Direct reprogramming of fibroblasts into functional hepatocytes via CRISPRa activation of endogenous Gata4 and Foxa3.

BACKGROUND: The ability to generate functional hepatocytes without relying on donor liver organs holds significant therapeutic promise in the fields of regenerative medicine and potential liver disease treatments. Clustered regularly interspaced short palindromic repeats (CRISPR) activator (CRISPRa) is a powerful tool that can conveniently and efficiently activate the expression of multiple endogenous genes simultaneously, providing a new strategy for cell fate determination. The main purpose of this study is to explore the feasibility of applying CRISPRa for hepatocyte reprogramming and its application in the treatment of mouse liver fibrosis. METHOD: The differentiation of mouse embryonic fibroblasts (MEFs) into functional induced hepatocyte-like cells (iHeps) was achieved by utilizing the CRISPRa synergistic activation mediator (SAM) system, which drove the combined expression of three endogenous transcription factors-Gata4, Foxa3, and Hnf1a-or alternatively, the expression of two transcription factors, Gata4 and Foxa3. In vivo, we injected adeno-associated virus serotype 6 (AAV6) carrying the CRISPRa SAM system into liver fibrotic Col1a1-CreER; Cas9fl/fl mice, effectively activating the expression of endogenous Gata4 and Foxa3 in fibroblasts. The endogenous transcriptional activation of genes was confirmed using real-time quantitative polymerase chain reaction (RT-qPCR) and RNA-seq, and the morphology and characteristics of the induced hepatocytes were observed through microscopy. The level of hepatocyte reprogramming in vivo is detected by immunofluorescence staining, while the improvement of liver fibrosis is evaluated through Sirius red staining, alpha-smooth muscle actin (α-SMA) immunofluorescence staining, and blood alanine aminotransferase (ALT) examination. RESULTS: Activation of only two factors, Gata4 and Foxa3, via CRISPRa was sufficient to successfully induce the transformation of MEFs into iHeps. These iHeps could be expanded in vitro and displayed functional characteristics similar to those of mature hepatocytes, such as drug metabolism and glycogen storage. Additionally, AAV6-based delivery of the CRISPRa SAM system effectively induced the hepatic reprogramming from fibroblasts in mice with live fibrosis. After 8 weeks of induction, the reprogrammed hepatocytes comprised 0.87% of the total hepatocyte population in the mice, significantly reducing liver fibrosis. CONCLUSION: CRISPRa-induced hepatocyte reprogramming may be a promising strategy for generating functional hepatocytes and treating liver fibrosis caused by hepatic diseases.

The digital health divide: Understanding telehealth adoption across racial lines in rural Illinois.

Due to long-standing barriers to healthcare access in rural areas, telehealth has been promoted as an effective means of delivering healthcare services. However, there is a general absence of quantitative data showing how geographic residence and race affect telehealth adoption. This study examines variations in telehealth adoption based on race and geographic residence in Southern Illinois using a mail survey. It finds that residents of urban Carbondale, compared to those in rural Cairo, have better access to broadband and are more likely to use telehealth. Respondents significantly differ from each other based on their geographic location of residence and race when it came to using telehealth to save money on travel and to save money on childcare. A significant barrier to telehealth adoption identified across all groups is privacy protection concern. The findings highlight the crucial role of broadband infrastructure in healthcare access and the need for trust in telehealth systems to ensure data privacy.

IL-6/gp130 signaling: a key unlocking regeneration.

Liver is an organ with notable capacity of regeneration. Reprogramming of hepatocytes towards an immature state is one of the important mechanisms for hepatocyte replenishment. Inflammatory response mediated by IL-6 and its family cytokines has been widely reported closely related with tissue regeneration in myriads of organs. Recently Hui and colleagues reported that the dedifferentiation of hepatocytes depends upon IL-6 signaling from Kupffer cells and the reprogramming of gene expression under the inflammatory condition is different from the regulation of gene expression during embryo hepatocyte specification, highlighting a tight linkage between extracellular microenvironment and parenchymal cell plasticity during tissue regenerative repair.

Targeting immunoregulation for cardiac regeneration.

Inducing endogenous cardiomyocyte proliferation and heart regeneration is a promising strategy to treat ischemic heart failure. The immune response has recently been considered critical in cardiac regeneration. Thus, targeting the immune response is a potent strategy to improve cardiac regeneration and repair after myocardial infarction. Here we reviewed the characteristics of the relationship between the postinjury immune response and heart regenerative capacity and summarized the latest studies focusing on inflammation and heart regeneration to identify potent targets of the immune response and strategies in the immune response to promote cardiac regeneration.

Antibacterial activity of γFe2 O3 /TiO2 nanoparticles on toxic cyanobacteria from a lake in Southern Illinois.

Frequent outbreaks of harmful algal blooms (HABs) have brought adverse impacts on human health, economic viability, and recreational activities in many communities in the United States. Cyanobacteria (or blue-green algae) blooms are the most common type of HABs in surface water. Current bactericides for controlling the blooms are disadvantageous due to the recycling difficulty. In this study, an innovative magnetic nanomaterial-γFe2 O3 /TiO2 nanoparticle-was used to inactivate toxic cyanobacteria species found in a lake in Southern Illinois that frequently experienced HABs. Cyanotoxin genes of mcy, nda, cyr, and sxt were used for targeting microcystin-, nodularin-, cylindrospermopsin-, and saxitoxin-producing cyanobacteria, respectively, by quantitative polymerase chain reaction (PCR) method. It was found that the concentration of chlorophyll a presents a strong correlation (R2 = 0.6024) with the gene copy obtained from 16S rRNA targeted for all cyanobacteria, but not with that from individual toxigenic cyanobacteria. The inactivation efficiencies of the nanomaterials under visible light were as high as 5-log and 1-log for cyanobacteria species containing mcyE/ndaF and sxtA genes, respectively, an improvement over the treatment under darkness. These nanomaterials can be recycled by their magnetic properties for reuse. Communities susceptible to HAB outbreaks are expected to benefit from the developed method for mitigating the blooms. PRACTITIONER POINTS: Lab-made γFe2 O3 /TiO2 nanoparticles can be used to inactivate microcystin/nodularin- and saxitoxin-producing cyanobacteria species. qPCR method can be used for targeting toxic cyanobacteria; Chl a cannot be used as a standalone indicator for HABs. Better inactivation efficiency under visible light indicated possible application of the technology under sunlight for HAB mitigation from surface water.

Aerobic Cross-Dehydrogenative Coupling Reactions for Selective Mono- and Dithiolation of Phenols.

A highly efficient strategy for the direct thiolation of phenols under transition metal-free and solvent-free conditions has been developed. These reactions are operationally simple with employing air (molecular oxygen) as an ideal oxidant and can selectively provide mono- and dithiolation products in good to excellent yields under basic conditions. The reaction tolerates a broad range of aryl thiols and arenes and is especially applicable for large-scale synthesis.

A review on drone-based harmful algae blooms monitoring.

Rapid development and applications of unmanned aerial vehicles (UAVs) provide promising solutions to and new opportunities for environmental monitoring. Owing to their flexibility in flight scheduling, high spatial resolution, and costs-effectiveness, UAVs have become a popular tool for monitoring dynamic environmental processes, such as emergence and outbreak of harmful algae blooms (HABs). The HABs outbreak, often linked with anthropogenic eutrophication, has become a serious environmental health problem that threats our communities. Existing studies show that UAV-based HABs monitoring is a cost-effective means of assisting environmental managers in developing precautionary warning system and coping strategies. This article summarized the state-of-the-art of using UAVs and lightweight onboard multispectral sensors for HABs monitoring from the perspective of quantitative remote sensing. It culminates in a discussion of challenges and opportunities for future research and applications on drone-based HABs monitoring.

OxLDL promotes lymphangiogenesis and lymphatic metastasis in gastric cancer by upregulating VEGF­C expression and secretion.

Gastric cancer is one of the most malignant tumor types, and its metastasis is a notable cause of mortality. Among the methods of tumor metastasis, lymphatic metastasis is the predominant one in gastric cancer. A previous study reported that the plasma oxidized low­density lipoprotein (oxLDL) is the risk factor associated with the development of tumors in patients with abnormal lipid metabolism, but the influence of plasma oxLDL in the lymphatic metastasis of gastric cancer remains unclear. In the present study, the concentration of plasma oxLDL from patients with gastric cancer was detected with an ELISA kit, and the lymphatic vessel density in gastric cancer tissues was determined by D2­40 staining. The correlation analysis of oxLDL concentration and lymphatic vessel density demonstrated that plasma oxLDL was positively correlated with lymphatic metastasis in patients with gastric cancer. Subsequently, the popliteal lymph node metastasis animal experiment with nude mice confirmed that oxLDL could promote the lymphatic metastasis of gastric cancer. Following this, the western blotting and ELISA data demonstrated that oxLDL promoted the expression and secretion of vascular endothelia growth factor (VEGF)­C in gastric cancer cell lines. Finally, blocking the lectin­like oxLDL­1 (LOX­1) receptor, a specific receptor for oxLDL, and the nuclear factor (NF)­κB signaling pathway following oxLDL (50 µg/ml) treatment in HGC­27 cells revealed that oxLDL could activate the NF­κB signaling pathway mediated by LOX­1, with subsequent upregulation of VEGF­C expression, and secretion in and from gastric cancer cells, and finally that it could promote the lymphatic metastasis of gastric cancer. These data indicate the association between the plasma oxLDL and the lymphatic metastasis of gastric cancer, and indicate that oxLDL elimination may be a potential therapeutic target for the prevention and intervention of early lymph node metastasis in gastric cancer.

Assessing Nebraska playa wetland inundation status during 1985-2015 using Landsat data and Google Earth Engine.

Playa wetlands in Nebraska provide globally important habitats for migratory waterfowl. Inundation condition is an important indicator of playa wetland functionality. However, there is a lack of long-term continuous monitoring records for playa wetlands. The objective of this study was to determine a suitable index for Landsat images to map the playa inundation status in March and April during 1985-2015. Four types of spectral indices-negative normalized vegetation index, Normalized Difference Water Index (NDWI), modified NDWI, and Tasseled Cap Wetness-Greenness Difference (TCWGD)-were evaluated to detect playa inundation conditions from Landsat images. The results indicate that the TCWGD is the most suitable index for distinguishing playa inundation status. By using Landsat images and Google Earth Engine, we mapped the spring inundation condition of Nebraska playas during 1985-2015. The results show that the total inundated areas were 176.79 km2 in spring migratory season, representing 18.92% of the total area of playa wetlands. There were 9898 wetlands inundated at least once in either March or April during the past 30 years, representing 29.41% of a total of 33,659 historical wetlands. After comparing the historical hydric soil footprints and the inundated areas, the results indicate that the hydrological conditions of the majority of playas in Nebraska have changed. The inundated wetlands are candidates for protection and/or partial restoration, and the un-inundated wetlands need more attention for wetland restoration. Wetlands in areas enrolled in conservation easements had a significantly high level of playa inundation status than non-conserved wetlands during spring migratory seasons in the past decades.These conservation easements only count for 4.29% of the total footprint areas, but they have contributed 20.82% of the inundation areas in Nebraska during the past 30 years.

Modeling vulnerability of groundwater to pollution under future scenarios of climate change and biofuels-related land use change: a case study in North Dakota, USA.

Modeling groundwater vulnerability to pollution is critical for implementing programs to protect groundwater quality. Most groundwater vulnerability modeling has been based on current hydrogeology and land use conditions. However, groundwater vulnerability is strongly dependent on factors such as depth-to-water, recharge and land use conditions that may change in response to future changes in climate and/or socio-economic conditions. In this research, a modeling framework, which employs three sets of models linked within a geographic information system (GIS) environment, was used to evaluate groundwater pollution risks under future climate and land use changes in North Dakota. The results showed that areas with high vulnerability will expand northward and/or northwestward in Eastern North Dakota under different scenarios. GIS-based models that account for future changes in climate and land use can help decision-makers identify potential future threats to groundwater quality and take early steps to protect this critical resource.