In situ reprogramming of cardiac fibroblasts into cardiomyocytes in mouse heart with chemicals.

Cardiomyocytes are terminal differentiated cells and have limited ability to proliferate or regenerate. Condition like myocardial infarction causes massive death of cardiomyocytes and is the leading cause of death. Previous studies have demonstrated that cardiac fibroblasts can be induced to transdifferentiate into cardiomyocytes in vitro and in vivo by forced expression of cardiac transcription factors and microRNAs. Our previous study have demonstrated that full chemical cocktails could also induce fibroblast to cardiomyocyte transdifferentiation both in vitro and in vivo. With the development of tissue clearing techniques, it is possible to visualize the reprogramming at the whole-organ level. In this study, we investigated the effect of the chemical cocktail CRFVPTM in inducing in situ fibroblast to cardiomyocyte transdifferentiation with two strains of genetic tracing mice, and the reprogramming was observed at whole-heart level with CUBIC tissue clearing technique and 3D imaging. In addition, single-cell RNA sequencing (scRNA-seq) confirmed the generation of cardiomyocytes from cardiac fibroblasts which carries the tracing marker. Our study confirms the use of small molecule cocktails in inducing in situ fibroblast to cardiomyocyte reprogramming at the whole-heart level and proof-of-conceptly providing a new source of naturally incorporated cardiomyocytes to help heart regeneration.

Decryption for nitrogen removal in Anammox-based coupled systems: Nitrite-induced mechanisms.

This study investigated the effects of NO2- on synergetic interactions between Anammox bacteria (AnAOB) and sulfur-oxidizing bacteria (SOB) in an autotrophic denitrification-Anammox system. The presence of NO2- (0-75 mg-N/L) was shown to significantly enhance NH4+ and NO3- conversion rates, achieving intensified synergy between AnAOB and SOB. However, once NO2- exceed a threshold concentration (100 mg-N/L), both NH4+ and NO3- conversion rates decreased with increased NO2- consumption via autotrophic denitrification. The cooperation between AnAOB and SOB was decoupled due to the NO2- inhibition. Improved system reliability and nitrogen removal performance was achieved in a long-term reactor operation with NO2- in the influent; reverse transcription-quantitative polymerase chain reaction analysis showed elevated hydrazine synthase gene transcription levels (5.00-fold), comparing to these in the reactor without NO2-. This study elucidated the mechanism of NO2- induced synergetic interactions between AnAOB and SOB, providing theoretical guidance for engineering applications of Anammox-based coupled systems.

Simultaneous partial nitrification, Anammox and nitrate-dependent Fe(II) oxidation (NDFO) for total nitrogen removal under limited dissolved oxygen and completely autotrophic conditions.

Sustainable nitrogen removal from wastewater at reduced energy and/or chemical consumptions is challenging. This paper investigated, for the first time, the feasibility of coupled partial nitrification, Anammox and nitrate-dependent Fe(II) oxidation (NDFO) for sustainable autotrophic nitrogen removal. With NH4+-N as the only nitrogen-containing compound in the influent, near-complete nitrogen removal (a total of 97.5 % with a maximal total nitrogen removal rate of 6.64 ± 2.68 mgN/L/d) was achieved in a sequencing batch reactor for a 203-d operation without organic carbon source addition and forced aeration. Anammox (predominated by Candidatus Brocadia) and NDFO bacteria (such as Denitratisoma) were successfully enriched, with total relative abundances up to 11.54 % and 10.19 %, respectively. Dissolved oxygen (DO) concentration was a key factor affecting the coupling of multi (ammonia oxidization, Anammox, NDFO, iron-reduction, etc.) bacterial communities, resulting in different total nitrogen removal efficiencies and rates. In batch tests, the optimal DO concentration was 0.50-0.68 mg/L with a maximal total nitrogen removal efficiency of 98.7 %. Fe(II) in the sludge not only competed with nitrite oxidizing bacteria for DO to prevent complete nitrification, but promoted the transcription of NarG and NirK genes (10.5 and 3.5 times higher than the group without Fe(II) addition) as indicated by the reverse transcription quantitative polymerase chain reaction (RT-qPCR), resulting in increased NDFO rate (by 2.7 times) and promoted NO2--N generated from NO3--N, which back fed the Anammox process, achieving near-complete nitrogen removal. The reduction of Fe(III) by iron-reducing bacteria (IRB) and hydrolytic and fermentative anaerobes enabled a sustainable Fe(II)/Fe(III) recycling, avoiding the need in continuous Fe(II) or Fe (III) dosage. The coupled system is expected to benefit the development of novel autotrophic nitrogen removal processes with neglectable energy and material consumptions for the treatment of wastewater with low organic carbon and NH4+-N contents in underdeveloped regions, such as decentralized rural wastewaters.

The Aurora Kinase Inhibitor CYC116 Promotes the Maturation of Cardiomyocytes Derived from Human Pluripotent Stem Cells.

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have great potential in applications such as regenerative medicine, cardiac disease modeling, and in vitro drug evaluation. However, hPSC-CMs are immature, which limits their applications. During development, the maturation of CMs is accompanied by a decline in their proliferative capacity. This phenomenon suggests that regulating the cell cycle may facilitate the maturation of hPSC-CMs. Aurora kinases are essential kinases that regulate the cell cycle, the role of which is not well studied in hPSC-CM maturation. Here, we demonstrate that CYC116, an inhibitor of Aurora kinases, significantly promotes the maturation of CMs derived from both human embryonic stem cells (H1 and H9) and iPSCs (induced PSCs) (UC013), resulting in increased expression of genes related to cardiomyocyte function, better organization of the sarcomere, increased sarcomere length, increased number of mitochondria, and enhanced physiological function of the cells. In addition, a number of other Aurora kinase inhibitors have also been found to promote the maturation of hPSC-CMs. Our data suggest that blocking aurora kinase activity and regulating cell cycle progression may promote the maturation of hPSC-CMs.