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Abstract While nano‐enabled biotechnology is offering great promise, in certain areas providing fundamental paradigm shift for disease management, we are rapidly moving into a new era of nanotechnology developmental stage, in which chemistry, manufacturing, and controls of nanotherapeutics (nano‐CMC) become a critical step for translational nanomedicine. This is not a trivial task, in fact, nano‐CMC requires a long list of considerations. Minimally, it includes raw materials, scale‐up synthesis routes, batch sizes, stability check, analytical methods, and documentation. The growing items on the list could collectively ensure the reproducible production of nanotherapeutics that are safe and efficacious and meet new drug application (NDA) specifications. This article begins from the lesson learned from liposome, such as Doxil® and other FDA‐approved liposomes, then continues to provide a literature summary on the scale‐up synthesis of nanoparticles that were designed for therapeutic or diagnostic purposes. We outlined the key challenges during the scale‐up activities, followed by the case studies to illustrate the impact of emerging strategies that may improve the reproducible synthesis of large‐batch nanoparticles. The availability of quality‐controlled large‐batch size opens the possibility to conduct robust preclinical studies and clinical trials. This includes the recent advances of using microfluidics system, implementation of multiparameters iterative CMC optimization, and use of computer software, allowing the successful preparation of different organic and inorganic nanoparticles at desired batch sizes. The authors also share personal insights with respect to how nano‐CMC research would facilitate “personalized and just‐in‐time” nanomedicine.

Autophagy plays an essential role in maintaining cellular homeostasis by responding to certain stressed conditions such as nutrient deprivation, organelle damage, pathogen infection, and exposure to certain nanomaterials. Through deliberate tuning of the physicochemical properties, the fate of nanomaterial-treated cells is subjected to various cellular toxicity, stress responses, and immune responses, some of which involve various autophagic mechanisms. Beginning from the molecular basis of the autophagy machinery, we briefly introduce the major understanding of autophagy in inflammation, immunity, cancer metabolism, and therapy. Different working mechanisms will be discussed to classify the impact of physicochemical characteristics on autophagy induction or inhibition by engineered nanomaterials. From the perspective of autophagy-targeting cancer therapeutics, we will further delineate the advanced nano-formulations for improved drug delivery to impact autophagy in the setting of cancer diseases and designing co-delivery nanomedicine that targets autophagy along with another major cancer pathway to achieve in vivo synergy. Moreover, cancer immunotherapy, aiming at immune cells or checkpoints, is also integrated with autophagy-regulatory components using multiple nano-platforms as an emerging strategy for cancer treatment. Considering the recent breakthroughs of nanotechnology targeting autophagy in cancer cells, antigen-presenting cells, or other cell types within tumor microenvironment by precisely designed nanomedicine may provide additional solutions for cancer treatment through autophagy-dependent metabolic regulation or immune pathways. However, one should closely monitor the extent of autophagy alternation and side effects from certain nanoparticles to avoid severe toxic responses.

Abstract There is an urgent need to develop new life‐prolonging therapy for pancreatic ductal adenocarcinoma (PDAC). It is demonstrated that improved irinotecan delivery by a lipid bilayer coated mesoporous silica nanoparticle, also known as a silicasome, can improve PDAC survival through a chemo‐immunotherapy response in an orthotopic Kras‐dependent pancreatic cancer model. This discovery is premised on the weak‐basic properties of irinotecan, which neutralizes the acidic lysosomal pH in PDAC cells. This effect triggers a linked downstream cascade of events that include autophagy inhibition, endoplasmic reticulum stress, immunogenic cell death (ICD), and programmed death‐ligand 1 (PD‐L1) expression. ICD is characterized by calreticulin expression and high‐mobility group box 1 (HMGB1) release in dying Kras‐induced pancreatic cancer (KPC) cells, which is demonstrated in a vaccination experiment to prevent KPC tumor growth on the contralateral site. The improved delivery of irinotecan by the silicasome is accompanied by robust antitumor immunity, which can be synergistically enhanced by anti‐PD‐1 in the orthotopic model. Immunophenotyping confirms the expression of calreticulin, HMGB1, PD‐L1, and an autophagy marker, in addition to perforin and granzyme B deposition. The chemo‐immunotherapy response elicited by the silicasome is more robust than free or a liposomal drug, Onivyde. The silicasome plus anti‐PD‐1 leads to significantly enhanced survival improvement, and is far superior to anti‐PD‐1 plus either free irinotecan or Onivyde.

Owing to a series of air pollution prevention and control policies, China’s PM2.5 pollution has greatly improved; however, the long-term spatial contiguous products that facilitate the analysis of the distribution and variation of PM2.5 pollution are insufficient. Due to the limitations of missing values in aerosol optical depth (AOD) products, the reconstruction of full-coverage PM2.5 concentration remains challenging. In this study, we present a two-stage daily adaptive modeling framework, based on machine learning, to solve this problem. We built the annual models in the first stage, then daily models were constructed in the second stage based on the output of the annual models, which incorporated the parameter and feature adaptive tuning strategy. Within this study, PM2.5 concentrations were adaptively modeled and reconstructed daily based on the multi-angle implementation of atmospheric correction (MAIAC) AOD products and other ancillary data, such as meteorological factors, population, and elevation. Our model validation showed excellent performance with an overall R2 = 0.91 and RMSE = 9.91 μg/m3 for the daily models, along with the site-based cross-validation R2s and RMSEs of 0.86–0.87 and 12–12.33 μg/m3; these results indicated the reliability and feasibility of the proposed approach. The daily full-coverage PM2.5 concentrations at 1 km resolution across China during the Three-Year Blue-Sky Action Plan were reconstructed in this study. We analyzed the distribution and variations of reconstructed PM2.5 at three different time scales. Overall, national PM2.5 pollution has significantly improved with the annual average concentration dropping from 33.67–28.03 μg/m3, which demonstrated that air pollution control policies are effective and beneficial. However, some areas still have severe PM2.5 pollution problems that cannot be ignored. In conclusion, the approach proposed in this study can accurately present daily full-coverage PM2.5 concentrations and the research outcomes could provide a reference for subsequent air pollution prevention and control decision-making.

Introduction Neuromyelitis optica spectrum disorders (NMOSD) is an inflammatory and heterogeneous astrocyte disorder of the central nervous system with the characteristic of higher incidence in women and Asian people. Most patients with NMOSD have a course of recurrence and remission that is prone to cause paralysis and blindness. Several studies have confirmed the efficacy and promising prospect of mycophenolate mofetil (MMF) in the treatment of NMOSD. Yet its therapeutic effect and safety are controversial. Although there has been two published literature that is relevant to the topic of this study, both of them have certain defects, and they can only provide answers about the efficacy or safety of MMF in the treatment of NMOSD from partial perspectives or conclusions. This research aims to perform a direct and comprehensive systematic review and meta-analysis to evaluate MMF’s effectiveness and safety in treating NMOSD.Methods and analysis This systematic review will cover all comparative researches, from randomised controlled trials to cohort studies, and case–control study. A relevant literature search will be conducted in PubMed, Web of Science, EMBASE, the Cochrane Library, China National Knowledge Infrastructure, Wanfang Database, China Science and Technology Journal Database and Chinese Biomedical Literature Database from their inception to 31 June 2020. We will also search registers of clinical trials, potential grey literature and abstracts from conferences. There are no limits on language and publication status. The reporting quality and risk of bias will be assessed by two researchers independently. Expanded Disability Status Scales and annualised relapse rate will be evaluated as the primary outcome. The secondary outcomes will consist of the frequency and severity of adverse events, best-corrected visual acuity, relapse-free rate and time to the next attack. A meta-analysis will be performed using RevMan V.5.3 software provided by the Cochrane Collaboration and Stata V.12.0.Ethics and dissemination Because the data used for this systematic review will be exclusively extracted from published studies, ethical approval and informed consent of patients will not be required. The systematic review will be published in a peer-reviewed journal, presented at conferences and will be shared on social media platforms.PROSPERO registration number CRD42020164179.

Summary: Mutations in CAPN3 cause limb girdle muscular dystrophy R1 (LGMDR1, formerly LGMD2A) and lead to progressive and debilitating muscle wasting. Calpain 3 deficiency is associated with impaired CaMKIIβ signaling and blunted transcriptional programs that encode the slow-oxidative muscle phenotype. We conducted a high-throughput screen on a target of CaMKII (Myl2) to identify compounds to override this signaling defect; 4 were tested in vivo in the Capn3 knockout (C3KO) model of LGMDR1. The leading compound, AMBMP, showed good exposure and was able to reverse the LGMDR1 phenotype in vivo, including improved oxidative properties, increased slow fiber size, and enhanced exercise performance. AMBMP also activated CaMKIIβ signaling, but it did not alter other pathways known to be associated with muscle growth. Thus, AMBMP treatment activates CaMKII and metabolically reprograms skeletal muscle toward a slow muscle phenotype. These proof-of-concept studies lend support for an approach to the development of therapeutics for LGMDR1.

Summary: Chromatographic-based approaches are most widely used for the analysis of enantiomers. Unambiguous results are achieved through the visualization of each enantiomer, while the separation process is often time consuming. By endowing 19F probes with suitable dynamic recognition properties, we develop here a detection scheme that produces chromatogram-like output in the analysis of enantiomeric mixtures without separation. A wide range of chiral compounds, including alcohols, ethers, amides, carbamates, oxazolidinones, sulfoxides, and sulfoximines, are successfully resolved, and the simultaneous enantiodifferentiation of different classes of chiral compounds is achieved. The method is also amenable for the determination of the enantiopurity of crude reaction products and the assignment of absolute configurations. The potential utility in high-throughput screening is demonstrated by the analysis of 24 nonracemic samples in 30 min with an NMR spectrometer equipped with an autosampler.

Street space quality assessment refers to the extraction and appropriate evaluation of the space quality information of urban streets, which is usually employed to improve the quality of urban planning and management. Compared to traditional approaches relying on expert knowledge, the advances of big data collection and analysis technologies provide an alternative for assessing street space more precisely. With street view imagery (SVI), points of interest (POI) and comment data from social media, this study evaluates street space quality from the perspective of exploring and discussing the relationship among street vitality, service facilities and built environment. Firstly, a transfer-learning-based framework is employed for SVI semantic segmentation to quantify the street built environment. Then, we use POI data to identify different urban functions that streets serve, and comment data are utilized to investigate urban vitality composition and integrate it with different urban functions associated with streets. Finally, a function-driven street space quality assessment approach is established. To examine its applicability and performance, the proposed method is experimented with data from part area in Xiamen, China. The output is compared to results based on expert opinion using the correlation analysis method. Results show that the proposed assessment approach designed in this study is in accordance with the validation data, with the overall R2 value being greater than 0.6. In particular, the proposed method shows better performance in scenic land and mixed functional streets with R2 value being greater than 0.8. This method is expected to be an efficient tool for discovering problems and optimizing urban planning and management.

Abstract Background Calcitonin gene-related peptide (CGRP) contributes to bone formation by stimulating bone marrow stromal cell (BMSC) proliferation and differentiation. However, the proliferative and apoptotic effects of CGRP on bone marrow-derived endothelial progenitor cells (EPCs) have not been investigated. Methods We tested the effects of CGRP on EPC proliferation and apoptosis by Cell Counting Kit-8, flow cytometry, and studied the effects of CGRP on the expression of proliferation- and apoptosis-related markers in EPCs and the underlying mitogen-activated protein kinase (MAPK) signalling pathway by quantitative polymerase chain reaction and western blotting. Results We detected EPC markers (CD34, CD133 and VEGFR-2) in 7-day cultures and found that CGRP (10− 10–10− 12 M) promoted the proliferation of cultured EPCs, with a peak increase of 30% at 10− 10 M CGRP. CGRP also upregulated the expression of proliferation-associated genes, including cyclin D1 and cyclin E, and increased the percentages of G2/M-phase and S-phase cells after incubation 72 h. CGRP inhibited serum deprivation (SD)-induced apoptosis in EPCs after 24 and 48 h and downregulated the expression of apoptosis-related genes, including caspase-3, caspase-8, caspase-9 and Bax. Phosphorylated (p-)ERK1/2, p-p38 and p-JNK protein levels in EPCs treated with CGRP were significantly lower than those in untreated EPCs. Pre-treatment with the calcitonin receptor-like receptor (CRLR) antagonist CGRP8–37 or a MAPK pathway inhibitor (PD98059, SB203580 or SP600125) completely or partially reversed the pro-proliferative and anti-apoptotic effects and the reduced p-ERK1/2, p-p38 and p-JNK expression induced by CGRP. Conclusion Our results show that CGRP exerts pro-proliferative and anti-apoptotic effects on EPCs and may act by inhibiting MAPK pathways.

Dust emission is an important corollary of the soil degradation process in arid and semi-arid areas worldwide. Soil organic carbon (SOC) is the main terrestrial pool in the carbon cycle, and dust emission redistributes SOC within terrestrial ecosystems and to the atmosphere and oceans. This redistribution plays an important role in the global carbon cycle. Herein, we present a systematic review of dust modelling, global dust budgets, and the effects of dust emission on SOC dynamics. Focusing on selected dust models developed in the past five decades at different spatio-temporal scales, we discuss the global dust sources, sinks, and budgets identified by these models and the effect of dust emissions on SOC dynamics. We obtain the following conclusions: (1) dust models have made considerable progress, but there are still some uncertainties; (2) a set of parameters should be developed for the use of dust models in different regions, and direct anthropogenic dust should be considered in dust emission estimations; and (3) the involvement of dust emission in the carbon cycle models is crucial for improving the accuracy of carbon assessment.

Pancreatic cancer remains difficult to treat mainly due to the drug delivery challenges posed by a strong stromal component. Here the authors develop nanocarriers that improve drug delivery efficiency and engage the host immune system against the tumor resulting in reduction of tumor growth and metastasis.

Neuropeptide Y (NPY) exhibits a critical but poorly understood regulatory signaling function and has been shown to promote proliferation, vascularization and migration in several types of cells and tissues. However, little is known about the specific role of NPY in the proliferation and apoptosis of bone marrow stromal cells (also known as bone marrow-derived mesenchymal stem cells, BMSCs), which contain a subpopulation of multipotent skeletal stem cells. Based on BrdU incorporation tests, Cell Counting Kit-8, flow cytometry, quantitative polymerase chain reaction and western blotting, we showed that NPY significantly promoted the proliferation of BMSCs in a concentration-dependent manner, with a maximal effect observed at a concentration of 10−10 M for pro-proliferative and 10−12 M for anti-apoptotic activities. Furthermore, NPY significantly increased the percentage of cells in S and G2/M phases. In addition, NPY exhibited a protective effect after 24 h of serum starvation as illustrated by a reduction in the apoptosis rate, degree of nuclear condensation, and expression of apoptosis markers, including caspase-3, caspase-9 and Bax mRNA expression. NPY also increased the mRNA and protein expression levels of canonical Wnt signaling pathway proteins, including β-catenin and c-myc, during the induced proliferative and anti-apoptotic processes. However, the proliferative and anti-apoptotic activities of NPY were partially blocked by both PD160170 (1 μM) and DKK1 (0.2 μg/mL). These compounds also blocked the mRNA and protein expression of β-catenin, p-GSK-3β and c-myc. Therefore, the results of the present study demonstrated that NPY exerts a proliferative and protective effect on BMSCs in a dose- and time-dependent manner in vitro, and importantly, these effects may be mediated via its Y1 receptor and involved in activation of the canonical Wnt signaling pathway.

Based on cores from tight oil reservoirs in Ordos Basin, water flooding experiments with both low and high displacement pressures were carried out. Combined with NMR, quantitative analysis approaches for produced oil under different microscopic effects were established for quantitative research of the microscopic mechanism of water flooding in tight reservoirs. The research indicated that under low displacement pressure, oil recovery mechanisms of hydrophilic cores mainly include displacement, imbibition, and denudation, and those of wetting cores mainly include displacement and imbibition. After increasing the displacement pressure, both hydrophilic and neutral wetting cores have a certain increase in oil recovery. The common point is that both oil controlled by small throats and remaining oil droplets controlled by traps in large pores have been activated, while the discrepancy is that the oil film of the neutral wetting core boundary layer becomes thinner, which improves oil recovery ratio, and these mechanisms have less effect on hydrophilic cores. The amount of oil produced by each type of oil recovery mode was quantitatively analyzed. Percentages of produced oil in hydrophilic cores by flooding and imbibition and denudation are 15% and 12%, respectively, which are the main oil recovery mechanisms; percentages of produced oil in neutral wetting cores by displacement and imbibition are 25% and 2%, respectively, with displacement as the main oil recovery mechanism. After increasing the displacement pressure, oil produced by hydrophilic and neutral wetting cores increased by 6% and 9%, respectively, indicating that with increasing the displacement pressure, a part of the boundary layer of oil could be produced in neutral wetting cores.

Our group recently characterized a cell-autonomous mammalian 12-h clock independent from the circadian clock, but its function and mechanism of regulation remain poorly understood. Here, we show that in mouse liver, transcriptional regulation significantly contributes to the establishment of 12-h rhythms of mRNA expression in a manner dependent on Spliced Form of X-box Binding Protein 1 (XBP1s). Mechanistically, the motif stringency of XBP1s promoter binding sites dictates XBP1s's ability to drive 12-h rhythms of nascent mRNA transcription at dawn and dusk, which are enriched for basal transcription regulation, mRNA processing and export, ribosome biogenesis, translation initiation, and protein processing/sorting in the Endoplasmic Reticulum (ER)-Golgi in a temporal order consistent with the progressive molecular processing sequence described by the central dogma information flow (CEDIF). We further identified GA-binding proteins (GABPs) as putative novel transcriptional regulators driving 12-h rhythms of gene expression with more diverse phases. These 12-h rhythms of gene expression are cell autonomous and evolutionarily conserved in marine animals possessing a circatidal clock. Our results demonstrate an evolutionarily conserved, intricate network of transcriptional control of the mammalian 12-h clock that mediates diverse biological pathways. We speculate that the 12-h clock is coopted to accommodate elevated gene expression and processing in mammals at the two rush hours, with the particular genes processed at each rush hour regulated by the circadian and/or tissue-specific pathways.

This paper explores the capability of high frequency microwave measurements at vertical and horizontal polarizations in detecting snowfall over land. Surface in-situ meteorological data were collected over Conterminous US during two winter seasons in 2014–2015 and 2015–2016. Statistical analysis of the in-situ data, matched with Global Precipitation Measurement (GPM) Microwave Imager (GMI) measurements on board NASA/JAXA Core Observatory, showed that the polarization difference at 166 GHz had the highest correlation to measured snowfall rate compared to the single channel high frequency measurements and the polarization difference at 89 GHz. A logistic regression model applied to the match-up data, using the polarization difference at 166 and 89 GHz as predictors, yielded an overall snowfall classification rate of 69.0%, with the largest contribution coming from the polarization difference at 166 GHz. Logistic regression using the four single channels as predictors (at 89 and 166 GHz, horizontal and vertical polarizations) further indicated that the horizontal polarization at 166 GHz was the most important contributor. An overall classification rate of 73% was achieved by including the 183.31 ± 3 GHz and 183.31 ± 7 GHz vertical polarization channels in the final logistic regression model. Evaluation of the final algorithm demonstrated skill in snowfall detection of two significant events.

More than one decade of observations from the Advanced Microwave Sounding Unit-A (AMSU-A) onboard the polar-orbiting satellites NOAA-15 to NOAA-19 and European Meteorological Operational satellite program-A (MetOp-A) provided global information on atmospheric temperature profiles, water vapor, cloud, precipitation, etc. These observations were primarily intended for weather related prediction and applications, however, in order to meet the requirements for climate application, further reprocessing must be conducted to first eliminate any potential satellites biases. After the geolocation and cross-scan bias corrections were applied to the dataset, follow-on research focused on the comparison amongst AMSU-A window channels (e.g., 23.8, 31.4, 50.3 and 89.0 GHz) from the six different satellites to remove any inter-satellite inconsistency. Inter-satellite differences can arise from many error sources, such as bias drift, sun-heating-induced instrument variability in brightness temperatures, radiance dependent biases due to inaccurate calibration nonlinearity, etc. The Integrated microwave inter-calibration approach (IMICA) approach was adopted in this study for inter-satellite calibration of AMSU-A window channels after the appropriate standard deviation (STD) thresholds were identified to restrict Simultaneous Nadir Overpass (SNO) data for window channels. This was a critical step towards the development of a set of fundamental and thematic climate data records (CDRs) for hydrological and climatological applications. NOAA-15 served as the main reference satellite for this study. For ensuing studies that expand to beyond 2015, however, it is recommended that a different satellite be adopted as the reference due to concerns over potential degradation of NOAA-15 AMSU-A.

We demonstrated that activated transcytosis is a major mechanism to complement the classic enhanced permeability and retention effect in pancreatic cancer. This was achieved by using an iRGD peptide that triggers transcytosis pathway at the tumor site. Co-administration of unconjugated iRGD substantially improved the effect of the chemotherapeutics delivering nanocarrier, and resulted in survival improvement in mice. Since the iRGD effect is commensurate with neuropilin-1 expression on tumor vasculature, it is necessary to contemplate a personalized approach to implement this technology.

The mechanisms contributing to an increased risk of thrombosis in uremia are complex and require clarification. There is scant morphological evidence of membrane-dependent binding of factor Xa (FXa) and factor Va (FVa) on endothelial cells (EC) in vitro. Our objectives were to confirm that exposed phosphatidylserine (PS) on microparticle (MP), EC, and peripheral blood cell (PBC) has a prothrombotic role in uremic patients and to provide visible and morphological evidence of PS-dependent prothrombinase assembly in vitro. We found that uremic patients had more circulating MP (derived from PBC and EC) than controls. Additionally, patients had more exposed PS on their MPs and PBCs, especially in the hemodialysis group. In vitro, EC exposed more PS in uremic toxins or serum. Moreover, reconstitution experiments showed that at the early stages, PS exposure was partially reversible. Using confocal microscopy, we observed that PS-rich membranes of EC and MP provided binding sites for FVa and FXa. Further, exposure of PS in uremia resulted in increased generation of FXa, thrombin, and fibrin and significantly shortened coagulation time. Lactadherin, a protein that blocks PS, reduced 80% of procoagulant activity on PBC, EC, and MP. Our results suggest that PBC and EC in uremic milieu are easily injured or activated, which exposes PS and causes a release of MP, providing abundant procoagulant membrane surfaces and thus facilitating thrombus formation. Blocking PS binding sites could become a new therapeutic target for preventing thrombosis.

Polo-like kinase 1 (PLK1), one of the key regulators of mitosis, is a target for cancer therapy due to its abnormally high activity in several tumors. Plk1 is highly conserved and shares a nearly identical 3-D structure between zebrafish and humans. The initial 10 mitoses of zebrafish embryonic cleavages occur every∼30 minutes, and therefore provide a rapid assay to evaluate mitosis inhibitors including those targeting Plk1. To increase efficiency and specificity, we first performed a computational virtual screen of∼60000 compounds against the human Plk1 3-D structure docked to both its kinase and Polo box domain. 370 candidates with the top free-energy scores were subjected to zebrafish assay and 3 were shown to inhibit cell division. Compared to general screen for compounds inhibiting zebrafish embryonic cleavage, computation increased the efficiency by 11 folds. One of the 3 compounds, named I2, was further demonstrated to effectively inhibit multiple tumor cell proliferation in vitro and PC3 prostate cancer growth in Xenograft mouse model in vivo. Furthermore, I2 inhibited Plk1 enzyme activity in a dose dependent manner. The IC50 values of I2 in these assays are compatible to those of ON-01910, a Plk1 inhibitor currently in Phase III clinic trials. Our studies demonstrate that zebrafish assays coupled with computational screening significantly improves the efficiency of identifying specific regulators of biological targets. The PLK1 inhibitor I2, and its analogs, may have potential in cancer therapeutics.

Keratoconus, historically viewed as a non-inflammatory disease, is an ectatic corneal disorder associated with progressive thinning of the corneal stroma. Recently, a few inflammatory mediators have been reported to be elevated in the tear fluid of keratoconus patients. Consequently, we investigated a wide range of inflammation regulating cytokines in the tears and sera of keratoconus and control subjects. Interleukin (IL)-1β, IL-4, IL-6, IL-10, IL-12, IL-13, IL-17, interferon (IFN)-γ, chemokine C-C motif ligand 5 (CCL5) and tumor necrosis factor (TNF)-α were tested in tear samples and sera of keratoconus and control individuals by multiplex immuno-bead assays. Selected cytokines were further tested by standard ELISA on pooled tear samples. All cytokines in the sera were generally low, with no significant changes between keratoconus and control subjects. However, in tear fluids, clear differences were detected between the two groups. These differences include increased IL-6, and decreased IL-12, TNF-α, IFN-γ, IL-4, IL-13 and CCL5 in keratoconus compared to control tear fluids. The decreases in IL-12, TNF-α and CCL5 were statistically significant, while the IL-13 decrease was statistically significant in the severe keratoconus group only. IL-17 could not be detected by multiplex immuno-bead assay, but showed an increase in keratoconus by conventional ELISA on a limited number of pooled tear samples. Our findings confirm increased IL-6, but dispute earlier reports of increased TNF-α, and suggest a cytokine imbalance in keratoconus disrupting corneal homeostasis. Moreover, an increase in IL-17 suggests tissue degenerative processes at work, contributing to the thinning and weakening of the corneal connective tissue in keratoconus.