Bladder cancer (BC) is the 12th most commonly diagnosed cancer worldwide. Although there are several well-established molecular and immunological classifications, prognostic and predictive markers for tumor cells and immune cells are still needed. Using a tissue microarray, we analyzed the expression of the chemokine CC motif ligand 5 (CCL5) by immunohistochemistry (IHC) in 175 muscle-invasive BC samples. The application of a single cutoff for the staining status of tumor cells (TCs; positive vs. negative) and immune cells (ICs; positive vs. negative) revealed 75 patients (42.9%) and 123 patients (70.3%) with CCL5-positive TCs or ICs, respectively. IHC results were associated with prognostic and predictive data. Multivariate Cox regression analysis revealed that positive CCL5 staining in TCs was associated with significantly shorter disease-specific survival (DSS; RR = 1.51; p = 0.047), but CCL5-negative ICs were associated with significantly shorter overall survival (OS; RR = 1.66; p = 0.005), DSS (RR = 2.02; p = 0.001) and recurrence-free survival (RFS; RR = 1.94; p = 0.002). Adjuvant chemotherapy was favorable for patients with CCL5-negative ICs for OS (RR = 0.30; p = 0.006), DSS (RR = 0.36; p = 0.022) and RFS (RR = 0.41; p = 0.046) but not for patients with CCL5-positive ICs, except in the subgroup of N1 + N2 patients, where it was associated with better OS. We suggest that CCL5 expression can be a prognostic and predictive marker for muscle-invasive bladder cancer patients. Full article

Bowel strictures are well recognized as one of the most severe complications in Crohn’s disease, with variable impacts on the prognosis and often needing surgical or endoscopic treatment. Distinguishing inflammatory strictures from fibrotic ones is of primary importance due to the different therapeutic approaches required. Indeed, to better understand the pathogenesis of fibrosis, it is crucial to investigate molecular processes involving genetic factors, cytokines, alteration of the intestinal barrier, and epithelial and endothelial damage, leading to an increase in extracellular matrix synthesis, which ultimately ends in fibrosis. In such a complex mechanism, the gut microbiota also seems to play a role. A better comprehension of molecular processes underlying bowel fibrosis, in addition to radiological and histopathological findings, has led to the identification of high-risk patients for personalized follow-up and testing of new therapies, primarily in preclinical models, targeting specific pathways involving Transforming Growth Factor-β, interleukins, extracellular matrix balance, and gut microbiota. Our review aims to summarize current evidence about molecular factors involved in intestinal fibrosis’ pathogenesis, paving the way for potential diagnostic biomarkers or anti-fibrotic treatments for stricturing Crohn’s disease. Full article

A vast amount of research has been conducted on path planning over recent decades, driven by the complexity of achieving optimal solutions. This paper reviews multi-robot path planning approaches and presents the path planning algorithms for various types of robots. Multi-robot path planning approaches have been classified as deterministic approaches, artificial intelligence (AI)-based approaches, and hybrid approaches. Bio-inspired techniques are the most employed approaches, and artificial intelligence approaches have gained more attention recently. However, multi-robot systems suffer from well-known problems such as the number of robots in the system, energy efficiency, fault tolerance and robustness, and dynamic targets. Deploying systems with multiple interacting robots offers numerous advantages. The aim of this review paper is to provide a comprehensive assessment and an insightful look into various path planning techniques developed in multi-robot systems, in addition to highlighting the basic problems involved in this field. This will allow the reader to discover the research gaps that must be solved for a better path planning experience for multi-robot systems. Full article

Inflammatory bowel disease (IBD) is a clinical condition of the gastrointestinal tract that has significant incidence in childhood. Major symptoms include abdominal pain, dyspepsia, delayed gastric emptying, anorexia, diarrhea and weight loss. IBD etiopathogenesis is multifactorial, with a proven involvement of cytokines. In this regard, cytokines like resistin and adiponectin produced by adipose tissue play a crucial role in inflammation. Particularly, resistin seems related to IBD severity and is considered a promising marker of disease occurrence and progression. Unraveling its mechanism of action and downstream effectors is mandatory when designing novel therapies. This preclinical study aims to further elucidate the action of resistin in causing functional gastrointestinal alterations, comparing it with the well-defined effect of adiponectin. To this end, we carried out electrophysiological analysis on murine gastric fundus. We found that resistin, similarly to adiponectin, increases smooth muscle cell (SMC) capacitance, indicative of cell surface remodeling, which is consistent with relaxation. However, contrary to adiponectin, resistin unalters membrane potential and inward Ca²⁺ entry and scarcely affects outward current, suggesting its inefficacy in markedly modifying electrical phenomena on the SMC membrane. This outcome, supporting the role of resistin in gastrointestinal distention, as observed in IBD, rules out a strikingly direct effect on SMCs. Full article

Transversely isotropic rocks, which are special anisotropic materials, are widely encountered in civil, mining, petroleum, geothermal, and radioactive waste-disposal engineering. Rock is frequently subject to cyclic loads resulting from natural and human-caused events. However, to date, the fracture mechanism of transversely isotropic rocks under cyclic loading remains poorly understood. To address this gap, uniaxial monotonic-loading and cyclic-loading tests were performed on slate specimens by the MTS815 system, during which acoustic emission (AE) signals inside the rock were monitored, and finally the fracture surfaces of the tested rock were scanned by scanning electron microscopy (SEM). Through these tests, the anisotropic mechanical properties, damage evolution, AE characteristics and fracture pattern of slate as a transversely isotropic rock were studied. The results show that the peak strength of specimens varies with the loading–foliation angle under monotonic and cyclic loading, following a U-shaped trend. The deformation modulus during unloading is more capable of characterizing the damage inside the specimen than that during loading. By defining the damage degree based on dissipation energy, it is found that the damage variable is influenced by the loading–foliation angle and the cyclic stress step. The AE characteristics of specimens exhibit significant anisotropy, closely correlated to the loading condition and loading–foliation angle. Regardless of cyclic stress step, the AE counts of specimens with a loading–foliation angle of 0° are mainly distributed near the peak region, whereas those of specimens with other loading–foliation angles occur primarily in the early stage of each cyclic loading. Finally, it is revealed that the fracture mechanism of slate specimens is determined by the loading–foliation angle, loading condition, and cyclic stress step. Full article

Recent advances in neoadjuvant systemic therapy (NST) have significantly improved pathologic complete response rates in early breast cancer, challenging the role of axillary lymph node dissection in nose-positive patients. Targeted axillary dissection (TAD) integrates marked lymph node biopsy (MLNB) and tracer-guided sentinel lymph node biopsy (SLNB). The introduction of new wire-free localisation markers (LMs) has streamlined TAD and increased its adoption. The primary endpoints include the successful localisation and retrieval rates of LMs. The secondary endpoints include the pathological complete response (pCR), SLNB, and MLNB concordance, as well as false-negative rates. Seventeen studies encompassing 1358 TAD procedures in 1355 met the inclusion criteria. The localisation and retrieval rate of LMs were 97% and 99%. A concordance rate of 67% (95% CI: 64–70) between SLNB and MLNB was demonstrated. Notably, 49 days (range: 0–272) was the average LM deployment time to surgery. pCR was observed in 46% (95% CI: 43–49) of cases, with no significant procedure-related complications. Omitting MLNB or SLNB would have under-staged the axilla in 15.2% or 5.4% (p = 0.0001) of cases, respectively. MLNB inclusion in axillary staging post-NST for initially node-positive patients is crucial. The radiation-free Savi Scout, with its minimal MRI artefacts, is the preferred technology for TAD. Full article

The bacterial agent of Lyme disease, Borrelia burgdorferi, exists in an enzootic cycle by adapting to dissimilar mammalian and tick environments. The genetic elements necessary for host and vector adaptation are spread across a bacterial genome comprised of a linear chromosome and essential linear and circular plasmids. The promoter trap system, In Vivo Expression Technology (IVET), has been used to identify promoters of B. burgdorferi that are transcriptionally active specifically during infection of a murine host. However, an observed infection bottleneck effect in mice prevented the application of this system to study promoters induced in a tick environment. In this study, we adapted a membrane-based in vitro feeding system as a novel method to infect the Ixodes spp. vector with B. burgdorferi. Once adapted, we performed IVET screens as a proof of principle via an infected bloodmeal on the system. The screen yielded B. burgdorferi promoters that are induced during tick infection and verified relative expression levels using qRT-PCR. The results of our study demonstrate the potential of our developed in vitro tick feeding system and IVET systems to gain insight into the adaptive gene expression of the Lyme disease bacteria to the tick vector. Full article

Background: The COVID-19 pandemic brought significant changes to daily life in Poland, with restrictions affecting various sectors, including entertainment, education, and travel. The pandemic’s impact extended to intimate aspects of life. This study aimed to compare the sexual functioning of young adults in Poland before and during the pandemic, using the Changes in Sexual Functioning Questionnaire (CSFQ-14). Methods: The research involved an online survey with demographic questions, the CSFQ-14 for pre-pandemic sexual functioning, and modified CSFQ-14 questions for the pandemic period. Sexual dysfunction was determined using predefined cutoff scores. Results: Overall, the study found no significant difference in the sexual functioning of young Poles during the COVID-19 pandemic compared to before. However, there were gender-specific trends. Women reported enhanced pleasure, satisfaction, and orgasm during lockdown, while men faced challenges with erection and ejaculation. A higher proportion of women experienced overall sexual dysfunction compared to men, both before and during the pandemic. This research provides insights into the impact of the COVID-19 pandemic on the sexual lives of young Poles. While overall sexual functioning remained relatively stable, specific aspects varied by gender. Conclusions: The study emphasizes the need to consider demographic factors, such as age and gender, when assessing the effects of external stressors like a pandemic on sexual health. Further research is essential to fully grasp these complexities and their potential long-term consequences. Full article

The booming growth of the internet of things has brought about widespread deployment of devices and massive amounts of sensing data to be processed. Federated learning (FL)-empowered mobile edge computing, known for pushing artificial intelligence to the network edge while preserving data privacy in learning cooperation, is a promising way to unleash the potential information of the data. However, FL’s multi-server collaborative operating architecture inevitably results in communication energy consumption between edge servers, which poses great challenges to servers with constrained energy budgets, especially wireless communication servers that rely on battery power. The device-to-device (D2D) communication mode developed for FL turns high-cost and long-distance server interactions into energy-efficient proximity delivery and multi-level aggregations, effectively alleviating the server energy constraints. A few studies have been devoted to D2D-enabled FL management, but most of them have neglected to investigate server computing power for FL operation, and they have all ignored the impact of dataset characteristics on model training, thus failing to fully exploit the data processing capabilities of energy-constrained edge servers. To fill this gap, in this paper we propose a D2D-assisted FL mechanism for energy-constrained edge computing, which jointly incorporates computing power allocation and dataset correlation into FL scheduling. In view of the variable impacts of computational power on model accuracy at different training stages, we design a partite graph-based FL scheme with adaptive D2D pairing and aperiodic variable local iterations of heterogeneous edge servers. Moreover, we leverage graph learning to exploit the performance gain of the dataset correlation between the edge servers in the model aggregation process, and we propose a graph-and-deep reinforcement learning-based D2D server pairing algorithm, which effectively reduces FL model error. The numerical results demonstrate that our designed FL schemes have great advantages in improving FL training accuracy under edge servers’ energy constraints. Full article

(1) Background: Retained placenta (RP) and mastitis are relevant diseases in dairy cows. This study mainly aimed to evaluate the contamination of uterine fluid by mastitis pathogens in dairy cows presenting RP. (2) Methods: Uterine fluids were sampled at RP diagnosis (89 ± 15 h after calving) from 5 primiparous and 10 (parity: 2–5) multiparous cows. The real-time PCR methodology was used to identify 15 mastitis and uterine pathogens. Results were analyzed using multivariate logistic regression, including the factors fever and parity. (3) Results: The prevalence of Escherichia coli was 93.3% (95% CI: 70.2–98.9%); Staphylococcus spp. (93.3%; 95% CI: 70.2–98.9%), yeasts (92.9%; 95% CI: 68.5–98.7%), Trueperella pyogenes/Peptoniphilus indolicus (80.0%; 95% CI: 54.8–93.0%), Streptococcus uberis (78.6%; 95% CI: 52.4–92.4%) and Streptococcus dysgalactiae (57.1%; 95% CI: 32.6–78.6%) comprised the largest proportions of pathogens in uterine contamination. Strep. uberis was related to the presence of fever (relative risk: 1.6; 95% CI: 1.0–2.8; p = 0.05). (4) Conclusions: Dairy cows with RP can be a relevant source of mastitis pathogens in farms. Only Strep. uberis was linked to clinical signs of infection. A high proportion of yeasts was observed in uterine fluids. Further research is needed to evaluate the real impact of RP on mastitis prevalence in dairy herds. Full article

A deeper understanding of gas emissions in milk production is crucial for promoting productive efficiency, sustainable resource use, and animal welfare. This paper aims to analyze ammonia and greenhouse gas emissions in dairy farming using bibliometric methods. A total of 187 English-language articles with experimental data from the Scopus and Web of Science databases (January 1987 to April 2024) were reviewed. Publications notably increased from 1997, with the highest number of papers published in 2022. Research mainly focuses on ammonia and methane emissions, including quantification, volatilization, and mitigation strategies. Other gases like carbon dioxide, nitrous oxide, and hydrogen sulfide were also studied. Key institutions include the University of California–Davis and Aarhus University. Bibliometric analysis revealed research evolution, identifying trends, gaps, and future research opportunities. This bibliometric analysis offers insights into emissions, air quality, sustainability, and animal welfare in dairy farming, highlighting areas for innovative mitigation strategies to enhance production sustainability. This research contributes to academia, enhancing agricultural practices, and informing environmental policies. It is possible to conclude that this research is a valuable tool for understanding the evolution of research on gas emissions in dairy cattle facilities, providing guidance for future studies and interventions to promote more sustainable production. Full article

The air-supported membrane coal shed is widely used in bulk cargo terminals. It not only effectively protects goods from adverse weather conditions but also helps reduce coal dust and harmful gas emissions, promoting the green and sustainable development of ports. However, in practical engineering, the design parameters of the coal shed are often based on experience, making it difficult to accurately assess the quality of the structural design. The flexibility of the membrane material also makes the structure susceptible to deformation or tearing. This paper mainly focuses on modeling and solving the optimization design issues of air-supported membrane coal shed structures. According to the evaluation criteria for the form of air-supported membrane coal sheds, a multi-objective structural optimization model is established to minimize the maximum stress on the membrane surface, ensure uniform stress distribution, maximize structural stiffness, and minimize costs. The study utilizes a combined optimization approach using ANSYS 19.0 and MATLAB 2016a, incorporating an improved NSGA-II algorithm program developed in MATLAB into ANSYS for structural form analysis and load calculation. The research results indicate that the optimal solution reduces the maximum stress on the loaded membrane surface by 5.36%, shortens the maximum displacement by 30.3%, and saves on economic costs by 9.85%. Compared to traditional empirical design methods, the joint use of MATLAB and ANSYS for optimization design can provide more superior solutions, helping ports to achieve environmental protection and economic efficiency goals. Full article

Wastewater Treatment Plants (WWTPs) play a crucial role in maintaining ecological balance, a cornerstone of environmental health for thriving biodiversity and undisturbed natural processes. This balance is crucial for the sustainability of ecosystems, directly influencing human health, biodiversity, and the overall quality of our natural environment. WWTPs contribute to this equilibrium by efficiently removing pollutants and harmful substances from wastewater, thus averting the degradation of water bodies that are essential for numerous ecological processes. WWTPs encompass multiple stages of wastewater and sludge treatment and are significant energy consumers globally, especially in secondary treatment, particularly the activated sludge method which is the most common method. With an upcoming directive from the European Union aiming to reduce energy consumption in WWTPs, this paper focuses on a literature review examining global practices implemented across all stages of WWTP treatment processes. It summarizes the key points of each study, focusing primarily on the outcomes of each application. This document concludes with an in-depth review of each study and provides general conclusions for each group of studies. The objective is to identify methods that have effectively reduced energy consumption and enhanced the overall energy efficiency of WWTPs. The main conclusions indicate that the studies encompass a wide range of applications that achieve significant reductions in energy consumption. However, additional testing of these applications in more diverse operating environments through trials could further enhance their reliability and increase acceptance among WWTP operators. Full article

The electron beam cold hearth melting (EBCHM) process is one of the key processes for titanium alloy production. The unique characteristic of this pyrometallurgy process is the application of the cold hearth, which is responsible for controlling the Low-Density Inclusions (LDIs) and High-Density Inclusions (HDIs) in the melt. As a key process of inclusion removal, the information such as melt residence time in the cold hearth is directly related to the control of metallurgical defects in the ingot, and may also affect the composition distribution of the ingot. In this paper, the details for the physical phenomena, namely the evolution of the pool, the evolution of the flow, and the evolution of the component in the cold hearth during EBCHM are investigated using a modified multi-physical numerical model. The effects of melting temperature and melting speed on these phenomena were investigated. The purpose is to provide more fundamental knowledge and to further enhance the applications of EBCHM for more titanium alloys. Full article

Musculoskeletal disorders, which are epidemiologically related to exposure to whole-body vibration (WBV), are frequently self-reported by workers in the construction sector. Several activities during building construction and demolition expose workers to this physical agent. Directive 2002/44/CE defined a method of assessing WBV exposure that was limited to an eight-hour working day, and did not consider the cumulative and long-term effects on the health of drivers. This study aims to propose a methodology for generating individualised models for vehicle drivers exposed to WBV that are easy to implement by companies, to ensure that the health of workers is not compromised in the short or long term. A measurement campaign was conducted with a professional driver, and the collected data were used to formulate six artificial neural networks to predict the daily compressive dose on the lumbar spine and to assess the short- and long-term WBV exposure. Accurate results were obtained from the developed artificial neural network models, with R² values above 0.90 for training, cross-validation, and testing. The approach proposed in this study offers a new tool that can be applied in the assessment of short- and long-term WBV to ensure that workers’ health is not compromised during their working life and subsequent retirement. Full article

This work examines the potential of some of the Gulf Cooperation Council countries (GCC) (Saudi Arabia (KSA), the United Arab Emirates (UAE), Qatar (QA), Bahrain (BH), Oman (OM)), Yemen (YE), Iraq (IQ), and Jordan (JO) to use their abundant solar radiation to generate electricity through PV technology. The study is structured to help decision-makers access the necessary data related to the status of solar-energy infrastructure and power production in the study region. The study investigates current efforts to establish PV technology and the challenges hindering the development of this technology. These efforts and challenges are then benchmarked against their status in Australia, which has climate and landscape conditions similar to those of the countries in the study region. It was found that Australia is successfully adopting solar energy in households and industrial locations despite its historical reliance on fossil fuels for energy production. This offers a potential avenue for replicating the Australian model of PV development in the study region. This work also addresses the effect of natural and anthropogenic aerosols on the performance of the PV panels. Meanwhile, it also proposes a conceptual model to help local governments and decision-makers in adopting solar-energy projects in the study region. Additionally, a preliminary carbon-footprint analysis of avoided emissions from PV energy utilization compared to national grid intensity was performed for each country. Findings show that the countries in the study region have great potential for using solar energy to gradually replace fossil fuels and protect the environment. It is observed that more hours of daylight and clear-to-scattered cloud coverage help increase solar irradiance near the ground all year around. Dust and aerosol loadings, however, were found to greatly reduce solar irradiance over the GCC area, especially during large dust events. Despite the high potential for harvesting solar energy in the study region, only a handful of PV plants and infrastructural facilities have been established, mostly in the KSA, the UAE, and Jordan. It was found that there is a critical need to put in place regulations, policies, and near-future vision to support solar energy generation and reduce reliance on fossil fuels for electricity production. Full article

A wide-angle scanning phased array with dual circular polarization in the Ka-band is presented in this paper. To improve the scanning capability of the phased array, the microstrip element is modified by loading many metal posts at its center and periphery. In addition, a stripline coupler is designed to achieve dual circularly polarized (CP) radiation, and the inner conductor of the subminiature micro-push-on (SSMP) connectors for feeding the coupler is extended to the top layer of the multilayer element by introducing an open stub, which simplifies the assembly process between the SSMP connector and multilayer printed circuit board (PCB) due to through-hole soldering instead of blind-hole soldering. The proposed element can cover a frequency range from 28 GHz to 30.5 GHz with a relative bandwidth of 8.5% in the Ka-band. An 8 × 8 phased array is constructed based on this proposed element, and a wide-angle scanning range from −65° to +65° is obtained for the dual circular polarization. The proposed array has a gain fluctuation of 5.1 dB and an axial ratio (AR) of less than 3.3 dB during beam-steering. The prototype is fabricated and measured, with a good agreement between the measured and simulated results. The proposed phased array can be applied in a Ka-band millimeter-wave (MMW) communication system. Full article

The dependence of photochemistry on excitation wavelength is not a recently observed phenomenon; nonetheless, it has, surprisingly enough, been largely ignored in the field. The reasons for this situation are not fully understood but might be related to a provisional extension of Kasha’s rule to photochemistry, or perhaps to a difficulty to justify the kind of short time-scales implied in such photochemistry, that challenges the usually held view giving predominance to fast internal conversion and vibrational relaxation. Regardless of the reasons, it is still a matter of fact that a complete and satisfactory interpretation for experimentally proven wavelength-dependent photochemistry is not yet available and the community endeavor to build a holistic understanding and a comprehensive view of the phenomenon. The present review is a non-exhaustive overview of the published data in the field, reporting on some of the most prominent features, issues, and interpretations. Full article

The purpose of this study was to determine Turkish physical education (PE) teachers’ level of technological innovation use and attitudes regarding their applications and diffusion. This study consisted of 678 Turkish PE teachers. In total, 265 (39.1%) of the PE teachers were female, and 413 (60.9%) were male. A sequential explanatory mixed-methods design was used in this study. “The Applying and Diffusing Technological Innovations Scale for Teachers” was used for data collection. The results showed that the main obstacles the PE teachers face in using technological innovations were the cost of products; security and privacy problems; the need for new versions and technical support; the lack of time; the lack of interest; difficulty in accessing technology; and the lack of understanding. Moreover, there was a significant difference in the tendency to apply and diffuse technological innovations in terms of gender, school type, and school level. We found that the “ability to use innovations” sub-dimension affects the PE teachers’ tendencies to apply and diffuse technology. As a result, we recommend providing various training programs on technological innovations to the older/longer-serving PE teachers, especially those working in public schools and secondary schools. Full article

Sesame is an important oilseed crop grown for human consumption in many countries, with a high commercial value due to its high oleic/linoleic acid ratio (O/L ratio). However, its properties may vary among different accessions. In the current study, 282 sesame accessions were evaluated to determine the effects of agronomic traits and genotypes on the O/L ratio. The O/L ratio was positively correlated with the oleic acid (C18:1), stearic acid (C18:0), and myristic acid (C14:0) concentrations, as well as the capsule zone length (CZL), capsule width (CW), and capsule length (CL), and negatively correlated with the linoleic acid (C18:2) and linolenic acid (C18:3) concentrations, the days to maturity (DTM), days to flowering (DTF), and the height of the first capsule-bearing node (HFC) (p < 0.05). In addition, the O/L ratio was affected by the FAD2 haplotype, as the Hap2 and Hap3 sesame accessions had lower O/L ratios. Therefore, we suggest that the increase and decrease in the contents of C18:1 and C18:2 are associated with the FAD2 haplotype. A total of 25 agronomic traits and fatty acid compositions were compared via statistical analysis, and accessions with a high O/L ratio were selected. The results of this study can be used as a basis for further research on the development of new sesame varieties through enhancing nutritional functionality. Full article

Over recent decades, Landsat satellite data has evolved into a highly valuable resource across diverse fields. Long-term satellite data records with integrity and consistency, such as the Landsat series, provide indispensable data for many applications. However, the malfunction of the Scan Line Corrector (SLC) on the Landsat 7 satellite in 2003 resulted in stripping in subsequent images, compromising the temporal consistency and data quality of Landsat time-series data. While various methods have been proposed to improve the quality of Landsat 7 SLC-off data, existing gap-filling methods fail to enhance the temporal resolution of reconstructed images, and spatiotemporal fusion methods encounter challenges in managing large-scale datasets. Therefore, we propose a method for reconstructing dense time series from SLC-off data. This method utilizes the Neighborhood Similar Pixel Interpolator to fill in missing values and leverages the time-series information to reconstruct high-resolution images. Taking the blue band as an example, the surface reflectance verification results show that the Mean Absolute Error (MAE) and BIAS reach minimum values of 0.0069 and 0.0014, respectively, with the Correlation Coefficient (CC) and Structural Similarity Index Metric (SSIM) reaching 0.93 and 0.94. The proposed method exhibits advantages in repairing SLC-off data and reconstructing dense time-series data, enabling enhanced remote sensing applications and reliable Earth’s surface reflectance data reconstruction. Full article

To characterize transforming growth factor-β (TGF-β) isoform (TGF-β1~3)-b’s biological effects on the human retinal pigment epithelium (RPE) under normoxia and hypoxia conditions, ARPE19 cells cultured by 2D (two-dimensional) and 3D (three-dimensional) conditions were subjected to various analyses, including (1) an analysis of barrier function by trans-epithelial electrical resistance (TEER) measurements; (2) qPCR analysis of major ECM molecules including collagen 1 (COL1), COL4, and COL6; α-smooth muscle actin (αSMA); hypoxia-inducible factor 1α (HIF1α); and peroxisome proliferator-activated receptor-gamma coactivator (PGC1α), a master regulator for mitochondrial respiration;, tight junction-related molecules, Zonula occludens-1 (ZO1) and E-cadherin; and vascular endothelial growth factor (VEGF); (3) physical property measurements of 3D spheroids; and (4) cellular metabolic analysis. Diverse effects among TGF-β isoforms were observed, and those effects were also different between normoxia and hypoxia conditions: (1) TGF-β1 and TGF-β3 caused a marked increase in TEER values, and TGF-β2 caused a substantial increase in TEER values under normoxia conditions and hypoxia conditions, respectively; (2) the results of qPCR analysis supported data obtained by TEER; (3) 3D spheroid sizes were decreased by TGF-β isoforms, among which TGF-β1 had the most potent effect under both oxygen conditions; (4) 3D spheroid stiffness was increased by TGF-β2 and TGF-β3 or by TGF-β1 and TGF-β3 under normoxia conditions and hypoxia conditions, respectively; and (5) the TGF-β isoform altered mitochondrial and glycolytic functions differently under oxygen conditions and/or culture conditions. These collective findings indicate that the TGF-β-induced biological effects of 2D and 3D cultures of ARPE19 cells were substantially diverse depending on the three TGF-β isoforms and oxygen levels, suggesting that pathological conditions including epithelial–mesenchymal transition (EMT) of the RPE may be exclusively modulated by both factors. Full article