The Relationship among Hypoxia,Proliferation, and Outcome in Patients with De Novo Glioblastoma: A Pilot Study

The hypoxia and proliferation index increase with grade in human glial tumors, but there is no agreement whether either has prognostic importance in glioblastomas. We evaluated these end points individually and together in 16 de novo human glioblastomas using antibodies against the 2-nitroimidazole hypoxia detection agent EF5 and the proliferation detection agent Ki-67. Frozen tumor tissue sections were fluorescence-stained for nuclei (Hoechst 33342), hypoxia (anti-EF5 antibodies), and proliferation (anti-Ki-67 antibodies). EF5 binding adjacent to Ki-67+ cells, overall EF5 binding, the ratio of these values, and the proliferation index were evaluated. Patients were classified using recursive partitioning analysis and followed up until recurrence and/or death. Recursive partitioning analysis was statistically significant for survival (P = .0026). Overall EF5 binding, EF5 binding near Ki-67+ cells, and proliferation index did not predict recurrence. Two additional survival analyses based on ratios of the overall EF5 binding to EF5 binding near Ki-67+ cells were performed. High and low ratio values were determined by two cutoff points: (a) the 50% value for the ratio [EF5/Ki-67_Binding]/[Tumor_binding] = Ratio_{EF5 50%} and (b) the median EF5 value (75.6%) of the ratio [EF5/Ki-67_Binding]/[Tumor_binding] = Ratio_{patients median}. On the basis of the Ratio_{EF5 50%}, recurrence (P = .0074) and survival (P = .0196) could be predicted. Using the Ratio_{patients median}, only survival could be predicted (P = .0291). In summary, patients had a worse prognosis if the [EF5/Ki-67_Binding]/[Tumor_binding] ratio was high. A hypothesis for the mechanisms and translational significance of these findings is discussed.     

Fibroblast growth factor-10 promotes cardiomyocyte differentiation from embryonic and induced pluripotent stem cells

Background

The fibroblast growth factor (FGF) family is essential to normal heart development. Yet, its contribution to cardiomyocyte differentiation from stem cells has not been systemically studied. In this study, we examined the mechanisms and characters of cardiomyocyte differentiation from FGF family protein treated embryonic stem (ES) cells and induced pluripotent stem (iPS) cells.

Methodology/Principal Findings

We used mouse ES cells stably transfected with a cardiac-specific α-myosin heavy chain (αMHC) promoter-driven enhanced green fluorescent protein (EGFP) and mouse iPS cells to investigate cardiomyocyte differentiation. During cardiomyocyte differentiation from mouse ES cells, FGF-3, -8, -10, -11, -13 and -15 showed an expression pattern similar to the mesodermal marker Brachyury and the cardiovascular progenitor marker Flk-1. Among them, FGF-10 induced cardiomyocyte differentiation in a time- and concentration-dependent manner. FGF-10 neutralizing antibody, small molecule FGF receptor antagonist PD173074 and FGF-10 and FGF receptor-2 short hairpin RNAs inhibited cardiomyocyte differentiation. FGF-10 also increased mouse iPS cell differentiation into cardiomyocyte lineage, and this effect was abolished by FGF-10 neutralizing antibody or PD173074. Following Gene Ontology analysis, microarray data indicated that genes involved in cardiac development were upregulated after FGF-10 treatment. In vivo, intramyocardial co-administration of FGF-10 and ES cells demonstrated that FGF-10 also promoted cardiomyocyte differentiation.

Conclusion/Significance

FGF-10 induced cardiomyocyte differentiation from ES cells and iPS cells, which may have potential for translation into clinical applications.     

Multi-locus sequence typing of enteroaggregative Escherichia coli isolates from Nigerian children uncovers multiple lineages

Background

Enteroaggregative Escherichia coli (EAEC) are defined by their stacked-brick adherence pattern to human epithelial cells. There is no all-encompassing genetic marker for EAEC. The category is commonly implicated in diarrhea but research is hampered by perplexing heterogeneity.

Methodology/Principal Findings

To identify key EAEC lineages, we applied multilocus sequence typing to 126 E. coli isolates from a Nigerian case-control study that showed aggregative adherence in the HEp-2 adherence assay, and 24 other EAEC strains from diverse locations. EAEC largely belonged to the A, B1 and D phylogenetic groups and only 7 (4.6%) isolates were in the B2 cluster. As many as 96 sequence types (STs) were identified but 60 (40%) of the EAEC strains belong to or are double locus variants of STs 10, 31, and 394. The remainder did not belong to predominant complexes. The most common ST complex, with predicted ancestor ST10, included 32 (21.3%) of the isolates. Significant age-related distribution suggests that weaned children in Nigeria are at risk for diarrhea from of ST10-complex EAEC. Phylogenetic group D EAEC strains, predominantly from ST31- and ST394 complexes, represented 38 (25.3%) of all isolates, include genome-sequenced strain 042, and possessed conserved chromosomal loci.

Conclusions/Significance

We have developed a molecular phylogenetic framework, which demonstrates that although grouped by a shared phenotype, the category of ‘EAEC’ encompasses multiple pathogenic lineages. Principal among isolates from Nigeria were ST10-complex EAEC that were associated with diarrhea in children over one year and ECOR D strains that share horizontally acquired loci.     

Synthesis and antitumor activity of bithienyl-pyrimidine derivatives with electrostatic binding side chains.

A series of bithienyl-pyrimidines having cationic side chain have been developed as antitumor agents. This work illustrates the overwhelming importance of the bithienyl unit for efficient DNA binding. The X-ray structure of 4-(2',2"-thien-5-yl)2-chloropyrimidine was obtained for postulating the conformation of the bithienyl-pyrimidine moiety.     

Orally bioavailable nonpeptide vitronectin receptor antagonists containing 2-aminopyridine arginine mimetics.

A peptide RGD analog containing a novel 2-aminopyridine arginine mimetic was discovered to have good affinity and selectivity for the vitronectin receptor. Incorporation of the 2-aminopyridine arginine mimetic into the 3-oxo-1,4-benzodiazepine-2-acetic acid integrin antagonist series led to novel and potent nonpeptide vitronectin receptor antagonists with promising levels of oral bioavailability.     

Thrombin inhibitors based on a propargylglycine template

A series of novel arylsulfonylpropargylglycinamide derivatives was investigated as thrombin inhibitors in which the SAR was focused on substituents at the acetylenic terminus. Several compounds in this series were identified as potent thrombin inhibitors (Ki up to 5 nM) that are highly selective over trypsin and other serine proteases as well.     

Micro RNAs as mediators of cardiovascular disease: Targets to be manipulated

Cardiovascular disease has been the leading cause of death worldwide for the last few decades. Even with therapid progression of the biomedical field, conquering/managing cardiovascular disease is not an easy task because it is multifactorial disease. One of the key players of the development and progression of numerous diseases is micro RNA(mi RNA). These small, non-coding RNAs bind to target m RNAs to inhibit translations of and/or degrade the target m RNAs, thus acting as negative regulators of gene expressions. Accumulating evidence indicates that non-physiological expressions of mi RNAs contribute to both development and progression of cardiovascular diseases. Since even a single mi RNA can have multiple targets, dysregulation of mi RNAs can lead to catastrophic changes of proteins that may be important for maintaining physiologic conditions of cells, tissues, and organs. Current knowledge on the role of mi RNAs in cardiovascular disease is mostly based on the observational data such as microarray of mi RNAs in animal disease models, thus relatively lacking insight of how such dysregulation of mi RNAs is initiated and regulated. Consequently, future research should aim to elucidate the more comprehensive mechanisms of mi RNA dysregulation during pathogenesis of the cardiovascular system so that appropriate countermeasures to prevent/manage cardiovascular disease can be developed.     

Development of recombinant <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis> producing virus-like particles of a fish nervous necrosis virus

Nervous necrosis virus (NNV) causes viral encephalopathy and retinopathy, a devastating disease of many species of cultured marine fish worldwide. In this study, we used the dimorphic non-pathogenic yeast Yarrowia lipolytica as a host to express the capsid protein of red-spotted grouper nervous necrosis virus (RGNNV-CP) and evaluated its potential as a platform for vaccine production. An initial attempt was made to express the codon-optimized synthetic genes encoding intact and N-terminal truncated forms of RGNNV-CP under the strong constitutive TEF1 promoter using autonomously replicating sequence (ARS)-based vectors. The full-length recombinant capsid proteins expressed in Y. lipolytica were detected not only as monomers and but also as trimers, which is a basic unit for formation of NNV virus-like particles (VLPs). Oral immunization of mice with whole recombinant Y. lipolytica harboring the ARS-based plasmids was shown to efficiently induce the formation of IgG against RGNNV-CP. To increase the number of integrated copies of the RGNNV-CP expression cassette, a set of 26S ribosomal DNA-based multiple integrative vectors was constructed in combination with a series of defective Ylura3 with truncated promoters as selection markers, resulting in integrants harboring up to eight copies of the RGNNV-CP cassette. Sucrose gradient centrifugation and transmission electron microscopy of this high-copy integrant were carried out to confirm the expression of RGNNV-CPs as VLPs. This is the first report on efficient expression of viral capsid proteins as VLPs in Y. lipolytica, demonstrating high potential for the Y. lipolytica expression system as a platform for recombinant vaccine production based on VLPs.     

Deoxynivalenol impair skin barrier function through the down regulation of filaggrin and claudin 1/8 in HaCaT keratinocyte

Deoxynivalenol (DON), a typical mycotoxin, is a substance that is biosynthesized mainly by the Fusarium species. It is usually found in wheat and other grains grown in the field. When it enters the human body, it causes severe diarrhea, abdominal pain, vomiting, and even death. In addition, DON is known to induce inflammation of the small and large intestine, and is also associated with the occurrence of cancer. However, until recently, the effects of DON on the human skin were unknown. To investigate how DON affects HaCaT, human immortalized keratinocytes, we used CCK-8 assay and a quantitative real-time RT-PCR method to detect changes in the expression of tight junctions and skin cell regulatory proteins. The CCK-8 assay was performed to determine the growth inhibitory concentration of keratinocytes by DON. DON affected the cell survival rate from 1 μM in a concentration dependent manner, with the minimum set as 1 μM and the maximum as 4 μM for all experiments. DON inhibited the mRNA expression of filaggrin by up to 71% and SERPINA1 up to 75%. The expression of AQP3 was reduced by up to 93% compared to the untreated control group. This may cause problems in the pH control function of the skin and weaken the function of moisturizing. In addition, in the presence of DON, the gene expression of claudin 1 and claudin 8, which are important proteins in the regulation of intercellular skin barrier, decreased by up to 47 and 80%, respectively. Snail/ Slug, suppressors of the claudin gene expression, each increased up to 625 and 974%, respectively. Also, the MMP9 gene increased by up to 515% in a concentrationdependent manner, perhaps causing a weakness of the barrier function of the skin. These results suggest that DON may causing the development of atopic skin by impairing the skin barrier and pH control of skin, as well as intestinal inflammation diseases. Therefore, particular attention should be paid to DON contamination during the development of cosmetic ingredients using grains.     

Multiple male morphs in the leaf‐footed bug Mictis longicornis (Hemiptera: Coreidae)

Species within the coreid clade (Hemiptera: Coreidae) can often be observed competing in intrasexual competitions over access to mates and territories. Coreids that partake in these competitions typically possess sexually dimorphic hind legs that are used to strike and squeeze their rivals. In addition to their weaponized legs, some coreid species also possess sexually dimorphic abdominal tubercles, which are assumed to be sexually selected weapons. Still, much remains unknown about the morphology of these structures. Here, using the species Mictis longicornis Westwood, we investigate the frequency distribution and static allometry of abdominal thickness, a measure that includes tubercle length. Furthermore, we also investigate the morphological relationship between abdominal tubercles and weaponized hind legs. We find that male abdominal thickness is best explained by a bimodal distribution, thereby describing the first observed male polymorphism in the coreid clade; a phenomenon typically associated with alternative reproductive tactics. Additionally, we find that major males are characterized primarily by having large weaponized legs and abdominal tubercles, which further suggests that abdominal tubercles are used in male–male competition.