TACSTD2 upregulation is an early reaction to lung infection.

TACSTD2 encodes a transmembrane glycoprotein Trop2 commonly overexpressed in carcinomas. While the Trop2 protein was discovered already in 1981 and first antibody-drug conjugate targeting Trop2 were recently approved for cancer therapy, the physiological role of Trop2 is still not fully understood. In this article, we show that TACSTD2/Trop2 expression is evolutionarily conserved in lungs of various vertebrates. By analysis of publicly available transcriptomic data we demonstrate that TACSTD2 level consistently increases in lungs infected with miscellaneous, but mainly viral pathogens. Single cell and subpopulation based transcriptomic data revealed that the major source of TACSTD2 transcript are lung epithelial cells and their progenitors and that TACSTD2 is induced directly in lung epithelial cells following infection. Increase in TACSTD2 expression may represent a mechanism to maintain/restore epithelial barrier function and contribute to regeneration process in infected/damaged lungs

Soluble collagen dissolution and assembling in pressurized carbon dioxide water solutions

Dissolution and gelation procedures have a great influence on gelation time, microstructure and mechanical properties of reconstituted collagen products. We have investigated the dissolution of atelocollagen in CO2/water solutions at low temperature (4 degrees C) at different CO2 pressures (0.3-0.9 MPa), as well as gelation kinetics and physico-chemical properties of the hydrogel obtained after CO2 removal. Compared to conventional methods, the CO2-assisted technique resulted in faster soluble collagen dissolution and faster gelation into transparent gels characterized by thin 10 nm fibrils. Electrophoresis and CD spectroscopy demonstrated that the process did not denature the soluble collagen. The possibility to obtain collagen dissolution and gelation without the use of chemical agent other than water and CO2 makes this process particularly appealing for biomedical applications

Macroporous bioceramic scaffolds based on tricalcium phosphates reinforced with silica: microstructural, mechanical, and biological evaluation

The positive effect of silica on microstructural, mechanical and biological properties of calcium phosphate scaffolds was investigated in this study. Scaffolds containing 3D interconnected spherical macropores with diameters in the range of 300-770 mu m were prepared by the polymer replica technique. Reinforcement was achieved by incorporating 5 to 20 wt % of colloidal silica into the initial hydroxyapatite (HA) powder. The HA was fully decomposed into alpha and beta-tricalcium phosphate, and silica was transformed into cristobalite at 1200 degrees C. Silica reinforced scaffolds exhibited compressive strength in the range of 0.3 to 30 MPa at the total porosity of 98-40%. At a nominal porosity of 75%, the compressive strength was doubled compared to scaffolds without silica. When immersed into a cultivation medium, the formation of an apatite layer on the surfaces of scaffolds indicated their bioactivity. The supportive effect of the silicon enriched scaffolds was examined using three different types of cells (human adipose-derived stromal cells, L929, and ARPE-19 cells). The cells firmly adhered to the surfaces of composite scaffolds with no sign of induced cell death. Scaffolds were non-cytotoxic and had good biocompatibility in vitro. They are promising candidates for therapeutic applications in regenerative medicine

Intact Cell Mass Spectrometry for Embryonic Stem Cell Biotyping

Stem cells represent a unique cell type that is capable of self-renewal and differentiation into somatic cell types. Since the derivation of human embryonic stem cells and induced pluripotent stem cells, enormous potential has been recognized for disease modeling, drug development and regenerative medicine. Both embryonic stem cells and induced pluripotent stem cells possess the ability to differentiate into all three germ layers, hence they are naturally prone to respond to various differentiation stimuli. These inherent cellular fluctuations, which can result in risky phenotypic instability, must be addressed prior to introduction of these cells to human medicine, since they represent one of the major biosafety obstacles in the development of bio-industrial or clinical-grade stem cell cultures. Therefore, there is an ongoing need for novel robust, feasible and sensitive methods for determination and confirmation of the otherwise identical cells status, as well as for the detection of hidden divergences from their optimal state. A method of choice can be the intact cell mass spectrometry. Here we show how it can be applied in routine quality control of embryonic stem cell cultures

The CHK1 inhibitor MU380 significantly increases the sensitivity of human docetaxel-resistant prostate cancer cells to gemcitabine through the induction of mitotic catastrophe

As treatment options for patients with incurable metastatic castration-resistant prostate cancer (mCRPC) are considerably limited, novel effective therapeutic options are needed. Checkpoint kinase 1 (CHK1) is a highly conserved protein kinase implicated in the DNA damage response (DDR) pathway that prevents the accumulation of DNA damage and controls regular genome duplication. CHK1 has been associated with prostate cancer (PCa) induction, progression, and lethality; hence, CHK1 inhibitors SCH900776 (also known as MK-8776) and the more effective SCH900776 analog MU380 may have clinical applications in the therapy of PCa. Synergistic induction of DNA damage with CHK1 inhibition represents a promising therapeutic approach that has been tested in many types of malignancies, but not in chemoresistant mCRPC. Here, we report that such therapeutic approach may be exploited using the synergistic action of the antimetabolite gemcitabine (GEM) and CHK1 inhibitors SCH900776 and MU380 in docetaxel-resistant (DR) mCRPC. Given the results, both CHK1 inhibitors significantly potentiated the sensitivity to GEM in a panel of chemo-naïve and matched DR PCa cell lines under 2D conditions. MU380 exhibited a stronger synergistic effect with GEM than clinical candidate SCH900776. MU380 alone or in combination with GEM significantly reduced spheroid size and increased apoptosis in all patient-derived xenograft 3D cultures, with a higher impact in DR models. Combined treatment induced premature mitosis from G1 phase resulting in the mitotic catastrophe as a prestage of apoptosis. Finally, treatment by MU380 alone, or in combination with GEM, significantly inhibited tumor growth of both PC339-DOC and PC346C-DOC xenograft models in mice. Taken together, our data suggest that metabolically robust and selective CHK1 inhibitor MU380 can bypass docetaxel resistance and improve the effectiveness of GEM in DR mCRPC models. This approach might allow for dose reduction of GEM and thereby minimize undesired toxicity and may represent a therapeutic o

Human embryonic stem cell line originating from blastocyst produced by intracytoplasmic sperm injection

The biological properties of the AL1 ES cell line will be assessed by introducing the cells into the brains of adult Wistar rats following a unilateral cortical photochemical lesion