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Transparency of the human cornea is responsible for clear vision, which is maintained by a monolayer of non-proliferative human corneal endothelial cells (HCEnCs). Dysfunction of these cells can result in irreversible corneal blindness. It is important to identify key factors that limit the proliferation of HCEnCs and thus attempt to reverse them. Extracellular vesicles contain cargo which includes microRNAs (miRNAs) that can modulate a cellular function. In non small cell lung cancer, expression of miR-195-5p has been shown to inhibit proliferation; therefore, we aimed to investigate the inhibitory effect of miR-195-5p in inducing the proliferation of HCEnCs. Human corneal endothelial cell line (HCEC-12) and primary HCEnCs were cultured with miR-195-5p scramble, mimic or inhibitor. Corneal tissues from human cadaveric and FECD donors, and from pigs, mice and rabbits, were used for RT-PCR. miR-195-5p showed an abundance value of 11,363.31 a.u. When normalized against HCEnCs from cadaveric donors, FECD tissues showed a significant upregulation of miR-195-5p (p < 0.05) but was significantly downregulated in pig (p < 0.001), mouse (p < 0.01) and rabbit (p < 0.001) CEnCs, which have known proliferative capacity. Proliferation, cell doubling, and wound healing rates were significantly higher when miR-195-5p was inhibited. Inhibiting miR-195-5p showed a significant improvement in viability (HEC staining), decreased cell apoptosis (TdT-dNTP staining) and expression of ZO-1, NA+/K+-ATPase and Ki-67 markers. Expression of miR-195-5p is found in HCEnCs and FECD cells, which restricts the proliferation of these cells. However, inhibiting miR-195-5p can induce the proliferation of HCEnCs, which opens exciting directions for future research in prolonging FECD pathogenesis by increasing the proliferative capacity of HCEnCs using anti-miR therapy in vivo. [ABSTRACT FROM AUTHOR]

Citrus is one of the most valuable crops in Syria, with the largest production areas being in the coastal provinces of Tartus and Latakia, where this study was performed. A companion paper reported on the basal crop coefficients derived from the field water balance and on the performance assessment of various irrigation methods used in a citrus orchard located in the same region. That study evidenced the need for the improved management of irrigation water, mainly reducing water applications and increasing productivity, thus leading to the current research. The main objectives consisted of (i) providing a set of reliable basal (Kcb) and average (Kc) crop coefficients to be used in practice in the citrus orchards of the Syrian coastal area, while accounting for the diversity of characteristics observed; (ii) to estimate the seasonal consumptive use of typical orchards under different climate-demand and deficit-irrigation scenarios; and (iii) to assess possible water savings and related yield reductions. The previously calibrated water balance model SIMDualKc was used for these purposes. The computed Kcb values for the mid-season and average demand for water ranged from 0.52, when the plant density was low, to 0.84, when plant density was very high. The corresponding Kc values, which further reflected the impact of drip irrigation on controlling soil evaporation, were 0.72 and 0.97, respectively. Overall, the consumptive use of water was estimated to range from 867 to 1573 mm. The assessed water-saving scenarios consisted of adopting increased management-allowed depletion (MAD) thresholds relative to the p depletion fraction for no stress: MAD = 1.05, 1.10, 1.20, and 1.30 p. For trees under a very high climatic demand, water savings ranged from 12 to 34%, but the yield losses induced by the water deficits ranged from 8 to 48%. Although the selection of optimal strategies should be based upon economic terms, these may only be used when the Syrian economy recovers from civil war and the current crisis. The present results show the feasibility of adopting such MAD thresholds for building an irrigation management platform. The data provided by the current study are valuable because they can be efficiently used to support of the irrigation management of Syrian citrus production systems. [ABSTRACT FROM AUTHOR]

Extracellular vesicles (EVs), specifically exosomes, carry a cell-type dependent cargo that is transported to the recipient cell and translated in the presence of a required machinery. Differences in the cargo carried by the corneal and conjunctival-derived EVs could be the agent that triggers the transdifferentiation of these two cell populations. Therefore, this study investigates the role of EVs in triggering the plasticity of corneal and conjunctival epithelial cells and identifies prospective miRNA and genes responsible for maintaining ocular surface homeostasis. The EVs were extracted from the conditioned media (after starving) of corneal epithelial (hTCEpi) and conjunctival (HCjE-Gi) cell lines using ultracentrifugation. HCjE-Gi cells were cultured with hTCEpi-derived EVs and vice-versa. The EVs were characterized as exosomes using Nanosight and Flow cytometry. KRT3 and KRT12 were used as associated corneal markers, whereas KRT7 and KRT13 were used as associated conjunctival markers with ΔNp63 as a differentiation marker. Shift of these markers was an indication of transdifferentiation. The cargo of the extracted exosomes from both the cell types was explored using next-generation sequencing. The hTCEpi-derived EVs induced conjunctival epithelial cells to express the corneal-associated markers KRT3 and KRT12, losing their conjunctival phenotype at both the mRNA and protein level. Simultaneously, HCjE-Gi-derived EVs induced corneal epithelial cells to express the conjunctival associated markers KRT7 and KRT13, losing their corneal phenotype. This process of differentiation was accompanied by an intermediate step of cell de-differentiation showed by up-regulation in the expression of epithelial stem cell marker ΔNp63, also shown on the ex vivo human cadaveric donor corneas. miRNA molecules (total of 11 including precursor and mature) with significant differences in their relative abundance between the two populations (p < 0.05) were found and investigated. miR-9-5p expression was higher in HCjE-Gi cells and HCjE-Gi-derived EVs when compared to hTCEpi cells and hTCEPi-derived EVs (p < 0.001). The results suggest that EVs released by the two cell types have the ability to influence the transdifferentiation of human conjunctival and corneal epithelial cells. miR-9-5p could have a role in stem cell homeostasis and cell differentiation via HES-1 gene. [ABSTRACT FROM AUTHOR]

The success of an irrigation decision support system (DSS) much depends on the reliability of the information provided to farmers. Remote sensing data can expectably help validate that information at the field scale. In this study, the MOHID-Land model, the core engine of the IrrigaSys DSS, was used to simulate the soil water balance in an irrigated vineyard located in southern Portugal during three growing seasons. Modeled actual basal crop coefficients and transpiration rates were then compared with the corresponding estimates derived from the normalized difference vegetation index (NDVI) computed from Sentinel-2 imagery. On one hand, the hydrological model was able to successfully estimate the soil water balance during the monitored seasons, exposing the need for improved irrigation schedules to minimize percolation losses. On the other hand, remote sensing products found correspondence with model outputs despite the conceptual differences between both approaches. With the necessary precautions, those products can be used to complement the information provided to farmers for irrigation of vine crop, further contributing to the regular validation of model estimates in the absence of field datasets. [ABSTRACT FROM AUTHOR]

The IrrigaSys decision support system (DSS) has supported farmers' decision-making regarding irrigation scheduling in the Sorraia Valley irrigation district in Southern Portugal over a span of six years (2017–2022). This study aims to conduct a postevaluation of farmers' adherence to the DSS, employing a multicriteria analysis (MCA) approach with data from the 2019 (driest year) and 2020 (average year) growing seasons. Two distinct scenarios were taken into consideration: the first focused on water conservation, and the second centered on farmers' economic returns. The outcomes of the first scenario revealed that farmers exhibited a reasonable level of expertise, particularly during the driest season. They achieved water-saving indicators comparable to those obtained when adhering to optimized irrigation schedules generated weekly by the DSS. In the wetter season, discrepancies emerged between farmers' and model indicators, primarily attributed to challenges in integrating reliable information from precipitation forecasts into the decision-making process. In the second scenario, both farmers' and model results exhibited close economic indicators throughout both seasons. While IrrigaSys requires further developments, these results show that the DSS has effectively contributed to supporting irrigation water management in the study region. [ABSTRACT FROM AUTHOR]