bluuebunny/crossref_metadata_2025 huggingface dataset:
Sildenafil Reduces Expression and Release of IL-6 and IL-8 Induced by Reactive Oxygen Species in Systemic Sclerosis Fibroblasts
Authors: Di Luigi Luigi , Sgrò Paolo , Duranti Guglielmo , Sabatini Stefania , Caporossi Daniela , Del Galdo Francesco , Dimauro Ivan , Antinozzi Cristina
Published: May 2020

Oxidative stress linked to vascular damage plays an important role in the pathogenesis of systemic sclerosis (SSc). Indeed, vascular damage at nailfold capillaroscopy in patients with Raynaud’s Phenomenon (RP) is a major risk factor for the development of SSc together with the presence of specific autoantiobodies. Here, we investigated the effects of the phosphodiesterase type 5 inhibitor (PDE5i) sildenafil, currently used in the management of RP, in modulating the proinflammatory response of dermal fibroblasts to oxidative stress in vitro. Human fibroblasts isolated from SSc patients and healthy controls were exposed to exogenous reactive oxygen species (ROS) (100 µM H2O2), in the presence or absence of sildenafil (1 µM). Treatment with sildenafil significantly reduced dermal fibroblast gene expression and cellular release of IL-6, known to play a central role in the pathogenesis of tissue damage in SSc and IL-8, directly induced by ROS. This reduction was associated with suppression of STAT3-, ERK-, NF-κB-, and PKB/AKT-dependent pathways. Our findings support the notion that the employment of PDE5i in the management of RP may be explored for its efficacy in modulating the oxidative stress-induced proinflammatory activation of dermal fibroblasts in vivo and may ultimately aid in the prevention of tissue damage caused by SSc.
Investigating Programmed Cell Death and Tumor Invasion in a Three-Dimensional (3D) Microfluidic Model of Glioblastoma
Authors: Samiei Ehsan , Seyfoori Amir , Toyota Brian , Ghavami Saeid , Akbari Mohsen
Published: May 2020

Glioblastoma multiforme (GBM) is a rapidly progressive and deadly form of brain tumor with a median survival rate of ~15 months. GBMs are hard to treat and significantly affect the patient’s physical and cognitive abilities and quality of life. Temozolomide (TMZ)—an alkylating agent that causes DNA damage—is the only chemotherapy choice for the treatment of GBM. However, TMZ also induces autophagy and causes tumor cell resistance and thus fails to improve the survival rate among patients. Here, we studied the drug-induced programmed cell death and invasion inhibition capacity of TMZ and a mevalonate cascade inhibitor, simvastatin (Simva), in a three-dimensional (3D) microfluidic model of GBM. We elucidate the role of autophagy in apoptotic cell death by comparing apoptosis in autophagy knockdown cells (Atg7 KD) against their scrambled counterparts. Our results show that the cells were significantly less sensitive to drugs in the 3D model as compared to monolayer culture systems. An immunofluorescence analysis confirmed that apoptosis is the mechanism of cell death in TMZ- and Simva-treated glioma cells. However, the induction of apoptosis in the 3D model is significantly lower than in monolayer cultures. We have also shown that autophagy inhibition (Atg7 KD) did not change TMZ and Simva-induced apoptosis in the 3D microfluidic model. Overall, for the first time in this study we have established the simultaneous detection of drug induced apoptosis and autophagy in a 3D microfluidic model of GBM. Our study presents a potential ex vivo platform for developing novel therapeutic strategies tailored toward disrupting key molecular pathways involved in programmed cell death and tumor invasion in glioblastoma.
Type 3 Diabetes and Its Role Implications in Alzheimer’s Disease
Authors: Nguyen Thuy Trang , Ta Qui Thanh Hoai , Nguyen Thi Kim Oanh , Nguyen Thi Thuy Dung , Van Giau Vo
Published: May 2020

The exact connection between Alzheimer’s disease (AD) and type 2 diabetes is still in debate. However, poorly controlled blood sugar may increase the risk of developing Alzheimer’s. This relationship is so strong that some have called Alzheimer’s “diabetes of the brain” or “type 3 diabetes (T3D)”. Given more recent studies continue to indicate evidence linking T3D with AD, this review aims to demonstrate the relationship between T3D and AD based on the fact that both the processing of amyloid-β (Aβ) precursor protein toxicity and the clearance of Aβ are attributed to impaired insulin signaling, and that insulin resistance mediates the dysregulation of bioenergetics and progress to AD. Furthermore, insulin-related therapeutic strategies are suggested to succeed in the development of therapies for AD by slowing down their progressive nature or even halting their future complications.
Influence of Cannabinoid Receptor Deficiency on Parameters Involved in Blood Glucose Regulation in Mice
Authors: Zibolka Juliane , Wolf Anja , Rieger Lisa , Rothgänger Candy , Jörns Anne , Lutz Beat , Zimmer Andreas , Dehghani Faramarz , Bazwinsky-Wutschke Ivonne
Published: May 2020

Cannabinoids are known to influence hormone secretion of pancreatic islets via G protein-coupled cannabinoid receptor type 1 and 2 (CB1 and CB2). The present study was designed to further investigate the impact of cannabinoid receptors on the parameters involved in insulin secretion and blood glucose recognition. To this end, CB1 and CB2 receptor knockout mice (10–12 week old, both sexes) were characterised at basal state and compared to wild-type mice. The elimination of cannabinoid receptor signalling resulted in alterations of blood glucose concentrations, body weights and insulin levels. Changes were dependent on the deleted receptor type and on the sex. Analyses at mRNA and protein levels provided evidence for the impact of cannabinoid receptor deficiency on the glucose sensing apparatus in the pancreas. Both receptor knockout mouse lines showed decreased mRNA and protein amounts of glucose transporters Glut1 and Glut2, combined with alterations in immunostaining. In addition, pancreatic glucokinase expression was elevated and immunohistochemical labelling was modified in the pancreatic islets. Taken together, CB1 and CB2 signalling pathways seem to influence glucose sensing in β-cells by affecting glucose transporters and glucokinase. These alterations were more pronounced in CB2 knockout mice, resulting in higher blood glucose and lower plasma insulin levels.
The Expression and Function of CD300 Molecules in the Main Players of Allergic Responses: Mast Cells, Basophils and Eosinophils
Authors: Vitallé Joana , Terrén Iñigo , Orrantia Ane , Bilbao Agurtzane , Gamboa Pedro M. , Borrego Francisco , Zenarruzabeitia Olatz
Published: May 2020

Allergy is the host immune response against non-infectious substances called allergens. The prevalence of allergic diseases is increasing worldwide. However, while some drugs counteract the symptomatology caused by allergic reactions, no completely effective treatments for allergic diseases have been developed yet. In this sense, the ability of surface activating and inhibitory receptors to modulate the function of the main effector cells of allergic responses makes these molecules potential pharmacological targets. The CD300 receptor family consists of members with activating and inhibitory capabilities mainly expressed on the surface of immune cells. Multiple studies in the last few years have highlighted the importance of CD300 molecules in several pathological conditions. This review summarizes the literature on CD300 receptor expression, regulation and function in mast cells, basophils and eosinophils, the main players of allergic responses. Moreover, we review the involvement of CD300 receptors in the pathogenesis of certain allergic diseases, as well as their prospective use as therapeutic targets for the treatment of IgE-dependent allergic responses.
Exploring the Enzymatic and Antibacterial Activities of Novel Mycobacteriophage Lysin B Enzymes
Authors: Abouhmad Adel , Korany Ahmed H. , Grey Carl , Dishisha Tarek , Hatti-Kaul Rajni
Published: May 2020

Mycobacteriophages possess different sets of lytic enzymes for disruption of the complex cell envelope of the mycobacteria host cells and release of the viral progeny. Lysin B (LysB) enzymes are mycolylarabinogalactan esterases that cleave the ester bond between the arabinogalactan and mycolic acids in the mycolylarabinogalactan-peptidoglycan (mAGP) complex in the cell envelope of mycobacteria. In the present study, four LysB enzymes were produced recombinantly and characterized with respect to their enzymatic and antibacterial activities. Examination of the kinetic parameters for the hydrolysis of para-nitrophenyl ester substrates, shows LysB-His6 enzymes to be active against a range of substrates (C4–C16), with a catalytic preference towards p-nitrophenyl laurate (C12). With p-nitrophenyl butyrate as substrate, LysB-His6 enzymes showed highest activity at 37 °C. LysB-His6 enzymes also hydrolyzed different Tween substrates with highest activity against Tween 20 and 80. Metal ions like Ca2+ and Mn2+ enhanced the enzymatic activity of LysB-His6 enzymes, while transition metal ions like Zn2+ and Cu2+ inhibited the enzymatic activity. The mycolylarabinogalactan esterase activity of LysB-His6 enzymes against mAGP complex was confirmed by LC-MS. LysB-His6 enzymes showed marginal antibacterial activity when tested alone against Mycobacterium smegmatis, however a synergetic activity was noticed when combined with outer membrane permealizers. These results confirm that LysB enzymes are lipolytic enzymes with potential application as antimycobacterials.
Synthetic Retinoid Seletinoid G Improves Skin Barrier Function through Wound Healing and Collagen Realignment in Human Skin Equivalents
Authors: Lee Eun-Soo , Ahn Yujin , Bae Il-Hong , Min Daejin , Park Nok Hyun , Jung Woonggyu , Kim Se-Hwa , Hong Yong Deog , Park Won Seok , Lee Chang Seok
Published: May 2020

The outer epidermal skin is a primary barrier that protects the body from extrinsic factors, such as ultraviolet (UV) radiation, chemicals and pollutants. The complete epithelialization of a wound by keratinocytes is essential for restoring the barrier function of the skin. However, age-related alterations predispose the elderly to impaired wound healing. Therefore, wound-healing efficacy could be also considered as a potent function of an anti-aging reagent. Here, we examine the epidermal wound-healing efficacy of the fourth-generation retinoid, seletinoid G, using HaCaT keratinocytes and skin tissues. We found that seletinoid G promoted the proliferation and migration of keratinocytes in scratch assays and time-lapse imaging. It also increased the gene expression levels of several keratinocyte proliferation-regulating factors. In human skin equivalents, seletinoid G accelerated epidermal wound closure, as assessed using optical coherence tomography (OCT) imaging. Moreover, second harmonic generation (SHG) imaging revealed that seletinoid G recovered the reduced dermal collagen deposition seen in ultraviolet B (UVB)-irradiated human skin equivalents. Taken together, these results indicate that seletinoid G protects the skin barrier by accelerating wound healing in the epidermis and by repairing collagen deficiency in the dermis. Thus, seletinoid G could be a potent anti-aging agent for protecting the skin barrier.
New Insights into Molecular Links Between Microbiota and Gastrointestinal Cancers: A Literature Review
Authors: Rastogi Yash Raj , Saini Adesh K. , Thakur Vijay Kumar , Saini Reena V.
Published: May 2020

Despite decades of exhaustive research on cancer, questions about cancer initiation, development, recurrence, and metastasis have still not been completely answered. One of the reasons is the plethora of factors acting simultaneously in a tumour microenvironment, of which not all have garnered attention. One such factor that has long remained understudied and has only recently received due attention is the host microbiota. Our sheer-sized microbiota exists in a state of symbiosis with the body and exerts significant impact on our body’s physiology, ranging from immune-system development and regulation to neurological and cognitive development. The presence of our microbiota is integral to our development, but a change in its composition (microbiota dysbiosis) can often lead to adverse effects, increasing the propensity of serious diseases like cancers. In the present review, we discuss environmental and genetic factors that cause changes in microbiota composition, disposing of the host towards cancer, and the molecular mechanisms (such as β-catenin signalling) and biochemical pathways (like the generation of oncogenic metabolites like N-nitrosamines and hydrogen sulphide) that the microbiota uses to initiate or accelerate cancers, with emphasis on gastrointestinal cancers. Moreover, we discuss how microbiota can adversely influence the success of colorectal-cancer chemotherapy, and its role in tumour metastasis. We also attempted to resolve conflicting results obtained for the butyrate effect on tumour suppression in the colon, often referred to as the ‘butyrate paradox’. In addition, we suggest the development of microbiota-based biomarkers for early cancer diagnosis, and a few target molecules of which the inhibition can increase the overall chances of cancer cure.
Targeting the Src Pathway Enhances the Efficacy of Selective FGFR Inhibitors in Urothelial Cancers with FGFR3 Alterations
Authors: Lima Nadia Carvalho , Atkinson Eliza , Bunney Tom D. , Katan Matilda , Huang Paul H.
Published: May 2020

Selective FGFR inhibitors such as infigratinib (BGJ398) and erdafitinib (JNJ-42756493) have been evaluated in clinical trials for cancers with FGFR3 molecular alterations, particularly in urothelial carcinoma patients. However, a substantial proportion of these patients (up to 50%) display intrinsic resistance to these drugs and receive minimal clinical benefit. There is thus an unmet need for alternative therapeutic strategies to overcome primary resistance to selective FGFR inhibitors. In this study, we demonstrate that cells expressing cancer-associated activating FGFR3 mutants and the FGFR3-TACC3 fusion showed primary resistance to infigratinib in long-term colony formation assays in both NIH-3T3 and urothelial carcinoma models. We find that expression of these FGFR3 molecular alterations resulted in elevated constitutive Src activation compared to wildtype FGFR3 and that cells co-opted this pathway as a means to achieve intrinsic resistance to infigratinib. Targeting the Src pathway with low doses of the kinase inhibitor dasatinib synergistically sensitized multiple urothelial carcinoma lines harbouring endogenous FGFR3 alterations to infigratinib. Our data provide preclinical rationale that supports the use of dasatinib in combination with selective FGFR inhibitors as a means to overcome intrinsic drug resistance in the salvage therapy setting in urothelial cancer patients with FGFR3 molecular alterations
Safeguarding Female Reproductive Health Against Endocrine Disrupting Chemicals—The FREIA Project
Authors: van Duursen Majorie B. M. , Boberg Julie , Christiansen Sofie , Connolly Lisa , Damdimopoulou Pauliina , Filis Panagiotis , Fowler Paul A. , Gadella Bart M. , Holte Jan , Jääger Kersti , Johansson Hanna K. L. , Li Tianyi , Mazaud-Guittot Séverine , Parent Anne-Simone , Salumets Andres , Soto Ana M. , Svingen Terje , Velthut-Meikas Agne , Bay Wedebye Eva , Xie Yuling , van den Berg Martin
Published: May 2020

Currently available test methods are not well-suited for the identification of chemicals that disturb hormonal processes involved in female reproductive development and function. This renders women’s reproductive health at increasing risk globally, which, coupled with increasing incidence rates of reproductive disorders, is of great concern. A woman’s reproductive health is largely established during embryonic and fetal development and subsequently matures during puberty. The endocrine system influences development, maturation, and function of the female reproductive system, thereby making appropriate hormone levels imperative for correct functioning of reproductive processes. It is concerning that the effects of human-made chemicals on the endocrine system and female reproductive health are poorly addressed in regulatory chemical safety assessment, partly because adequate test methods are lacking. Our EU-funded project FREIA aims to address this need by increasing understanding of how endocrine disrupting chemicals (EDCs) can impact female reproductive health. We will use this information to provide better test methods that enable fit-for-purpose chemical regulation and then share our knowledge, promote a sustainable society, and improve the reproductive health of women globally.
Pancreatic Cancer and Its Microenvironment—Recent Advances and Current Controversies
Authors: Stopa Kinga B. , Kusiak Agnieszka A. , Szopa Mateusz D. , Ferdek Pawel E. , Jakubowska Monika A.
Published: May 2020

Pancreatic ductal adenocarcinoma (PDAC) causes annually well over 400,000 deaths world-wide and remains one of the major unresolved health problems. This exocrine pancreatic cancer originates from the mutated epithelial cells: acinar and ductal cells. However, the epithelia-derived cancer component forms only a relatively small fraction of the tumor mass. The majority of the tumor consists of acellular fibrous stroma and diverse populations of the non-neoplastic cancer-associated cells. Importantly, the tumor microenvironment is maintained by dynamic cell-cell and cell-matrix interactions. In this article, we aim to review the most common drivers of PDAC. Then we summarize the current knowledge on PDAC microenvironment, particularly in relation to pancreatic cancer therapy. The focus is placed on the acellular stroma as well as cell populations that inhabit the matrix. We also describe the altered metabolism of PDAC and characterize cellular signaling in this cancer.
Diabetes and Hyperglycemia Affect Platelet GPIIIa Expression: Effects on Adhesion Potential of Blood Platelets from Diabetic Patients under In Vitro Flow Conditions
Authors: Przygodzki Tomasz , Luzak Boguslawa , Kassassir Hassan , Mnich Ewelina , Boncler Magdalena , Siewiera Karolina , Kosmalski Marcin , Szymanski Jacek , Watala Cezary
Published: May 2020

Blood platelets play a crucial role in the early stages of atherosclerosis development. The process is believed to require firm adhesion of platelets to atherosclerosis-prone sites of the artery. However, little evidence exists regarding whether the blood platelets of individuals with pathological conditions associated with atherosclerosis have higher potential for adhesion. This process is to a large extent dependent on receptors present on the platelet membrane. Therefore, the aim of the presented study was to determine whether blood platelets from diabetic patients have higher capacity of adhesion under flow conditions and how diabetes affects one of the crucial platelet receptors involved in the process of adhesion—GPIIIa. The study compares the ability of platelets from non-diabetic and diabetic humans to interact with fibrinogen and von Willebrand factor, two proteins found in abundance on an inflamed endothelium, under flow conditions. The activation and reactivity of the blood platelets were also characterized by flow cytometry. Platelets from diabetic patients did not demonstrate enhanced adhesion to either studied protein, although they presented increased basal activation and responsiveness towards low concentrations of agonists. Platelets from diabetic patients were characterized by lower expression of GPIIIa, most likely due to an enhanced formation of platelet-derived microparticles PMPs, as supported by the observation of elevated concentration of this integrin and of GPIIIa-positive PMPs in plasma. We conclude that altered functionality of blood platelets in diabetes does not increase their adhesive potential. Increased glycation and decrease in the amount of GPIIIa on platelets may be partially responsible for this effect. Therefore, higher frequency of interactions of platelets with the endothelium, which is observed in animal models of diabetes, is caused by other factors. A primary cause may be a dysfunctional vascular wall.
Inflammatory and Infectious Processes Serve as Links between Atrial Fibrillation and Alzheimer’s Disease
Authors: Martins Gabriela Lopes , Duarte Rita Carolina Figueiredo , Mukhamedyarov Marat Alexandrovich , Palotás András , Ferreira Cláudia Natália , Reis Helton José
Published: May 2020

Atrial fibrillation (AF) is one of the most prevalent forms of arrhythmia that carries an increased risk of stroke which, in turn, is strongly associated with cognitive decline. The majority of dementia cases are caused by Alzheimer’s disease (AD) with obscure pathogenesis. While the exact mechanisms are unknown, the role of inflammatory processes and infectious agents have recently been implicated in both AD and AF, suggesting a common link between these maladies. Here, we present the main shared pathways underlying arrhythmia and memory loss. The overlapping predictive biomarkers and emerging joint pharmacological approaches are also discussed.
Glycosylation Tunes Neuroserpin Physiological and Pathological Properties
Authors: Visentin Cristina , Broggini Luca , Sala Benedetta Maria , Russo Rosaria , Barbiroli Alberto , Santambrogio Carlo , Nonnis Simona , Dubnovitsky Anatoly , Bolognesi Martino , Miranda Elena , Achour Adnane , Ricagno Stefano
Published: May 2020

Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. Thus, our results support the relevance of gNS as a new in vitro tool to study the molecular bases of FENIB.
Targeting ERK-Hippo Interplay in Cancer Therapy
Authors: Vališ Karel , Novák Petr
Published: May 2020

Extracellular signal-regulated kinase (ERK) is a part of the mitogen-activated protein kinase (MAPK) signaling pathway which allows the transduction of various cellular signals to final effectors and regulation of elementary cellular processes. Deregulation of the MAPK signaling occurs under many pathological conditions including neurodegenerative disorders, metabolic syndromes and cancers. Targeted inhibition of individual kinases of the MAPK signaling pathway using synthetic compounds represents a promising way to effective anti-cancer therapy. Cross-talk of the MAPK signaling pathway with other proteins and signaling pathways have a crucial impact on clinical outcomes of targeted therapies and plays important role during development of drug resistance in cancers. We discuss cross-talk of the MAPK/ERK signaling pathway with other signaling pathways, in particular interplay with the Hippo/MST pathway. We demonstrate the mechanism of cell death induction shared between MAPK/ERK and Hippo/MST signaling pathways and discuss the potential of combination targeting of these pathways in the development of more effective anti-cancer therapies.
Kaempferol and Its Glycoside, Kaempferol 7-O-rhamnoside, Inhibit PD-1/PD-L1 Interaction In Vitro
Authors: Kim Ji Hye , Kim Young Soo , Choi Jang-Gi , Li Wei , Lee Eun Jin , Park Jin-Wan , Song Jaeyoung , Chung Hwan-Suck
Published: May 2020

Kaempferol (KO) and kaempferol 7-O-rhamnoside (KR) are natural products from various oriental herbs such as Geranii Herba. Previous studies have reported some biological activities of KO and KR; however, their effects on PD-1/PD-L1 interaction have not been reported yet. To elucidate their inhibitory activities on PD-1/PD-L1 protein–protein interaction (PPI), biochemical assays including competitive ELISA and biolayer interferometry (BLI) systems were performed. Cellular PD-1/PD-L1 blocking activity was measured in a co-culture system with PD-1 Jurkat and PD-L1/aAPC CHO-K1 cells by T-cell receptor (TCR) activation-induced nuclear factor of activated T cells (NFAT)-luciferase reporter assay. The detailed binding mode of action was simulated by an in silico docking study and pharmacophore analysis. Competitive ELISA revealed that KO and its glycoside KR significantly inhibited PD-1/PD-L1 interaction. Cellular PD-1/PD-L1 blocking activity was monitored by KO and KR at non-cytotoxic concentration. Surface plasmon resonance (SPR) and biolayer interferometry (BLI) analysis suggested the binding affinity and direct inhibition of KR against PD-1/PD-L1. An in silico docking simulation determined the detailed mode of binding of KR to PD-1/PD-L1. Collectively, these results suggest that KR could be developed as a potent small molecule inhibitor for PD-1/PD-L1 blockade.
Functional Relationship between Osteogenesis and Angiogenesis in Tissue Regeneration
Authors: Diomede Francesca , Marconi Guya Diletta , Fonticoli Luigia , Pizzicanella Jacopo , Merciaro Ilaria , Bramanti Placido , Mazzon Emanuela , Trubiani Oriana
Published: May 2020

Bone tissue renewal can be outlined as a complicated mechanism centered on the interaction between osteogenic and angiogenic events capable of leading to bone formation and tissue renovation. The achievement or debacle of bone regeneration is focused on the primary role of vascularization occurrence; in particular, the turning point is the opportunity to vascularize the bulk scaffolds, in order to deliver enough nutrients, growth factors, minerals and oxygen for tissue restoration. The optimal scaffolds should ensure the development of vascular networks to warrant a positive suitable microenvironment for tissue engineering and renewal. Vascular Endothelial Growth Factor (VEGF), a main player in angiogenesis, is capable of provoking the migration and proliferation of endothelial cells and indirectly stimulating osteogenesis, through the regulation of the osteogenic growth factors released and through paracrine signaling. For this reason, we concentrated our attention on two principal groups involved in the renewal of bone tissue defects: the cells and the scaffold that should guarantee an effective vascularization process. The application of Mesenchymal Stem Cells (MSCs), an excellent cell source for tissue restoration, evidences a crucial role in tissue engineering and bone development strategies. This review aims to provide an overview of the intimate connection between blood vessels and bone formation that appear during bone regeneration when MSCs, their secretome—Extracellular Vesicles (EVs) and microRNAs (miRNAs) —and bone substitutes are used in combination.
Aptamers and Antisense Oligonucleotides for Diagnosis and Treatment of Hematological Diseases
Authors: Giudice Valentina , Mensitieri Francesca , Izzo Viviana , Filippelli Amelia , Selleri Carmine
Published: May 2020

Aptamers or chemical antibodies are single-stranded DNA or RNA oligonucleotides that bind proteins and small molecules with high affinity and specificity by recognizing tertiary or quaternary structures as antibodies. Aptamers can be easily produced in vitro through a process known as systemic evolution of ligands by exponential enrichment (SELEX) or a cell-based SELEX procedure. Aptamers and modified aptamers, such as slow, off-rate, modified aptamers (SOMAmers), can bind to target molecules with less polar and more hydrophobic interactions showing slower dissociation rates, higher stability, and resistance to nuclease degradation. Aptamers and SOMAmers are largely employed for multiplex high-throughput proteomics analysis with high reproducibility and reliability, for tumor cell detection by flow cytometry or microscopy for research and clinical purposes. In addition, aptamers are increasingly used for novel drug delivery systems specifically targeting tumor cells, and as new anticancer molecules. In this review, we summarize current preclinical and clinical applications of aptamers in malignant and non-malignant hematological diseases.
MicroRNA Nanotherapeutics for Lung Targeting. Insights into Pulmonary Hypertension
Authors: Carregal-Romero Susana , Fadón Lucía , Berra Edurne , Ruíz-Cabello Jesús
Published: May 2020

In this review, the potential future role of microRNA-based therapies and their specific application in lung diseases is reported with special attention to pulmonary hypertension. Current limitations of these therapies will be pointed out in order to address the challenges that they need to face to reach clinical applications. In this context, the encapsulation of microRNA-based therapies in nanovectors has shown improvements as compared to chemically modified microRNAs toward enhanced stability, efficacy, reduced side effects, and local administration. All these concepts will contextualize in this review the recent achievements and expectations reported for the treatment of pulmonary hypertension.
Assessing the Performance of Dried-Blood-Spot DNA Extraction Methods in Next Generation Sequencing
Authors: Hendrix Miyono M. , Cuthbert Carla D. , Cordovado Suzanne K.
Published: May 2020

An increasing number of newborn screening laboratories in the United States and abroad are moving towards incorporating next-generation sequencing technology, or NGS, into routine screening, particularly for cystic fibrosis. As more programs utilize this technology for both cystic fibrosis and beyond, it is critical to identify appropriate DNA extraction methods that can be used with dried blood spots that will result in consistent, high-quality sequencing results. To provide comprehensive quality assurance and technical assistance to newborn screening laboratories wishing to incorporate NGS assays, CDC’s Newborn Screening and Molecular Biology Branch designed a study to evaluate the performance of nine commercial or laboratory-developed DNA extraction methods that range from a highly purified column extraction to a crude detergent-based no-wash boil prep. The DNA from these nine methods was used in two NGS library preparations that interrogate the CFTR gene. All DNA extraction methods including the cruder preps performed reasonably well with both library preps. One lower-concentration, older sample was excluded from one of the assay evaluations due to poor performance across all DNA extraction methods. When 84 samples, versus eight, were run on a flow cell, the DNA quality and quantity were more significant variables.
Habitat Preferences and Trophic Position of Brachydiplax chalybea flavovittata Ris, 1911 (Insecta: Odonata) Larvae in Youngsan River Wetlands of South Korea
Authors: Choi Jong-Yun , Kim Seong-Ki , Kim Jeong-Cheol , Kwon Soon-Jik
Published: May 2020

In freshwater ecosystems, habitat heterogeneity supports high invertebrate density and diversity, and it contributes to the introduction and settlement of non-native species. In the present study, we identified the habitat preferences and trophic level of Brachydiplax chalybea flavovittata larvae, which were distributed in four of the 17 wetlands we examined in the Yeongsan River basin, South Korea. Larval density varied across four microhabitat types: open water area, and microhabitats dominated by Myriophyllum aquaticum, Paspalum distichum, and Zizania latifolia. Microhabitats dominated by M. aquaticum had the highest larval density, followed by those dominated by P. distichum. The larvae were more prevalent in silt sediments than in plant debris or sand. Stable isotope analysis showed that B. chalybea flavovittata is likely to consume, as a food source, other species of Odonata larvae. We conclude that successful settlement of B. chalybea flavovittata can be attributed to their habitat preferences. As temperature increases due to climate change, the likelihood of B. chalybea flavovittata spreading throughout South Korea increases. We, therefore, recommend continued monitoring of the spread and ecological impacts of B. chalybea flavovittata.
Using a Two-Sex Life Table Tool to Calculate the Fitness of Orius strigicollis as a Predator of Pectinophora gossypiella
Authors: Ali Shahzaib , Li Sizhe , Jaleel Waqar , Musa Khan Muhammad , Wang Jintao , Zhou Xingmiao
Published: May 2020

A two-sex life table is a useful tool for studying the fitness of predators. Previous studies of Orius strigicollis Poppius (Heteroptera: Anthocoridae) fitness have not been done on Pectinophora gossypiella (Lepidoptera: Gelechiidae) using a two-sex life table tool. This study reports the fitness of the minute predatory flower bug, O. strigicollis when feeding on the cotton pest P. gossypiella using a two-sex life table tool. Different densities (5, 10, and 15 eggs) of P. gossypiella eggs were used to calculate the feeding capacity and fitness of O. strigicollis in the laboratory at 28 °C ± 1, 75 ± 5% RH and 16:8 (L:D). The results concluded that O. strigicollis is an efficient predator of P. gossypiella. The maximum growth capacity of the predatory bug O. strigicollis was attained when it was fed on 10 and 15 P. gossypiella eggs. Furthermore, shorter generation and development time were also observed in the case of 15 eggs of P. gossypiella. These results suggest that O. strigicollis has considerable predatory potential and prefers feeding on P. gossypiella eggs than on the first instar larvae at the fourth instar or the female stage. Although the field potential of O. strigicollis is still unknown, this study will support future investigations in terms of field applications.
Transcriptional Response of ATP-Binding Cassette (ABC) Transporters to Insecticide in the Brown Planthopper, Nilaparvata lugens (Stål)
Authors: Li Zhao , Cai Tingwei , Qin Yao , Zhang Yunhua , Jin Ruoheng , Mao Kaikai , Liao Xun , Wan Hu , Li Jianhong
Published: May 2020

The ATP-binding cassette (ABC) transporter superfamily is one of the largest groups of proteins and plays a non-negligible role in phase III of the detoxification process, which is highly involved in the response of insects to environmental stress (plant secondary metabolites and insecticides). In the present study, in Nilaparvata lugens, we identified 32 ABC transporters, which are grouped into eight subfamilies (ABCA–H) based on phylogenetic analysis. The temporal and spatial expression profiles suggested that the nymphal stages (1st–5th) and adult males showed similarity, which was different from eggs and adult females, and NlABCA1, NlABCA2, NlABCB6, NlABCD2, NlABCG4, NlABCG12, NlABCG15, and NlABCH1 were highly expressed in the midgut and Malpighian tubules. In addition, ABCG12, which belongs to the ABC transporter G subfamily, was significantly upregulated after exposure to sulfoxaflor, nitenpyram, clothianidin, etofenprox, chlorpyrifos, and isoprocarb. Moreover, verapamil significantly increased the sensitivity of N. lugens to nitenpyram, clothianidin, etofenprox, chlorpyrifos, and isoprocarb. These results provide a basis for further research on ABC transporters involved in detoxification in N. lugens, and for a more comprehensive understanding of the response of N. lugens to environmental stress.
Nonlinear-Elastic Orthotropic Material Modeling of an Epoxy-Based Polymer for Predicting the Material Behavior of Transversely Loaded Fiber-Reinforced Composites
Authors: Lüders Caroline
Published: May 2020

Micromechanical analyses of transversely loaded fiber-reinforced composites are conducted to gain a better understanding of the damage behavior and to predict the composite behavior from known parameters of the fibers and the matrix. Currently, purely elastic material models for the epoxy-based polymeric matrix do not capture the nonlinearity and the tension/compression-asymmetry of the resin’s material behavior. In the present contribution, a purely elastic material model is presented that captures these effects. To this end, a nonlinear-elastic orthotropic material modeling is proposed. Using this matrix material model, finite element-based simulations are performed to predict the composite behavior under transverse tension, transverse compression and shear. Therefore, the composite’s cross-section is modeled by a representative volume element. To evaluate the matrix modeling approach, the simulation results are compared to experimental data and the prediction error is computed. Furthermore, the accuracy of the prediction is compared to that of selected literature models. Compared to both experimental and literature data, the proposed modeling approach gives a good prediction of the composite behavior under matrix-dominated load cases.
Low-Power Embedded System for Gait Classification Using Neural Networks
Authors: Luna-Perejón Francisco , Domínguez-Morales Manuel , Gutiérrez-Galán Daniel , Civit-Balcells Antón
Published: May 2020

Abnormal foot postures can be measured during the march by plantar pressures in both dynamic and static conditions. These detections may prevent possible injuries to the lower limbs like fractures, ankle sprain or plantar fasciitis. This information can be obtained by an embedded instrumented insole with pressure sensors and a low-power microcontroller. However, these sensors are placed in sparse locations inside the insole, so it is not easy to correlate manually its values with the gait type; that is why a machine learning system is needed. In this work, we analyse the feasibility of integrating a machine learning classifier inside a low-power embedded system in order to obtain information from the user’s gait in real-time and prevent future injuries. Moreover, we analyse the execution times, the power consumption and the model effectiveness. The machine learning classifier is trained using an acquired dataset of 3000+ steps from 6 different users. Results prove that this system provides an accuracy over 99% and the power consumption tests obtains a battery autonomy over 25 days.
Damage in Rubble Mound Breakwaters. Part I: Historical Review of Damage Models
Authors: Campos Álvaro , Castillo Carmen , Molina-Sanchez Rafael
Published: May 2020

The term “damage” in rubble mound breakwaters is usually related to the foremost failure mode of this kind of coastal structures: their hydraulic instability. The characterization of the breakwater response against wave action was and will be the goal of hundreds of studies. Because of the large amount of information, the present review on damage in rubble mound breakwaters is divided in two papers, which are closely linked but conceptually different; whereas Part II is focused on the various approaches for defining and measuring damage, Part I summarizes the diverse strategies for modelling damage development and progression. The present paper compiles 146 references on this topic, chronologically discussed over almost a century of history: from 1933 to 2020. It includes 23 formulations of hydraulic stability models and 11 formulations of damage progression models, together with main advances and shortcomings up to date. The future of rubble mound design is linked to risk-based tools and advanced management strategies, for which deeper comprehension about the spatial and temporal evolution of damage during the useful life of each particular structure is required. For this aim, damage progression probabilistic models, full-scale monitoring and standardization will presumably be some of the key challenges in the upcoming years.
Synthetic Biology Approaches to Engineer Saccharomyces cerevisiae towards the Industrial Production of Valuable Polyphenolic Compounds
Authors: Rainha João , Gomes Daniela , Rodrigues Lígia R. , Rodrigues Joana L.
Published: May 2020

Polyphenols are plant secondary metabolites with diverse biological and potential therapeutic activities such as antioxidant, anti-inflammatory and anticancer, among others. However, their extraction from the native plants is not enough to satisfy the increasing demand for this type of compounds. The development of microbial cell factories to effectively produce polyphenols may represent the most attractive solution to overcome this limitation and produce high amounts of these bioactive molecules. With the advances in the synthetic biology field, the development of efficient microbial cell factories has become easier, largely due to the development of the molecular biology techniques and by the identification of novel isoenzymes in plants or simpler organisms to construct the heterologous pathways. Furthermore, efforts have been made to make the process more profitable through improvements in the host chassis. In this review, advances in the production of polyphenols by genetically engineered Saccharomyces cerevisiae as well as by synthetic biology and metabolic engineering approaches to improve the production of these compounds at industrial settings are discussed.
Antibacterial PEGylated Solid Lipid Microparticles for Cosmeceutical Purpose: Formulation, Characterization, and Efficacy Evaluation
Authors: Angellotti Giuseppe , Murgia Denise , Presentato Alessandro , D’Oca Maria Cristina , Scarpaci Amalia Giulia , Alduina Rosa , Raimondi Maria Valeria , De Caro Viviana
Published: May 2020

The development of efficacious means of delivering antioxidant polyphenols from natural sources for the treatment of skin diseases is of great interest for many cosmetic and pharmaceutical companies. Resveratrol (RSV) and Limonene (LIM) have been shown to possess good anti-inflammatory and antibacterial properties against Staphylococcus aureus infections responsible for many skin disorders, such as acne vulgaris. In this study, solid lipid microparticles are designed as composite vehicles capable of encapsulating a high amount of trans-RSV and enhancing its absorption through the stratum corneum. A microparticulate system based on mixture of PEGylate lipids, long-chain alcohols and LIM is able to entrap RSV in an amorphous state, increasing its half-life and avoiding inactivation due to isomerization phenomena, which represents the main drawback in topical formulations. Particles have been characterized in term of shape, size distribution and drug loading. Antimicrobial tests against S. aureus have highlighted that empty microspheres possess per se antimicrobial activity, which is enhanced by the presence of LIM, demonstrating that they can represent an interesting bactericide vehicle for RSV administration on the skin.
Synthetic Hydroxyapatite Inhibits Bisphosphonate Toxicity to the Oral Mucosa In Vitro
Authors: Bullock George , Miller Cheryl , McKechnie Alasdair , Hearnden Vanessa
Published: May 2020

Medication-related osteonecrosis of the jaw (MRONJ) is a side effect of bisphosphonate therapy, characterised by exposed necrotic bone. The soft tissues of the oral mucosa no longer provide a protective barrier and MRONJ patients experience pain, infections and difficulties eating. We hypothesised that hydroxyapatite (Ca5(PO4)3(OH)) could reduce bisphosphonate concentrations and protect the oral mucosa by exploiting bisphosphonate’s calcium binding affinity. The effect of zoledronic acid (ZA) and pamidronic acid (PA) on the metabolism of oral fibroblasts, oral keratinocytes and three-dimensional oral mucosa models was investigated and then repeated in the presence of hydroxyapatite granules. Without hydroxyapatite, ZA and PA significantly reduced the metabolic activity of oral cells in a dose-dependent manner. Both drugs reduced epithelial thickness and 30 µM ZA resulted in loss of the epithelium. Hydroxyapatite granules had a protective effect on oral cells, with metabolic activity retained. Oral mucosa models retained their multi-layered epithelium when treated with ZA in the presence of hydroxyapatite granules and metabolic activity was comparable to controls. These results demonstrate hydroxyapatite granules protected oral soft tissues from damage caused by bisphosphonate exposure. Porous hydroxyapatite granules are currently used for socket preservation and this data suggests their potential to prevent MRONJ in at-risk patients.
Thermo-Mechanical Simulation of Hybrid Welding of DP/AISI 316 and TWIP/AISI 316 Dissimilar Weld
Authors: Perulli Patrizia , Dassisti Michele , Casalino Giuseppe
Published: May 2020

In this paper, hybrid laser-MAG (metal active gas) welding of twinning-induced plasticity (TWIP) and dual-phase (DP) steels with austenitic stainless steel (AISI316) was simulated by means of the finite element method. A thermo-mechanical model, which uses a 3D heat sources, was developed using the software Simufact Welding. The calculated dimensions, shape and distortion of the weld were compared with the experimental results, thence the model was validated. The metallurgical transformations for the DP steel were evaluated using the continuous cooling transformation (CCT) diagram and the calculated cooling rate. The numerical model predicted accurately the shape of the molten pool, the thermal cycles as well as the geometrical distortion of the butt weld. Therefore, the numerical model showed a good reliability and its potential for further development.
Effect of Different Compatibilizers on the Properties of Poly (Lactic Acid)/Poly (Butylene Adipate-Co-Terephthalate) Blends Prepared under Intense Shear Flow Field
Authors: He Hezhi , Wang Guozhen , Chen Ming , Xiong Chengtian , Li Yi , Tong Yi
Published: May 2020

In this report, poly(lactic acid) (PLA) and Poly(butylene adipate-co-terephthalate) (PBAT) with three kinds of compatibilizers were melt blended under intensive shear flow. A self-made parallel three-screw extruder was developed to generate such flow during the process. Mechanical properties, chemical reactions among PLA, PBAT and compatibilizers, rheological behavior and morphology were investigated. The mechanical tests showed that the notched impact strength of super-tough composite with 10 wt% EGMA is about 20 times than that of pure PLA. The Fourier transform infrared spectroscopy (FT-IR) results showed that the epoxy functional groups or maleic anhydride functional groups of KT-20, KT-915 and EGMA reacted with the hydroxyl groups of PLA or PBAT macromolecules, resulting in a bridge of PLA and PBAT. About rheological properties, the tan δ—angular frequency curves and the η’’- η’ curves confirmed the chemical reactions mentioned above and indicated better compatibility of η’’- η’ between PLA and PBAT, respectively. Meanwhile, the loss modulus and storage modulus—angular frequency curves demonstrated the discrepancy of different compatibilizer components. In particular, from scanning electron microscopy (SEM) images, it can be seen that the phase size and dispersion uniformity of PBAT adjusted by compatibilizer, corresponding to better compatibility that is described in the η’’- η’ curves. The approach for producing super-tough PLA/PBAT/compatibilizer by intensive shear flow provides a viable direction for further improving PLA performance.