Pharmacological inhibition of cyclin-dependent kinases triggers anti-fibrotic effects in hepatic stellate cells in vitro

Liver fibrosis is a wound healing process in response to chronic liver injury, which is characterized by the accumulation of extracellular collagen produced by Hepatic Stellate Cells (HSCs). This process involves cell cycle re-entry and proliferation of normally quiescent HSCs controlled by cyclins and associated cyclin-dependent kinases (Cdks). Cdk2 mediates the entry and progression through S-phase in complex with E-and A-type cyclins. We have demonstrated that cyclin E1 is essential for liver fibrogenesis in mice, but it is not known if this is dependent on Cdk2 or related Cdks. Here, we aimed to evaluate the benefit of the pan-Cdk inhibitor CR8 for treatment of liver fibrosis in vitro. CR8-treatment reduced proliferation and survival in immortalized HSC lines and in addition attenuated pro-fibrotic properties in primary murine HSCs. Importantly, primary murine hepatocytes were much more tolerant against the cytotoxic and anti-proliferative effects of CR8. We identified CR8 dosages mediating anti-fibrotic effects in primary HSCs without affecting cell cycle activity and survival in primary hepatocytes. In conclusion, the pharmacological pan-Cdk inhibitor CR8 restricts the pro-fibrotic properties of HSCs, while preserving proliferation and viability of hepatocytes at least in vitro. Therefore, CR8 and related drugs might be beneficial for the treatment of liver fibrosis

Onset and cause of increased seismic activity near Pecos, West Texas, USA from observations at the Lajitas TXAR Seismic Array

In recent years, numerous small earthquakes have occurred near the town of Pecos in West Texas; however, when this activity began and whether it was caused by increased petroleum industry activity has been uncertain because prior to 2017 there were few permanent seismograph stations in the region. We identify and locate earthquakes using data recorded since 2000 at TXAR, a sensitive 10‐station seismic array situated about 240 km south of Pecos. We thus show that in 2007 one earthquake occurred near Pecos, in 2009 several more occurred, and subsequently activity has increased considerably, with more than 2000 events identified in 2017. A time‐of‐day and year‐by‐year analysis identifies geographic areas in West Texas where events are likely to be natural earthquakes and quarry blasts. However, for the Pecos events, annual seismicity rates increase along with annual volumes of petroleum production and fluid waste disposal, suggesting a causal link. Analysis of seismograms collected by the EarthScope Transportable Array indicates the 2009 earthquakes had focal depths of 4.0‐5.2 km below sea level, within or just below strata where petroleum is produced and/or wastewater is injected. The largest earthquake to date had magnitude ML3.7, but the recent high activity rates suggest that greater magnitudes may be possible. For the years 2000‐2017, we provide a catalog of 10,753 epicenters of seismic events recorded at TXAR. Plain Language Summary Petroleum production in the Permian Basin of West Texas has been accelerating since 2007, and by 2023 it is anticipated Permian Basin production will exceed the production of every nation in the world other than Saudi Arabia. Developing this domestic source of energy has profound economic and political implications, especially since protecting vital foreign sources of energy has been a major factor affecting U.S. foreign policy. In recent years, numerous small earthquakes have occurred in the Delaware Basin (a subregion of the Permian Basin), but when this seismicity began has been uncertain because there were few seismographs in this region before 2017. We show that these anomalous earthquakes first occurred in 2009, and that many of them are probably induced by petroleum production in the Delaware Basin. Understanding the relationship between production and earthquake activity is a critical first step towards mitigating seismic hazards that could affect local populations and compromise the development of these vital petroleum resources

Artificial Insemination as an Alternative Transmission Route for African Swine Fever Virus

The rapid spread of the African swine fever virus (ASFV), causing severe disease with often high fatality rates in Eurasian suids, prevails as a threat for pig populations and dependent industries worldwide. Although advancing scientific progress continually enhances our understanding of ASFV pathogenesis, alternative transmission routes for ASFV have yet to be assessed. Here, we demonstrate that ASFV can efficiently be transferred from infected boars to naïve recipient gilts through artificial insemination (AI). In modern pig production, semen from boar studs often supplies many sow herds. Thus, the infection of a boar stud presents the risk of rapidly and widely distributing ASFV within or between countries. Daily blood and semen collection from four boars after intramuscular inoculation with ASFV strain ‘Estonia 2014’ resulted in the detection of ASFV genomes in the semen as early as 2 dpi, in blood at 1 dpi while semen quality remained largely unaffected. Ultimately, after insemination with extended semen, 7 of 14 gilts were ASFV positive by 7 days post insemination, and all gilts were ASFV positive by 35 days post insemination. Twelve out of 13 pregnant gilts aborted or resorbed at the onset of fever. A proportion of fetuses originating from the remaining gilt showed both abnormalities and replication of ASFV in fetal tissues. Thus, we present evidence for the efficient transmission of ASFV to gilts via AI and also to implanted embryos. These results underline the critical role that boar semen could play in ASFV transmission

The Na+/H+ Exchanger Nhe1 Modulates Network Excitability via GABA Release

Brain functions are extremely sensitive to pH changes because of the pH-dependence of proteins involved in neuronal excitability and synaptic transmission. Here, we show that the Na+/H+ exchanger Nhe1, which uses the Na+ gradient to extrude H+, is expressed at both inhibitory and excitatory presynapses. We disrupted Nhe1 specifically in mice either in Emx1- positive glutamatergic neurons or in parvalbumin-positive cells, mainly GABAergic interneurons. While Nhe1 disruption in excitatory neurons had no effect on overall network excitability, mice with disruption of Nhe1 in parvalbumin-positive neurons displayed epileptic activity. From our electrophysiological analyses in the CA1 of the hippocampus, we conclude that the disruption in parvalbumin-positive neurons impairs the release of GABA-loaded vesicles, but increases the size of GABA quanta. The latter is most likely an indirect pH-dependent effect, as Nhe1 was not expressed in purified synaptic vesicles itself. Conclusively, our data provide first evidence that Nhe1 affects network excitability via modulation of inhibitory interneurons

Langerhans Cells Facilitate Epithelial DNA Damage and Squamous Cell Carcinoma

Polyaromatic hydrocarbons (PAHs) are prevalent, potent carcinogens, and 7,12-dimethylbenz[a]anthracene (DMBA) is a model PAH widely used to study tumorigenesis. Mice lacking Langerhans cells (LCs), a signatory epidermal dendritic cell (DC), are protected from cutaneous chemical carcinogenesis, independent of T cell immunity. Investigation of the underlying mechanism revealed that LC-deficient skin was relatively resistant to DMBA-induced DNA damage. LCs efficiently metabolized DMBA to DMBA-trans-3,4-diol, an intermediate proximal to oncogenic Hras mutation, and DMBA-treated LC-deficient skin contained significantly fewer Hras mutations. Moreover, DMBA-trans-3,4-diol application bypassed tumor resistance in LC-deficient mice. Additionally, the genotoxic impact of DMBA on human keratinocytes was significantly increased by prior incubation with human-derived LC. Thus, tissue-associated DC can enhance chemical carcinogenesis via PAH metabolism, highlighting the complex relation between immune cells and carcinogenesis