Background Ovarian cancer is a highly aggressive malignant disease in gynecologic cancer. It is an urgent task to develop three-dimensional (3D) cell models in vitro and dissect the cell progression-related drug resistance mechanisms in vivo. In the present study, RADA16-I peptide has the reticulated nanofiber scaffold networks in hydrogel, which is utilized to develop robust 3D cell culture of a high metastatic human ovarian cancer HO-8910PM cell line accompanied with the counterparts of Matrigel and collagen I. Results Consequently, HO-8910PM cells were successfully cultivated in three types of hydrogel biomaterials, such as RADA16-I hydrogel, Matrigel, and collagen I, according to 3D cell culture protocols. Designer RADA16-I peptide had well-defined nanofiber networks architecture in hydrogel, which provided nanofiber cell microenvironments analogous to Matrigel and collagen I. 3D-cultured HO-8910PM cells in RADA16-I hydrogel, Matrigel, and collagen I showed viable cell proliferation, proper cell growth, and diverse cell shapes in morphology at the desired time points. For a long 3D cell culture period, HO-8910PM cells showed distinct cell aggregate growth patterns in RADA16-I hydrogel, Matrigel, and collagen I, such as cell aggregates, cell colonies, cell clusters, cell strips, and multicellular tumor spheroids (MCTS). The cell distribution and alignment were described vigorously. Moreover, the molecular expression of integrin β1, E-cadherin and N-cadherin were quantitatively analyzed in 3D-cultured MCTS of HO-8910PM cells by immunohistochemistry and western blotting assays. The chemosensitivity assay for clinical drug responses in 3D context indicated that HO-8910PM cells in three types of hydrogels showed significantly higher chemoresistance to cisplatin and paclitaxel compared to 2D flat cell culture, including IC50 values and inhibition rates. Conclusion Based on these results, RADA16-I hydrogel is a highly competent, high-profile, and proactive nanofiber scaffold to maintain viable cell proliferation and high cell vitality in 3D cell models, which may be particularly utilized to develop useful clinical drug screening platform in vitro.

To construct a three-dimensional (3D) culture model of adenovirus in vitro using the nanoself-assembling peptide RADA16-I as a 3D cell culture scaffold combined with virology experimental technology to provide a novel research method for virus isolation and culture, pathogenesis research, antiviral drug screening and vaccine preparation. Methods: The nanoself-assembling peptide RADA16-I was used as a 3D scaffold material for 293T cell culture, and adenovirus was cultured in the cells. The growth, morphological characteristics and pathological effects of 3D-cultured 293T cells after adenovirus infection were observed with an inverted microscope and MTS. The proliferation of adenovirus in 293T cells was observed by TEM and detected by qPCR. The levels of TNF-α and IL-8 secreted by adenovirus-infected 293T cells in the RADA16-I 3D culture system were detected by ELISA. Results: The 293T cells grew well in the RADA16-I 3D culture system for a prolonged period of time. The adenovirus infection persisted for a long time with multiple proliferation peaks, which closely resembled those of in vivo infections. The adenovirus virions amplified in the 3D system remained infectious. There were multiple secretion peaks of TNF-α and IL-8 secretion levels in adenovirus-infected 293T cells cultured in 3D culture systems. Conclusion: The nanoself-assembling peptide RADA16-I can be used as a 3D scaffold for adenovirus isolation, culture and research. The 3D culture system shows more realistic in vivo effects than two-dimensional (2D) culture.

M. 2008. Differential expression of dehydrin genes in hull-less barley (Hordeum vulgare ssp. vulgare) depending on duration of dehydration stress. Can. J. Plant Sci. 88: 899Á906. Dehydrins (DHN) are water-soluble lipid-associating proteins that accumulate during low-temperature and water-stress conditions, and are thought to play a protective role in freezing and drought tolerance in plants. The synthesis of the dehydrins, representing a link between environment and nuclear activity through dehydrin genes (Dhn) expression, was known to be a common response to drought in plants. The association of mRNA accumulations of Dhn genes (Dhn6, Dhn11 and Dhn13) with phenotypically diverse drought-tolerant genotypes was investigated in hull-less barley (Hordeum vulgare subsp. vulgare). Notably, differences between genotypes were observed, mainly in Dhn6 and Dhn13 gene, depending on the duration of the dehydration stress. Compared with drought-sensitive genotypes, the relative expression levels of Dhn6 gene were significantly higher in drought-tolerant lines subjected to 8 h water deficit. By control with continuative ascending accumulations in sensitive plants during drought, the highest transcript level of Dhn13 was observed in tolerant genotypes after 8 h of dehydration, and tended to descend the same levels as sensitive lines subjected to 12 h of dehydration. Based on decreased accumulations of Dhn11 during drought stress, it was concluded that Dhn11 was not up-regulated by water stress between genotypes. The observed differences in the present study may indicate an association of physiological response to water stress with differential accumulations of Dhn genes between drought-contrasting genotypes. The authors suggest different DHNs may play variable functional roles in structural protection in plants subjected to progressive water stress, associated with drought-contrasting genotypes.Key words: Dhydrin genes, quantitative reverse transcription polymerase chain reaction, water deficit, Tibetan hull-less barley Qian, G., Liu, Y., Ao, D., Yang, F. et Yu, M. 2008. Variation de l'expression des ge`nes de la de´hydrine dans une orge nue (Hordeum vulgare ssp. vulgare) selon la dure´e de la de´shydratation. Can. J. Plant Sci. 88: 899Á906. La de´hydrine (DHN) est une prote´ine lipophile hydrosoluble qui s'accumule quand la tempe´rature descend et qu'il y a stress hydrique. On pense qu'elle prote`ge la plante contre le gel et lui permet de tole´rer la se´cheresse. La synthe`se des de´hydrines, qui repre´sente un lien entre l'environnement et l'activite´du noyau par le biais des ge`nes qui la codent (Dhn), est une re´action commune a`la se´cheresse chez les ve´ge´taux. Les auteurs ont e´tudie´la relation entre l'accumulation de l'ARNm des ge`nes Dhn (Dhn6, Dhn11 et Dhn13) et des varie´te´s de phe´notype variable tole´rant la se´cheresse chez l'orge nue (Hordeum vulgare subsp. vulgare). Ils ont ainsi observe´des variations entre les ge´notypes, principalement au niveau des ge`nes Dhn6 et Dhn13, selon la dure´e du stress engend...

Viral infectious diseases seriously threaten human health. At present, the prevention and treatment of viral diseases depends primarily on vaccines and drugs. Commonly used research method include animal models or simple traditional two-dimensional (2D) isolation culture, but 2D cell behaviour is different from the human physiological microenvironment. Therefore, a new model that can simulate the human microenvironment is needed. Here, the nanoself-assembled peptide KLD-12 was used as a culture scaffold for 293T cells in threedimensional (3D) adenovirus culture. As a new 3D virus culture model, it simulates in vivo virus infection, and the model can produce infectious particles. In addition, the antiviral drug sensitivity between the 3D and 2D cultures was significantly different. We established a 3D adenovirus culture model that can be used for adenovirus proliferation and antiviral drug screening, as well as for gene therapy, vaccine research and other research. Additionally, it can partially replace animal models.

Background Multiple host factors are involved in modulating type I interferon expression induced by viruses; however, the mechanism is not fully elucidated. Influenza A virus infection causes severe respiratory symptoms and triggers a series of signaling cascades and host innate immune responses, including interferon production. The co-IP/MS technology was used to screen several antiviral factors in the early stage. Among these factors, ariadne-1 homolog (ARIH1) caught our attention. Methods Western blot assay was performed to detect the level of proteins and software ImageJ was used to analyze the band intensities. Polymerase activity assay was conducted to evaluate the polymerase activity of influenza A virus. Tissue culture infective dose (TCID50) assay was performed to measure influenza A virus titers, and quantitative RT-PCR assay was applied to test the mRNA level of IFN-β, ISG56, and CXCL10. Luciferase reporter assay was used to confirm the target of ARIH1 in RIG-I signaling. Immunoprecipitation assay was performed to detect the interaction and the ubiquitination of the proteins. All data were analyzed by biostatistical methods and presented as means ± standard deviation from three independent experiments. Statistical significance was determined using two-tailed student’s t test. A P value of less than 0.05 was considered statistically significant, and a P value of less than 0.01 was considered highly significant (ns, P ≥ 0.05; *, P < 0.05; and **, P < 0.01). Results We found that ARIH1, a member of E3 ubiquitin ligases, enhanced cellular antiviral responses. Subsequent study showed that ARIH1 was up-regulated during influenza A virus infection. Further analysis showed that ARIH1 enhanced IFN-β and downstream gene expression by affecting the degradation of RIG-I through the SQSTM1/p62 signaling pathway. Conclusion This newly revealed mechanism shows that cellular response increases of ARIH1 and promotes IFN-β expression to boost host survival during viral infection.