Consistent expansion of human corneal endothelial cells (hCECs) is critical in the development of tissue engineered endothelial constructs. However, a wide range of complex culture media, developed ...from different basal media have been reported in the propagation of hCECs, some with more success than others. These results are further confounded by donor-to-donor variability. The aim of this study is to evaluate four culture media in the isolation and propagation of hCECs isolated from a series of paired donor corneas in order to negate donor variability. Isolated primary hCECs were cultured in four previously published medium coded in this study as: M1-DMEM; M2-OptiMEM-I; M3-DMEM/F12, & M4-Ham's F12/M199. Primary hCECs established in these conditions were expanded for two passages and analyzed for (1) their propensity to adhere and proliferate; (2) their expression of characteristic corneal endothelium markers: Na+K+/ATPase and ZO-1; and (3) their cellular morphology throughout the study. We found that hCECs isolated in all four media showed rapid attachment when cultured on FNC-coated dishes. However, hCECs established in the four media exhibited different proliferation profiles with striking morphological differences. Corneal endothelial cells cultured in M1 and M3 could not be propagated beyond the first and second passage respectively. The hCECs cultured in M2 and M4 were significantly more proliferative and expressed markers characteristics of human corneal endothelium: Na+K+/ATPase and ZO-1. However, the unique morphological characteristics of cultivated hCECs were not maintained in either M2 or M4 beyond the third passage.The proliferative capacity and morphology of hCECs are vastly affected by the four culture media. For the development of tissue engineered graft materials using cultured hCECs derived from the isolation methodology described in this study, we propose the use of proliferative media M2 or M4 up to the third passage, or before the cultured hCECs lose their unique cellular morphology.
The gut microbiota plays a key role in modulating host immune response. We conducted a prospective, observational study to examine gut microbiota composition in association with immune responses and ...adverse events in adults who have received the inactivated vaccine (CoronaVac; Sinovac) or the mRNA vaccine (BNT162b2; BioNTech; Comirnaty).
We performed shotgun metagenomic sequencing in stool samples of 138 COVID-19 vaccinees (37 CoronaVac and 101 BNT162b2 vaccinees) collected at baseline and 1 month after second dose of vaccination. Immune markers were measured by SARS-CoV-2 surrogate virus neutralisation test and spike receptor-binding domain IgG ELISA.
We found a significantly lower immune response in recipients of CoronaVac than BNT162b2 vaccines (p<0.05).
was persistently higher in subjects with high neutralising antibodies to CoronaVac vaccine (p=0.023) and their baseline gut microbiome was enriched in pathways related to carbohydrate metabolism (linear discriminant analysis (LDA) scores >2 and p<0.05). Neutralising antibodies in BNT162b2 vaccinees showed a positive correlation with the total abundance of bacteria with flagella and fimbriae including
(p=0.028). The abundance of
and two
species were enriched in individuals with fewer adverse events following either of the vaccines indicating that these bacteria may play an anti-inflammatory role in host immune response (LDA scores>3 and p<0.05).
Our study has identified specific gut microbiota markers in association with improved immune response and reduced adverse events following COVID-19 vaccines. Microbiota-targeted interventions have the potential to complement effectiveness of COVID-19 vaccines.
We previously demonstrated that fermitin family member 1 (FERMT1) was significantly overexpressed in colon cancer (CC) and associated with poor metastasis-free survival. This study aimed to ...investigate the precise role of FERMT1 in CC metastasis and the mechanism by which FERMT1 is involved in the epithelial-mesenchymal transition (EMT). Correlations between FERMT1 and EMT markers (E-cadherin, Slug, N-cadherin and β-catenin) were examined via immunohistochemistry in a cohort of CC tissues and adjacent normal colon mucosae. A series of in vitro and in vivo assays were performed to elucidate the function of FERMT1 in CC metastasis and underlying mechanisms. The upregulated expression of FERMT1 in CC tissues correlated positively with that of Slug, N-cadherin and β-catenin, but correlated inversely with E-cadherin expression. Altered FERMT1 expression led to marked changes in the proliferation, migration, invasion and EMT markers of CC cells both in vitro and in vivo. Investigations of underlying mechanisms found that FERMT1 interacted directly with β-catenin and activated the Wnt/β-catenin signaling pathway by decreasing the phosphorylation level of β-catenin, enhancing β-catenin nuclear translocation and increasing the transcriptional activity of β-catenin/TCF/LEF. Activation of the Wnt/β-catenin pathway by CHIR99021 reversed the effect of FERMT1 knockdown, whereas inhibition of the Wnt/β-catenin pathway by XAV939 impaired the effect of FERMT1 overexpression on EMT and cell motility. In conclusion, findings of this study suggest that FERMT1 activates the β-catenin transcriptional activity to promote EMT in CC metastasis.
As agricultural land and freshwater inextricably interrelate and interact with each other, the conventional water and land policy in “silos” should give way to nexus thinking when formulating the ...land and water management strategies. This study constructs a systems multi-regional input-output (MRIO) model to expound global land-water nexus by simultaneously tracking agricultural land and freshwater use flows along the global supply chains. Furthermore, land productivity and irrigation water requirements of 160 crops in different regions are investigated to reflect the land-water linkage. Results show that developed economies (e.g., USA and Japan) and major large developing economies (e.g., mainland China and India) are the overriding drivers of agricultural land and freshwater use globally. In general, significant net transfers of these two resources are identified from resource-rich and less-developed economies to resource-poor and more-developed economies. For some crops, blue water productivity is inversely related to land productivity, indicating that irrigation water consumption is sometimes at odds with land use. The results could stimulus international cooperation for sustainable land and freshwater management targeting on original suppliers and final consumers along the global supply chains. Moreover, crop-specific land-water linkage could provide insights for trade-off decisions on minimizing the environmental impacts on local land and water resources.
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•Agricultural land and freshwater use within global supply chains are tracked simultaneously.•Land productivity and irrigation water requirements of 160 crops in different regions are investigated.•Approximately one-third of land use and water withdrawal is attributed to interregional trade.•Trade-off decisions should be made for both local agricultural production and global supply chain management.
We explore an analogue of optical frequency combs in driven nonlinear phononic systems, and present a mechanism for generating phononic frequency combs through nonlinear resonances. In the underlying ...process, a set of phonon modes is simultaneously excited by the external driving which yields frequency combs with an array of discrete and equidistant spectral lines of each nonlinearly excited phonon mode. Frequency combs through nonlinear resonance of different orders are investigated, and in particular the possibility of correlation tailoring in higher-order cases is revealed. We suggest that our results can be applied in various nonlinear acoustic processes, such as phonon harvesting, and can also be generalized to other nonlinear systems.
Current treatments for reversible blindness caused by corneal endothelial cell failure involve replacing the failed endothelium with donor tissue using a one donor-one recipient strategy. Due to the ...increasing pressure of a worldwide donor cornea shortage there has been considerable interest in developing alternative strategies to treat endothelial disorders using expanded cell replacement therapy. Protocols have been developed which allow successful expansion of endothelial cells in vitro but this approach requires a supporting material that would allow easy transfer of cells to the recipient. We describe the first use of plastic compressed collagen as a highly effective, novel carrier for human corneal endothelial cells. A human corneal endothelial cell line and primary human corneal endothelial cells retained their characteristic cobblestone morphology and expression of tight junction protein ZO-1 and pump protein Na+/K+ ATPase α1 after culture on collagen constructs for up to 14 days. Additionally, ultrastructural analysis suggested a well-integrated endothelial layer with tightly opposed cells and apical microvilli. Plastic compressed collagen is a superior biomaterial in terms of its speed and ease of production and its ability to be manipulated in a clinically relevant manner without breakage. This method provides expanded endothelial cells with a substrate that could be suitable for transplantation allowing one donor cornea to potentially treat multiple patients.
In this paper, a closed-loop logistics system with manufacturing and remanufacturing cycle is analyzed using three optimization methods. They are: (i) sequential optimization, (ii) centralized ...optimization without benefit sharing, and (iii) centralized optimization with benefit sharing. In the first method, the decisions of sequential optimization are made initially by the retailers, and then by the manufacturer. In the second method, the decision of the centralized optimization is made simultaneously by the whole logistics system. The second method is not favorable to the retailers because the retailers' profit will become smaller as a result of collaboration. Therefore a third method with benefit sharing is investigated. The logistics system includes a manufacturer and multi-retailer considering price-sensitive demand and deterioration. Industrial examples of such products are IC chips, computers and mobile phones that usually outdate or decrease in value due to technological innovation. The decrease in value is considered as a form of deterioration. In order to add value, these outdated products can be remanufactured and resold to market. Four key effects are investigated in this study: (i) recycling in supply chain, (ii) deterioration, (iii) collaboration effect between manufacturer and retailers using three optimization methods, and (iv) price sensitive demand.
Boreal fires have immediate effects on regional carbon budgets by emitting CO2 into the atmosphere at the time of burning, but they also have legacy effects by initiating a long-term carbon sink ...during post-fire vegetation recovery. Quantifying these different effects on the current-day pan-boreal (44–84° N) carbon balance and quantifying relative contributions of legacy sinks by past fires is important for understanding and predicting the carbon dynamics in this region. Here we used the global dynamic vegetation model ORCHIDEE–SPITFIRE (Organising Carbon and Hydrology In Dynamic Ecosystems – SPread and InTensity of FIRE) to attribute the contributions by fires in different decades between 1850 and 2009 to the carbon balance of 2000–2009, taking into account the atmospheric CO2 change and climate change since 1850. The fire module of ORCHIDEE–SPITFIRE was turned off for each decade in turn and was also turned off before and after the decade in question in order to model the legacy carbon trajectory by fires in each past decade. We found that, unsurprisingly, fires that occurred in 2000–2009 are a carbon source (−0.17 Pg C yr−1) for the carbon balance of 2000–2009, whereas fires in all decades before 2000 contribute carbon sinks with a collective contribution of 0.23 Pg C yr−1. This leaves a net fire sink effect of 0.06 Pg C yr−1, or 6.3 % of the simulated regional carbon sink (0.95 Pg C yr−1). Further, fires with an age of 10–40 years (i.e., those that occurred during 1960–1999) contribute more than half of the total sink effect of fires. The small net sink effect of fires indicates that current-day fire emissions are roughly balanced out by legacy sinks. The future role of fires in the regional carbon balance remains uncertain and will depend on whether changes in fires and associated carbon emissions will exceed the enhanced sink effects of previous fires, both being strongly affected by global change.
The Pediatric Interagency Registry for Mechanical Circulatory Support (Pedimacs), originally a National Institutes of Health-sponsored U.S. database, provides a platform to understand the population ...of children supported with ventricular assist devices (VADs) during this time of increasing numbers, new devices, expanding indications, and improved outcomes.
Between September 19, 2012, and December 31, 2019, 44 hospitals implanted 1031 devices in 856 patients under 19 years of age.
Overall, diagnosis was cardiomyopathy in 497 (58%) patients, congenital heart disease (CHD) in 216 (25%), myocarditis in 85 (10%), and other in 58 (7%). Positive outcome (alive on device or bridge to transplantation and recovery) occurred in 82% at 6 months. The patient cohort for implantable continuous (IC) pumps (n = 365) (age 13.2 ± 3.9 years, 18% Interagency Registry for Mechanically Assisted Circulatory Support Intermacs profile 1, 23% intubated at implantation, 16% with CHD) was significantly different from the paracorporeal continuous (PC) pump cohort (n = 212) (age 3.6 ± 4.9 years, 46% Intermacs profile 1, 81% intubated, 42% CHD) and the paracorporeal pulsatile (PP) pump cohort (n = 230) (age 2.7 ± 3.5 years, 31% Intermacs profile 1, 76% intubated, 26% CHD). Consistent with their cohort composition, positive outcomes at 6 months based on device type were the following: IC, 92%; PC, 68%; and PP, 81%. The incidence of cerebrovascular accidents in the IC, PC, and PP cohorts was 7%, 14%, and 15%, respectively.
IC VADs, the most common VAD type placed in children, are associated with improved outcomes compared with PP and PC devices, though PP and PC devices are limited to supporting our most challenging patients. Noteworthy, the incidence of cerebrovascular accidents for pediatric VADs has significantly decreased and is now 11% overall. This report demonstrates again that although often attributed to age, size, or device type, much of the burden in mortality and adverse events is correlated to the patient's overall state at VAD implantation.
In addition to their key roles in cellular survival, death, proliferation and metabolism, the Foxo subfamily of forkhead (Fox) transcription factors play critical roles in the homeostasis of ...immune-relevant cells, including T cells, B cells, neutrophils and other non-lymphoid lineages that modulate inflammation in disease states such as inflammatory arthritis and systemic lupus erythematosus. This review summarizes such current and expanding knowledge of the Foxo family members in immunity, and their potential as therapeutic targets in inflammatory disease.
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