The development and characterization of a human mesothelioma in vitro 3D model to investigate immunotoxin therapy.

BackgroundTumor microenvironments present significant barriers to penetration by antibodies and immunoconjugates. Tumor microenvironments, however, are difficult to study in vitro. Cells cultured as monolayers exhibit less resistance to therapy than those grown in vivo and an alternative research model more representative of the in vivo tumor is more desirable. SS1P is an immunotoxin composed of the Fv portion of a mesothelin-specific antibody fused to a bacterial toxin that is presently undergoing clinical trials in mesothelioma.Methodology/principal findingsHere, we examined how the tumor microenvironment affects the penetration and killing activity of SS1P in a new three-dimensional (3D) spheroid model cultured in vitro using the human mesothelioma cell line (NCI-H226) and two primary cell lines isolated from the ascites of malignant mesothelioma patients. Mesothelioma cells grown as monolayers or as spheroids expressed comparable levels of mesothelin; however, spheroids were at least 100 times less affected by SS1P. To understand this disparity in cytotoxicity, we made fluorescence-labeled SS1P molecules and used confocal microscopy to examine the time course of SS1P penetration within spheroids. The penetration was limited after 4 hours. Interestingly, we found a significant increase in the number of tight junctions in the core area of spheroids by electron microscopy. Expression of E-Cadherin, a protein involved in the assembly and sealing of tight junctions and highly expressed in malignant mesothelioma, was found significantly increased in spheroids as compared to monolayers. Moreover, we found that siRNA silencing and antibody inhibition targeting E-Cadherin could enhance SS1P immunotoxin therapy in vitro.Conclusion/significanceThis work is one of the first to investigate immunotoxins in 3D tumor spheroids in vitro. This initial description of an in vitro tumor model may offer a simple and more representative model of in vivo tumors and will allow for further investigations of the microenvironmental effects on drug penetration and tumor cell killing. We believe that the methods developed here may apply to the studies of other tumor-targeting antibodies and immunoconjugates in vitro

Loss of T Cell Progenitor Checkpoint Control Underlies Leukemia Initiation in Rag1-Deficient Nonobese Diabetic Mice

NOD mice exhibit major defects in the earliest stages of T cell development in the thymus. Genome-wide genetic and transcriptome analyses were used to investigate the origins and consequences of an early T cell developmental checkpoint breakthrough in Rag1-deficient NOD mice. Quantitative trait locus analysis mapped the presence of checkpoint breakthrough cells to several known NOD diabetes susceptibility regions, particularly insulin-dependent diabetes susceptibility genes (Idd)9/11 on chromosome 4, suggesting common genetic origins for T cell defects affecting this trait and autoimmunity. Genome-wide RNA deep-sequencing of NOD and B6 Rag1-deficient thymocytes revealed the effects of genetic background prior to breakthrough, as well as the cellular consequences of the breakthrough. Transcriptome comparison between the two strains showed enrichment in differentially expressed signal transduction genes, prominently tyrosine kinase and actin-binding genes, in accord with their divergent sensitivities to activating signals. Emerging NOD breakthrough cells aberrantly expressed both stem cell–associated proto-oncogenes, such as Lmo2, Hhex, Lyl1, and Kit, which are normally repressed at the commitment checkpoint, and post–β-selection checkpoint genes, including Cd2 and Cd5. Coexpression of genes characteristic of multipotent progenitors and more mature T cells persists in the expanding population of thymocytes and in the thymic leukemias that emerge with age in these mice. These results show that Rag1-deficient NOD thymocytes have T cell defects that can collapse regulatory boundaries at two early T cell checkpoints, which may predispose them to both leukemia and autoimmunity

One-dimensional coordination polymers containing penta-supertetrahedral sulfide clusters linked by dipyridyl ligands

通讯作者地址: Feng, PY (通讯作者), Univ Calif Riverside, Dept Chem, Riverside, CA 92521 USA 地址: 1. Univ Calif Riverside, Dept Chem, Riverside, CA 92521 USA 2. Calif State Univ Long Beach, Dept Chem & Biochem, Long Beach, CA 90840 USA 电子邮件地址: [email protected] new one-dimensional coordination polymers [Zn8S(SC6H5)(14).C12H10N2](1), [Zn7CoS(SC6H5)14(.)C(13)H(14)N(2)](2) and [Zn8S(SC6H5)14(.)C(13)H(14)N(2)](3) have been prepared containing penta-supertetrahedral clusters and linear crosslinking dipyridyl ligands; the two complexes show optical transitions with band gaps of similar to 3.44 eV (1) and similar to 3.54 eV (2)

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Development of an Instrument to Measure Flow Experience in Computer Game Play

Effective measurement of the enjoyment of computer game play will assist game designers to understand the strength and flaw of the game from players’ perspective. We argue that flow experience in computer game play contributes to enjoyment. This paper reports on the development of an instrument for measuring flow experience in computer game play. The instrument was developed based on the flow theory (Csikszentmihalyi, 1993) and a rigorous method introduced by Moore and Benbasat (1991). The results show that the validity and reliability of the instrument are satisfactory. This instrument will help IS researchers to further investigate the application of flow theory in computer games

Research on Energy Trading Mechanism of Zero carbon Park Based on Blockchain Technology

In the context of global climate change and various environmental problems, stabilizing the global average temperature, implementing near zero carbon emissions, and finally achieving absolute zero carbon emissions in the future have become an important task of China’s ecological civilization construction. As an important population and industrial cluster, the park is the carrier of important economic and social activities, so the park is the most important form to achieve the dual carbon goal. Based on blockchain technology, this paper comprehensively uses carbon neutral mechanisms such as zero carbon energy substitution, low carbon technology, carbon sequestration and carbon sink, and purchase of voluntary emission reductions to gradually reduce the carbon emissions in the park to zero, and finally realizes intelligent and automatic management of carbon emission reduction management in the park

Research on Energy Trading Mechanism of Zero carbon Park Based on Blockchain Technology

In the context of global climate change and various environmental problems, stabilizing the global average temperature, implementing near zero carbon emissions, and finally achieving absolute zero carbon emissions in the future have become an important task of China’s ecological civilization construction. As an important population and industrial cluster, the park is the carrier of important economic and social activities, so the park is the most important form to achieve the dual carbon goal. Based on blockchain technology, this paper comprehensively uses carbon neutral mechanisms such as zero carbon energy substitution, low carbon technology, carbon sequestration and carbon sink, and purchase of voluntary emission reductions to gradually reduce the carbon emissions in the park to zero, and finally realizes intelligent and automatic management of carbon emission reduction management in the park

A novel wideband, low-profile and second-order miniaturized band-pass frequency selective surfaces

A novel wideband, low-profile and second-order miniaturized band-pass frequency selective surface (FSS) made of metallic mesh and its complementary structures with skewed arrays of modified triples is presented in this paper. Compared with traditional second-order bandpass FSSs obtained using λ/4 apart from one another, the novel FSS with an overall thickness of λ/18 is composed of three metallic layers (the outside and middle layers are complementary) separated by two electric thin dielectric substrates. This arrangement can shorten the inter-element spacing and increase the bandwidth, while the up and bottom metallic layers can constitute a symmetric biplanar FSS and thus realize ability of maximally flat second-order bandpass response. The novel FSS has a −3 dB bandwidth about 8.2 GHz (6.9 -15.1 GHz) and a fractional bandwidth exceeds 75%. Moreover, such an FSS has the merits of stable performance for incident angles within 50° and different polarizations. The principles of operation along with guidelines for the design of the proposed FSS, the simulated results by vector modal matching method, and the experimental values of the fabricated prototype are also presented and discussed

Silene vanchingshanensis (Caryophyllaceae) a new species from Southwest China

Silene vanchingshanensis (Caryophyllaceae), a new species from Fanjingshan Mountain in Guizhou (southwest China) is described and illustrated. It is morphologically similar to S. morrisonmontana and S. hupehensis, from which it can be easily distinguished by having pubescent stems usually 10–15 cm long, linear-oblanceolate leaves 3–6 cm × 3–6 mm, often 2–5-flowered cymes, pink or violet petals and narrowly ovoid capsules