Active eosinophilic esophagitis is characterized by epithelial barrier defects and eosinophil extracellular trap formation.

BACKGROUND Eosinophilic esophagitis (EoE) exhibits esophageal dysfunction owing to an eosinophil-predominant inflammation. Activated eosinophils generate eosinophil extracellular traps (EETs) able to kill bacteria. There is evidence of an impaired barrier function in EoE that might allow pathogens to invade the esophagus. This study aimed to investigate the presence and distribution of EETs in esophageal tissues from EoE patients and their association with possible epithelial barrier defects. METHODS Anonymized tissue samples from 18 patients with active EoE were analyzed. The presence of DNA nets associated with eosinophil granule proteins forming EETs and the expression of filaggrin, the protease inhibitor lympho-epithelial Kazal-type-related inhibitor (LEKTI), antimicrobial peptides, and cytokines were evaluated by confocal microscopy following immune fluorescence staining techniques. RESULTS Eosinophil extracellular trap formation occurred frequently and was detected in all EoE samples correlating with the numbers of infiltrating eosinophils. While the expression of both filaggrin and LEKTI was reduced, epithelial antimicrobial peptides (human beta-defensin-2, human beta-defensin-3, cathelicidin LL-37, psoriasin) and cytokines (TSLP, IL-25, IL-32, IL-33) were elevated in EoE as compared to normal esophageal tissues. There was a significant correlation between EET formation and TSLP expression (P = 0.02) as well as psoriasin expression (P = 0.016). On the other hand, a significant negative correlation was found between EET formation and LEKTI expression (P = 0.016). CONCLUSION Active EoE exhibits the presence of EETs. Indications of epithelial barrier defects in association with epithelial cytokines are also present which may have contributed to the activation of eosinophils. The formation of EETs could serve as a firewall against the invasion of pathogens.

Paleohydraulic reconstruction of a 40 kyr-old terrace sequence implies that water discharge was larger than today

The evolution of landscapes crucially depends on the climate history. This is particularly evident in South America where landscape responses to orbital climate shifts have been well documented. However, while most studies have focused on inferring temperature variations from paleoclimate proxy data, estimates of water budget changes have been complicated because of a lack of adequate physical information. Here, we present a methodology and related results, which allowed us to extract water discharge values from the sedimentary record of the 40 Ka-old fluvial terrace deposits in the Pisco valley, western Peru. In particular, this valley hosts a Quaternary cut-and-fill succession that we used, in combination with beryllium-10 (10Be)-based sediment flux, gauging records, channel geometries and grain size measurements, to quantitatively assess sediment and water discharge values c. 40 Ka ago in relation to present-day conditions. We compare these discharge estimates to the discharge regime of the modern Pisco River and find that the water discharge of the paleo-Pisco River, during the Minchin pluvial period c. 40 Ka ago, was c. 7–8 times greater than the modern Pisco River if considering the mean and the maximum water discharge. In addition, the calculations show that inferred water discharge estimates are mainly dependent on channel gradients and grain size values, and to a lesser extent on channel width measures. Finally, we found that the c. 40 Ka-old Minchin terrace material was poorer sorted than the modern deposits, which might reflect that sediment transport during the past period was characterized by a larger divergence from equal mobility compared to the modern situation. In summary, the differences in grain size distribution and inferred water discharge estimates between the modern and the paleo-Pisco River suggests that the 40 Ka-old Minchin period was characterized by a wetter climate and more powerful flood events.

Leptomeningeal contrast enhancement and blood-CSF barrier dysfunction in aseptic meningitis

OBJECTIVE To investigate the blood-CSF barrier (BCSFB) dysfunction in aseptic meningitis. METHODS In our case series of 14 patients with acute aseptic meningitis, we compared MRI characteristics with CSF findings. RESULTS Contrast enhancement in the sulcal space in a leptomeningeal pattern was visualized in 7 patients with BCSFB dysfunction categorized as moderate to severe as evidenced by the CSF/serum albumin ratio (Qalb) but was not present in those with mild or no barrier disturbance (p = 0.001). The Qalb as a marker for the leakiness of the BCSFB and, more indirectly, of the blood-brain barrier (BBB) was positively correlated with the incidence of leptomeningeal contrast enhancement seen on postcontrast fluid-attenuated inversion recovery (FLAIR) MRI (p = 0.003). Patients with a more pronounced brain barrier dysfunction recovered more slowly and stayed longer in the hospital. CONCLUSIONS The severity of meningeal BBB disturbance can be estimated on postcontrast FLAIR MRI, which may be of diagnostic value in patients with aseptic meningitis.

Engineering an in vitro air-blood barrier by 3D bioprinting

Intensive efforts in recent years to develop and commercialize in vitro alternatives in the field of risk assessment have yielded new promising two- and three dimensional (3D) cell culture models. Nevertheless, a realistic 3D in vitro alveolar model is not available yet. Here we report on the biofabrication of the human air-blood tissue barrier analogue composed of an endothelial cell, basement membrane and epithelial cell layer by using a bioprinting technology. In contrary to the manual method, we demonstrate that this technique enables automatized and reproducible creation of thinner and more homogeneous cell layers, which is required for an optimal air-blood tissue barrier. This bioprinting platform will offer an excellent tool to engineer an advanced 3D lung model for high-throughput screening for safety assessment and drug efficacy testing.

Comparing CFSR and conventional weather data for discharge and soil loss modelling with SWAT in small catchments in the Ethiopian Highlands

Accurate rainfall data are the key input parameter for modelling river discharge and soil loss. Remote areas of Ethiopia often lack adequate precipitation data and where these data are available, there might be substantial temporal or spatial gaps. To counter this challenge, the Climate Forecast System Reanalysis (CFSR) of the National Centers for Environmental Prediction (NCEP) readily provides weather data for any geographic location on earth between 1979 and 2014. This study assesses the applicability of CFSR weather data to three watersheds in the Blue Nile Basin in Ethiopia. To this end, the Soil and Water Assessment Tool (SWAT) was set up to simulate discharge and soil loss, using CFSR and conventional weather data, in three small-scale watersheds ranging from 112 to 477 ha. Calibrated simulation results were compared to observed river discharge and observed soil loss over a period of 32 years. The conventional weather data resulted in very good discharge outputs for all three watersheds, while the CFSR weather data resulted in unsatisfactory discharge outputs for all of the three gauging stations. Soil loss simulation with conventional weather inputs yielded satisfactory outputs for two of three watersheds, while the CFSR weather input resulted in three unsatisfactory results. Overall, the simulations with the conventional data resulted in far better results for discharge and soil loss than simulations with CFSR data. The simulations with CFSR data were unable to adequately represent the specific regional climate for the three watersheds, performing even worse in climatic areas with two rainy seasons. Hence, CFSR data should not be used lightly in remote areas with no conventional weather data where no prior analysis is possible.

Modeling immune functions of the mouse blood-cerebrospinal fluid barrier in vitro: primary rather than immortalized mouse choroid plexus epithelial cells are suited to study immune cell migration across this brain barrier.

BACKGROUND The blood-cerebrospinal fluid barrier (BCSFB) established by the choroid plexus (CP) epithelium has been recognized as a potential entry site of immune cells into the central nervous system during immunosurveillance and neuroinflammation. The location of the choroid plexus impedes in vivo analysis of immune cell trafficking across the BCSFB. Thus, research on cellular and molecular mechanisms of immune cell migration across the BCSFB is largely limited to in vitro models. In addition to forming contact-inhibited epithelial monolayers that express adhesion molecules, the optimal in vitro model must establish a tight permeability barrier as this influences immune cell diapedesis. METHODS We compared cell line models of the mouse BCSFB derived from the Immortomouse(®) and the ECPC4 line to primary mouse choroid plexus epithelial cell (pmCPEC) cultures for their ability to establish differentiated and tight in vitro models of the BCSFB. RESULTS We found that inducible cell line models established from the Immortomouse(®) or the ECPC4 tumor cell line did not express characteristic epithelial proteins such as cytokeratin and E-cadherin and failed to reproducibly establish contact-inhibited epithelial monolayers that formed a tight permeability barrier. In contrast, cultures of highly-purified pmCPECs expressed cytokeratin and displayed mature BCSFB characteristic junctional complexes as visualized by the junctional localization of E-cadherin, β-catenin and claudins-1, -2, -3 and -11. pmCPECs formed a tight barrier with low permeability and high electrical resistance. When grown in inverted filter cultures, pmCPECs were suitable to study T cell migration from the basolateral to the apical side of the BCSFB, thus correctly modelling in vivo migration of immune cells from the blood to the CSF. CONCLUSIONS Our study excludes inducible and tumor cell line mouse models as suitable to study immune functions of the BCSFB in vitro. Rather, we introduce here an in vitro inverted filter model of the primary mouse BCSFB suited to study the cellular and molecular mechanisms mediating immune cell migration across the BCSFB during immunosurveillance and neuroinflammation.