Effect of partial desulfation and oversulfation of sodium spirulan on the potency of anti-herpetic activities

The aim of the present study is to evaluate the effect of partial desulfation and oversulfation of sodium spirulan (Na-SP) isolated from Spirulina platensis on the exhibition of anti-herpes simplex viruses type 1 and 2 (HSV-1 and -2) activities. Partially desulfated (PDS-SPs) and oversulfated derivatives (OS-SPs) were obtained by solvolytic desulfation and sulfation with dicyclocarbodiimide-sulfuric acid, respectively. When PDS-SPs were subjected to anti-HSV-1 assay, antiviral potency was dependent on their sulfate content, and PDS-SPs with lower sulfate content than 8.6% were found to be inactive. Some derivatives showing anti-HSV-1 effect also showed anti-HSV-2 activity. Anti-HSV-1 effect of OS-SPs was equivalent to that of Na-SP when they were added to the medium during viral infection and throughout the incubation thereafter, while they were enhanced as compared with Na-SP when they were added to the medium immediately after viral infection. The results of time-of-addition experiments suggested that the most sensitive phase of OS-SP-2 and -5 might be the early steps of viral adsorption and penetration.     

TGF-β1 increases cellular invasion of colorectal neuroendocrine carcinoma cell line through partial epithelial-mesenchymal transition

Epithelial–mesenchymal transition (EMT) plays a pivotal role in cancer progression and metastasis in many types of malignancies, including colorectal cancer. Although the importance of EMT is also considered in colorectal neuroendocrine carcinoma (NEC), its regulatory mechanisms have not been elucidated. We recently established a human colorectal NEC cell line, SS-2. In this study, we aimed to clarify whether these cells were sensitive to transforming growth factor beta 1 (TGF-β1) and whether EMT could be induced through TGF-β1/Smad signaling, with the corresponding NEC cell-specific changes in invasiveness. In SS-2 cells, activation of TGF-β1 signaling, as indicated by phosphorylation of Smad2/3, was dose-dependent, demonstrating that SS-2 cells were responsive to TGF-β1. Analysis of EMT markers showed that mRNA levels changed with TGF-β1 treatment and that E-cadherin, an EMT marker, was expressed in cell-cell junctions even after TGF-β1 treatment. Invasion assays showed that TGF-β1-treated SS-2 cells invaded more rapidly than non-treated cells, and these cells demonstrated increased metalloproteinase activity and cell adhesion. Among integrins involved in cell-to-matrix adhesion, α2-integrin was exclusively upregulated in TGF-β1-treated SS-2 cells, but not in other colon cancer cell lines, and adhesion and invasion were inhibited by an anti-α2-integrin blocking antibody. Our findings suggest that α2-integrin may represent a novel therapeutic target for the metastasis of colorectal NEC cells.     

CD23 molecule acts as a galactose-binding lectin in the cell aggregation of EBV-transformed human B-cell lines

Epstein-Barr virus (EBV)-transformed human B-cell lines, L-KT9and DH3 cells express CD23 antigen, and grow in a mixture ofsingle and aggregated cells. The CD23 molecule has high aminoacid sequence homology with C-type lectin and recently we haveshown that the solubilized CD23 molecule can really interactwith galactose residues on glycoproteins. In this study, therefore,we tested whether CD23 antigen on the cell surface really actsas a galactose-binding lectin in the aggregation of these cells.The EBV-transformed cells (L-KT9) were separated into an aggregated-cell-richfraction and a single-cell-rich fraction. Aggregated cells disaggregatedafter removal of galactose by ß-galactosidase treatment,whereas single cells made large aggregation on sialidase treatment,and this aggregation was inhibited in the presence of asialo-fetuin.On the other hand, naturally aggregated cells become singlecells with anti-CD23 monoclonal antibody (mAB) as well as thesoluble form of CD23, but not with anti-CD21 mAB. In addition,L-KT9 and DH3 cells bound to asialo-fetuin-coupled Sepharose(ASF-Sepharose) and this binding was significantly inhibitedby pre-treatment of cells with anti-CD23, but not with anti-CD21or other antiadhesion molecules. From these results, we concludethat the naturally aggregated state of EBV-transformed cellsoccurs mainly through the interaction of CD23 as a lectin moleculeand galactose residues as its ligand. CD23 molecule cell aggregation EBV-transformed B cells glycosidase treatment low-affinity IgE receptor     

Sample size determination in clinical trials with multiple co‐primary endpoints including mixed continuous and binary variables

In the field of pharmaceutical drug development, there have been extensive discussions on the establishment of statistically significant results that demonstrate the efficacy of a new treatment with multiple co‐primary endpoints. When designing a clinical trial with such multiple co‐primary endpoints, it is critical to determine the appropriate sample size for indicating the statistical significance of all the co‐primary endpoints with preserving the desired overall power because the type II error rate increases with the number of co‐primary endpoints. We consider overall power functions and sample size determinations with multiple co‐primary endpoints that consist of mixed continuous and binary variables, and provide numerical examples to illustrate the behavior of the overall power functions and sample sizes. In formulating the problem, we assume that response variables follow a multivariate normal distribution, where binary variables are observed in a dichotomized normal distribution with a certain point of dichotomy. Numerical examples show that the sample size decreases as the correlation increases when the individual powers of each endpoint are approximately and mutually equal.     

Coprinopsis novorugosobispora sp. nov., an agaric ammonia fungus from Beijing,China

Coprinopsis novorugosobispora (Basidiomycota, Psathyrellaceae), a new ammonia fungus, fructificated from the soil sample collected in Beijing, China. In macro- and micro-morphology, this species is similar to C. rugosobispora reported from Belgium, but differs in having smaller basidiospore and rougher basidiospore ornamentation. This is the first report of ammonia fungi from continental China.