bluuebunny/crossref_metadata_2025 huggingface dataset:
Deficiency of the Lysosomal Protein CLN5 Alters Lysosomal Function and Movement
Authors: Basak Indranil , Hansen Rachel A. , Ward Michael E. , Hughes Stephanie M.
Published: September 2021

Batten disease is a devastating, childhood, rare neurodegenerative disease characterised by the rapid deterioration of cognition and movement, leading to death within ten to thirty years of age. One of the thirteen Batten disease forms, CLN5 Batten disease, is caused by mutations in the CLN5 gene, leading to motor deficits, mental deterioration, cognitive impairment, visual impairment, and epileptic seizures in children. A characteristic pathology in CLN5 Batten disease is the defects in lysosomes, leading to neuronal dysfunction. In this study, we aimed to investigate the lysosomal changes in CLN5-deficient human neurons. We used an induced pluripotent stem cell system, which generates pure human cortical-like glutamatergic neurons. Using CRISPRi, we inhibited the expression of CLN5 in human neurons. The CLN5-deficient human neurons showed reduced acidic organelles and reduced lysosomal enzyme activity measured by microscopy and flow cytometry. Furthermore, the CLN5-deficient human neurons also showed impaired lysosomal movement—a phenotype that has never been reported in CLN5 Batten disease. Lysosomal trafficking is key to maintain local degradation of cellular wastes, especially in long neuronal projections, and our results from the human neuronal model present a key finding to understand the underlying lysosomal pathology in neurodegenerative diseases.
Inflammation, Anxiety, and Stress in Attention-Deficit/Hyperactivity Disorder
Authors: Saccaro Luigi F. , Schilliger Zoé , Perroud Nader , Piguet Camille
Published: September 2021

Attention-deficit/hyperactivity disorder (ADHD) is a prevalent and serious neurodevelopmental disorder characterized by symptoms of inattention and/or hyperactivity/impulsivity. Chronic and childhood stress is involved in ADHD development, and ADHD is highly comorbid with anxiety. Similarly, inflammatory diseases and a pro-inflammatory state have been associated with ADHD. However, while several works have studied the relationship between peripheral inflammation and stress in affective disorders such as depression or bipolar disorder, fewer have explored this association in ADHD. In this narrative review we synthetize evidence showing an interplay between stress, anxiety, and immune dysregulation in ADHD, and we discuss the implications of a potential disrupted neuroendocrine stress response in ADHD. Moreover, we highlight confounding factors and limitations of existing studies on this topic and critically debate multidirectional hypotheses that either suggest inflammation, stress, or anxiety as a cause in ADHD pathophysiology or inflammation as a consequence of this disease. Untangling these relationships will have diagnostic, therapeutic and prognostic implications for ADHD patients.
What Do We Know about Theory of Mind Impairment in Parkinson’s Disease?
Authors: Trompeta Clara , Fernández Rodríguez Beatriz , Gasca-Salas Carmen
Published: September 2021

Theory of mind (ToM) is a social cognitive skill that involves the ability to attribute mental states to self and others (what they think (cognitive ToM) and feel (affective ToM)). We aim to provide an overview of previous knowledge of ToM in Parkinson’s disease (PD). In the last few years more attention has been paid to the study of this construct as a non-motor manifestation of PD. In advanced stages, both components of ToM (cognitive and affective) are commonly impaired, although in early PD results remain controversial. Executive dysfunction correlates with ToM deficits and other cognitive domains such as language and visuospatial function have also been related to ToM. Recent studies have demonstrated that PD patients with mild cognitive impairment show ToM deficits more frequently in comparison with cognitively normal PD patients. In addition to the heterogeneity of ToM tests administered in different studies, depression and dopaminergic medication may also be acting as confounding factors, but there are still insufficient data to support this. Neuroimaging studies conducted to understand the underlying networks of cognitive and affective ToM deficits in PD are lacking. The study of ToM deficit in PD continues to be important, as this may worsen quality of life and favor social stigma. Future studies should be considered, including assessment of the patients’ cognitive state, associated mood disorders, and the role of dopaminergic deficit.
Saudi Women and Vision 2030: Bridging the Gap?
Authors: Saleh Wafaa , Malibari Areej
Published: September 2021

(1) Background, Travel characteristics of Saudi women contrast significantly from those in the west. This is not only because they have different culture, attitudes and preferences but also until recently, Saudi women were not allowed to drive. In 2018, they were granted the right to drive. It has been anticipated that enabling women to drive will improve their mobility and employability. (2) Methods: This study presents a qualitative study into factors affecting Saudi women’s travel decisions “before” and “after” enabling women to drive in the Kingdom. Two six “before” and “after” focus groups have been carried out to investigate the decision-making process associated with Saudi women’s travel, available options of travel and perception of Saudi women towards private car driving. (3) Results: The results reveal that main travelling options for professional and high-income women is a private driver in the “before” scenario and a ride-share option with a family member. In the “after” scenario, high income professional women prefer “drive own car” option. Moreover, many of the participants indicated that it is likely that they might keep private drivers as well. (4) Conclusion. The results from this research indicate that there has been significant change in travel characteristics, attitudes and behaviour of Saudi women since they were granted the right to drive. This is likely to have significant implications for decision and policy makers. Further research into potential impacts of the current situation on car ownership and use, impacts on public transport system, environmental impacts and sustainability is needed.
Automated Raman Micro-Spectroscopy of Epithelial Cell Nuclei for High-Throughput Classification
Authors: O’Dwyer Kevin , Domijan Katarina , Dignam Adam , Butler Marion , Hennelly Bryan M.
Published: September 2021

Raman micro-spectroscopy is a powerful technique for the identification and classification of cancer cells and tissues. In recent years, the application of Raman spectroscopy to detect bladder, cervical, and oral cytological samples has been reported to have an accuracy greater than that of standard pathology. However, despite being entirely non-invasive and relatively inexpensive, the slow recording time, and lack of reproducibility have prevented the clinical adoption of the technology. Here, we present an automated Raman cytology system that can facilitate high-throughput screening and improve reproducibility. The proposed system is designed to be integrated directly into the standard pathology clinic, taking into account their methodologies and consumables. The system employs image processing algorithms and integrated hardware/software architectures in order to achieve automation and is tested using the ThinPrep standard, including the use of glass slides, and a number of bladder cancer cell lines. The entire automation process is implemented, using the open source Micro-Manager platform and is made freely available. We believe that this code can be readily integrated into existing commercial Raman micro-spectrometers.
Alpha-Lipoic Acid Prevents Side Effects of Therapeutic Nanosilver without Compromising Cytotoxicity in Experimental Pancreatic Cancer
Authors: An Xuefeng , Liu Li , Schaefer Michael , Yan Bin , Scholz Christian , Hillmer Stefan , Wang Kangtao , Luo Yiqiao , Ji Huihui , Gladkich Jury , Herr Ingrid
Published: September 2021

Silver nanoparticles (AgNPs) have attracted attention in cancer therapy and might support the treatment of pancreatic ductal adenocarcinoma (PDAC). Silver is in clinical use in wound dressings, catheters, stents and implants. However, the side effects of systemic AgNP treatment due to silver accumulation limit its therapeutic application. We evaluated whether the antioxidant and natural agent α-lipoic acid might prevent these side effects. We synthesized AgNPs using an Ionic-Pulser® Pro silver generator and determined the concentration by inductively coupled plasma–optical emission spectrometry. The effect of α-lipoic acid was examined in four PDAC and two nonmalignant cell lines by MTT, FACS analysis, TEM, xenotransplantation and immunohistochemistry. The viability of PDAC cells was nearly totally abolished by AgNP treatment, whereas nonmalignant cells largely resisted. α-Lipoic acid prevented AgNP-induced cytotoxicity in nonmalignant cells but not in PDAC cells, which might be due to the higher sensitivity of malignant cells to silver-induced cytotoxicity. α-Lipoic acid protected mitochondria from AgNP-induced damage and led to precipitation of AgNPs. AgNPs reduced the growth of tumor xenografts, and cotreatment with α-lipoic acid protected chick embryos from AgNP-induced liver damage. Together, α-lipoic acid strongly reduced AgNP-induced side effects without weakening the therapeutic efficacy.
Processing Speed and Time since Diagnosis Predict Adaptive Functioning Measured with WeeFIM in Pediatric Brain Tumor Survivors
Authors: Oprandi Maria Chiara , Oldrati Viola , delle Fave Morena , Panzeri Daniele , Gandola Lorenza , Massimino Maura , Bardoni Alessandra , Poggi Geraldina
Published: September 2021

(1) Background: Brain tumor (BT) survivors show difficulties in the acquisition of developmental milestones, related to academic achievement, vocational employment, social relationships, and autonomy. The skills underlying adaptive functioning (AF) are usually damaged in BT survivors due to the presence of the brain tumor, treatment-related factors, and other neurological sequelae. In this study, we aimed to explore the contribution of different cognitive factors in children with BT to AF, considering diagnosis-related variables. (2) Methods: Standardized cognitive assessment was undertaken and clinical information was collected from a retrospective cohort of 78 children with a BT, aged between 6 and 18 year old at the time of the assessment. Regression models were computed to investigate the influence of the selected variables on daily functional skills as measured by the Functional Independence Measure for Children (WeeFIM). (3) Results: The analyses showed that the main explanatory variables are processing speed and time since diagnosis. Other clinical variables, such as age at diagnosis and hydrocephalus, differentially influence functional skills according to distinct domains (i.e., self-care, mobility, and cognition). (4) Conclusions: The main explanatory variables of AF that emerged in our models point to a potential target of improving AF management in pediatric BT survivors.
Dynamic 11C-Methionine PET-CT: Prognostic Factors for Disease Progression and Survival in Patients with Suspected Glioma Recurrence
Authors: Mattoli Maria Vittoria , Trevisi Gianluca , Scolozzi Valentina , Capotosti Amedeo , Cocciolillo Fabrizio , Marini Irene , Mare Valerio , Indovina Luca , Caulo Massimo , Saponiero Antonella , Balducci Mario , Taralli Silvia , Calcagni Maria Lucia
Published: September 2021

Purpose: The prognostic evaluation of glioma recurrence patients is important in the therapeutic management. We investigated the prognostic value of 11C-methionine PET-CT (MET-PET) dynamic and semiquantitative parameters in patients with suspected glioma recurrence. Methods: Sixty-seven consecutive patients who underwent MET-PET for suspected glioma recurrence at MR were retrospectively included. Twenty-one patients underwent static MET-PET; 46/67 underwent dynamic MET-PET. In all patients, SUVmax, SUVmean and tumour-to-background ratio (T/B) were calculated. From dynamic acquisition, the shape and slope of time-activity curves, time-to-peak and its SUVmax (SUVmaxTTP) were extrapolated. The prognostic value of PET parameters on progression-free (PFS) and overall survival (OS) was evaluated using Kaplan–Meier survival estimates and Cox regression. Results: The overall median follow-up was 19 months from MET-PET. Recurrence patients (38/67) had higher SUVmax (p = 0.001), SUVmean (p = 0.002) and T/B (p < 0.001); deceased patients (16/67) showed higher SUVmax (p = 0.03), SUVmean (p = 0.03) and T/B (p = 0.006). All static parameters were associated with PFS (all p < 0.001); T/B was associated with OS (p = 0.031). Regarding kinetic analyses, recurrence (27/46) and deceased (14/46) patients had higher SUVmaxTTP (p = 0.02, p = 0.01, respectively). SUVmaxTTP was the only dynamic parameter associated with PFS (p = 0.02) and OS (p = 0.006). At univariate analysis, SUVmax, SUVmean, T/B and SUVmaxTTP were predictive for PFS (all p < 0.05); SUVmaxTTP was predictive for OS (p = 0.02). At multivariate analysis, SUVmaxTTP remained significant for PFS (p = 0.03). Conclusion: Semiquantitative parameters and SUVmaxTTP were associated with clinical outcomes in patients with suspected glioma recurrence. Dynamic PET-CT acquisition, with static and kinetic parameters, can be a valuable non-invasive prognostic marker, identifying patients with worse prognosis who require personalised therapy.
Quantifying the Effects of Medical Examination and Possible Risk Factors against the Incidence of Cervical Cancer in a Low Human Papillomavirus Vaccination Coverage: An Ecological Study in Japan
Authors: Yu Yueming , Matsuyama Ryota , Tsunematsu Miwako , Kakehashi Masayuki
Published: September 2021

Cervical cancer (CC) is one of the most common gynecological malignancies in females, mainly caused by human papillomavirus (HPV). In countries with lower HPV vaccine coverage, such as Japan, medical examination may play a key role in decreasing CC incidence. This study aimed to quantify the effect of medical examination on cervical cancer (CC) incidence in Japan, considering the effects of possible risk factors. By collecting Japan’s Prefectural data on CC incidence (2013–2017), incidence of sexually transmitted diseases (STDs; Chlamydia, Herpes, Condyloma, and Gonorrhea; 1993–2012), screening and detailed examination rate against CC (2013–2016), smoking rate (2001–2013), economic status (disposable income and economic surplus; 2014–2015), and education status (2015), we analyzed associations among them using Pearson’s correlation coefficients. Additionally, assuming that the incidence of STDs reflects the frequency of risky sexual behavior at the co-infection point with HPV, we constructed generalized linear models to predict CC incidence, taking a 5–20-year time-lag between incidences of STDs and the CC incidence. Against CC incidence, Chlamydia in females and Gonorrhea in males with a 15-year time-lag showed positive associations, while Condyloma in both genders with a 15-year time-lag, screening rate, economic status, and smoking rate showed negative associations. An increase in screening test rate by 10% was estimated to decrease CC incidence by 9.6%. This means that screening tests decrease CC incidence effectively, but not drastically, suggesting the need for additional countermeasures for CC prevention.
Catalytic Oxidations with Meta-Chloroperoxybenzoic Acid (m-CPBA) and Mono- and Polynuclear Complexes of Nickel: A Mechanistic Outlook
Authors: Nesterov Dmytro S. , Nesterova Oksana V.
Published: September 2021

Selective catalytic functionalization of organic substrates using peroxides as terminal oxidants remains a challenge in modern chemistry. The high complexity of interactions between metal catalysts and organic peroxide compounds complicates the targeted construction of efficient catalytic systems. Among the members of the peroxide family, m-chloroperoxybenzoic acid (m-CPBA) exhibits quite complex behavior, where numerous reactive species could be formed upon reaction with a metal complex catalyst. Although m-CPBA finds plenty of applications in fine organic synthesis and catalysis, the factors that discriminate its decomposition routes under catalytic conditions are still poorly understood. The present review covers the advances in catalytic C–H oxidation and olefine epoxidation with m-CPBA catalyzed by mono- and polynuclear complexes of nickel, a cheap and abundant first-row transition metal. The reaction mechanisms are critically discussed, with special attention to the O–O bond splitting route. Selectivity parameters using recognized model hydrocarbon substrates are summarized and important factors that could improve further catalytic studies are outlined.
Entrapment of Phenylalanine Ammonia-Lyase in Nanofibrous Polylactic Acid Matrices by Emulsion Electrospinning
Authors: Koplányi Gábor , Sánta-Bell Evelin , Molnár Zsófia , Tóth Gergő Dániel , Józó Muriel , Szilágyi András , Ender Ferenc , Pukánszky Béla , Vértessy Beáta G. , Poppe László , Balogh-Weiser Diána
Published: September 2021

Immobilization of the recombinant, plant-derived Petroselinum crispum phenylalanine ammonia lyase (PcPAL) in electrospun matrices have the potential to create promising, easy-to-use biocatalysts. Polylactic acid (PLA) a biologically inert, commercial biopolymer, was chosen as the material of the carrier system. PLA could be electrospun properly only from water-immiscible organic solvents, which limits its application as a carrier of sensitive biological objects. The emulsion electrospinning is a proper solution to overcome this issue using non-ionic emulsifiers with different hydrophilic-lipophilic balance (HLB) values. The stabilized emulsion could protect the sensitive PcPAL dissolved in the aqueous buffer phase and improve fiber formation, plus help to keep the biocatalytic activity of enzymes. In this study, the first approach is described to produce PLA nanofibers containing PcPAL enzymes by emulsion electrospinning and to use the resulted biocatalyst in the ammonia elimination reaction from l-phenylalanine.
Structure-Function and Industrial Relevance of Bacterial Aminopeptidase P
Authors: Omar Muhamad Nadzmi , Raja Abd Rahman Raja Noor Zaliha , Muhd Noor Noor Dina , Latip Wahhida , Knight Victor Feizal , Ali Mohd Shukuri Mohamad
Published: September 2021

Aminopeptidase P (APPro, E.C 3.4.11.9) cleaves N-terminal amino acids from peptides and proteins where the penultimate residue is proline. This metal-ion-dependent enzyme shares a similar fold, catalytic mechanism, and substrate specificity with methionine aminopeptidase and prolidase. It adopts a canonical pita bread fold that serves as a structural basis for the metal-dependent catalysis and assembles as a tetramer in crystals. Similar to other metalloaminopeptidase, APPro requires metal ions for its maximal enzymatic activity, with manganese being the most preferred cation. Microbial aminopeptidase possesses unique characteristics compared with aminopeptidase from other sources, making it a great industrial enzyme for various applications. This review provides a summary of recent progress in the study of the structure and function of aminopeptidase P and describes its various applications in different industries as well as its significance in the environment.
Bioethanol Upgrading to Renewable Monomers Using Hierarchical Zeolites: Catalyst Preparation, Characterization, and Catalytic Studies
Authors: Iadrat Ploychanok , Wattanakit Chularat
Published: September 2021

Bioethanol is one of the most promising renewable resources for the production of important monomers. To date, there have been various processes proposed for bioethanol conversion to renewable monomers. In this review, the catalytic bioethanol upgrading to various types of monomers using hierarchical zeolites as catalysts is illustrated, including the recent design and preparation of hierarchical zeolites for these catalytic processes. The characterizations of catalysts including textural properties, pore architectures, acidic properties, and active species are also exemplified. Moreover, the catalytic studies with various processes of monomer production from bioethanol including bioethanol dehydration, bioethanol to hydrocarbons, and bioethanol to butadiene are revealed in terms of catalytic activities and mechanistic studies. In addition, the future perspectives of these catalytic circumstances are proposed in both economic and sustainable development contexts.
Meristematic Connectome: A Cellular Coordinator of Plant Responses to Environmental Signals?
Authors: Chiatante Donato , Montagnoli Antonio , Trupiano Dalila , Sferra Gabriella , Bryant John , Rost Thomas L. , Scippa Gabriella S.
Published: September 2021

Mechanical stress in tree roots induces the production of reaction wood (RW) and the formation of new branch roots, both functioning to avoid anchorage failure and limb damage. The vascular cambium (VC) is the factor responsible for the onset of these responses as shown by their occurrence when all primary tissues and the root tips are removed. The data presented confirm that the VC is able to evaluate both the direction and magnitude of the mechanical forces experienced before coordinating the most fitting responses along the root axis whenever and wherever these are necessary. The coordination of these responses requires intense crosstalk between meristematic cells of the VC which may be very distant from the place where the mechanical stress is first detected. Signaling could be facilitated through plasmodesmata between meristematic cells. The mechanism of RW production also seems to be well conserved in the stem and this fact suggests that the VC could behave as a single structure spread along the plant body axis as a means to control the relationship between the plant and its environment. The observation that there are numerous morphological and functional similarities between different meristems and that some important regulatory mechanisms of meristem activity, such as homeostasis, are common to several meristems, supports the hypothesis that not only the VC but all apical, primary and secondary meristems present in the plant body behave as a single interconnected structure. We propose to name this structure “meristematic connectome” given the possibility that the sequence of meristems from root apex to shoot apex could represent a pluricellular network that facilitates long-distance signaling in the plant body. The possibility that the “meristematic connectome” could act as a single structure active in adjusting the plant body to its surrounding environment throughout the life of a plant is now proposed.
Neutrophil Extracellular Traps Contribute to COVID-19 Hyperinflammation and Humoral Autoimmunity
Authors: Torres-Ruiz Jiram , Absalón-Aguilar Abdiel , Nuñez-Aguirre Miroslava , Pérez-Fragoso Alfredo , Carrillo-Vázquez Daniel Alberto , Maravillas-Montero José Luis , Mejía-Domínguez Nancy R. , Llorente Luis , Alcalá-Carmona Beatriz , Lira-Luna Jaquelin , Núñez-Álvarez Carlos , Juárez-Vega Guillermo , Meza-Sánchez David , Hernández-Gilsoul Thierry , Tapia-Rodríguez Miguel , Gómez-Martín Diana
Published: September 2021

The coronavirus disease 2019 (COVID-19) is related to enhanced production of NETs, and autoimmune/autoinflammatory phenomena. We evaluated the proportion of low-density granulocytes (LDG) by flow cytometry, and their capacity to produce NETs was compared with that of conventional neutrophils. NETs and their protein cargo were quantified by confocal microscopy and ELISA. Antinuclear antibodies (ANA), anti-neutrophil cytoplasmic antibodies (ANCA) and the degradation capacity of NETs were addressed in serum. MILLIPLEX assay was used to assess the cytokine levels in macrophages’ supernatant and serum. We found a higher proportion of LDG in severe and critical COVID-19 which correlated with severity and inflammatory markers. Severe/critical COVID-19 patients had higher plasmatic NE, LL-37 and HMGB1-DNA complexes, whilst ISG-15-DNA complexes were lower in severe patients. Sera from severe/critical COVID-19 patients had lower degradation capacity of NETs, which was reverted after adding hrDNase. Anti-NET antibodies were found in COVID-19, which correlated with ANA and ANCA positivity. NET stimuli enhanced the secretion of cytokines in macrophages. This study unveils the role of COVID-19 NETs as inducers of pro-inflammatory and autoimmune responses. The deficient degradation capacity of NETs may contribute to the accumulation of these structures and anti-NET antibodies are related to the presence of autoantibodies.
New Insights into the Pivotal Role of Iron/Heme Metabolism in TLR4/NF-κB Signaling-Mediated Inflammatory Responses in Human Monocytes
Authors: Kang Dong Young , Sp Nipin , Jo Eun Seong , Lee Jin-Moo , Jang Kyoung-Jin
Published: September 2021

Iron metabolism and heme biosynthesis are essential processes in cells during the energy cycle. Alteration in these processes could create an inflammatory condition, which results in tumorigenesis. Studies are conducted on the exact role of iron/heme metabolism in induced inflammatory conditions. This study used lipopolysaccharide (LPS)- or high-glucose-induced inflammation conditions in THP-1 cells to study how iron/heme metabolism participates in inflammatory responses. Here, we used iron and heme assays for measuring total iron and heme. We also used flow cytometry and Western blotting to analyze molecular responses. Our results demonstrated that adding LPS or high-glucose induced iron formation and heme synthesis and elevated the expression levels of proteins responsible for iron metabolism and heme synthesis. We then found that further addition of heme or 5-aminolevulinic acid (ALA) increased heme biosynthesis and promoted inflammatory responses by upregulating TLR4/NF-κB and inflammatory cytokine expressions. We also demonstrated the inhibition of heme synthesis using succinylacetone (SA). Moreover, N-MMP inhibited LPS- or high-glucose-induced inflammatory responses by inhibiting TLR4/NF-κB signaling. Hence, iron/heme metabolism checkpoints could be considered a target for treating inflammatory conditions.
Comparative Review of Microglia and Monocytes in CNS Phagocytosis
Authors: Andoh Megumi , Koyama Ryuta
Published: September 2021

Macrophages maintain tissue homeostasis by phagocytosing and removing unwanted materials such as dead cells and cell debris. Microglia, the resident macrophages of the central nervous system (CNS), are no exception. In addition, a series of recent studies have shown that microglia phagocytose the neuronal synapses that form the basis of neural circuit function. This discovery has spurred many neuroscientists to study microglia. Importantly, in the CNS parenchyma, not only microglia but also blood-derived monocytes, which essentially differentiate into macrophages after infiltration, exert phagocytic ability, making the study of phagocytosis in the CNS even more interesting and complex. In particular, in the diseased brain, the phagocytosis of tissue-damaging substances, such as myelin debris in multiple sclerosis (MS), has been shown to be carried out by both microglia and blood-derived monocytes. However, it remains largely unclear why blood-derived monocytes need to invade the parenchyma, where microglia are already abundant, to assist in phagocytosis. We will also discuss whether this phagocytosis can affect the fate of the phagocytosing cell itself as well as the substance being phagocytosed and the surrounding environment in addition to future research directions. In this review, we will introduce recent studies to answer a question that often arises when studying microglial phagocytosis: under what circumstances and to what extent blood-derived monocytes infiltrate the CNS and contribute to phagocytosis. In addition, the readers will learn how recent studies have experimentally distinguished between microglia and infiltrating monocytes. Finally, we aim to contribute to the progress of phagocytosis research by discussing the effects of phagocytosis on phagocytic cells.
Contractile Activity of Myotubes Derived from Human Induced Pluripotent Stem Cells: A Model of Duchenne Muscular Dystrophy
Authors: Yoshioka Kantaro , Ito Akira , Horie Masanobu , Ikeda Kazushi , Kataoka Sho , Sato Keiichiro , Yoshigai Taichi , Sakurai Hidetoshi , Hotta Akitsu , Kawabe Yoshinori , Kamihira Masamichi
Published: September 2021

Duchenne muscular dystrophy (DMD) is a genetic disorder that results from deficiency of the dystrophin protein. In recent years, DMD pathological models have been created using induced pluripotent stem (iPS) cells derived from DMD patients. In addition, gene therapy using CRISPR-Cas9 technology to repair the dystrophin gene has been proposed as a new treatment method for DMD. However, it is not known whether the contractile function of myotubes derived from gene-repaired iPS cells can be restored. We therefore investigated the maturation of myotubes in electrical pulse stimulation culture and examined the effect of gene repair by observing the contractile behaviour of myotubes. The contraction activity of myotubes derived from dystrophin-gene repaired iPS cells was improved by electrical pulse stimulation culture. The iPS cell method used in this study for evaluating muscle contractile activity is a useful technique for analysing the mechanism of hereditary muscular disease pathogenesis and for evaluating the efficacy of new drugs and gene therapy.
Synthesis and Characterization of Ce3+-Doped Ni0.5Cd0.5Fe2O4 Nanoparticles by Sol–Gel Auto-Combustion Method
Authors: Ahmad Danyal , Mehboob Nasir , Zaman Abid , Ahmed Nabeel , Ahmed Kashif , Mushtaq Muhammad , Althubeiti Khaled , Ali Asad , Sultana Fozia , Bashir Khalid
Published: September 2021

Cerium (Ce)-doped Ni0.5Cd0.5CexFe2−xO4 (0.0 ≤ x ≤ 0.20) was synthesized using the sol–gel auto-combustion method. X-ray diffraction (XRD) analysis revealed that all the samples retained spinel cubic crystal structure with space group Fd3m at 800 °C. Crystal structure parameters, such as lattice constant, average crystallite size, and X-ray density were estimated from the major XRD (311) peak. Bulk density and porosity were also calculated. The average crystallite size was estimated to be in the range of 20–24 nm. SEM images displayed agglomerated particles with a porous morphology. The dielectric constant (ε′) increased and the dielectric loss tangent (tanδ) decreased with rising Ce concentration. The hysteresis loop (M–H loop) was measured at room temperature using a vibrating-sample magnetometer (VSM), which showed a nonlinear decrease in magnetization and coercivity with increasing Ce concentration.
Experimental and Numerical Investigation of Load Failure at the Interface Joint of Repaired Potholes Using Hot Mix Asphalt with Steel Fiber Additive
Authors: Eisa Mohamed S. , Abdelhaleem Fahmy S. , Khater Vivian A.
Published: September 2021

The maintenance of potholes is a long-standing problem. Previous studies focused on pothole patching materials and methods but not on bonding at the interface joint. In this study, the influence of the patching shape and depth on the bonding at the interface joint using two patching materials: hot mix asphalt (HMA) and hot mix asphalt containing 5% (by volume) steel fiber (HMA+) was investigated. Slabs with circular and square potholes in the middle with different depths (35, 50 and 70 mm) were prepared. The two shapes of potholes were patched with two patching materials: HMA and HMA+, at different depths. The slabs were tested after patching using a rigid steel frame. The experimental results were compared with those obtained from finite element analysis using the ABAQUS software, applying the same model of slabs with the same dimensions and properties of the materials used. The results indicated that the bonding at joint interface for circular-patched potholes slightly improved using HMA+ and this was independent of patching depth. As for the square-patched potholes, the bonding at the interface joint was better than for the circular-patched ones; the bonding increased with increasing depth. Using HMA+ for patching the square-patched potholes, the bonding at the interface joint slightly increased, only for the 3.5 cm depth.
Effect of Cross-Linking Modification on Structural and Film-Forming Characteristics of Pearl Millet (Pennisetum glaucum L.) Starch
Authors: Siroha Anil Kumar , Bangar Sneh Punia , Sandhu Kawaljit Singh , Trif Monica , Kumar Manoj , Guleria Prixit
Published: September 2021

Pearl millet starch was modified using epichlorohydrin (EPI) at different concentrations (0.1%; 0.3%; 0.5%; and 0.8%) and evaluated for physicochemical, rheological, in vitro digestibility, and film-forming characteristics. The degree of cross-linking was observed at higher levels (0.5% and 0.8%) of EPI. Upon cross-linking, breakdown and setback viscosity reduced whereas pasting temperature was increased. Storage modulus (G′) and loss modulus (G″) value of cross-linked (CL) starches ranged between 2877 to 5744 Pa and 168 to 237 Pa, respectively, during the frequency sweep test. A drastic decrease was observed for steady shear (yield stress and consistency index) characteristics of CL starches. Resistant starch (RS) content was increased after starch modification, which imparts its nutritional values and starch modified at 0.8% had the highest RS content. Modifications of starch at different levels had significant effects on the moisture, opacity, solubility and mechanical properties of films. Outcomes of this study will be helpful to understand the properties of native and CL starches for their potential applications in preparation of edible films.
Enhanced Properties of Micro Arc Oxidation Coating with Cu Addition on TC4 Alloy in Marine Environment
Authors: Gao Wei , Liu Jiangnan , Wei Jingpeng , Yao Yuhong , Ma Xiqun , Yang Wei
Published: September 2021

By contrast with the traditional method of adding hard particles into micro arc oxidation (MAO) coating to improve its wear performance, this study introduced copper into the MAO coating on TC4 alloy by adding copper pyrophosphate to enhance the wear property in a marine environment and the antibacterial property. The results demonstrated that the MAO coating with copper pyrophosphate addition showed a porous structure, and Cu was mainly concentrated around micropores. CuO and Cu2O were formed in this MAO coating. This MAO coating with Cu had a high bonding strength to the substrate. Although the hardness of the coating with Cu had been reduced, it could reduce the friction coefficient and enhance the wear property in simulated seawater due to the lubrication of Cu. Furthermore, this MAO coating with Cu addition had obvious antibacterial and bactericidal effects due to the antibacterial effect of Cu.
Strength, Fracture and Durability Characteristics of Ambient Cured Alkali—Activated Mortars Incorporating High Calcium Industrial Wastes and Powdered Reagents
Authors: Sood Dhruv , Hossain Khandaker M. A.
Published: September 2021

Alkali-activated mortars (AAMs) are developed incorporating binary/ternary combinations of industrial wastes comprising of fly ash class C (FA-C), fly ash class F (FA-F) and ground granulated blast furnace slag (GGBFS) with alkaline reagents and silica sand. The use of high calcium precursors, calcium-based powder form reagents, dry mixing method, and ambient curing with performance characterization based on chemical ratios and fracture properties are some novel aspects of the study. The mechanical (dry density, compressive strength, ultrasonic pulse velocity, elastic modulus, fracture/crack tip toughness and fracture energy), durability (shrinkage/expansion and mass change in water and ambient curing conditions, water absorption and freeze-thaw resistance) and microstructural (SEM/EDS and XRD analyses) characteristics of eight AAMs are investigated. The binary (FA-C + GGBFS) mortars obtained higher compressive strengths (between 35 MPa and 42.6 MPa), dry densities (between 2032 kg/m3 and 2088 kg/m3) and ultrasonic pulse velocities (between 3240 m/s and 4049 m/s) than their ternary (FA-C + FA-F + GGBFS) counterparts. The elastic modulus and fracture toughness for mortars incorporating reagent 2 (calcium hydroxide: sodium sulphate = 2.5:1) were up to 1.7 and five times higher than those with reagent 1 (calcium hydroxide: sodium metasilicate = 1:2.5). This can be attributed to the additional formation of C-S-H with C-A-S-H/N-C-A-S-H binding phases in mortars with reagent 2. Ternary mortars exhibited comparatively lower shrinkage/expansion and initial sorptivity indices than their binary counterparts due to the lower geopolymerisation potential of fly ash class F that facilitated the reduction of matrix porosity. All mortar specimens demonstrated 100% or more relative dynamic modulus of elasticity after 60 freeze-thaw cycles, indicating the damage recovery and satisfactory durability due to probable micro-level re-arrangement of the binding phases. This study confirmed the viability of producing cement-free AAMs with satisfactory mechanical and durability characteristics.
Comprehensive Genomic Profiling of Circulating Cell-Free DNA Distinguishes Focal MET Amplification from Aneuploidy in Diverse Advanced Cancers
Authors: Kumaki Yuichi , Olsen Steve , Suenaga Mitsukuni , Nakagawa Tsuyoshi , Uetake Hiroyuki , Ikeda Sadakatsu
Published: September 2021

Amplification (amp) of MET can be observed in cases of focal gene copy number gain, such as MET-driven amp, or with a gain of chromosome 7, such as aneuploidy. Several studies have shown that only high-level focal MET amp (MET/CEP7 ratio ≥5) is oncogenic, with such tumors responding to targeted therapy. However, there are few reports on how to distinguish between focal amplification and aneuploidy using next-generation sequencing (NGS). A total of 1025 patients with advanced solid tumors (typically pre-treated) were tested with a non-invasive comprehensive cfDNA NGS panel (Guardant360) from July 2014 to June 2019. Since bioinformatics upgrades of Guardant360 were undergoing in September 2018, focal MET amp was determined by our independent algorithm using the cohorts tested before September 2018 (291 patients), and validation was performed in the remaining cohort (734 patients). MET alterations (alts) associated with aberrant signaling were found in 110 patients (10.7%) among nine different cancer types, most commonly in non-small cell (12.2%, 62/510) and small cell (33.3%, 3/9) lung cancers, gastroesophageal cancer (19.4%, 7/36), and prostate adenocarcinoma (15.6%; 5/32). Among 291 patients tested before September 2018, 37 (12.7%) had MET alts. Among these, 24 (64.9%) had amps, 5 (13.5%) had exon 14 skipping, and 13 (35.1%) had single nucleotide variants (SNVs). Co-alterations, such as amp + SNVs, were found in four samples (10.8%). Among 24 MET amps, 29.2% (7/24) were focal according to our algorithm. MET copy number was significantly higher with focal amp compared to non-focal amp (mean copy number 3.26 vs. 2.44, respectively, p = 0.00304). In 734 patients tested after September 2018, our definition of focal MET amp was detected in 4.2% (31/734). Overall, focal amplification based on our algorithm was 3.7% (=38/1025). This study describes an approach to distinguish focal and non-focal MET amplification using comprehensive genomic profiling of cfDNA in advanced cancer patients. Focal MET amp accounted for ~30% of all MET amp, which was found in 3.7% of patients with diverse cancers and was associated with a higher plasma copy number. Clinical studies are warranted to assess the clinical utility of targeted therapies for tumors with focal MET amplification detected by NGS of cfDNA.
Balancing Speed and Accuracy in Cardiac Magnetic Resonance Function Post-Processing: Comparing 2 Levels of Automation in 3 Vendors to Manual Assessment
Authors: Snel Gert J.H. , Poort Sharon , Velthuis Birgitta K. , van Deursen Vincent M. , Nguyen Christopher T. , Sosnovik David , Dierckx Rudi A.J.O. , Slart Riemer H.J.A. , Borra Ronald J.H. , Prakken Niek H.J.
Published: September 2021

Automating cardiac function assessment on cardiac magnetic resonance short-axis cines is faster and more reproducible than manual contour-tracing; however, accurately tracing basal contours remains challenging. Three automated post-processing software packages (Level 1) were compared to manual assessment. Subsequently, automated basal tracings were manually adjusted using a standardized protocol combined with software package-specific relative-to-manual standard error correction (Level 2). All post-processing was performed in 65 healthy subjects. Manual contour-tracing was performed separately from Level 1 and 2 automated analysis. Automated measurements were considered accurate when the difference was equal or less than the maximum manual inter-observer disagreement percentage. Level 1 (2.1 ± 1.0 min) and Level 2 automated (5.2 ± 1.3 min) were faster and more reproducible than manual (21.1 ± 2.9 min) post-processing, the maximum inter-observer disagreement was 6%. Compared to manual, Level 1 automation had wide limits of agreement. The most reliable software package obtained more accurate measurements in Level 2 compared to Level 1 automation: left ventricular end-diastolic volume, 98% and 53%; ejection fraction, 98% and 60%; mass, 70% and 3%; right ventricular end-diastolic volume, 98% and 28%; ejection fraction, 80% and 40%, respectively. Level 1 automated cardiac function post-processing is fast and highly reproducible with varying accuracy. Level 2 automation balances speed and accuracy.
Diagnostic Performance of KLCA-NCC 2018 Criteria for Hepatocellular Carcinoma Using Magnetic Resonance Imaging: A Systematic Review and Meta-Analysis
Authors: Kim Dong Hwan , Kim Bohyun , Youn Seo Yeon , Kim Hokun , Choi Joon-Il
Published: September 2021

Several imaging-based systems have been proposed for the diagnosis of hepatocellular carcinoma (HCC) using magnetic resonance imaging (MRI), reflecting geographical differences in the clinical environment for HCC. We conducted a systematic review and meta-analysis to determine the performance of the Korean Liver Cancer Association-National Cancer Center (KLCA-NCC) 2018 criteria for the MRI diagnosis of HCC. Original studies reporting the performance of the KLCA-NCC 2018 criteria for the diagnosis of HCC using MRI were identified in MEDLINE and EMBASE until 29 March 2021. The meta-analytic pooled sensitivity and specificity of the KLCA-NCC 2018 criteria for diagnosing HCC were calculated using a bivariate random-effects model. A meta-regression analysis was performed to explore study heterogeneity further. Eight studies involving 1690 HCCs reported the accuracy of the KLCA-NCC 2018 imaging criteria. The pooled sensitivity and specificity of the definite HCC criteria for diagnosing HCC were 81% (95% confidence interval, 76–85%; I2 = 86%) and 90% (86–93%; I2 = 23%), respectively. For five available studies, the pooled sensitivity and specificity of the definite HCC criteria for diagnosing HCCs smaller than 20 mm were 80% (72–86%; I2 = 76%) and 91% (86–94%; I2 = 0%), respectively. A considerable threshold effect with a correlation coefficient of 0.667 was observed. The results of the meta-regression analysis revealed that the accuracy of the definite HCC criteria differed significantly depending on the type of MRI contrast agent (p = 0.01). In conclusion, the KLCA-NCC 2018 criteria had good overall diagnostic performance in diagnosing HCC. Substantial study heterogeneity was observed for sensitivity, which was significantly influenced by the type of contrast agent and by a threshold effect.
The Use of Saliva as a Biosample in the Light of COVID-19
Authors: Duś-Ilnicka Irena , Krala Elżbieta , Cholewińska Paulina , Radwan-Oczko Małgorzata
Published: September 2021

Saliva is easy to collect and a biofluid that is readily available without the need for special equipment for its collection. The collection process, which is non-invasive and inexpensive, leads to obtaining a biomaterial that can serve as a source of information for molecular diagnostics of diseases in general medicine, genetics and dentistry. Unfortunately, many of the salivary methodologies are lacking important parameters to provide for not only the safety of the operator, but also the quality and reproducibility of the research. Since the COVID-19 pandemic, salivary diagnostics demonstrate a great potential for research of SARS-CoV 2. In this review, good practice for unstimulated saliva collection and patient preparation was provided, based on the latest literature and available guidelines. Schemes for saliva collection procedures were presented following an extended literature search. Descriptions of salivary probes/cups, techniques of saliva collection, and the use of specific buffering solutions for the stability of collected samples for SARS-CoV-2 detection were also evaluated.
Depth of Stromal Invasion as the Most Prognostically Relevant Regression System in Locally Advanced Cervical Cancer after Neoadjuvant Treatment: A Systematic Review and Meta-Analysis Grading
Authors: Zannoni Gian Franco , Travaglino Antonio , Raffone Antonio , Arciuolo Damiano , D’Alessandris Nicoletta , Scaglione Giulia , Tralongo Pietro , Inzani Frediano , Angelico Giuseppe , Santoro Angela
Published: September 2021

Background: several different criteria have been proposed to categorize the pathological response in cervical cancer after neoadjuvant therapy; although it is unclear what the most prognostically valuable one is. Objective: to assess the prognostic value of pathological criteria for categorizing the response in cervical cancer after neoadjuvant therapy, through a systematic review and meta-analysis. Methods: four electronic databases were searched from January to December 2020 for all studies, assessing the prognostic value of pathological response in cervical cancer after neoadjuvant therapy. Hazard ratio (HR) for overall survival (OS) was calculated with a significant p-value < 0.05. A meta-analysis was performed for each criteria assessed in at least three studies. Results: sixteen studies were included. Criteria for pathological response included (i) residual stromal invasion < vs. >3 mm; (ii) complete response vs. any residual; (iii) proportion of viable cells; (iv) residual tumor diameter; and (v) intracervical vs. extracervical residual. Criteria (i) and (ii) were suitable for meta-analysis. The presence of a residual tumor with stromal invasion > 3 mm showed a HR of 4.604 (95% CI; 3.229–6.565; p < 0.001), while the presence of any residual showed a HR of 1.610 (95% CI; 1.245–2.081; p < 0.001); statistical heterogeneity was absent in both analyses. Conclusions: dichotomizing the pathological response in cervical cancer after neoadjuvant therapy as < vs. >3 mm stromal invasion is more prognostically valuable than dichotomizing as complete response vs. any residual. Further studies are necessary to evaluate other systems.