bluuebunny/crossref_metadata_2025 huggingface dataset:
Is Skull-Vibration-Induced Nystagmus Modified with Aging?
Authors: Neri Giampiero , Neri Letizia , Xhepa Klajdi , Mazzatenta Andrea
Published: March 2022

Background: Despite clinical practice utilizing the Dumas test (SVINT), some questions remain unanswered, including the age-related changes in frequency (FN) and slow-phase angular velocity (SPAV). This study aims to retrospectively evaluate their variations in subjects affected by unilateral peripheral vestibular loss (UPVL). Methods: We evaluated the selected samples based on the results of the SVINT, the results of the vestibular-evoked potentials (C-VEMP and O-VEMP), and the results of the head impulse test (HIT) and we compared the results against the age of the patients. We calculated the timing between the onset of UPVL and clinical evaluation in days. The presence or absence of VEMP indicated the UPVL severity. UPVL and BPPV patients with spontaneous or pseudo-spontaneous nystagmus were compared. Results: Statistical analysis showed changes in the FN and SPAV depending on age and the side of the application of the stimulus. We also observed that, in the UPVL, the severity of the disease modifies the SPAV, but not the frequency. Conclusions: The SVINT is a simple, reliable, and straightforward test that, if evaluated instrumentally, can show significant differences with aging. Further studies need to be performed to refine the clinical significance of the test and clarify its physiological background.
Dynamical Analysis of a Predator-Prey Model Incorporating Predator Cannibalism and Refuge
Authors: Rayungsari Maya , Suryanto Agus , Kusumawinahyu Wuryansari Muharini , Darti Isnani
Published: March 2022

We consider a mathematical model to describe the interaction between predator and prey that includes predator cannibalism and refuge. We aim to study the dynamics and its long-term behavior of the proposed model, as well as to discuss the effects of crucial parameters associated with the model. We first show the boundedness and positivity of the solution of the model. Then, we study the existence and stability of all possible equilibrium points. The local stability of the model around each equilibrium point is studied via the linearized system, while the global stability is performed by defining a Lyapunov function. The model has four equilibrium points. It is found that the equilibrium point representing the extinction of both prey and predator populations is always unstable, while the other equilibrium points are conditionally stable. In addition, there is forward bifurcation phenomena that occur under certain condition. To support our analytical findings, we perform some numerical simulations.
A Multisensory Network Drives Nuclear Mechanoadaptation
Authors: Echarri Asier
Published: March 2022

Cells have adapted to mechanical forces early in evolution and have developed multiple mechanisms ensuring sensing of, and adaptation to, the diversity of forces operating outside and within organisms. The nucleus must necessarily adapt to all types of mechanical signals, as its functions are essential for virtually all cell processes, many of which are tuned by mechanical cues. To sense forces, the nucleus is physically connected with the cytoskeleton, which senses and transmits forces generated outside and inside the cell. The nuclear LINC complex bridges the cytoskeleton and the nuclear lamina to transmit mechanical information up to the chromatin. This system creates a force-sensing macromolecular complex that, however, is not sufficient to regulate all nuclear mechanoadaptation processes. Within the nucleus, additional mechanosensitive structures, including the nuclear envelope and the nuclear pore complex, function to regulate nuclear mechanoadaptation. Similarly, extra nuclear mechanosensitive systems based on plasma membrane dynamics, mechanotransduce information to the nucleus. Thus, the nucleus has the intrinsic structural components needed to receive and interpret mechanical inputs, but also rely on extra nuclear mechano-sensors that activate nuclear regulators in response to force. Thus, a network of mechanosensitive cell structures ensures that the nucleus has a tunable response to mechanical cues.
A Series of PSMA-Targeted Near-Infrared Fluorescent Imaging Agents
Authors: Chen Ying , Minn Il , Rowe Steven P. , Lisok Alla , Chatterjee Samit , Brummet Mary , Banerjee Sangeeta Ray , Mease Ronnie C. , Pomper Martin G.
Published: March 2022

We have synthesized a series of 10 new, PSMA-targeted, near-infrared imaging agents intended for use in vivo for fluorescence-guided surgery (FGS). Compounds were synthesized from the commercially available amine-reactive active NHS ester of DyLight800. We altered the linker between the PSMA-targeting urea moiety and the fluorophore with a view to improve the pharmacokinetics. Chemical yields for the conjugates ranged from 51% to 86%. The Ki values ranged from 0.10 to 2.19 nM. Inclusion of an N-bromobenzyl substituent at the ε-amino group of lysine enhanced PSMA+ PIP tumor uptake, as did hydrophilic substituents within the linker. The presence of a polyethylene glycol chain within the linker markedly decreased renal uptake. In particular, DyLight800-10 demonstrated high specific uptake relative to background signal within kidney, confirmed by immunohistochemistry. These compounds may be useful for FGS in prostate, renal or other PSMA-expressing cancers.
Mechanistic Investigation of GHS-R Mediated Glucose-Stimulated Insulin Secretion in Pancreatic Islets
Authors: Pradhan Geetali , Lee Jong Han , Wu Chia-Shan , Wang Hongying , Lin Ligen , Donti Taraka , Graham Brett H. , Rajan Arun S. , Balasubramanyam Ashok , Samson Susan L. , Guo Shaodong , Sun Yuxiang
Published: March 2022

Ghrelin receptor, a growth hormone secretagogue receptor (GHS-R), is expressed in the pancreas. Emerging evidence indicates that GHS-R is involved in the regulation of glucose-stimulated insulin secretion (GSIS), but the mechanism by which GHS-R regulates GSIS in the pancreas is unclear. In this study, we investigated the role of GHS-R on GSIS in detail using global Ghsr−/− mice (in vivo) and Ghsr-ablated pancreatic islets (ex vivo). GSIS was attenuated in both Ghsr−/− mice and Ghsr-ablated islets, while the islet morphology was similar between WT and Ghsr−/− mice. To elucidate the mechanism underpinning Ghsr-mediated GSIS, we investigated the key steps of the GSIS signaling cascade. The gene expression of glucose transporter 2 (Glut2) and the glucose-metabolic intermediate—glucose-6-phosphate (G6P) were reduced in Ghsr-ablated islets, supporting decreased glucose uptake. There was no difference in mitochondrial DNA content in the islets of WT and Ghsr−/− mice, but the ATP/ADP ratio in Ghsr−/− islets was significantly lower than that of WT islets. Moreover, the expression of pancreatic and duodenal homeobox 1 (Pdx1), as well as insulin signaling genes of insulin receptor (IR) and insulin receptor substrates 1 and 2 (IRS1/IRS2), was downregulated in Ghsr−/− islets. Akt is the key mediator of the insulin signaling cascade. Concurrently, Akt phosphorylation was reduced in the pancreas of Ghsr−/− mice under both insulin-stimulated and homeostatic conditions. These findings demonstrate that GHS-R ablation affects key components of the insulin signaling pathway in the pancreas, suggesting the existence of a cross-talk between GHS-R and the insulin signaling pathway in pancreatic islets, and GHS-R likely regulates GSIS via the Akt-Pdx1-GLUT2 pathway.
Emerging Roles for G Protein-Coupled Estrogen Receptor 1 in Cardio-Renal Health: Implications for Aging
Authors: Singh Ravneet , Nasci Victoria L. , Guthrie Ginger , Ertuglu Lale A. , Butt Maryam K. , Kirabo Annet , Gohar Eman Y.
Published: March 2022

Cardiovascular (CV) and renal diseases are increasingly prevalent in the United States and globally. CV-related mortality is the leading cause of death in the United States, while renal-related mortality is the 8th. Despite advanced therapeutics, both diseases persist, warranting continued exploration of disease mechanisms to develop novel therapeutics and advance clinical outcomes for cardio-renal health. CV and renal diseases increase with age, and there are sex differences evident in both the prevalence and progression of CV and renal disease. These age and sex differences seen in cardio-renal health implicate sex hormones as potentially important regulators to be studied. One such regulator is G protein-coupled estrogen receptor 1 (GPER1). GPER1 has been implicated in estrogen signaling and is expressed in a variety of tissues including the heart, vasculature, and kidney. GPER1 has been shown to be protective against CV and renal diseases in different experimental animal models. GPER1 actions involve multiple signaling pathways: interaction with aldosterone and endothelin-1 signaling, stimulation of the release of nitric oxide, and reduction in oxidative stress, inflammation, and immune infiltration. This review will discuss the current literature regarding GPER1 and cardio-renal health, particularly in the context of aging. Improving our understanding of GPER1-evoked mechanisms may reveal novel therapeutics aimed at improving cardio-renal health and clinical outcomes in the elderly.
Recent Progress on Micro-Fabricated Alkali Metal Vapor Cells
Authors: Wang Xuelei , Ye Mao , Lu Fei , Mao Yunkai , Tian Hao , Li Jianli
Published: March 2022

Alkali vapor cells are the core components of atomic sensing instruments such as atomic gyroscopes, atomic magnetometers, atomic clocks, etc. Emerging integrated atomic sensing devices require high-performance miniaturized alkali vapor cells, especially micro-fabricated vapor cells. In this review, bonding methods for vapor cells of this kind are summarized in detail, including anodic bonding, sacrificial micro-channel bonding, and metal thermocompression bonding. Compared with traditional through-lighting schemes, researchers have developed novel methods for micro-fabricated vapor cells under both single- and double-beam schemes. In addition, emerging packaging methods for alkali metals in micro-fabricated vapor cells can be categorized as physical or chemical approaches. Physical methods include liquid transfer and wax pack filling. Chemical methods include the reaction of barium azide with rubidium chloride, ultraviolet light decomposition (of rubidium azide), and the high-temperature electrolysis of rubidium-rich glass. Finally, the application trend of micro-fabricated alkali vapor cells in the field of micro-scale gyroscopes, micro-scale atomic clocks, and especially micro-scale biomagnetometers is reviewed. Currently, the sensing industry has become a major driving force for the miniaturization of atomic sensing devices, and in the near future, the micro-fabricated alkali vapor cell technology of atomic sensing devices may experience extensive developments.
Challenges for the Implementation of BIM Methodology in the Execution of Underground Works
Authors: Baraibar José-Manuel , de-Paz Jesús , Rico Jokin
Published: March 2022

After a few years of the coexistence of the building information modelling (BIM) methodology with the architecture, engineering, and construction professions, its main uses are often limited to 3D modelling and collision checking between different disciplines. However, while this way of working demonstrates opportunities for optimization and clear benefits, there is still much potential for the BIM methodology to be explored. In the scope of a particular underground work, the Arnotegi tunnels of the Bilbao Metropolitan Southern Bypass, a specific contractual framework favouring the collaboration among stakeholders has been defined to implement the use of this methodology by the main participants in the project, encouraging more advanced uses, such as the use of the model as an integrator of the information contained in the common data environment. Due to the very essence of tunnel construction and the relative geotechnical uncertainty of the terrain, the tunnel model evolves day by day during the course of the work, with information being shared in real time between all those involved. This approach has made it possible to improve the quality of decisions and the perception of important information by presenting it in a transparent and easily interpretable way.
Integrated EDM and 4D BIM-Based Decision Support System for Construction Projects Control
Authors: Ayman Hassan Mohamed , Mahfouz Sameh Youssef , Alhady Ahmed
Published: March 2022

Project schedule monitoring and controlling are critical challenges of construction project management that are not adequately implemented, likely due to the predominance of earned value management and the lack of utilizing technology, such as BIM tools. Unlike earned value, earned duration management (EDM) was developed, which includes several indices to track schedule progress and measure the performance of a schedule. The goal of this research was to establish a decision support system to track and monitor construction project activities during construction, with better performance and accuracy. A survey was conducted and distributed among ten site engineers, selected from different construction sites. The survey asked the site engineers about the possible durations of certain activities; based on their answers, the authors started the proposed system. In this study, we aimed to develop a decision support system (DSS), which combines BIM with EDM to help calculate probabilistic total project duration, visually detecting critical activities, monitoring visually risky activities subjected to delay and visually categorizing the accuracy of estimated duration for delayed activities.
Extracorporeal Removal of Thermosensitive Liposomal Doxorubicin from Systemic Circulation after Tumor Delivery to Reduce Toxicities
Authors: Motamarry Anjan , Wolfe A. Marissa , Ramajayam Krishna K. , Pattanaik Sanket , Benton Thomas , Peterson Yuri , Faridi Pegah , Prakash Punit , Twombley Katherine , Haemmerich Dieter
Published: March 2022

Thermosensitive liposomal doxorubicin (TSL-Dox) combined with localized hyperthermia enables targeted drug delivery. Tumor drug uptake occurs only during hyperthermia. We developed a novel method for removal of systemic TSL-Dox remaining after hyperthermia-triggered delivery to reduce toxicities. The carotid artery and jugular vein of Norway brown rats carrying two subcutaneous BN-175 tumors were catheterized. After allowing the animals to recover, TSL-Dox was infused at 7 mg/kg dose. Drug delivery to one of the tumors was performed by inducing 15 min microwave hyperthermia (43 °C). At the end of hyperthermia, an extracorporeal circuit (ECC) comprising a heating module to release drug from TSL-Dox followed by an activated carbon filter to remove free drug was established for 1 h (n = 3). A computational model simulated TSL-Dox pharmacokinetics, including ECC filtration, and predicted cardiac Dox uptake. In animals receiving ECC, we were able to remove 576 ± 65 mg of Dox (29.7 ± 3.7% of the infused dose) within 1 h, with a 2.9-fold reduction of plasma AUC. Fluorescent monitoring enabled real-time quantification of blood concentration and removed drug. Computational modeling predicted that up to 59% of drug could be removed with an ideal filter, and that cardiac uptake can be reduced up to 7×. We demonstrated removal of drug remaining after tumor delivery, reduced plasma AUC, and reduced cardiac uptake, suggesting reduced toxicity.
Quantification of Tumor Hypoxia through Unsupervised Modelling of Consumption and Supply Hypoxia MR Imaging in Breast Cancer
Authors: Mo Torgeir , Brandal Siri Helene Bertelsen , Köhn-Luque Alvaro , Engebraaten Olav , Kristensen Vessela N. , Fleischer Thomas , Hompland Tord , Seierstad Therese
Published: March 2022

The purpose of the present study is to investigate if consumption and supply hypoxia (CSH) MR-imaging can depict breast cancer hypoxia, using the CSH-method initially developed for prostate cancer. Furthermore, to develop a generalized pan-cancer application of the CSH-method that doesn’t require a hypoxia reference standard for training the CSH-parameters. In a cohort of 69 breast cancer patients, we generated, based on the principles of intravoxel incoherent motion modelling, images reflecting cellular density (apparent diffusion coefficient; ADC) and vascular density (perfusion fraction; fp). Combinations of the information in these images were compared to a molecular hypoxia score made from gene expression data, aiming to identify a way to apply the CSH-methodology in breast cancer. Attempts to adapt previously proposed models for prostate cancer included direct transfers and model parameter rescaling. A novel approach, based on rescaling ADC and fp data to give more nuanced response in the relevant physiologic range, was also introduced. The new CSH-method was validated in a prostate cancer cohort with known hypoxia status. The proposed CSH-method gave estimates of hypoxia that was strongly correlated to the molecular hypoxia score in breast cancer, and hypoxia as measured in pathology slices stained with pimonidazole in prostate cancer. The generalized approach to CSH-imaging depicted hypoxia in both breast and prostate cancers and requires no model training. It is easy to implement using readily available technology and encourages further investigation of CSH-imaging in other cancer entities and in other settings, with the goal being to overcome hypoxia-induced resistance to treatment.
Impact of Three-Month Androgen Deprivation Therapy on [68Ga]Ga-PSMA-11 PET/CT Indices in Men with Advanced Prostate Cancer—Results from a Pilot Prospective Study
Authors: Tseng Jing-Ren , Chang Szu-Han , Wu Yao-Yu , Fan Kang-Hsing , Yu Kai-Jie , Yang Lan-Yan , Hsiao Ing-Tsung , Liu Feng-Yuan , Pang See-Tong
Published: March 2022

Purpose: The purpose of this pilot prospective study is to examine the gallium-68-prostate-specific membrane antigen-11 ([68Ga]Ga-PSMA-11) positron emission tomography/computed tomography (PET/CT) imaging response in patients with advanced or metastatic hormone-naïve prostate cancer (PC) after 3 months of androgen deprivation therapy (ADT). Methods: We prospectively included men with untreated, clinical stage III or IV PC scheduled to receive ADT for at least 6 months. [68Ga]Ga-PSMA-11 PET/CT images were obtained before the start of ADT and 10–14 weeks thereafter. The following indices were examined: maximum standardized uptake value (SUVmax), mean SUV, PSMA total volume, and PSMA total lesion values of the prostate, nodes, bones, and whole-body. The therapeutic response was assessed using the modified PET response criteria in solid tumors 1.0. A subgroup analysis of patients with the International Society of Urological Pathology (ISUP) grade group 5 versus <5 was also performed. Results: A total of 30 patients were eligible. All PSMA PET/CT indices were significantly reduced (p < 0.001) after 3 months of ADT. Twenty-four (80%) patients showed partial response. Complete response, stable disease, and disease progression were observed in two patients each. Sixteen patients with ISUP grade group 5 showed a less prominent SUVmax reduction (p = 0.006), and none of them reached complete response. Conclusions: Three months of ADT in patients with untreated, advanced PC significantly reduced PSMA PET/CT indices. While most participants partially responded to ADT, patients with ISUP grade group 5 showed a less prominent SUVmax reduction. Collectively, our pilot results indicate that [68Ga]Ga-PSMA-11 PET/CT imaging holds promise to monitor treatment response after the first three months of ADT.
The B-Cell-Specific Ablation of B4GALT1 Reduces Cancer Formation and Reverses the Changes in Serum IgG Glycans during the Induction of Mouse Hepatocellular Carcinoma
Authors: Sha Jichen , Zhang Rongrong , Fan Jiteng , Gu Yong , Pan Yiqing , Han Jing , Xu Xiaoyan , Ren Shifang , Gu Jianxin
Published: March 2022

Serum immunoglobulin G (IgG) glycosylation, especially galactosylation, has been found to be related to a variety of tumors, including hepatocellular carcinoma (HCC). However, whether IgG glycan changes occur in the early stages of HCC formation remains unclear. We found that the galactosylation level increased and that the related individual glycans showed regular changes over the course of HCC induction. Then, the effect of the B-cell-specific ablation of β1,4galactosyltransferase 1 (CKO B4GALT1) and B4GALT1 defects on the IgG glycans that were modified during the model induction process and HCC formation is investigated in this study. CKO B4GALT1 reduces serum IgG galactosylation levels and reduces cancer formation. Furthermore, insignificant changes in the B-cell B4GALT1 and unchanged serum IgG galactosylation levels were found during cancer induction in female mice, which might contribute to the lower cancer incidence in female mice than in male mice. The gender differences observed during glycan and B4GALT1 modification also add more evidence that the B4GALT1 in B cells and in serum IgG galactosylation may play an important role in HCC. Therefore, the findings of the present research can be used to determine the methods for the early detection of HCC as well as for prevention.
Fluorescence Polarization Imaging of Methylene Blue Facilitates Quantitative Detection of Thyroid Cancer in Single Cells
Authors: Jermain Peter R. , Fischer Andrew H. , Joseph Lija , Muzikansky Alona , Yaroslavsky Anna N.
Published: March 2022

Background: Diagnostic accuracy of the standard of care fine-needle aspiration cytology (FNAC) remains a significant problem in thyroid oncology. Therefore, a robust and accurate method for reducing uncertainty of cytopathological evaluation would be invaluable. Methods: In this double-blind study, we employed fluorescence emission and quantitative fluorescence polarization (Fpol) confocal imaging for sorting thyroid cells into benign/malignant categories. Samples were collected from malignant tumors, benign nodules, and normal thyroid epithelial tissues. Results: A total of 32 samples, including 12 from cytologically indeterminate categories, were stained using aqueous methylene blue (MB) solution, imaged, and analyzed. Fluorescence emission images yielded diagnostically relevant information on cytomorphology. Significantly higher MB Fpol was measured in thyroid cancer as compared to benign and normal cells. The results obtained from 12 indeterminate samples revealed that MB Fpol accurately differentiated benign and malignant thyroid nodules. Conclusions: The developed imaging approach holds the potential to provide an accurate and objective biomarker for thyroid cancer, improve diagnostic accuracy of cytopathology, and decrease the number of lobectomy and near-total thyroidectomy procedures.
Reproducibility of Gene Expression Signatures in Diffuse Large B-Cell Lymphoma
Authors: Plaça Jessica Rodrigues , Diepstra Arjan , Los Tjitske , Mendeville Matías , Seitz Annika , Lugtenburg Pieternella J. , Zijlstra Josée , Lam King , da Silva Wilson Araújo , Ylstra Bauke , de Jong Daphne , van den Berg Anke , Nijland Marcel
Published: March 2022

Multiple gene expression profiles have been identified in diffuse large B-cell lymphoma (DLBCL). Besides the cell of origin (COO) classifier, no signatures have been reproduced in independent studies or evaluated for capturing distinct aspects of DLBCL biology. We reproduced 4 signatures in 175 samples of the HOVON-84 trial on a panel of 117 genes using the NanoString platform. The four gene signatures capture the COO, MYC activity, B-cell receptor signaling, oxidative phosphorylation, and immune response. Performance of our classification algorithms were confirmed in the original datasets. We were able to validate three of the four GEP signatures. The COO algorithm resulted in 94 (54%) germinal center B-cell (GCB) type, 58 (33%) activated B-cell (ABC) type, and 23 (13%) unclassified cases. The MYC-classifier revealed 77 cases with a high MYC-activity score (44%) and this MYC-high signature was observed more frequently in ABC as compared to GCB DLBCL (68% vs. 32%, p < 0.00001). The host response (HR) signature of the consensus clustering was present in 55 (31%) patients, while the B-cell receptor signaling, and oxidative phosphorylation clusters could not be reproduced. The overlap of COO, consensus cluster and MYC activity score differentiated six gene expression clusters: GCB/MYC-high (12%), GCB/HR (16%), GCB/non-HR (27%), COO-Unclassified (13%), ABC/MYC-high (25%), and ABC/MYC-low (7%). In conclusion, the three validated signatures identify distinct subgroups based on different aspects of DLBCL biology, emphasizing that each classifier captures distinct molecular profiles.
Anticancer Activity of (S)-5-Chloro-3-((3,5-dimethylphenyl)sulfonyl)-N-(1-oxo-1-((pyridin-4-ylmethyl)amino)propan-2-yl)-1H-indole-2-carboxamide (RS4690), a New Dishevelled 1 Inhibitor
Authors: Coluccia Antonio , Bufano Marianna , La Regina Giuseppe , Puxeddu Michela , Toto Angelo , Paone Alessio , Bouzidi Amani , Musto Giorgia , Badolati Nadia , Orlando Viviana , Biagioni Stefano , Masci Domiziana , Cantatore Chiara , Cirilli Roberto , Cutruzzolà Francesca , Gianni Stefano , Stornaiuolo Mariano , Silvestri Romano
Published: March 2022

Wingless/integrase-11 (WNT)/β-catenin pathway is a crucial upstream regulator of a huge array of cellular functions. Its dysregulation is correlated to neoplastic cellular transition and cancer proliferation. Members of the Dishevelled (DVL) family of proteins play an important role in the transduction of WNT signaling by contacting its cognate receptor, Frizzled, via a shared PDZ domain. Thus, negative modulators of DVL1 are able to impair the binding to Frizzled receptors, turning off the aberrant activation of the WNT pathway and leading to anti-cancer activity. Through structure-based virtual screening studies, we identified racemic compound RS4690 (1), which showed a promising selective DVL1 binding inhibition with an EC50 of 0.74 ± 0.08 μM. Molecular dynamic simulations suggested a different binding mode for the enantiomers. In the in vitro assays, enantiomer (S)-1 showed better inhibition of DVL1 with an EC50 of 0.49 ± 0.11 μM compared to the (R)-enantiomer. Compound (S)-1 inhibited the growth of HCT116 cells expressing wild-type APC with an EC50 of 7.1 ± 0.6 μM and caused a high level of ROS production. These results highlight (S)-1 as a lead compound for the development of new therapeutic agents against WNT-dependent colon cancer.
Effect of Binding Modules Fused to Cutinase on the Enzymatic Synthesis of Polyesters
Authors: Ferrario Valerio , Todea Anamaria , Wolansky Lisa , Piovesan Nicola , Guarneri Alice , Ribitsch Doris , Guebitz Georg M. , Gardossi Lucia , Pellis Alessandro
Published: March 2022

In relation to the development of environmentally-friendly processing technologies for the continuously growing market of plastics, enzymes play an important role as green and sustainable biocatalysts. The present study reports the use of heterogeneous immobilized biocatalysts in solvent-free systems for the synthesis of aliphatic oligoesters with Mws and monomer conversions up to 1500 Da and 74%, respectively. To improve the accessibility of hydrophilic and hydrophobic substrates to the surface of the biocatalyst and improve the reaction kinetic and the chain elongation, two different binding modules were fused on the surface of cutinase 1 from Thermobifida cellulosilytica. The fusion enzymes were successfully immobilized (>99% of bound protein) via covalent bonding onto epoxy-activated beads. To the best of our knowledge, this is the first example where fused enzymes are used to catalyze transesterification reactions for polymer synthesis purposes.
A Coculture Model Mimicking the Tumor Microenvironment Unveils Mutual Interactions between Immune Cell Subtypes and the Human Seminoma Cell Line TCam-2
Authors: Gayer Fabian A. , Fichtner Alexander , Legler Tobias J. , Reichardt Holger M.
Published: March 2022

Testicular germ cell cancer (TGCC) is the most common type of cancer in young men. Seminomas account for around half of them and are characterized by a pronounced infiltration of immune cells. So far, the impact of the tumor microenvironment (TME) on disease progression, especially the interaction of individual immune cell subtypes with the tumor cells, remains unclear. To address this question, we used an in vitro TME model involving the seminoma-derived cell line Tcam-2 and immune cell subsets purified from human peripheral blood. T cells and monocytes were strongly activated when individually cocultured with Tcam-2 cells as revealed by increased expression of activation markers and pro-inflammatory cytokines both on the mRNA and protein level. Importantly, the interaction between tumor and immune cells was mutual. Gene expression of pluripotency markers as well as markers of proliferation and cell cycle activity were upregulated in Tcam-2 cells in cocultures with T cells, whereas gene expression of SOX17, a marker for seminomas, was unaltered. Interestingly, the impact of monocytes on gene expression of Tcam-2 cells was less pronounced, indicating that the effects of individual immune cell subsets on tumor cells in the TME are highly specific. Collectively, our data indicate that seminoma cells induce immune cell activation and thereby generate a strong pro-inflammatory milieu, whereas T cells conversely increase the proliferation, metastatic potential, and stemness of tumor cells. Although the employed model does not fully mimic the physiological situation found in TGCC in vivo, it provides new insights potentially explaining the connection between inflammatory infiltrates in seminomas and their tendency to burn out and metastasize.
Plant E3 Ligases and Their Role in Abiotic Stress Response
Authors: Al-Saharin Raed , Hellmann Hanjo , Mooney Sutton
Published: March 2022

Plants, as sessile organisms, have limited means to cope with environmental changes. Consequently, they have developed complex regulatory systems to ameliorate abiotic stresses im-posed by environmental changes. One such system is the ubiquitin proteasome pathway, which utilizes E3 ligases to target proteins for proteolytic degradation via the 26S proteasome. Plants ex-press a plethora of E3 ligases that are categorized into four major groups depending on their structure. They are involved in many biological and developmental processes in plants, such as DNA repair, photomorphogenesis, phytohormones signaling, and biotic stress. Moreover, many E3 ligase targets are proteins involved in abiotic stress responses, such as salt, drought, heat, and cold. In this review, we will provide a comprehensive overview of E3 ligases and their substrates that have been connected with abiotic stress in order to illustrate the diversity and complexity of how this pathway enables plant survival under stress conditions.
Pathological Relevance of Post-Translationally Modified Alpha-Synuclein (pSer87, pSer129, nTyr39) in Idiopathic Parkinson’s Disease and Multiple System Atrophy
Authors: Sonustun Berkiye , Altay Melek Firat , Strand Catherine , Ebanks Kirsten , Hondhamuni Geshanthi , Warner Thomas T. , Lashuel Hilal A. , Bandopadhyay Rina
Published: March 2022

Aggregated alpha-synuclein (α-synuclein) is the main component of Lewy bodies (LBs), Lewy neurites (LNs), and glial cytoplasmic inclusions (GCIs), which are pathological hallmarks of idiopathic Parkinson’s disease (IPD) and multiple system atrophy (MSA). Initiating factors that culminate in forming LBs/LNs/GCIs remain elusive. Several species of α-synuclein exist, including phosphorylated and nitrated forms. It is unclear which α-synuclein post-translational modifications (PTMs) appear within aggregates throughout disease pathology. Herein we aimed to establish the predominant α-synuclein PTMs in postmortem IPD and MSA pathology using immunohistochemistry. We examined the patterns of three α-synuclein PTMs (pS87, pS129, nY39) simultaneously in pathology-affected regions of 15 IPD cases, 5 MSA cases, and 6 neurologically normal controls. All antibodies recognized LBs, LNs, and GCIs, albeit to a variable extent. pS129 α-synuclein antibody was particularly immunopositive for LNs and synaptic dot-like structures, followed by nY39 α-synuclein antibody. GCIs, neuronal inclusions, and small threads were positive for nY39 α-synuclein in MSA. Quantification of the LB scores revealed that pS129 α-synuclein was the dominant and earliest α-synuclein PTM, followed by nY39 α-synuclein, while lower amounts of pSer87 α-synuclein appeared later in disease progression in PD. These results may have implications for novel biomarker and therapeutic developments.
Direct Interaction of Mitochondrial Cytochrome c Oxidase with Thyroid Hormones: Evidence for Two Binding Sites
Authors: Oleynikov Ilya P. , Sudakov Roman V. , Azarkina Natalia V. , Vygodina Tatiana V.
Published: March 2022

Thyroid hormones regulate tissue metabolism to establish an energy balance in the cell, in particular, by affecting oxidative phosphorylation. Their long-term impact is mainly associated with changes in gene expression, while the short-term effects may differ in their mechanisms. Our work was devoted to studying the short-term effects of hormones T2, T3 and T4 on mitochondrial cytochrome c oxidase (CcO) mediated by direct contact with the enzyme. The data obtained indicate the existence of two separate sites of CcO interaction with thyroid hormones, differing in their location, affinity and specificity to hormone binding. First, we show that T3 and T4 but not T2 inhibit the oxidase activity of CcO in solution and on membrane preparations with Ki ≈ 100–200 μM. In solution, T3 and T4 compete in a 1:1 ratio with the detergent dodecyl-maltoside to bind to the enzyme. The peroxidase and catalase partial activities of CcO are not sensitive to hormones, but electron transfer from heme a to the oxidized binuclear center is affected. We believe that T3 and T4 could be ligands of the bile acid-binding site found in the 3D structure of CcO by Ferguson-Miller’s group, and hormone-induced inhibition is associated with dysfunction of the K-proton channel. A possible role of this interaction in the physiological regulation of the enzyme is discussed. Second, we find that T2, T3, and T4 inhibit superoxide generation by oxidized CcO in the presence of excess H2O2. Inhibition is characterized by Ki values of 0.3–5 μM and apparently affects the formation of O2●− at the protein surface. The second binding site for thyroid hormones presumably coincides with the point of tight T2 binding on the Va subunit described in the literature.
Overexpression of HVA1 Enhances Drought and Heat Stress Tolerance in Triticum aestivum Doubled Haploid Plants
Authors: Samtani Harsha , Sharma Aishwarye , Khurana Paramjit
Published: March 2022

Plant responses to multiple environmental stresses include various signaling pathways that allow plant acclimation and survival. Amongst different stresses, drought and heat stress severely affect growth and productivity of wheat. HVA1, a member of the group 3 LEA protein, has been well known to provide protection against drought stress. However, its mechanism of action and its role in other stresses such as heat remain unexplored. In this study, doubled haploid (DH) wheat plants overexpressing the HVA1 gene were analyzed and found to be both drought-and heat stress-tolerant. The transcriptome analysis revealed the upregulation of transcription factors such as DREB and HsfA6 under drought and heat stress, respectively, which contribute toward the tolerance mechanism. Particularly under heat stress conditions, the transgenic plants had a lower oxidative load and showed enhanced yield. The overexpression lines were found to be ABA-sensitive, therefore suggesting the role of HsfA6 in providing heat tolerance via the ABA-mediated pathway. Thus, apart from its known involvement in drought stress, this study highlights the potential role of HVA1 in the heat stress signaling pathway. This can further facilitate the engineering of multiple stress tolerance in crop plants, such as wheat.
Waste Clearance in the Brain and Neuroinflammation: A Novel Perspective on Biomarker and Drug Target Discovery in Alzheimer’s Disease
Authors: Uchida Kazuhiko
Published: March 2022

Alzheimer’s disease (AD) is a multifactorial disease with a heterogeneous etiology. The pathology of Alzheimer’s disease is characterized by amyloid-beta and hyperphosphorylated tau, which are necessary for disease progression. Many clinical trials on disease-modifying drugs for AD have failed to indicate their clinical benefits. Recent advances in fundamental research have indicated that neuroinflammation plays an important pathological role in AD. Damage- and pathogen-associated molecular patterns in the brain induce neuroinflammation and inflammasome activation, causing caspase-1-dependent glial and neuronal cell death. These waste products in the brain are eliminated by the glymphatic system via perivascular spaces, the blood-brain barrier, and the blood–cerebrospinal fluid barrier. Age-related vascular dysfunction is associated with an impairment of clearance and barrier functions, leading to neuroinflammation. The proteins involved in waste clearance in the brain and peripheral circulation may be potential biomarkers and drug targets in the early stages of cognitive impairment. This short review focuses on waste clearance dysfunction in AD pathobiology and discusses the improvement of waste clearance as an early intervention in prodromal AD and preclinical stages of dementia.
Investigations of TiO2/NanoTiO2 Bimodal Coatings Obtained by a Hybrid PVD/ALD Method on Al-Si-Cu Alloy Substrate
Authors: Staszuk Marcin , Reimann Łukasz , Pakuła Daniel , Pawlyta Mirosława , Musztyfaga-Staszuk Małgorzata , Czaja Paweł , Beneš Petr
Published: March 2022

This study aimed to investigate the influence of bimodal TiO2/nanoTiO2 coatings obtained in the PVD/ALD hybrid process on an Al-Si-Cu-type aluminium alloy on the physicochemical properties of the investigated materials. The reference materials were uncoated substrates and samples coated with TiO2 coatings in single PVD and ALD processes. Tests were carried out on the morphology of coatings using scanning electron microscopy (SEM) and atomic force microscopy (AFM) to determine the structure of the tested coatings and their influence on physicochemical properties. The tests of physicochemical properties were carried out using the potentiodynamic and electrochemical impedance spectroscopy methods. The analysis of corrosion products obtained during the study of physicochemical properties was performed using SEM and EDS analysis. Based on the analysis of anode polarisation curves of the tested materials and Tafel analysis, it was found that a sample demonstrated the highest corrosion resistance with a bimodal coating-type TiO2/nanoTiO2, which had the lowest value of corrosion current and the highest value of polarisation resistance. The slightest corrosion resistance was characteristic for uncovered samples. Moreover, the influence of the tested coatings on the improvement of tribological contact of the surfaces of the coated materials with the counter-specimen made of cemented carbides was demonstrated.
Comparison of Aqueous and Gelled 3.5% NaCl Electrolytes for Assessing the Corrosion Resistance of Thermal Spray Stainless-Steel Coatings in Electrochemical Corrosion Tests
Authors: Kutschmann Pia , Grimm Maximilian , Lindner Thomas , Ernst Kerstin Raffaela , Schwabe Olga , Pluta Christian , Lampke Thomas
Published: March 2022

Corrosion testing with gel electrolytes gained attention in the past decade due to the advantage of almost non-destructive and in situ electrochemical measurements of bulk materials. Regarding thermal spray coatings, gel electrolytes offered the opportunity to prevent the infiltration of the typical microstructural features such as pores and microcracks. Using the example of stainless-steel AISI 316L coatings deposited by high velocity air fuel (HVAF) spraying on mild and stainless-steel substrates, the electrochemical corrosion behavior was analyzed in 3.5% NaCl electrolytes in an aqueous and gelled state. In this context, potentiodynamic polarization tests were carried out in a three-electrode corrosion cell, which was adapted for gel electrolyte testing. Gelling was realized with a technical gelatin. The characteristic corrosion values, such as open circuit potential, corrosion potential, and corrosion current density, revealed for the gelled state that the influence of the substrate material used could be eliminated and thus, the coatings itself could be characterized. In contrast, the coating specific microstructure and substrate material significantly affected the potentiodynamic polarization curve in the 3.5% NaCl aqueous electrolyte. Optical microscopy of the coating surfaces and cross-sections proved that the corrosion attack caused by aqueous electrolytes could be mimicked with the gel electrolyte.
NDN Content Store and Caching Policies: Performance Evaluation
Authors: Silva Elídio Tomás da , Macedo Joaquim Melo Henriques de , Costa António Luís Duarte
Published: March 2022

Among various factors contributing to performance of named data networking (NDN), the organization of caching is a key factor and has benefited from intense studies by the networking research community. The performed studies aimed at (1) finding the best strategy to adopt for content caching; (2) specifying the best location, and number of content stores (CS) in the network; and (3) defining the best cache replacement policy. Accessing and comparing the performance of the proposed solutions is as essential as the development of the proposals themselves. The present work aims at evaluating and comparing the behavior of four caching policies (i.e., random, least recently used (LRU), least frequently used (LFU), and first in first out (FIFO)) applied to NDN. Several network scenarios are used for simulation (2 topologies, varying the percentage of nodes of the content stores (5–100), 1 and 10 producers, 32 and 41 consumers). Five metrics are considered for the performance evaluation: cache hit ratio (CHR), network traffic, retrieval delay, interest re-transmissions, and the number of upstream hops. The content request follows the Zipf–Mandelbrot distribution (with skewness factor α=1.1 and α=0.75). LFU presents better performance in all considered metrics, except on the NDN testbed, with 41 consumers, 1 producer and a content request rate of 100 packets/s. For the level of content store from 50% to 100%, LRU presents a notably higher performance. Although the network behavior is similar for both skewness factors, when α=0.75, the CHR is significantly reduced, as expected.
Synthesis, X-ray Structure, Antimicrobial and Anticancer Activity of a Novel [Ag(ethyl-3-quinolate)2(citrate)] Complex
Authors: Altowyan Mezna , El-Naggar Mostafa , Abu-Youssef Morsy , Soliman Saied , Haukka Matti , Barakat Assem , Badr Ahmed
Published: March 2022

A novel Ag(I) citrate complex with ethyl-3-quinolate (Et3qu) was synthesized. Its structure was confirmed using X-ray single crystal to be [Ag(Et3qu)2(citrate)]. It crystallized in the Triclinic crystal system and P-1 space group with unit cell parameters of a = 8.6475(2) Å, b = 11.4426(3) Å, c = 15.2256(3) Å, α = 73.636(2)°, β = 79.692(2)° and γ = 86.832(2)°, while the unit cell volume was 1422.19(6) Å3. In the unit cell, there are two [Ag(Et3qu)2(citrate)] molecules and one unit as the asymmetric formula. The molecular structure comprised one Ag(I) coordinated with two Et3qu molecules via two almost equidistant Ag-N bonds and one citrate ion acting as a mono-negative monodentate ligand via a short Ag-O bond (2.5401(14) Å). Hence, Ag(I) is tri-coordinated and has a highly distorted triangular planar coordination geometry which is more like to be described as a slightly distorted T-shape. The supramolecular structure of the [Ag(Et3qu)2(citrate)] complex was analyzed using Hirshfeld calculations. The H···H (39.3–40.1%), O···H (33.2-34.0%), C···C (9.1–9.5%) and C···H (7.2–7.4%) contacts shared significantly in the packing of the studied Ag(I) complex. The antimicrobial and anticancer activities of the Ag(I) complex were investigated. The [Ag(Et3qu)2(citrate)] complex has broad-spectrum antimicrobial activity specifically against the fungus A. fumigatus. In addition, the IC50 values of 1.87 ± 0.09 µg/mL and 0.95 ± 0.06 µg/mL against the breast MCF-7 and lung A-549 cell lines, respectively, revealed the potent anticancer activity of the [Ag(Et3qu)2(citrate)] complex compared to the free Et3qu (IC50 = 30.64 ± 1.98 and 22.89 ± 1.48 µg/mL, respectively).
Current Advances in the Management of Adult Craniopharyngiomas
Authors: Lara-Velazquez Montserrat , Mehkri Yusuf , Panther Eric , Hernandez Jairo , Rao Dinesh , Fiester Peter , Makary Raafat , Rutenberg Michael , Tavanaiepour Daryoush , Rahmathulla Gazanfar
Published: March 2022

Craniopharyngiomas (CPs) are slow growing, histologically benign intracranial tumors located in the sellar–suprasellar region. Although known to have low mortality, their location and relationship to the adjacent neural structures results in patients having significant neurologic, endocrine, and visual comorbidities. The invasive nature of this tumor makes complete resection a challenge and contributes to its recurrence. Additionally, these tumors are bimodally distributed, being treated with surgery, and are followed by other adjuncts, such as focused radiation therapy, e.g., Gamma knife. Advances in surgical techniques, imaging tools, and instrumentations have resulted in the evolution of surgery using endoscopic techniques, with residual components being treated by radiotherapy to target the residual tumor. Advances in molecular biology have elucidated the main pathways involved in tumor development and recurrence, but presently, no other treatments are offered to patients, besides surgery, radiation, and endocrine management, as the disease and tumor evolve. We review the contemporary management of these tumors, from the evolution of surgical treatments, utilizing standard open microscopic approaches to the more recent endoscopic surgery, and discuss the current recommendations for care of these patients. We discuss the developments in radiation therapy, such as radiosurgery, being used as treatment strategies for craniopharyngioma, highlighting their beneficial effects on tumor resections while decreasing the rates of adverse outcomes. We also outline the recent chemotherapy modalities, which help control tumor growth, and the immune landscape on craniopharyngiomas that allow the development of novel immunotherapies.
Electrochemotherapy Is Effective in the Treatment of Bone Metastases
Authors: Campanacci Laura , Cevolani Luca , De Terlizzi Francesca , Saenz Laura , Alì Nikolin , Bianchi Giuseppe , Donati Davide Maria
Published: March 2022

Bone metastases induce pain, risk of fracture, and neural compression, and reduced mobility and quality of life. Electrochemotherapy (ECT) is a minimally invasive local treatment based on a high-voltage electric pulse combined with an anticancer drug. Preclinical and clinical studies have supported the use of ECT in patients with metastatic bone disease, demonstrating that it does not damage the mineral structure of the bone and its regenerative capacity, and that is feasible and efficient for the treatment of bone metastases. Since 2009, 88 patients with bone metastasis have received ECT at the Rizzoli Institute. 2014 saw the start of a registry of patients with bone metastases treated with ECT, whose data are recorded in a shared database. We share the Rizzoli Institute experience of 38 patients treated with ECT for a bone metastasis, excluding patients not included in the registry (before 2014) and those treated with bone fixation. Mean follow-up was 2 months (1–52). Response to treatment using RECIST criteria was 29% objective responses, 59% stable disease, and 16% progressive disease. Using PERCIST, the response was 36% OR, 14% SD, and 50% PD with no significant differences between the two criteria. A significant decrease in pain and better quality of life was observed at FU.