/kg of colchicine for 3 h resulted in drastic improvement of the yield and quality of M-II figures. This treatment did not appear to increase the aneuploidy frequency in M-II's.Supported in part by research grant R-808582 from the US Environmental Protection Agency and research grant RR5511-21 from the National Institute of Health.     

Mutational hotspot in the p53 gene in human hepatocellular carcinomas.

Human hepatocellular carcinomas (HCC) from patients in Qidong, an area of high incidence in China, in which both hepatitis B virus and aflatoxin B1 are risk factors, were analysed for mutations in p53, a putative tumour-suppressor gene. Eight of the 16 HCC had a point mutation at the third base position of codon 249. The G----T transversion in seven HCC DNA samples and the G----C transversion in the other HCC are consistent with mutations caused by aflatoxin B1 in mutagenesis experiments. No mutations were found in exons 5,6,8 or the remainder of exon 7. These results contrast with p53 mutations previously reported in carcinomas and sarcomas of human lung, colon, oesophagus and breast; these are primarily scattered over four of the five evolutionarily conserved domains, which include codon 249 (refs 4-9). We suggest that the mutant p53 protein may be responsible for a selective clonal expansion of hepatocytes during carcinogenesis.     

Multipartite matroids and secret sharing

In a secret-sharing scheme, a secret value is distributed among a set of participants by giving each participant a share. The re-quirement is that only predefined subsets of participants can recover the secret from their shares. The family of the predefined authorized subsets is called the access structure. An access structure is ideal if there exists a secret-sharing scheme realizing it in which the shares have optimal length, that is, in which the shares are taken from the same domain as the secrets. Brickell and Davenport proved that ideal access structures are induced by matroids. Subsequently, ideal access structures and access structures induced by matroids have received a lot of attention. Seymour gave the first example of an access structure induced by a matroid namely the Vamos matroid, that is non-ideal. Since every matroid is multipartite and has the associated discrete polymatroid, in this paper, by dealing with the rank functions of discrete polymatroids, we obtain a sufficient condition for a multipartite access structure to be ideal. Furthermore, we give a new proof that all access structures related to bipartite and tripartite matroids coincide with the ideal ones. Our results give new contributions to the open problem, that is, which matroids induce ideal access structures.     

Advanced materials for aqueous supercapacitors in the asymmetric design

Supercapacitors have been recognized as one of the promising energy storage devices in the future energy technology. In this perspective, rapid progress is made in the development of fundamental and applied aspects of supercapacitors. Various techniques have been developed specifically to estimate the specific capacitance. Numerous efforts have been made in the literature to increase the specific capacitance of electrode materials. Recently, researchers pay more attention on designing supercapacitors of asymmetric type with extending cell voltage and dissimilar materials with complementary working potentials. Researchers try to increase the specific energy of asymmetric supercapacitors (ASCs). Conversely, it is still a challenge to find a suitable operation conditions for ASCs in various designs, especially for the one with battery type electrode. In this review, we describe our recent research works and other reports on the preparation of various nanostructured electrode materials and the performances of both symmetric and asymmetric supercapacitors. Finally, we demonstrate effects of charge balance on the capacitive performances of ASCs which consist of one electrode material of the battery type and one capacitive material. We also demonstrate how to evaluate the charge capacities of both positive and negative electrode materials for this ASC application.     

Pregnenolone stabilizes microtubules and promotes zebrafish embryonic cell movement

Embryonic cell movement is essential for morphogenesis and the establishment of body shapes, but little is known about its mechanism. Here we report that pregnenolone, which is produced from cholesterol by the steroidogenic enzyme Cyp11a1 (cholesterol side-chain cleavage enzyme, P450scc), functions in promoting cell migration during epiboly. Epiboly is a process in which embryonic cells spread from the animal pole to cover the underlying yolk. During epiboly, cyp11a1 is expressed in an extra-embryonic yolk syncytial layer. Reducing cyp11a1 expression in zebrafish using antisense morpholino oligonucleotides did not perturb cell fates, but caused epibolic delay. This epibolic defect was partially rescued by the injection of cyp11a1 RNA or the supplementation of pregnenolone. We show that the epibolic delay is accompanied by a decrease in the level of polymerized microtubules, and that pregnenolone can rescue this microtubule defect. Our results indicate that pregnenolone preserves microtubule abundance and promotes cell movement during epiboly.     

Selective development of CD4+ T cells in transgenic mice expressing a class II MHC-restricted antigen receptor

T lymphocytes are predisposed to recognition of foreign protein fragments bound to cell-surface molecules encoded by the major histocompatibility complex (MHC). There is now compelling evidence that this specificity is a consequence of a selection process operating on developing T lymphocytes in the thymus. As a result of this positive selection, thymocytes that express antigen receptors with a threshold affinity for self MHC-encoded glycoproteins preferentially emigrate from the thymus and seed peripheral lymphoid organs. The specificity for both foreign antigen and MHC molecules is imparted by the alpha and beta chains of the T-cell antigen receptor (TCR). Two other T-cell surface proteins, CD4 and CD8, which bind non-polymorphic regions of class II and class I MHC molecules respectively, are also involved in these recognition events and play an integral role in thymic selection. In order to elucidate the developmental pathways of class II MHC-restricted T cells in relation to these essential accessory molecules, we have produced TCR-transgenic mice expressing a receptor specific for a fragment of pigeon cytochrome c and the Ek (class II MHC) molecule. The transgenic TCR is expressed on virtually all T cells in mice expressing Ek. The thymuses of these mice contain an abnormally high percentage of mature CD4+CD8- cells. In addition, the peripheral T-cell population is almost exclusively CD4+, demonstrating that the MHC specificity of the TCR determines the phenotype of T cells during selection in the thymus.