Synthesis of new pyrazolone and pyrazole-based adamantyl chalcones and antimicrobial activity.

Chalcones and their derivatives are becoming increasingly popular due to their various pharmacological effects. Chalcone molecules may be extracted from natural resources, entirely synthesised, or biosynthesised by modifying the natural ones. In the present study, five pyrazole-based adamantyl heterocyclic compounds were synthesised by condensation of 1-adamantyl chalcone with substituted phenylhydrazine. The products were characterised by using ¹H NMR, ¹³C NMR and FT-IR spectroscopy. The microbiological activity of these compounds was investigated against bacteria and fungi. The new compounds showed good to moderate activity against the microbial species used for screening. All developed molecules showed antibacterial activity against Gram-negative and Gram-positive. These molecules showed antifungal activities against Fusarium oxysporum fungus and in a dose-dependent manner, apart from RS-1 molecules which showed compromised antifungal activity and even at a high dose.

MicroRNAs in combined spent culture media and sperm are associated with embryo quality and pregnancy outcome.

OBJECTIVE: To identify differentially abundant miRNAs in sperm samples and spent culture media (SCM) of embryos of different grade toward a prediction of pregnancy outcome. DESIGN: Array-based reverse-transcription quantitative polymerase chain reaction profiling and validation. SETTING: University research institute and in vitro fertilization center. PATIENT(S): Couples (n = 61) undergoing infertility treatment with the use of intracytoplasmic sperm injection. INTERVENTIONS(S): None. MAIN OUTCOME MEASURE(S): Abundance levels of miRNAs in combined SCM of embryos of different quality and in sperm samples associated with pregnancy outcome. RESULT(S): Out of 372 screened miRNAs, miR-19b-3p and let-7a-5p were detected consistently in all SCM and sperm samples. The abundance levels of miRNAs were significantly altered between SCM of embryos with different quality (G1, G2, and G3 grades). Specifically, miR-320a and miR-15a-5p were differentially abundant in G1 vs. G2, miR-21-5p in G1 vs. G3, and miR-20a-5p in G2 vs. G3. The abundance levels of combined SCM and sperm derived miRNAs were also significantly altered between different pregnancy outcomes. MiR-19b-3p showed the highest area under the receiver operating characteristic curve values between positive and negative outcomes, with lower abundance levels in both combined SCM and sperm samples associated with a positive pregnancy outcome. MiR-320a, miR-15a-5p, miR-21-5p, and miR-20a-5p showed similar results in combined SCM samples. CONCLUSION(S): miRNA abundance levels in combined SCM and sperm differed significantly depending on embryo quality and pregnancy outcome. MiR-19b-3p may serve as a potential biomarker to predict pregnancy outcome.

Mesoderm-derived stem cells: the link between the transcriptome and their differentiation potential.

Human adult stem cells (hASCs) have become an attractive source for autologous cell transplantation, tissue engineering, developmental biology, and the generation of human-based alternative in vitro models. Among the 3 germ cell layers, the mesoderm is the origin of today's most widely used and characterized hASC populations. A variety of isolated nonhematopoietic mesoderm-derived stem cell populations exist, and all of them show important differences in terms of function, efficacy, and differentiation potential both in vivo and in vitro. To better understand whether the intrinsic properties of these cells contribute to the overall differentiation potential of hASCs, we compared the global gene expression profiles of 4 mesoderm-derived stem cell populations: human adipose tissue-derived stromal cells, human bone marrow-derived stromal cells (hBMSCs), human (fore)skin-derived precursor cells (hSKPs), and human Wharton's jelly-derived mesenchymal stem cells (hWJs). Significant differences in gene expression profiles were detected between distinct stem cell types. hSKPs predominantly expressed genes involved in neurogenesis, skin, and bone development, whereas hWJs and, to some extent, hBMSCs showed an increased expression of genes involved in cardiovascular and liver development. Interestingly, the observed differential gene expression of distinct hASCs could be linked to existing differentiation data in which hASCs were differentiated toward specific cell types. As such, our data suggest that the intrinsic gene expression of the undifferentiated stem cells has an important impact on their overall differentiation potential as well as their application in stem cell-based research. Yet, the factors that define these intrinsic properties remain to be determined.

Successful isolation of liver progenitor cells by aldehyde dehydrogenase activity in naïve mice.

UNLABELLED: The role of progenitor cells in liver repair and fibrosis has been extensively described, but their purification remains a challenge, hampering their characterization and use in regenerative medicine. To address this issue, we developed an easy and reproducible liver progenitor cell (LPC) isolation strategy based on aldehyde dehydrogenase (ALDH) activity, a common feature shared by many progenitor cells. We demonstrate that a subset of nonparenchymal mouse liver cells displays high levels of ALDH activity, allowing the isolation of these cells by fluorescence-activated cell sorting. Immunocytochemistry and qPCR analyses on freshly isolated ALDH(+) cells reveal an enrichment in cells expressing liver stem cell markers such as EpCAM, CK19, CD133, and Sox9. In culture, the ALDH(+) population can give rise to functional hepatocyte-like cells as illustrated by albumin and urea secretion and cytochrome P450 activity. ALDH1A1 expression can be detected in canals of Hering and bile duct epithelial cells and is increased on liver injury. Finally, we showed that the isolation and differentiation toward hepatocyte-like cells of LPCs with high ALDH activity is also successfully applicable to human liver samples. CONCLUSION: High ALDH activity is a feature of LPCs that can be taken advantage of to isolate these cells from untreated mouse as well as human liver tissues. This novel protocol is practically relevant, because it provides an easy and nontoxic method to isolate liver stem cells from normal tissue for potential therapeutic purposes.

Evaluation of the multipotent character of human adipose tissue-derived stem cells isolated by Ficoll gradient centrifugation and red blood cell lysis treatment.

In the present study, the multipotent potential of two differential isolated human adipose-derived stem cell (hADSC) populations was evaluated. More specifically, hADSC isolated by means of classical Ficoll (F) gradient centrifugation were compared to hADSC isolated by means of red blood cell (RBC) lysis treatment and subsequent cultivation as 3D spheres. No significant difference in the genotypic expression of the multipotent markers Oct-4, Sox-2, Nanog, Klf-4 and cMyc could be observed between both isolation methods. Upon adipogenic and osteogenic differentiation, both hADSC populations showed lipid droplet accumulation and mineral deposition, respectively. Although, a more pronounced mineral deposition was observed in hADSC-RBC, suggesting a higher osteogenic potential. Upon exposure to keratinogenic media, both hADSC populations expressed the keratinocyte markers filaggrin and involucrin, evidencing a successful keratinogenic differentiation. Yet, no differences in expression were observed between the distinctive isolation procedures. Finally, upon exposure to neurogenic differentiation media, a significant difference in marker expression was observed. Indeed, hADSC-RBC only expressed vimentin and nestin, whereas hADSC-F expressed vimentin, nestin, NF-200, MBP and TH, suggesting a higher neurogenic potential. In summary, our data suggest that the choice of the most efficient isolation procedure of hADSC depends on the differentiated cell type ultimately required.

Current status of human adipose-derived stem cells: differentiation into hepatocyte-like cells.

The shortage of human organ donors and the low cell quality of available liver tissues represent major obstacles for the clinical application of orthotropic liver transplantation and hepatocyte transplantation, respectively. Therefore, worldwide research groups are investigating alternative extrahepatic cell sources. Recent in vitro studies have demonstrated that mesenchymal stem cells (MSCs) from various sources, including human bone marrow, adipose tissue, and umbilical cord, can be differentiated into hepatocyte-like cells when appropriate conditions are used. In particular, interest exists for human adipose-derived stems cells (hASCs) as an attractive cell source for generating hepatocyte-like cells. The hASCs are multipotent MSCs that reside in adipose tissue, with the ability to self-renew and differentiate into multiple cell lineages. Moreover, these cells can secrete multiple growth factors and cytokines that exert beneficial effects on organ or tissue injury. In this review, we will not only present recent data regarding hASC biology, their isolation, and differentiation capability towards hepatocytes, but also the potential application of hASC-derived hepatocytes to study drug toxicity. Additionally, this review will discuss the therapeutic potential of hASCs as undifferentiated cells in liver regeneration.

The quest for liver progenitor cells: a practical point of view.

Many chronic liver diseases can lead to hepatic dysfunction with organ failure. At present, orthotopic liver transplantation represents the benchmark therapy of terminal liver disease. However this practice is limited by shortage of donor grafts, the need for lifelong immunosuppression and very demanding state-of-the-art surgery. For this reason, new therapies have been developed to restore liver function, primarily in the form of hepatocyte transplantation and artificial liver support devices. While already offered in very specialized centers, both of these modalities still remain experimental. Recently, liver progenitor cells have shown great promise for cell therapy, and consequently they have attracted a lot of attention as an alternative or supportive tool for liver transplantation. These liver progenitor cells are quiescent in the healthy liver and become activated in certain liver diseases in which the regenerative capacity of mature hepatocytes and/or cholangiocytes is impaired. Although reports describing liver progenitor cells are numerous, they have not led to a consensus on the identity of the liver progenitor cell. In this review, we will discuss some of the characteristics of these cells and the different ways that have been used to obtain these from rodents. We will also highlight the challenges that researchers are facing in their quest to identify and use liver progenitor cells.

Evaluation of medicinal plant hepatotoxicity in co-cultures of hepatocytes and monocytes.

Non-parenchymal cells might play an important role in the modulation of xenobiotic metabolism in liver and its pharmacological and toxicological consequences. Therefore, the role of cell-to-cell interactions in herbal induced liver toxicity was investigated in monocultures of cells from the human hepatocyte cell line (HepG2) and in co-cultures of cells from the HepG2 cell line and cells from the human monocyte cell line (THP1). Cells were treated with various concentrations (1-500 microg ml(-1)) of extracts of Pistacia palaestina, Juglans regia and Quercus ithaburensis for 24 h. Extracts from Cleome droserifolia, a known toxic plant, were taken as positive control. In the co-culture system, toxic effects were observed after exposure to extracts of Pistacia palestina and C. droserifolia. These two extracts significantly reduced by cell viability as measured the MTT test and the LDH assay. Whereas in hepatocyte cultures, only extracts of C. droserifolia were found to affect the cell viability. The production levels of albumin from hepatocytes were not affected by treatment with plant extracts in both culture systems. It seems that the observed reduction in cell viability after exposure to extracts of P. palestina in co-cultures but not in monocultures is a result of monocyte-derived factors. The use of liver cell co-cultures is therefore a useful approach to investigate the influence of intercellular communication on xenobiotic metabolism in liver.