bluuebunny/crossref_metadata_2025 huggingface dataset:
Development and Biological Characterization of a Novel Selective TrkA Agonist with Neuroprotective Properties against Amyloid Toxicity
Authors: Rogdakis Thanasis , Charou Despoina , Latorrata Alessia , Papadimitriou Eleni , Tsengenes Alexandros , Athanasiou Christina , Papadopoulou Marianna , Chalikiopoulou Constantina , Katsila Theodora , Ramos Isbaal , Prousis Kyriakos C. , Wade Rebecca C. , Sidiropoulou Kyriaki , Calogeropoulou Theodora , Gravanis Achille , Charalampopoulos Ioannis
Published: March 2022

Neurotrophins are growth factors that exert important neuroprotective effects by preventing neuronal death and synaptic loss. Nerve Growth Factor (NGF) acts through the activation of its high-affinity, pro-survival TrkA and low-affinity, pro-apoptotic p75NTR receptors. NGF has been shown to slow or prevent neurodegenerative signals in Alzheimer’s Disease (AD) progression. However, its low bioavailability and its blood–brain-barrier impermeability limit the use of NGF as a potential therapeutic agent against AD. Based on our previous findings on synthetic dehydroepiandrosterone derivatives, we identified a novel NGF mimetic, named ENT-A013, which selectively activates TrkA and exerts neuroprotective, anti-amyloid-β actions. We now report the chemical synthesis, in silico modelling, metabolic stability, CYP-mediated reaction phenotyping and biological characterization of ENT-A013 under physiological and neurodegenerative conditions. We show that ENT-A013 selectively activates the TrkA receptor and its downstream kinases Akt and Erk1/2 in PC12 cells, protecting these cells from serum deprivation-induced cell death. Moreover, ENT-A013 promotes survival of primary Dorsal Root Ganglion (DRG) neurons upon NGF withdrawal and protects hippocampal neurons against Amyloid β-induced apoptosis and synaptic loss. Furthermore, this neurotrophin mimetic partially restores LTP impairment. In conclusion, ENT-A013 represents a promising new lead molecule for developing therapeutics against neurodegenerative disorders, such as Alzheimer’s Disease, selectively targeting TrkA-mediated pro-survival signals.
Exploiting Shock Waves to Trigger the Anticancer Sonodynamic Activity of 5-Aminolevulinc Acid-Derived Protoporphyrin IX on In Vitro 2D and 3D Cancer Models
Authors: Foglietta Federica , Panzanelli Patrizia , Serpe Loredana , Canaparo Roberto
Published: March 2022

Sonodynamic therapy (SDT) is a noninvasive method for cancer treatment based on selective activation of a sonosensitiser by ultrasound (US), which results in the generation of reactive oxygen species (ROS) and cancer cell death. SDT uses a similar approach to photodynamic therapy (PDT), but can overcome the main drawback of PDT, i.e., poor tissue penetration of light. This research work investigated the anticancer effect of SDT on various two- (2D) and three-dimensional (3D) in vitro tumour models, using PDT as a reference treatment. Sonodynamic experiments were performed with pulsed US, specifically with shock waves (SW) and the prodrug 5-aminolevulinic acid (Ala), which is converted—at the mitochondrial level—into the sonosensitiser protoporphyrin IX (PPIX). SW-mediated PPIX sonodynamic activation resulted in a significant decrease in cell proliferation, especially on human fibrosarcoma (HT-1080) cells, where PPIX accumulation was higher compared to human melanoma (A2058) and neuroblastoma (SH-SY5 Y) cells. Moreover, SW-mediated SDT showed significant ROS generation, cell line-dependent in its amount, probably due to differences in Ala-induced PPIX synthesis. In all cancer cell lines, apoptosis was highlighted as the main cancer cell death pathway determined by SW-mediated SDT, along with significant cytochrome c release, and a consequent increase in DNA damage. The efficacy of SDT with SW and Ala in halting cancer cell proliferation was also confirmed in 3D cancer spheroids. The present study suggests that SW-mediated SDT is a valuable approach to slow down tumour proliferation, thus opening an innovative scenario in cancer treatment.
Bezafibrate Reduces Elevated Hepatic Fumarate in Insulin-Deficient Mice
Authors: Franko Andras , Irmler Martin , Prehn Cornelia , Heinzmann Silke S. , Schmitt-Kopplin Philippe , Adamski Jerzy , Beckers Johannes , von Kleist-Retzow Jürgen-Christoph , Wiesner Rudolf , Häring Hans-Ulrich , Heni Martin , Birkenfeld Andreas L. , de Angelis Martin Hrabě
Published: March 2022

Glucotoxic metabolites and pathways play a crucial role in diabetic complications, and new treatment options which improve glucotoxicity are highly warranted. In this study, we analyzed bezafibrate (BEZ) treated, streptozotocin (STZ) injected mice, which showed an improved glucose metabolism compared to untreated STZ animals. In order to identify key molecules and pathways which participate in the beneficial effects of BEZ, we studied plasma, skeletal muscle, white adipose tissue (WAT) and liver samples using non-targeted metabolomics (NMR spectroscopy), targeted metabolomics (mass spectrometry), microarrays and mitochondrial enzyme activity measurements, with a particular focus on the liver. The analysis of muscle and WAT demonstrated that STZ treatment elevated inflammatory pathways and reduced insulin signaling and lipid pathways, whereas BEZ decreased inflammatory pathways and increased insulin signaling and lipid pathways, which can partly explain the beneficial effects of BEZ on glucose metabolism. Furthermore, lysophosphatidylcholine levels were lower in the liver and skeletal muscle of STZ mice, which were reverted in BEZ-treated animals. BEZ also improved circulating and hepatic glucose levels as well as lipid profiles. In the liver, BEZ treatment reduced elevated fumarate levels in STZ mice, which was probably due to a decreased expression of urea cycle genes. Since fumarate has been shown to participate in glucotoxic pathways, our data suggests that BEZ treatment attenuates the urea cycle in the liver, decreases fumarate levels and, in turn, ameliorates glucotoxicity and reduces insulin resistance in STZ mice.
Immune-Mediated Inflammatory Responses of Alveolar Epithelial Cells: Implications for COVID-19 Lung Pathology
Authors: Barilli Amelia , Visigalli Rossana , Ferrari Francesca , Bianchi Massimiliano G. , Dall’Asta Valeria , Rotoli Bianca Maria
Published: March 2022

Background. Clinical and experimental evidence point to a dysregulated immune response caused by SARS-CoV-2 as the primary mechanism of lung disease in COVID-19. However, the pathogenic mechanisms underlying COVID-19-associated ARDS (Acute Respiratory Distress Syndrome) remain incompletely understood. This study aims to explore the inflammatory responses of alveolar epithelial cells to either the spike S1 protein or to a mixture of cytokines secreted by S1-activated macrophages. Methods and Results. The exposure of alveolar A549 cells to supernatants from spike-activated macrophages caused a further release of inflammatory mediators, with IL-8 reaching massive concentrations. The investigation of the molecular pathways indicated that NF-kB is involved in the transcription of IP-10 and RANTES, while STATs drive the expression of all the cytokines/chemokines tested, with the exception of IL-8 which is regulated by AP-1. Cytokines/chemokines produced by spike-activated macrophages are also likely responsible for the observed dysfunction of barrier integrity in Human Alveolar Epithelial Lentivirus-immortalized cells (hAELVi), as demonstrated by an increased permeability of the monolayers to mannitol, a marked decrease of TEER and a disorganization of claudin-7 distribution. Conclusion. Upon exposure to supernatants from S1-activated macrophages, A549 cells act both as targets and sources of cytokines/chemokines, suggesting that alveolar epithelium along with activated macrophages may orchestrate lung inflammation and contribute to alveolar injury, a hallmark of ARDS.
Side-Directed Release of Differential Extracellular Vesicle-associated microRNA Profiles from Bronchial Epithelial Cells of Healthy and Asthmatic Subjects
Authors: Schindler Viktoria E. M. , Alhamdan Fahd , Preußer Christian , Hintz Lukas , Alashkar Alhamwe Bilal , Nist Andrea , Stiewe Thorsten , Pogge von Strandmann Elke , Potaczek Daniel P. , Thölken Clemens , Garn Holger
Published: March 2022

Extracellular vesicles (EVs) are released by virtually all cells and may serve as intercellular communication structures by transmitting molecules such as proteins, lipids, and nucleic acids between cells. MicroRNAs (miRNAs) are an abundant class of vesicular RNA playing a pivotal role in regulating intracellular processes. In this work, we aimed to characterize vesicular miRNA profiles released in a side-directed manner by bronchial epithelial cells from healthy and asthmatic subjects using an air−liquid interface cell culture model. EVs were isolated from a culture medium collected from either the basolateral or apical cell side of the epithelial cell cultures and characterized by nano-flow cytometry (NanoFCM) and bead-based flow cytometry. EV-associated RNA profiles were assessed by small RNA sequencing and subsequent bioinformatic analyses. Furthermore, miRNA-associated functions and targets were predicted and miRNA network analyses were performed. EVs were released at higher numbers to the apical cell side of the epithelial cells and were considerably smaller in the apical compared to the basolateral compartment. EVs from both compartments showed a differential tetraspanins surface marker expression. Furthermore, 236 miRNAs were differentially expressed depending on the EV secretion side, regardless of the disease phenotype. On the apical cell side, 32 miRNAs were significantly altered in asthmatic versus healthy conditions, while on the basolateral cell side, 23 differentially expressed miRNAs could be detected. Downstream KEGG pathway analysis predicted mTOR and MAPK signaling pathways as potential downstream targets of apically secreted miRNAs. In contrast, miRNAs specifically detected at the basolateral side were associated with processes of T and B cell receptor signaling. The study proves a compartmentalized packaging of EVs by bronchial epithelial cells supposedly associated with site-specific functions of cargo miRNAs, which are considerably affected by disease conditions such as asthma.
Neurodevelopmental Disorders: From Pathophysiology to Novel Therapeutic Approaches
Authors: Kosel Markus , Perroud Nader
Published: March 2022

This special issue of Biomedicines on Neurodevelopmental Disorders (NDD): “From Pathophysiology to Novel Therapeutic Approaches”, is a precursor of what we hope will develop into a thriving and inspiring transdisciplinary field, including genetics, psychiatry, neurology, as well as basic and applied neurosciences and molecular biology in the research area [...]
A Wearable Breath Sensor Based on Fiber-Tip Microcantilever
Authors: Zhao Cong , Liu Dan , Cai Zhihao , Du Bin , Zou Mengqiang , Tang Shuo , Li Bozhe , Xiong Cong , Ji Peng , Zhang Lichao , Gong Yuan , Xu Gaixia , Liao Changrui , Wang Yiping
Published: March 2022

Respiration rate is an essential vital sign that requires monitoring under various conditions, including in strong electromagnetic environments such as in magnetic resonance imaging systems. To provide an electromagnetically-immune breath-sensing system, we propose an all-fiber-optic wearable breath sensor based on a fiber-tip microcantilever. The microcantilever was fabricated on a fiber-tip by two-photon polymerization microfabrication based on femtosecond laser, so that a micro Fabry–Pérot (FP) interferometer was formed between the microcantilever and the end-face of the fiber. The cavity length of the micro FP interferometer was reduced as a result of the bending of the microcantilever induced by breath airflow. The signal of breath rate was rebuilt by detecting power variations of the FP interferometer reflected light and applying dynamic thresholds. The breath sensor achieved a high sensitivity of 0.8 nm/(m/s) by detecting the reflection spectrum upon applied flow velocities from 0.53 to 5.31 m/s. This sensor was also shown to have excellent thermal stability as its cross-sensitivity of airflow with respect to the temperature response was only 0.095 (m/s)/°C. When mounted inside a wearable surgical mask, the sensor demonstrated the capability to detect various breath patterns, including normal, fast, random, and deep breaths. We anticipate the proposed wearable breath sensor could be a useful and reliable tool for respiration rate monitoring.
Impact of the COVID-19 Pandemic on Patients with Parkinson’s Disease from the Perspective of Treating Physicians—A Nationwide Cross-Sectional Study
Authors: Wolff Andreas Wolfgang , Haller Bernhard , Demleitner Antonia Franziska , Westenberg Erica , Lingor Paul
Published: March 2022

The COVID-19 pandemic has posed challenges to maintaining medical care for patients with Parkinson’s disease (PD). The Parkinson’s Disease during the COVID-19 Pandemic (ParCoPa) survey was conducted as an online, nationwide, cross-sectional survey from December 2020 to March 2021 and aimed to assess the impact of the pandemic on the medical care of PD patients from the physicians’ perspective. Invitations containing a randomly generated registration code were mailed to healthcare professionals from sixty-seven specialty centers in Germany. Confounders for the worsening of subjective treatment quality, perceived health risk due to the profession, and adequate protective measures against SARS-CoV-2 were assessed using logistic regression analysis. Of all forty physicians who responded, 87.5% reported a worsening of motor and nonmotor symptoms in their patients, 97.5% experienced cancellation of appointments, and difficulties in organizing advanced and supplementary therapies were reported by over 95%. Participants offered alternative consultation options, mostly in the form of telephone (77.5%) or online (64.1%) consultations, but telephone consultations were the most accepted by patients (“broadly accepted”, 40.0%). We identified pandemic-related deficits in providing care for patients with PD and areas of improvement to ensure continued care for this vulnerable patient population.
Disorder-Specific Profiles of Self-Perceived Emotional Abilities in Schizophrenia and Major Depressive Disorder
Authors: Weiss Elisabeth M. , Deisenhammer Eberhard A. , Fink Andreas , Marksteiner Josef , Canazei Markus , Papousek Ilona
Published: March 2022

Deficits in social cognition are a core feature of neuropsychiatric disorders. The purpose of this study was to compare profiles of self-perceived abilities across the core domains of emotional functioning between patients with schizophrenia (n = 22), major depressive disorder (n = 31) and healthy participants (n = 43) with the Self-report Emotional Ability Scale (SEAS). Profile analyses were used to explore group differences in the overall level of self-perceived effectiveness of emotional functioning and in the patterns in which the four functions of emotion perception and regulation in the intra- and inter-personal domains are arranged to each other. Both patient groups showed significantly lower overall levels of self-perceived emotional functioning compared to healthy controls. Most importantly, we found significant differences between patient groups in their profile patterns. Patients with schizophrenia indicated experiencing difficulties in all investigated domains, but the profile pattern largely matched that of healthy individuals. Instead, the profile of patients with depression was much more accentuated, showing lower perceived effectiveness of emotion perception and regulation in the intra-personal domain compared to inter-personal functions. Our results of disorder-specific emotional deficits may have profound implications for early screening and identification of at-risk populations as well as recovery-oriented interventions.
Neural Correlates of Numerical Estimation: The Role of Strategy Use
Authors: Ashkenazi Sarit , Tikochinski Refael , Ganor-Stern Dana
Published: March 2022

Introduction: Computation estimation is the ability to provide an approximate answer to a complex arithmetic problem without calculating it exactly. Despite its importance in daily life, the neuronal network underlying computation estimation is largely unknown. Methods: We looked at the neuronal correlates of two computational estimation strategies: approximated calculation and sense of magnitude (SOM)–intuitive representation of magnitude, without calculation. During an fMRI scan, thirty-one college students judged whether the result of a two-digit multiplication problem was larger or smaller than a given reference number. In two different blocks, they were asked to use a specific strategy (AC or SOM). Results: The two strategies activated brain regions related to calculation, numerical cognition, decision-making, and working memory. AC more than SOM elicited activations in multiple, domain-specific brain regions in the parietal lobule, including the left SMG (BA 40), the bilateral superior parietal lobule (BA 7), and the right inferior parietal lobule (BA 7). The activation level of the IFG was positively correlated to individual accuracy, indicating that the IFG has an essential role in both strategies. Conclusions: These finding suggest that the analogic code of magnitude is more involved in the AC than the SOM strategy.
The Environmental Profile of Clinker, Cement, and Concrete: A Life Cycle Perspective Study Based on Ecuadorian Data
Authors: Petroche Daniel M. , Ramirez Angel D.
Published: March 2022

Concrete is the most-used material in the construction industry, and the second most-used after water. Cement is the main component of concrete. A total of 8% of global CO2 emissions correspond to the cement industry; CO2 is the main greenhouse gas contributing to global warming. To mitigate climate change, it is necessary to design buildings with a lower environmental impact, and therefore, it is crucial to assess the environmental profile of the local production of construction materials. This study uses the life cycle assessment methodological framework to evaluate the environmental sustainability of the cement and concrete industry in Ecuador. The inventory accounts for 62.8% of national cement production, with data corresponding to 2019. The OpenLCA software was used to perform the life cycle inventory and impact assessment calculations. Eight impact categories were assessed, including Global Warming Potential (GWP). Clinker has a GWP result of 897.04 kg CO2-Eq/ton. Hydraulic cement types MH, GU, and HE have GWPs ranging from 465.89 to 696.81 kg CO2-Eq/ton. Results of ready-mixed concrete range from 126.02 to 442.14 kg CO2-Eq/m3. Reducing the content of clinker in cement and concrete should be the aim so as to improve their environmental profiles. This study contributes to the development of regional life cycle inventory data for Latin America. This research is the first to be developed regarding construction materials in Ecuador and contributes to the sustainable design of structures with pozzolan-lime cement and concrete.
Predicting Marshall Flow and Marshall Stability of Asphalt Pavements Using Multi Expression Programming
Authors: Awan Hamad Hassan , Hussain Arshad , Javed Muhammad Faisal , Qiu Yanjun , Alrowais Raid , Mohamed Abdeliazim Mustafa , Fathi Dina , Alzahrani Abdullah Mossa
Published: March 2022

The traditional method to obtain optimum bitumen content and the relevant parameters of asphalt pavements entails time-consuming, complicated and expensive laboratory procedures and requires skilled personnel. This research study uses innovative and advanced machine learning techniques, i.e., Multi-Expression Programming (MEP), to develop empirical predictive models for the Marshall parameters, i.e., Marshall Stability (MS) and Marshall Flow (MF) for Asphalt Base Course (ABC) and Asphalt Wearing Course (AWC) of flexible pavements. A comprehensive, reliable and wide range of datasets from various road projects in Pakistan were produced. The collected datasets contain 253 and 343 results for ABC and AWC, respectively. Eight input parameters were considered for modeling MS and MF. The overall performance of the developed models was assessed using various statistical measures in conjunction with external validation. The relationship between input and output parameters was determined by performing parametric analysis, and the results of trends were found to be consistent with earlier research findings stating that the developed predicted models are well trained. The results revealed that developed models are superior and efficient in terms of prediction and generalization capability for output parameters, as evident by the correlation coefficient (R) (in this case >0.90) for both ABC and AWC.
Tumor-Associated Macrophages in Gliomas—Basic Insights and Treatment Opportunities
Authors: Andersen Johannes K. , Miletic Hrvoje , Hossain Jubayer A.
Published: March 2022

Glioma refers to a group of primary brain tumors which includes glioblastoma (GBM), astrocytoma and oligodendroglioma as major entities. Among these, GBM is the most frequent and most malignant one. The highly infiltrative nature of gliomas, and their intrinsic intra- and intertumoral heterogeneity, pose challenges towards developing effective treatments. The glioma microenvironment, in addition, is also thought to play a critical role during tumor development and treatment course. Unlike most other solid tumors, the glioma microenvironment is dominated by macrophages and microglia—collectively known as tumor-associated macrophages (TAMs). TAMs, like their homeostatic counterparts, are plastic in nature and can polarize to either pro-inflammatory or immunosuppressive states. Many lines of evidence suggest that immunosuppressive TAMs dominate the glioma microenvironment, which fosters tumor development, contributes to tumor aggressiveness and recurrence and, very importantly, impedes the therapeutic effect of various treatment regimens. However, through the development of new therapeutic strategies, TAMs can potentially be shifted towards a proinflammatory state which is of great therapeutic interest. In this review, we will discuss various aspects of TAMs in the context of glioma. The focus will be on the basic biology of TAMs in the central nervous system (CNS), potential biomarkers, critical evaluation of model systems for studying TAMs and finally, special attention will be given to the potential targeted therapeutic options that involve the TAM compartment in gliomas.
Ipilimumab, Pembrolizumab, or Nivolumab in Combination with BBI608 in Patients with Advanced Cancers Treated at MD Anderson Cancer Center
Authors: Vo Henry Hiep , Cartwright Carrie , Song I-Wen , Karp Daniel D. , Nogueras Gonzalez Graciela M. , Xie Yuran , Karol Michael , Hitron Matthew , Vining David , Tsimberidou Apostolia-Maria
Published: March 2022

Background: BBI608 is an investigational reactive oxygen species generator that affects several molecular pathways. We investigated BBI608 combined with immune checkpoint inhibitors in patients with advanced cancers. Methods: BBI608 (orally twice daily) was combined with ipilimumab (3 mg/kg IV every 3 weeks); pembrolizumab (2 mg/kg IV every 3 weeks); or nivolumab (3 mg/kg IV every 4 weeks). We assessed the safety, antitumor activity and the pharmacokinetic profile of BBI combined with immunotherapy. Results: From 1/2017 to 3/2017, 12 patients were treated (median age, 54 years; range, 31–78; 6 men). Treatment was overall well tolerated. No dose-limiting toxicity was observed. The most common adverse events were diarrhea (5 patients: grade (G)1–2, n = 3; G3, n = 2) and nausea (4 patients, all G1). Prolonged disease stabilization was noted in five patients treated with BBI608/nivolumab lasting for 12.1, 10.1, 8.0, 7.7 and 7.4 months. The median progression-free survival was 2.73 months. The median overall survival was 7.56 months. Four patients had prolonged overall survival (53.0, 48.7, 51.9 and 48.2 months). Conclusions: Checkpoint inhibitors combined with BBI608 were well tolerated. Several patients had prolonged disease stabilization and overall survival. Prospective studies to elucidate the mechanisms of response and resistance to BBI608 are warranted.
Negative Relationship between Post-Treatment Stromal Tumor-Infiltrating Lymphocyte (TIL) and Survival in Triple-Negative Breast Cancer Patients Treated with Dose-Dense Dose-Intense NeoAdjuvant Chemotherapy
Authors: Giacchetti Sylvie , Faucheux Lilith , Gardair Charlotte , Cuvier Caroline , de Roquancourt Anne , Campedel Luca , Groheux David , de Bazelaire Cedric , Lehmann-Che Jacqueline , Miquel Catherine , Cahen Doidy Laurence , Amellou Malika , Madelaine Isabelle , Reyal Fabien , Someil Laetitia , Hocini Hamid , Hennequin Christophe , Teixeira Luis , Espié Marc , Chevret Sylvie , Soumelis Vassili , Hamy Anne-Sophie
Published: March 2022

Background: Patients with triple-negative breast cancers (TNBC) have a poor prognosis unless a pathological complete response (pCR) is achieved after neoadjuvant chemotherapy (NAC). Few studies have analyzed changes in TIL levels following dose-dense dose-intense (dd-di) NAC. Patients and methods: From 2009 to 2018, 117 patients with TNBC received dd-di NAC at our institution. We aimed to identify factors associated with pre- and post-NAC TIL levels, and oncological outcomes relapse-free survival (RFS), and overall survival (OS). Results: Median pre-NAC and post-NAC TIL levels were 15% and 3%, respectively. Change in TIL levels with treatment was significantly correlated with metabolic response (SUV) and pCR. High post-NAC TIL levels were associated with a weak metabolic response after two cycles of NAC, with the presence of residual disease and nodal involvement at NAC completion. In multivariate analyses, high post-NAC TIL levels independently predicted poor RFS and poor OS (HR = 1.4 per 10% increment, 95%CI (1.1; 1.9) p = 0.014 and HR = 1.8 per 10% increment 95%CI (1.3–2.3), p < 0.0001, respectively). Conclusion: Our results suggest that TNBC patients with TIL enrichment after NAC are at higher risk of relapse. These patients are potential candidates for adjuvant treatment, such as immunotherapy, in clinical trials.
Prediction of Early Response to Immunotherapy: DCE-US as a New Biomarker
Authors: Naccache Raphael , Belkouchi Younes , Lawrance Littisha , Benatsou Baya , Hadchiti Joya , Cournede Paul-Henry , Ammari Samy , Talbot Hugues , Lassau Nathalie
Published: March 2022

Purpose: The objective of our study is to propose fast, cost-effective, convenient, and effective biomarkers using the perfusion parameters from dynamic contrast-enhanced ultrasound (DCE-US) for the evaluation of immune checkpoint inhibitors (ICI) early response. Methods: The retrospective cohort used in this study included 63 patients with metastatic cancer eligible for immunotherapy. DCE-US was performed at baseline, day 8 (D8), and day 21 (D21) after treatment onset. A tumor perfusion curve was modeled on these three dates, and change in the seven perfusion parameters was measured between baseline, D8, and D21. These perfusion parameters were studied to show the impact of their variation on the overall survival (OS). Results: After the removal of missing or suboptimal DCE-US, the Baseline-D8, the Baseline-D21, and the D8-D21 groups included 37, 53, and 33 patients, respectively. A decrease of more than 45% in the area under the perfusion curve (AUC) between baseline and D21 was significantly associated with better OS (p = 0.0114). A decrease of any amount in the AUC between D8 and D21 was also significantly associated with better OS (p = 0.0370). Conclusion: AUC from DCE-US looks to be a promising new biomarker for fast, effective, and convenient immunotherapy response evaluation.
Persistent Large Granular Lymphocyte Clonal Expansions: “The Root of Many Evils”—And of Some Goodness
Authors: Bravo-Pérez Carlos , Carrillo-Tornel Salvador , García-Torralba Esmeralda , Jerez Andrés
Published: March 2022

Large granular lymphocyte leukemia (LGLL) is a chronic disease of either mature phenotype cytotoxic CD3+ T lymphocytes or CD3- NK cells. LGLL diagnosis is hampered by the fact that reactive persistent clonal LGL expansions may fulfill the current criteria for LGLL diagnoses. In addition to the presence of characteristic clinical and hematological signs such as anemia or neutropenia, LGLL/LGL clonal expansions have been associated with an array of conditions/disorders. We review here the presence of these persistent clonal expansions in autoimmune, hematological disorders and solid neoplasms and after hematopoietic stem cell transplantation. These associations are a unique translational research framework to discern whether these persistently expanded LGL clones are causes or consequences of the concomitant clinical settings and, more importantly, when they should be targeted.
Hemodynamic Imaging in Cerebral Diffuse Glioma—Part B: Molecular Correlates, Treatment Effect Monitoring, Prognosis, and Future Directions
Authors: Stumpo Vittorio , Guida Lelio , Bellomo Jacopo , Van Niftrik Christiaan Hendrik Bas , Sebök Martina , Berhouma Moncef , Bink Andrea , Weller Michael , Kulcsar Zsolt , Regli Luca , Fierstra Jorn
Published: March 2022

Gliomas, and glioblastoma in particular, exhibit an extensive intra- and inter-tumoral molecular heterogeneity which represents complex biological features correlating to the efficacy of treatment response and survival. From a neuroimaging point of view, these specific molecular and histopathological features may be used to yield imaging biomarkers as surrogates for distinct tumor genotypes and phenotypes. The development of comprehensive glioma imaging markers has potential for improved glioma characterization that would assist in the clinical work-up of preoperative treatment planning and treatment effect monitoring. In particular, the differentiation of tumor recurrence or true progression from pseudoprogression, pseudoresponse, and radiation-induced necrosis can still not reliably be made through standard neuroimaging only. Given the abundant vascular and hemodynamic alterations present in diffuse glioma, advanced hemodynamic imaging approaches constitute an attractive area of clinical imaging development. In this context, the inclusion of objective measurable glioma imaging features may have the potential to enhance the individualized care of diffuse glioma patients, better informing of standard-of-care treatment efficacy and of novel therapies, such as the immunotherapies that are currently increasingly investigated. In Part B of this two-review series, we assess the available evidence pertaining to hemodynamic imaging for molecular feature prediction, in particular focusing on isocitrate dehydrogenase (IDH) mutation status, MGMT promoter methylation, 1p19q codeletion, and EGFR alterations. The results for the differentiation of tumor progression/recurrence from treatment effects have also been the focus of active research and are presented together with the prognostic correlations identified by advanced hemodynamic imaging studies. Finally, the state-of-the-art concepts and advancements of hemodynamic imaging modalities are reviewed together with the advantages derived from the implementation of radiomics and machine learning analyses pipelines.
Dual Effect of Combined Metformin and 2-Deoxy-D-Glucose Treatment on Mitochondrial Biogenesis and PD-L1 Expression in Triple-Negative Breast Cancer Cells
Authors: Repas Jernej , Zupin Mateja , Vodlan Maja , Veranič Peter , Gole Boris , Potočnik Uroš , Pavlin Mojca
Published: March 2022

Metformin and 2-deoxy-D-glucose (2DG) exhibit multiple metabolic and immunomodulatory anti-cancer effects, such as suppressed proliferation or PD-L1 expression. Their combination or 2DG alone induce triple-negative breast cancer (TNBC) cell detachment, but their effects on mitochondria, crucial for anchorage-independent growth and metastasis formation, have not yet been evaluated. In the present study, we explored the effects of metformin, 2DG and their combination (metformin + 2DG) on TNBC cell mitochondria in vitro. Metformin + 2DG increased mitochondrial mass in TNBC cells. This was associated with an increased size but not number of morphologically normal mitochondria and driven by the induction of mitochondrial biogenesis rather than suppressed mitophagy. 2DG and metformin + 2DG strongly induced the unfolded protein response by inhibiting protein N-glycosylation. Together with adequate energy stress, this was one of the possible triggers of mitochondrial enlargement. Suppressed N-glycosylation by 2DG or metformin + 2DG also caused PD-L1 deglycosylation and reduced surface expression in MDA-MB-231 cells. PD-L1 was increased in low glucose and normalized by both drugs. 2DG and metformin + 2DG reduced PD-1 expression in Jurkat cells beyond the effects on activation, while cytokine secretion was mostly preserved. Despite increasing mitochondrial mass in TNBC cells, metformin and 2DG could therefore potentially be used as an adjunct therapy to improve anti-tumor immunity in TNBC.
Prevalence of Cognitive Impairment before Prostate Cancer Treatment
Authors: Araújo Natália , Costa Adriana , Lopes Catarina , Lopes-Conceição Luisa , Ferreira Augusto , Carneiro Filipa , Oliveira Jorge , Morais Samantha , Pacheco-Figueiredo Luís , Ruano Luis , Tedim Cruz Vítor , Pereira Susana , Lunet Nuno
Published: March 2022

Cognitive impairment is common among patients with different types of cancer, even before cancer treatment, but no data were reported among patients with prostate cancer (PCa), who may be at high risk due to advanced age. This study aims to estimate the prevalence of cognitive impairment before PCa treatment. Between February 2018 and April 2021, the NEON-PC cohort recruited 605 patients with PCa proposed for treatment at the Portuguese Institute of Oncology of Porto. The Montreal Cognitive Assessment (MoCA) was used to assess cognitive performance. Participants with a MoCA < 1.5 standard deviations (SD) of age- and education-specific normative values were considered to have probable cognitive impairment (PCI) and were referred for a comprehensive neuropsychological assessment. Data from the population-based cohort EPIPorto (n = 351 men aged ≥40 years, evaluated in 2013–2015) were used for comparison. The prevalence of PCI was 17.4% in EPIPorto and 14.7% in NEON-PC (age- and education-adjusted odds ratio: 0.82, 95%CI: 0.58,1.18). Neuropsychological assessment was performed in 63 patients with PCa: 54.0% had cognitive impairment. These results suggest that the impact of PCa on cognitive performance could be negligible in the short term, contrary to what other studies have reported regarding other types of cancer.
Utilization of CO2-Available Organocatalysts for Reactions with Industrially Important Epoxides
Authors: Weidlich Tomáš , Kamenická Barbora
Published: March 2022

Recent knowledge in chemistry has enabled the material utilization of greenhouse gas (CO2) for the production of organic carbonates using mild reaction conditions. Organic carbonates, especially cyclic carbonates, are applicable as green solvents, electrolytes in batteries, feedstock for fine chemicals and monomers for polycarbonate production. This review summarizes new developments in the ring opening of epoxides with subsequent CO2-based formation of cyclic carbonates. The review highlights recent and major developments for sustainable CO2 conversion from 2000 to the end of 2021 abstracted by Web of Science. The syntheses of epoxides, especially from bio-based raw materials, will be summarized, such as the types of raw material (vegetable oils or their esters) and the reaction conditions. The aim of this review is also to summarize and to compare the types of homogeneous non-metallic catalysts. The three reaction mechanisms for cyclic carbonate formation are presented, namely activation of the epoxide ring, CO2 activation and dual activation. Usually most effective catalysts described in the literature consist of powerful sources of nucleophile such as onium salt, of hydrogen bond donors and of tertiary amines used to combine epoxide activation for facile epoxide ring opening and CO2 activation for the subsequent smooth addition reaction and ring closure. The most active catalytic systems are capable of activating even internal epoxides such as epoxidized unsaturated fatty acid derivatives for the cycloaddition of CO2 under relatively mild conditions. In case of terminal epoxides such as epichlorohydrin, the effective utilization of diluted sources of CO2 such as flue gas is possible using the most active organocatalysts even at ambient pressure.
Retinoic Acid Receptor Alpha Is Essential in Postnatal Sertoli Cells but Not in Germ Cells
Authors: Condrea Diana , Souali-Crespo Sirine , Féret Betty , Klopfenstein Muriel , Faisan Sylvain , Mark Manuel , Ghyselinck Norbert B. , Vernet Nadège
Published: March 2022

Retinoic acid signaling is indispensable for the completion of spermatogenesis. It is known that loss of retinoic acid nuclear receptor alpha (RARA) induces male sterility due to seminiferous epithelium degeneration. Initial genetic studies established that RARA acts in Sertoli cells, but a recent paper proposed that RARA is also instrumental in germ cells. In the present study, we have re-assessed the function of RARA in germ cells by genetically ablating the Rara gene in spermatogonia and their progenies using a cell-specific conditional mutagenesis approach. We show that loss of Rara in postnatal male germ cells does not alter the histology of the seminiferous epithelium. Furthermore, RARA-deficient germ cells differentiate normally and give rise to normal, living pups. This establishes that RARA plays no crucial role in germ cells. We also tested whether RARA is required in Sertoli cells during the fetal period or after birth. For this purpose, we deleted the Rara gene in Sertoli cells at postnatal day 15 (PN15), i.e., after the onset of the first spermatogenic wave. To do so, we used temporally controlled cell-specific mutagenesis. By comparing the testis phenotypes generated when Rara is lost either at PN15 or at embryonic day 13, we show that RARA exerts all of its functions in Sertoli cells not at the fetal stage but from puberty.
Transcriptomics Reveals Discordant Lipid Metabolism Effects between In Vitro Models Exposed to Elafibranor and Liver Samples of NAFLD Patients after Bariatric Surgery
Authors: Boeckmans Joost , Gatzios Alexandra , Heymans Anja , Rombaut Matthias , Rogiers Vera , De Kock Joery , Vanhaecke Tamara , Rodrigues Robim M.
Published: March 2022

Background and aims: Non-alcoholic steatohepatitis (NASH) is a life-threatening stage of non-alcoholic fatty liver disease (NAFLD) for which no drugs have been approved. We have previously shown that human-derived hepatic in vitro models can be used to mimic key cellular mechanisms involved in the progression of NASH. In the present study, we first characterize the transcriptome of multiple in vitro NASH models. Subsequently, we investigate how elafibranor, which is a peroxisome proliferator-activated receptor (PPAR)-α/δ agonist that has recently failed a phase 3 clinical trial as a potential anti-NASH compound, modulates the transcriptome of these models. Finally, we compare the elafibranor-induced gene expression modulation to transcriptome data of patients with improved/resolved NAFLD/NASH upon bariatric surgery, which is the only proven clinical NASH therapy. Methods: Human whole genome microarrays were used for the transcriptomics evaluation of hepatic in vitro models. Comparison to publicly available clinical datasets was conducted using multiple bioinformatic application tools. Results: Primary human hepatocytes (PHH), HepaRG, and human skin stem cell-derived hepatic progenitors (hSKP-HPC) exposed to NASH-inducing triggers exhibit up to 35% overlap with datasets of liver samples from NASH patients. Exposure of the in vitro NASH models to elafibranor partially reversed the transcriptional modulations, predicting an inhibition of toll-like receptor (TLR)-2/4/9-mediated inflammatory responses, NFκB-signaling, hepatic fibrosis, and leukocyte migration. These transcriptomic changes were also observed in the datasets of liver samples of patients with resolved NASH. Peroxisome Proliferator Activated Receptor Alpha (PPARA), PPARG Coactivator 1 Alpha (PPARGC1A), and Sirtuin 1 (SIRT1) were identified as the major common upstream regulators upon exposure to elafibranor. Analysis of the downstream mechanistic networks further revealed that angiopoietin Like 4 (ANGPTL4), pyruvate dehydrogenase kinase 4 (PDK4), and perilipin 2 (PLIN2), which are involved in the promotion of hepatic lipid accumulation, were also commonly upregulated by elafibranor in all in vitro NASH models. Contrarily, these genes were not upregulated in liver samples of patients with resolved NASH. Conclusion: Transcriptomics comparison between in vitro NASH models exposed to elafibranor and clinical datasets of NAFLD patients after bariatric surgery reveals commonly modulated anti-inflammatory responses, but discordant modulations of key factors in lipid metabolism. This discordant adverse effect of elafibranor deserves further investigation when assessing PPAR-α/δ agonism as a potential anti-NASH therapy.
Isolation and Crystallization of the D156C Form of Optogenetic ChR2
Authors: Zhang Liying , Wang Kaituo , Ning Shuo , Pedersen Per Amstrup , Duelli Annette Susanne , Gourdon Pontus Emanuel
Published: March 2022

Channelrhodopsins (ChRs) are light-gated ion channels that are receiving increasing attention as optogenetic tools. Despite extensive efforts to gain understanding of how these channels function, the molecular events linking light absorption of the retinal cofactor to channel opening remain elusive. While dark-state structures of ChR2 or chimeric proteins have demonstrated the architecture of non-conducting states, there is a need for open- and desensitized-state structures to uncover the mechanistic principles underlying channel activity. To facilitate comprehensive structural studies of ChR2 in non-closed states, we report a production and purification procedure of the D156C form of ChR2, which displays prolonged channel opening compared to the wild type. We demonstrate considerable yields (0.45 mg/g fermenter cell culture) of recombinantly produced protein using S. cerevisiae, which is purified to high homogeneity both as opsin (retinal-free) and as functional ChR2 with added retinal. We also develop conditions that enable the growth of ChR2 crystals that scatter X-rays to 6 Å, and identify a molecular replacement solution that suggests that the packing is different from published structures. Consequently, our cost-effective production and purification pipeline opens the way for downstream structural studies of different ChR2 states, which may provide a foundation for further adaptation of this protein for optogenetic applications.
Redox Implications of Extreme Task Performance: The Case in Driver Athletes
Authors: Reid Michael B.
Published: March 2022

Redox homeostasis and redox-mediated signaling mechanisms are fundamental elements of human biology. Physiological levels of reactive oxygen species (ROS) and reactive nitrogen species (RNS) modulate a range of functional processes at the cellular, tissue, and systemic levels in healthy humans. Conversely, excess ROS or RNS activity can disrupt function, impairing the performance of daily activities. This article analyzes the impact of redox mechanisms on extreme task performance. Such activities (a) require complex motor skills, (b) are physically demanding, (c) are performed in an extreme environment, (d) require high-level executive function, and (e) pose an imminent risk of injury or death. The current analysis utilizes race car driving as a representative example. The physiological challenges of this extreme task include physical exertion, g loading, vibration, heat exposure, dehydration, noise, mental demands, and emotional factors. Each of these challenges stimulates ROS signaling, RNS signaling, or both, alters redox homeostasis, and exerts pro-oxidant effects at either the tissue or systemic levels. These redox mechanisms appear to promote physiological stress during race car driving and impair the performance of driver athletes.
Disruption of Claudin-Made Tight Junction Barriers by Clostridium perfringens Enterotoxin: Insights from Structural Biology
Authors: Ogbu Chinemerem P. , Roy Sourav , Vecchio Alex J.
Published: March 2022

Claudins are a family of integral membrane proteins that enable epithelial cell/cell interactions by localizing to and driving the formation of tight junctions. Via claudin self-assembly within the membranes of adjoining cells, their extracellular domains interact, forming barriers to the paracellular transport of small molecules and ions. The bacterium Clostridium perfringens causes prevalent gastrointestinal disorders in mammals by employing an enterotoxin (CpE) that targets claudins. CpE binds to claudins at or near tight junctions in the gut and disrupts their barrier function, potentially by disabling their assembly or via cell signaling means—the mechanism(s) remain unclear. CpE ultimately destroys claudin-expressing cells through the formation of a cytotoxic membrane-penetrating β-barrel pore. Structures obtained by X-ray crystallography of CpE, claudins, and claudins in complex with CpE fragments have provided the structural bases of claudin and CpE functions, revealing potential mechanisms for the CpE-mediated disruption of claudin-made tight junctions. This review highlights current progress in this space—what has been discovered and what remains unknown—toward efforts to elucidate the molecular mechanism of CpE disruption of tight junction barriers. It further underscores the key insights obtained through structure that are being applied to develop CpE-based therapeutics that combat claudin-overexpressing cancers or modulate tight junction barriers.
Influence of Mo and Fe on Photosynthetic and Nitrogenase Activities of Nitrogen-Fixing Cyanobacteria under Nitrogen Starvation
Authors: Sadvakasova Asemgul K. , Kossalbayev Bekzhan D. , Token Aziza I. , Bauenova Meruert O. , Wang Jingjing , Zayadan Bolatkhan K. , Balouch Huma , Alwasel Saleh , Leong Yoong Kit , Chang Jo-Shu , Allakhverdiev Suleyman I.
Published: March 2022

The potential of cyanobacteria to perform a variety of distinct roles vital for the biosphere, including nutrient cycling and environmental detoxification, drives interest in studying their biodiversity. Increasing soil erosion and the overuse of chemical fertilizers are global problems in developed countries. The option might be to switch to organic farming, which entails largely the use of biofertilisers. Cyanobacteria are prokaryotic, photosynthetic organisms with considerable potential, within agrobiotechnology, to produce biofertilisers. They contribute significantly to plant drought resistance and nitrogen enrichment in the soil. This study sought, isolated, and investigated nitrogen-fixing cyanobacterial strains in rice fields, and evaluated the effect of Mo and Fe on photosynthetic and nitrogenase activities under nitrogen starvation. Cyanobacterial isolates, isolated from rice paddies in Kazakhstan, were identified as Trichormus variabilis K-31 (MZ079356), Cylindrospermum badium J-8 (MZ079357), Nostoc sp. J-14 (MZ079360), Oscillatoria brevis SH-12 (MZ090011), and Tolypothrix tenuis J-1 (MZ079361). The study of the influence of various concentrations of Mo and Fe on photosynthetic and nitrogenase activities under conditions of nitrogen starvation revealed the optimal concentrations of metals that have a stimulating effect on the studied parameters.