bluuebunny/crossref_metadata_2025 huggingface dataset:
Equine Chorionic Gonadotropin as an Effective FSH Replacement for In Vitro Ovine Follicle and Oocyte Development
Authors: Di Berardino Chiara , Peserico Alessia , Capacchietti Giulia , Crociati Martina , Monaci Maurizio , Tosi Umberto , Mauro Annunziata , Russo Valentina , Bernabò Nicola , Barboni Barbara
Published: November 2021

The use of assisted reproductive technologies (ART) still requires strategies through which to maximize individual fertility chances. In vitro folliculogenesis (ivF) may represent a valid option to convey the large source of immature oocytes in ART. Several efforts have been made to set up ivF cultural protocols in medium-sized mammals, starting with the identification of the most suitable gonadotropic stimulus. In this study, Equine Chorionic Gonadotropin (eCG) is proposed as an alternative to Follicle Stimulating Hormone (FSH) based on its long superovulation use, trans-species validation, long half-life, and low costs. The use of 3D ivF on single-ovine preantral (PA) follicles allowed us to compare the hormonal effects and to validate their influence under two different cultural conditions. The use of eCG helped to stimulate the in vitro growth of ovine PA follicles by maximizing its influence under FBS-free medium. Higher performance of follicular growth, antrum formation, steroidogenic activity and gap junction marker expression were recorded. In addition, eCG, promoted a positive effect on the germinal compartment, leading to a higher incidence of meiotic competent oocytes. These findings should help to widen the use of eCG to ivF as a valid and largely available hormonal support enabling a synchronized in vitro follicle and oocyte development.
Drosophila Solute Carrier 5A5 Regulates Systemic Glucose Homeostasis by Mediating Glucose Absorption in the Midgut
Authors: Li Yue , Wang Weidong , Lim Hui-Ying
Published: November 2021

The small intestine is the initial site of glucose absorption and thus represents the first of a continuum of events that modulate normal systemic glucose homeostasis. A better understanding of the regulation of intestinal glucose transporters is therefore pertinent to our efforts in curbing metabolic disorders. Using molecular genetic approaches, we investigated the role of Drosophila Solute Carrier 5A5 (dSLC5A5) in regulating glucose homeostasis by mediating glucose uptake in the fly midgut. By genetically knocking down dSLC5A5 in flies, we found that systemic and circulating glucose and trehalose levels are significantly decreased, which correlates with an attenuation in glucose uptake in the enterocytes. Reciprocally, overexpression of dSLC5A5 significantly increases systemic and circulating glucose and trehalose levels and promotes glucose uptake in the enterocytes. We showed that dSLC5A5 undergoes apical endocytosis in a dynamin-dependent manner, which is essential for glucose uptake in the enterocytes. Furthermore, we showed that the dSLC5A5 level in the midgut is upregulated by glucose and that dSLC5A5 critically directs systemic glucose homeostasis on a high-sugar diet. Together, our studies have uncovered the first Drosophila glucose transporter in the midgut and revealed new mechanisms that regulate glucose transporter levels and activity in the enterocyte apical membrane.
Heterologous Expression of the Melatonin-Related Gene HIOMT Improves Salt Tolerance in Malus domestica
Authors: Tan Kexin , Zheng Jiangzhu , Liu Cheng , Liu Xianghan , Liu Xiaomin , Gao Tengteng , Song Xinyang , Wei Zhiwei , Ma Fengwang , Li Chao
Published: November 2021

Melatonin, a widely known indoleamine molecule that mediates various animal and plant physiological processes, is formed from N-acetyl serotonin via N-acetylserotonin methyltransferase (ASMT). ASMT is an enzyme that catalyzes melatonin synthesis in plants in the rate-determining step and is homologous to hydroxyindole-O-methyltransferase (HIOMT) melatonin synthase in animals. To date, little is known about the effect of HIOMT on salinity in apple plants. Here, we explored the melatonin physiological function in the salinity condition response by heterologous expressing the homologous human HIOMT gene in apple plants. We discovered that the expression of melatonin-related gene (MdASMT) in apple plants was induced by salinity. Most notably, compared with the wild type, three transgenic lines indicated higher melatonin levels, and the heterologous expression of HIOMT enhanced the expression of melatonin synthesis genes. The transgenic lines showed reduced salt damage symptoms, lower relative electrolyte leakage, and less total chlorophyll loss from leaves under salt stress. Meanwhile, through enhanced activity of antioxidant enzymes, transgenic lines decreased the reactive oxygen species accumulation, downregulated the expression of the abscisic acid synthesis gene (MdNCED3), accordingly reducing the accumulation of abscisic acid under salt stress. Both mechanisms regulated morphological changes in the stomata synergistically, thereby mitigating damage to the plants’ photosynthetic ability. In addition, transgenic plants also effectively stabilized their ion balance, raised the expression of salt stress–related genes, as well as alleviated osmotic stress through changes in amino acid metabolism. In summary, heterologous expression of HIOMT improved the adaptation of apple leaves to salt stress, primarily by increasing melatonin concentration, maintaining a high photosynthetic capacity, reducing reactive oxygen species accumulation, and maintaining normal ion homeostasis.
Tryptophan Side-Chain Oxidase Enzyme Suppresses Hepatocellular Carcinoma Growth through Degradation of Tryptophan
Authors: Ai Yang , Wang Ben , Xiao Shuai , Luo Sang , Wang Yefu
Published: November 2021

Tryptophan metabolism plays a role in the occurrence and development of hepatocellular carcinoma cells. By degrading certain amino acids, tumor growth can be limited while maintaining the body’s normal nutritional requirements. Tryptophan side-chain oxidase (TSO) enzyme can degrade tryptophan, and its inhibitory effect on hepatocellular carcinoma cells is worthy of further study. To investigate the degradation effect on tryptophan, TSO was isolated and purified from qq Pseudomonas. The reaction products were identified with high performance liquid chromatography (HPLC) and high-performance liquid chromatography tandem mass spectrometry (HPLC-MS). De novo sequencing provided the complete amino acid sequence of TSO. The results of CCK-8, colony formation, transwell, and qPCR confirmed that TSO had inhibitory effects on the proliferation and migration of HCCLM3 (human hepatocarcinoma cell line) and HepG2 cells. The results of flow cytometry confirmed its apoptotic activity. In animal experiments, we found that the tumor-suppressive effect was better in the oncotherapy group than the intraperitoneal injection group. The results of immunohistochemistry also suggested that TSO could inhibit proliferation and promote apoptosis. In conclusion, a specific enzyme that can degrade tryptophan and inhibit the growth of hepatoma cells was authenticated, and its basic information was obtained by extraction/purification and amino acid sequencing.
Aristolochic Acid Induces Renal Fibrosis and Senescence in Mice
Authors: Urate Shingo , Wakui Hiromichi , Azushima Kengo , Yamaji Takahiro , Suzuki Toru , Abe Eriko , Tanaka Shohei , Taguchi Shinya , Tsukamoto Shunichiro , Kinguchi Sho , Uneda Kazushi , Kanaoka Tomohiko , Atobe Yoshitoshi , Funakoshi Kengo , Yamashita Akio , Tamura Kouichi
Published: November 2021

The kidney is one of the most susceptible organs to age-related impairments. Generally, renal aging is accompanied by renal fibrosis, which is the final common pathway of chronic kidney diseases. Aristolochic acid (AA), a nephrotoxic agent, causes AA nephropathy (AAN), which is characterized by progressive renal fibrosis and functional decline. Although renal fibrosis is associated with renal aging, whether AA induces renal aging remains unclear. The aim of the present study is to investigate the potential use of AAN as a model of renal aging. Here, we examined senescence-related factors in AAN models by chronically administering AA to C57BL/6 mice. Compared with controls, the AA group demonstrated aging kidney phenotypes, such as renal atrophy, renal functional decline, and tubulointerstitial fibrosis. Additionally, AA promoted cellular senescence specifically in the kidneys, and increased renal p16 mRNA expression and senescence-associated β-galactosidase activity. Furthermore, AA-treated mice exhibited proximal tubular mitochondrial abnormalities, as well as reactive oxygen species accumulation. Klotho, an antiaging gene, was also significantly decreased in the kidneys of AA-treated mice. Collectively, the results of the present study indicate that AA alters senescence-related factors, and that renal fibrosis is closely related to renal aging.
In Vivo Renin Activity Imaging in the Kidney of Progeroid Ercc1 Mutant Mice
Authors: van Thiel Bibi S. , van der Linden Janette , Ridwan Yanto , Garrelds Ingrid M. , Vermeij Marcel , Clahsen-van Groningen Marian C. , Qadri Fatimunnisa , Alenina Natalia , Bader Michael , Roks Anton J. M. , Danser A. H. Jan , Essers Jeroen , van der Pluijm Ingrid
Published: November 2021

Changes in the renin–angiotensin system, known for its critical role in the regulation of blood pressure and sodium homeostasis, may contribute to aging and age-related diseases. While the renin–angiotensin system is suppressed during aging, little is known about its regulation and activity within tissues. However, this knowledge is required to successively treat or prevent renal disease in the elderly. Ercc1 is involved in important DNA repair pathways, and when mutated causes accelerated aging phenotypes in humans and mice. In this study, we hypothesized that unrepaired DNA damage contributes to accelerated kidney failure. We tested the use of the renin-activatable near-infrared fluorescent probe ReninSense680™ in progeroid Ercc1d/− mice and compared renin activity levels in vivo to wild-type mice. First, we validated the specificity of the probe by detecting increased intrarenal activity after losartan treatment and the virtual absence of fluorescence in renin knock-out mice. Second, age-related kidney pathology, tubular anisokaryosis, glomerulosclerosis and increased apoptosis were confirmed in the kidneys of 24-week-old Ercc1d/− mice, while initial renal development was normal. Next, we examined the in vivo renin activity in these Ercc1d/− mice. Interestingly, increased intrarenal renin activity was detected by ReninSense in Ercc1d/− compared to WT mice, while their plasma renin concentrations were lower. Hence, this study demonstrates that intrarenal RAS activity does not necessarily run in parallel with circulating renin in the aging mouse. In addition, our study supports the use of this probe for longitudinal imaging of altered RAS signaling in aging.
CNGB3 Missense Variant Causes Recessive Achromatopsia in Original Braunvieh Cattle
Authors: Häfliger Irene M. , Marchionatti Emma , Stengård Michele , Wolf-Hofstetter Sonja , Paris Julia M. , Jacinto Joana G. P. , Watté Christine , Voelter Katrin , Occelli Laurence M. , Komáromy András M. , Oevermann Anna , Goepfert Christine , Borgo Angelica , Roduit Raphaël , Spengeler Mirjam , Seefried Franz R. , Drögemüller Cord
Published: November 2021

Sporadic occurrence of inherited eye disorders has been reported in cattle but so far pathogenic variants were found only for rare forms of cataract but not for retinopathies. The aim of this study was to characterize the phenotype and the genetic aetiology of a recessive form of congenital day-blindness observed in several cases of purebred Original Braunvieh cattle. Electroretinography in an affected calf revealed absent cone-mediated function, whereas the rods continue to function normally. Brain areas involved in vision were morphologically normal. When targeting cones by immunofluorescence, a decrease in cone number and an accumulation of beta subunits of cone cyclic-nucleotide gated channel (CNGB3) in the outer plexiform layer of affected animals was obvious. Achromatopsia is a monogenic Mendelian disease characterized by the loss of cone photoreceptor function resulting in day-blindness, total color-blindness, and decreased central visual acuity. After SNP genotyping and subsequent homozygosity mapping with twelve affected cattle, we performed whole-genome sequencing and variant calling of three cases. We identified a single missense variant in the bovine CNGB3 gene situated in a ~2.5 Mb homozygous genome region on chromosome 14 shared between all cases. All affected cattle were homozygous carriers of the p.Asp251Asn mutation that was predicted to be deleterious, affecting an evolutionary conserved residue. In conclusion, we have evidence for the occurrence of a breed-specific novel CNGB3-related form of recessively inherited achromatopsia in Original Braunvieh cattle which we have designated OH1 showing an allele frequency of the deleterious allele of ~8%. The identification of carriers will enable selection against this inherited disorder. The studied cattle might serve as an animal model to further elucidate the function of CNGB3 in mammals.
Global Analysis of Transcriptome and Translatome Revealed That Coordinated WNT and FGF Regulate the Carapacial Ridge Development of Chinese Soft-Shell Turtle
Authors: Zhang Jun , Yu Peng , Zhao Yang , Zhou Qinyan , Yang Jiayu , Hu Qingtao , Liu Tiantian , Bao Chuanhe , Su Shiping , Gui Jian-Fang
Published: November 2021

The turtle carapace is composed of severely deformed fused dorsal vertebrae, ribs, and bone plates. In particular, the lateral growth in the superficial layer of turtle ribs in the dorsal trunk causes an encapsulation of the scapula and pelvis. The recent study suggested that the carapacial ridge (CR) is a new model of epithelial–mesenchymal transition which is essential for the arrangement of the ribs. Therefore, it is necessary to explore the regulatory mechanism of carapacial ridge development to analyze the formation of the turtle shell. However, the current understanding of the regulatory network underlying turtle carapacial ridge development is poor due to the lack of both systematic gene screening at different carapacial ridge development stages and gene function verification studies. In this study, we obtained genome-wide gene transcription and gene translation profiles using RNA sequencing and ribosome nascent-chain complex mRNA sequencing from carapacial ridge tissues of Chinese soft-shell turtle at different development stages. A correlation analysis of the transcriptome and translatome revealed that there were 129, 670, and 135 codifferentially expressed genes, including homodirection and opposite-direction differentially expressed genes, among three comparison groups, respectively. The pathway enrichment analysis of codifferentially expressed genes from the Kyoto Encyclopedia of Genes and Genomes showed dynamic changes in signaling pathways involved in carapacial ridge development. Especially, the results revealed that the Wnt signaling pathway and MAPK signaling pathway may play important roles in turtle carapacial ridge development. In addition, Wnt and Fgf were expressed during the carapacial ridge development. Furthermore, we discovered that Wnt5a regulated carapacial ridge development through the Wnt5a/JNK pathway. Therefore, our studies uncover that the morphogenesis of the turtle carapace might function through the co-operation between conserved WNT and FGF signaling pathways. Consequently, our findings revealed the dynamic signaling pathways acting on the carapacial ridge development of Chinese soft-shell turtle and provided new insights into uncover the molecular mechanism underlying turtle shell morphogenesis.
Iron Homeostasis Disorder and Alzheimer’s Disease
Authors: Peng Yu , Chang Xuejiao , Lang Minglin
Published: November 2021

Iron is an essential trace metal for almost all organisms, including human; however, oxidative stress can easily be caused when iron is in excess, producing toxicity to the human body due to its capability to be both an electron donor and an electron acceptor. Although there is a strict regulation mechanism for iron homeostasis in the human body and brain, it is usually inevitably disturbed by genetic and environmental factors, or disordered with aging, which leads to iron metabolism diseases, including many neurodegenerative diseases such as Alzheimer’s disease (AD). AD is one of the most common degenerative diseases of the central nervous system (CNS) threatening human health. However, the precise pathogenesis of AD is still unclear, which seriously restricts the design of interventions and treatment drugs based on the pathogenesis of AD. Many studies have observed abnormal iron accumulation in different regions of the AD brain, resulting in cognitive, memory, motor and other nerve damages. Understanding the metabolic balance mechanism of iron in the brain is crucial for the treatment of AD, which would provide new cures for the disease. This paper reviews the recent progress in the relationship between iron and AD from the aspects of iron absorption in intestinal cells, storage and regulation of iron in cells and organs, especially for the regulation of iron homeostasis in the human brain and prospects the future directions for AD treatments.
Beyond HAT Adaptor: TRRAP Liaisons with Sp1-Mediated Transcription
Authors: Yin Bo-Kun , Wang Zhao-Qi
Published: November 2021

The members of the phosphatidylinositol 3-kinase-related kinase (PIKK) family play vital roles in multiple biological processes, including DNA damage response, metabolism, cell growth, mRNA decay, and transcription. TRRAP, as the only member lacking the enzymatic activity in this family, is an adaptor protein for several histone acetyltransferase (HAT) complexes and a scaffold protein for multiple transcription factors. TRRAP has been demonstrated to regulate various cellular functions in cell cycle progression, cell stemness maintenance and differentiation, as well as neural homeostasis. TRRAP is known to be an important orchestrator of many molecular machineries in gene transcription by modulating the activity of some key transcription factors, including E2F1, c-Myc, p53, and recently, Sp1. This review summarizes the biological and biochemical studies on the action mode of TRRAP together with the transcription factors, focusing on how TRRAP-HAT mediates the transactivation of Sp1-governing biological processes, including neurodegeneration.
Keratins as an Inflammation Trigger Point in Epidermolysis Bullosa Simplex
Authors: Evtushenko Nadezhda A. , Beilin Arkadii K. , Kosykh Anastasiya V. , Vorotelyak Ekaterina A. , Gurskaya Nadya G.
Published: November 2021

Epidermolysis bullosa simplex (EBS) is a group of inherited keratinopathies that, in most cases, arise due to mutations in keratins and lead to intraepidermal ruptures. The cellular pathology of most EBS subtypes is associated with the fragility of the intermediate filament network, cytolysis of the basal layer of the epidermis, or attenuation of hemidesmosomal/desmosomal components. Mutations in keratins 5/14 or in other genes that encode associated proteins induce structural disarrangements of different strengths depending on their locations in the genes. Keratin aggregates display impaired dynamics of assembly and diminished solubility and appear to be the trigger for endoplasmic reticulum (ER) stress upon being phosphorylated by MAPKs. Global changes in cellular signaling mainly occur in cases of severe dominant EBS mutations. The spectrum of changes initiated by phosphorylation includes the inhibition of proteasome degradation, TNF-α signaling activation, deregulated proliferation, abnormal cell migration, and impaired adherence of keratinocytes. ER stress also leads to the release of proinflammatory danger-associated molecular pattern (DAMP) molecules, which enhance avalanche-like inflammation. Many instances of positive feedback in the course of cellular stress and the development of sterile inflammation led to systemic chronic inflammation in EBS. This highlights the role of keratin in the maintenance of epidermal and immune homeostasis.
Biochemical Characterisation of Human Transglutaminase 4
Authors: Csobán-Szabó Zsuzsa , Bécsi Bálint , El Alaoui Saïd , Fésüs László , Korponay-Szabó Ilma Rita , Király Róbert
Published: November 2021

Transglutaminases are protein-modifying enzymes involved in physiological and pathological processes with potent therapeutic possibilities. Human TG4, also called prostate transglutaminase, is involved in the development of autoimmune and tumour diseases. Although rodent TG4 is well characterised, biochemical characteristics of human TG4 that could help th e understanding of its way of action are not published. First, we analysed proteomics databases and found that TG4 protein is present in human tissues beyond the prostate. Then, we studied in vitro the transamidase activity of human TG4 and its regulation using the microtitre plate method. Human TG4 has low transamidase activity which prefers slightly acidic pH and a reducing environment. It is enhanced by submicellar concentrations of SDS suggesting that membrane proximity is an important regulatory event. Human TG4 does not bind GTP as tested by GTP-agarose and BODIPY-FL-GTPγS binding, and its proteolytic activation by dispase or when expressed in AD-293 cells was not observed either. We identified several potential human TG4 glutamine donor substrates in the AD-293 cell extract by biotin-pentylamine incorporation and mass spectrometry. Several of these potential substrates are involved in cell–cell interaction, adhesion and proliferation, suggesting that human TG4 could become an anticancer therapeutic target.
Molecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile Brassica napus
Authors: Xiao Qing , Wang Huadong , Chen Hui , Chen Xiaohan , Wen Jing , Dai Cheng , Ma Chaozhi , Tu Jinxing , Shen Jinxiong , Fu Tingdong , Yi Bin
Published: November 2021

Temperature-sensitive male sterility is a heritable agronomic trait affected by genotype-environment interactions. In rapeseed (Brassica napus), Polima (pol) temperature-sensitive cytoplasmic male sterility (TCMS) is commonly used for two-line breeding, as the fertility of pol TCMS lines can be partially restored at certain temperatures. However, little is known about the underlying molecular mechanism that controls fertility restoration. Therefore, we aimed to investigate the fertility conversion mechanism of the pol TCMS line at two different ambient temperatures (16 °C and 25 °C). Our results showed that the anthers developed and produced vigorous pollen at 16 °C but not at 25 °C. In addition, we identified a novel co-transcript of orf224-atp6 in the mitochondria that might lead to fertility conversion of the pol TCMS line. RNA-seq analysis showed that 1637 genes were significantly differentially expressed in the fertile flowers of 596-L when compared to the sterile flower of 1318 and 596-H. Detailed analysis revealed that differentially expressed genes were involved in temperature response, ROS accumulation, anther development, and mitochondrial function. Single-molecule long-read isoform sequencing combined with RNA sequencing revealed numerous genes produce alternative splicing transcripts at high temperatures. Here, we also found that alternative oxidase, type II NAD(P)H dehydrogenases, and transcription factor Hsfs might play a crucial role in male fertility under the low-temperature condition. RNA sequencing and bulked segregant analysis coupled with whole-genome sequencing identified the candidate genes involved in the post-transcriptional modification of orf224. Overall, our study described a putative mechanism of fertility restoration in a pol TCMS line controlled by ambient temperature that might help utilise TCMS in the two-line breeding of Brassica crops.
Natural Killer T Cells Are Involved in Atherosclerotic Plaque Instability in Apolipoprotein-E Knockout Mice
Authors: Ohmura Yoshinori , Ishimori Naoki , Saito Akimichi , Yokota Takashi , Horii Shunpei , Tokuhara Satoshi , Iwabuchi Kazuya , Tsutsui Hiroyuki
Published: November 2021

The infiltration and activation of macrophages as well as lymphocytes within atherosclerotic lesion contribute to the pathogenesis of plaque rupture. We have demonstrated that invariant natural killer T (iNKT) cells, a unique subset of T lymphocytes that recognize glycolipid antigens, play a crucial role in atherogenesis. However, it remained unclear whether iNKT cells are also involved in plaque instability. Apolipoprotein E (apoE) knockout mice were fed a standard diet (SD) or a high-fat diet (HFD) for 8 weeks. Moreover, the SD- and the HFD-fed mice were divided into two groups according to the intraperitoneal injection of α-galactosylceramide (αGC) that specifically activates iNKT cells or phosphate-buffered saline alone (PBS). ApoE/Jα18 double knockout mice, which lack iNKT cells, were also fed an SD or HFD. Plaque instability was assessed at the brachiocephalic artery by the histological analysis. In the HFD group, αGC significantly enhanced iNKT cell infiltration and exacerbated atherosclerotic plaque instability, whereas the depletion of iNKT cells attenuated plaque instability compared to PBS-treated mice. Real-time PCR analyses in the aortic tissues showed that αGC administration significantly increased expressional levels of inflammatory genes such as IFN-γ and MMP-2, while the depletion of iNKT cells attenuated these expression levels compared to those in the PBS-treated mice. Our findings suggested that iNKT cells are involved in the exacerbation of plaque instability via the activation of inflammatory cells and upregulation of MMP-2 in the vascular tissues.
Calciprotein Particles Link Disturbed Mineral Homeostasis with Cardiovascular Disease by Causing Endothelial Dysfunction and Vascular Inflammation
Authors: Shishkova Daria K. , Velikanova Elena A. , Bogdanov Leo A. , Sinitsky Maxim Yu. , Kostyunin Alexander E. , Tsepokina Anna V. , Gruzdeva Olga V. , Mironov Andrey V. , Mukhamadiyarov Rinat A. , Glushkova Tatiana V. , Krivkina Evgenia O. , Matveeva Vera G. , Hryachkova Oksana N. , Markova Victoria E. , Dyleva Yulia A. , Belik Ekaterina V. , Frolov Alexey V. , Shabaev Amin R. , Efimova Olga S. , Popova Anna N. , Malysheva Valentina Yu. , Kolmykov Roman P. , Sevostyanov Oleg G. , Russakov Dmitriy M. , Dolganyuk Viatcheslav F. , Gutakovsky Anton K. , Zhivodkov Yuriy A. , Kozhukhov Anton S. , Brusina Elena B. , Ismagilov Zinfer R. , Barbarash Olga L. , Yuzhalin Arseniy E. , Kutikhin Anton G.
Published: November 2021

An association between high serum calcium/phosphate and cardiovascular events or death is well-established. However, a mechanistic explanation of this correlation is lacking. Here, we examined the role of calciprotein particles (CPPs), nanoscale bodies forming in the human blood upon its supersaturation with calcium and phosphate, in cardiovascular disease. The serum of patients with coronary artery disease or cerebrovascular disease displayed an increased propensity to form CPPs in combination with elevated ionised calcium as well as reduced albumin levels, altogether indicative of reduced Ca2+-binding capacity. Intravenous administration of CPPs to normolipidemic and normotensive Wistar rats provoked intimal hyperplasia and adventitial/perivascular inflammation in both balloon-injured and intact aortas in the absence of other cardiovascular risk factors. Upon the addition to primary human arterial endothelial cells, CPPs induced lysosome-dependent cell death, promoted the release of pro-inflammatory cytokines, stimulated leukocyte adhesion, and triggered endothelial-to-mesenchymal transition. We concluded that CPPs, which are formed in the blood as a result of altered mineral homeostasis, cause endothelial dysfunction and vascular inflammation, thereby contributing to the development of cardiovascular disease.
DBS Screening for Glycogen Storage Disease Type 1a: Detection of c.648G>T Mutation in G6PC by Combination of Modified Competitive Oligonucleotide Priming-PCR and Melting Curve Analysis
Authors: Niba Emma Tabe Eko , Wijaya Yogik Onky Silvana , Awano Hiroyuki , Taniguchi Naoko , Takeshima Yasuhiro , Nishio Hisahide , Shinohara Masakazu
Published: November 2021

Glycogen storage disease type Ia (GSDIa) is an autosomal recessive disorder caused by glucose-6-phosphatase (G6PC) deficiency. GSDIa causes not only life-threatening hypoglycemia in infancy, but also hepatocellular adenoma as a long-term complication. Hepatocellular adenoma may undergo malignant transformation to hepatocellular carcinoma. New treatment approaches are keenly anticipated for the prevention of hepatic tumors. Gene replacement therapy (GRT) is a promising approach, although early treatment in infancy is essential for its safety and efficiency. Thus, GRT requires screening systems for early disease detection. In this study, we developed a screening system for GSDIa using dried blood spots (DBS) on filter paper, which can detect the most common causative mutation in the East-Asian population, c.648G>T in the G6PC gene. Our system consisted of nested PCR analysis with modified competitive oligonucleotide priming (mCOP)-PCR in the second round and melting curve analysis of the amplified products. Here, we tested 54 DBS samples from 50 c.648G (wild type) controls and four c.648T (mutant) patients. This system, using DBS samples, specifically amplified and clearly detected wild-type and mutant alleles from controls and patients, respectively. In conclusion, our system will be applicable to newborn screening for GSDIa in the real world.
Probiotics against Viral Infections: Current Clinical Trials and Future Perspectives
Authors: Reyes-Castillo Pedro , González-Vázquez Raquel , Torres-Maravilla Edgar , Tello Mario , Bermúdez-Humarán Luis , Mayorga-Reyes Lino
Published: November 2021

Viral infections represent a major health problem worldwide. Due to the wide variety of etiological agents and their increasing resistance to anti-virals and antibiotics treatments, new strategies for effective therapies need to be developed. Scientific evidence suggests that probiotics may have prophylactic and therapeutic effects in viral diseases. Indeed, these microorganisms interact harmoniously with the intestinal microbiota and protect the integrity of the intestinal barrier as well as modulate the host immune system. Currently, clinical trials with probiotics have been documented in respiratory tract infections, infections caused by human immunodeficiency viruses, herpes, human papillomavirus and hepatic encephalopathy. However, the benefits documented so far are difficult to extrapolate, due to the strain-dependent effect. In addition, the dose of the microorganism used as well as host characteristics are other parameters that should be consider when advocating the use of probiotics to treat viral infections. This review addresses the scientific evidence of the efficacy of probiotics in clinical strains perspective in viral infectious diseases in the last 10 years.
Literature Review of Deep Network Compression
Authors: Alqahtani Ali , Xie Xianghua , Jones Mark W.
Published: November 2021

Deep networks often possess a vast number of parameters, and their significant redundancy in parameterization has become a widely-recognized property. This presents significant challenges and restricts many deep learning applications, making the focus on reducing the complexity of models while maintaining their powerful performance. In this paper, we present an overview of popular methods and review recent works on compressing and accelerating deep neural networks. We consider not only pruning methods but also quantization methods, and low-rank factorization methods. This review also intends to clarify these major concepts, and highlights their characteristics, advantages, and shortcomings.
Hornets and Honey Bees: A Coevolutionary Arms Race between Ancient Adaptations and New Invasive Threats
Authors: Cappa Federico , Cini Alessandro , Bortolotti Laura , Poidatz Juliette , Cervo Rita
Published: November 2021

Hornets and honey bees have a long history of coevolution resulting in a plethora of captivating adaptations and counteradaptations between predator and prey. From simple physiological mechanisms to complex behavioral strategies, some Vespa hornets have specialized in hunting honey bees, while the latter have put in place effective defenses to counteract their attack. Both hornets and honey bees have evolved the ability to detect the odors and the pheromones emitted by the other to locate the prey or to spot foraging predators. Hornets often rely on their bigger size, heavily armored body and destructive attacks, while honey bees differentiated collective defense responses finely coordinated to deter or kill the hornet menace. However, when new species of hornets and honey bees come into contact, the absence of coevolution can have a heavy impact on the defenseless bees. The evolutionary arms race between hornets and honey bees provides not only compelling examples of adaptations and counteradaptations between predator and prey, but could also represent a starting point for the development of effective and sustainable strategies to protect honey bees and beekeeping activities and to control invasive alien species of hornets.
Calcium Carbonate Particles Filled Homopolymer Polypropylene at Different Loading Levels: Mechanical Properties Characterization and Materials Failure Analysis
Authors: Peng Yucheng , Musah Munkaila , Via Brian , Wang Xueqi
Published: November 2021

Calcium carbonate (CaCO3) particles have been widely used in filling thermoplastics for different applications in automotive, packaging, and construction. No agreement has been reached in the research community regarding the function of CaCO3 for enhancing toughness of homopolymer polypropylene (HPP). This study was to understand the effect of different loading levels of CaCO3 on HPP toughness, including notched and unnotched impact strength. A batch mixer was used to thermally compound CaCO3 particles with HPP at loading levels of 10, 20, 30, 40, and 50 wt.%, followed by specimen preparation using an injection molding process. The mechanical properties of the composites, including tensile, flexural, and impact were characterized. The results indicated that tensile strengths decreased significantly with increasing loading levels of CaCO3 particles while the tensile and flexural modulus increased significantly with increasing particle loadings. The composite tensile properties changed linearly with increasing CaCO3 loadings. The notched Izod impact strength of the composites was sustained by adding CaCO3 particles up to 40 wt.% while the unnotched impact strength decreased significantly with the addition of CaCO3 particles. Different deformation mechanisms between notched (fracture propagation) and unnotched (fracture initiation and propagation) impact tests were proposed to be the reason.
Structural Damage Detection Using Supervised Nonlinear Support Vector Machine
Authors: Sepahvand Kian K.
Published: November 2021

Damage detection, using vibrational properties, such as eigenfrequencies, is an efficient and straightforward method for detecting damage in structures, components, and machines. The method, however, is very inefficient when the values of the natural frequencies of damaged and undamaged specimens exhibit slight differences. This is particularly the case with lightweight structures, such as fiber-reinforced composites. The nonlinear support vector machine (SVM) provides enhanced results under such conditions by transforming the original features into a new space or applying a kernel trick. In this work, the natural frequencies of damaged and undamaged components are used for classification, employing the nonlinear SVM. The proposed methodology assumes that the frequencies are identified sequentially from an experimental modal analysis; for the study propose, however, the training data are generated from the FEM simulations for damaged and undamaged samples. It is shown that nonlinear SVM using kernel function yields in a clear classification boundary between damaged and undamaged specimens, even for minor variations in natural frequencies.
Metallization by Sputtering to Improve the Bond Strength between Zirconia Ceramics and Resin Cements
Authors: Kimura Tatsuya , Aoyagi Yujin , Taka Norimasa , Kanatani Mitsugu , Uoshima Katsumi
Published: November 2021

Zirconia has been used as a prosthesis material for over a decade because of its excellent mechanical properties and esthetics. The surface treatment for zirconia generally involves sandblasting and the application of primers for favorable bond strength between the surface and resin. However, sandblasting causes the microcracking and chipping of the zirconia surface. To overcome these challenges, the metallization of the zirconia surface was performed. Ti and Au were sputtered on yttria stabilized zirconia (YSZ) disks and heated to 800 °C for 15 min in air. These disks were bonded to stainless-steel rods using resin cement. Then, shear bond strength tests were performed using an Instron-type testing machine. The shear bond strength of the Ti sputtering group was significantly higher than that of the other groups. According to the results of X-ray photoelectron spectroscopy and electron probe microanalysis, the Ti-sputtered YSZ surface contained both sub-titanium oxide and titanium oxide before heating. Sub-titanium oxide was converted to titanium oxide by heating. These results suggest that metallization using Ti is effective for zirconia surface treatment to improve the shear bond strength between YSZ and resin cement. This metallization technique for YSZ has potential in clinical applications.
The Simulation of Sloped Bank Effect Influence on Container Ship Trajectory
Authors: Baric Mate , Mohovic Robert , Mohovic Djani , Pavic Vinko
Published: November 2021

The latest container vessel grounding in the Suez Canal, which occurred on 23 March 2021 (the Ever Given), raised many questions regarding the safety of navigation. The sudden concern about safety is due to fears that traffic flow through the Suez Canal could be blocked for longer periods of time. Besides external forces imposed by wind, in this case bank effect had a significant influence on the ship’s grounding. Bank effect occurs due to restricted water flow caused by narrow waterways. Many fairway design standards consider sloped banks such as those of the Suez Canal as unsubstantial in bank-effect forces. This paper analyses the impact of sloped banks on container ship trajectory and proposes minimal distances that may decrease bank-effect forces in order to reduce the risk of vessel grounding and increase the safety of navigation. However, this type of accident has happened before and may occur again due to a small sailing distance from the bank in cases where vessel speed is increased.
Transmission and Colonization of Pneumocystis jirovecii
Authors: Vera Cristian , Rueda Zulma Vanessa
Published: November 2021

Pneumocystis spp. was discovered in 1909 and was classified as a fungus in 1988. The species that infects humans is called P. jirovecii and important characteristics of its genome have recently been discovered. Important advances have been made to understand P. jirovecii, including aspects of its biology, evolution, lifecycle, and pathogenesis; it is now considered that the main route of transmission is airborne and that the infectious form is the asci (cyst), but it is unclear whether there is transmission by direct contact or droplet spread. On the other hand, P. jirovecii has been detected in respiratory secretions of hosts without causing disease, which has been termed asymptomatic carrier status or colonization (frequency in immunocompetent patients: 0–65%, pregnancy: 15.5%, children: 0–100%, HIV-positive patients: 20–69%, cystic fibrosis: 1–22%, and COPD: 16–55%). This article briefly describes the history of its discovery and the nomenclature of Pneumocystis spp., recently uncovered characteristics of its genome, and what research has been done on the transmission and colonization of P. jirovecii. Based on the literature, the authors of this review propose a hypothetical natural history of P. jirovecii infection in humans.
Cryptococcus neoformans VNII as the Main Cause of Cryptococcosis in Domestic Cats from Rio de Janeiro, Brazil
Authors: Reis Rosani Santos , Bonna Isabel Cristina Fábregas , Antonio Isabela Maria da Silva , Pereira Sandro Antonio , Nascimento Carlos Roberto Sobrinho do , Ferraris Fausto Klabund , Brito-Santos Fábio , Ferreira Gremião Isabella Dib , Trilles Luciana
Published: November 2021

Cryptococcosis is a systemic fungal disease acquired from contaminated environments with propagules of the basidiomycetous yeasts of the Cryptococcus neoformans and C. gattii species complexes. The C. neoformans species complex classically comprises four major molecular types (VNI, VNII, VNIII, and VNIV), and the C. gattii species complex comprises another four (VGI, VGII, VGIII, and VGIV) and the newly identified molecular type VGV. These major molecular types differ in their epidemiological and ecological features, clinical presentations, and therapeutic outcomes. Generally, the most common isolated types are VNI, VGI, and VGII. The epidemiological profile of cryptococcosis in domestic cats is poorly studied and cats can be the sentinels for human infections. Therefore, the present study aimed to determine the molecular characterization of Cryptococcus spp. isolated from domestic cats and their dwellings in the metropolitan area of Rio de Janeiro, Brazil. A total of 36 Cryptococcus spp. strains, both clinical and environmental, from 19 cats were subtyped using multilocus sequence typing (MLST). The ploidy was identified using flow cytometry and the mating type was determined through amplification with specific pheromone primers. All strains were mating type alpha and 6/36 were diploid (all VNII). Most isolates (63.88%) were identified as VNII, a rare molecular type, leading to the consideration that this genotype is more likely related to skin lesions, since there was a high percentage (68.75%) of cats with skin lesions, which is also considered rare. Further studies regarding the molecular epidemiology of cryptococcosis in felines are still needed to clarify the reason for the large proportion of the rare molecular type VNII causing infections in cats.
Two New Species of Fibrodontia (Trechisporales, Basidiomycota) with a Key to Worldwide Species
Authors: Liu Shi-Liang , He Shuang-Hui , Liu Dong-Mei , Zhou Li-Wei
Published: November 2021

Fibrodontia is a genus of wood-inhabiting fungi consisting of four species so far, including F. gossypina as generic type. Two new species, Fibrodontia austrosinensis and F. subalba, are described and illustrated from China. Fibrodontia austrosinensis from southwestern China is characterized by a grandinioid to odontioid hymenophore with numerous small aculei, a dimitic hyphal system with scattered, smooth skeletal hyphae and ellipsoid basidiospores measuring 4.2–5.2 × 3.5–4.5 μm. Fibrodontia subalba from the West Tianshan Mountain in northwestern China is distinguished by an odontioid to hydnoid hymenophore, a dimitic hyphal system, and ellipsoid basidiospores measuring 3.7–4.4 × 2.8–3.4 μm. The phylogenies inferred from the data set of nuc rDNA ITS1-5.8S-ITS2 (ITS) and D1–D2 domains of nuc 28S rDNA (28S), and that of ITS, 28S, translation elongation factor (tef1α), and RNA polymerase II second largest subunit (rpb2) supported Fibrodontia as a monophyletic genus in the Trechisporales, and F. austrosinensis and F. subalba as separate lineages within Fibrodontia. Multi-rate Poisson Tree Processes, Automatic Barcode Gap Discovery and genetic distance methods based on ITS sequences of Fibrodontia also supported F. austrosinensis and F. subalba as distinct species. The taxonomic status of F. fimbriata that was recently transferred from Cystidiodendron, is briefly discussed. A key to all six known species of Fibrodontia is provided.
Predicting Final User Satisfaction Using Momentary UX Data and Machine Learning Techniques
Authors: Koonsanit Kitti , Nishiuchi Nobuyuki
Published: November 2021

User experience (UX) evaluation investigates how people feel about using products or services and is considered an important factor in the design process. However, there is no comprehensive UX evaluation method for time-continuous situations during the use of products or services. Because user experience changes over time, it is difficult to discern the relationship between momentary UX and episodic or cumulative UX, which is related to final user satisfaction. This research aimed to predict final user satisfaction by using momentary UX data and machine learning techniques. The participants were 50 and 25 university students who were asked to evaluate a service (Experiment I) or a product (Experiment II), respectively, during usage by answering a satisfaction survey. Responses were used to draw a customized UX curve. Participants were also asked to complete a final satisfaction questionnaire about the product or service. Momentary UX data and participant satisfaction scores were used to build machine learning models, and the experimental results were compared with those obtained using seven built machine learning models. This study shows that participants’ momentary UX can be understood using a support vector machine (SVM) with a polynomial kernel and that momentary UX can be used to make more accurate predictions about final user satisfaction regarding product and service usage.
Accuracy in Cement Hydration Investigations: Combined X-ray Microtomography and Powder Diffraction Analyses
Authors: Salcedo Inés R. , Cuesta Ana , Shirani Shiva , León-Reina Laura , Aranda Miguel A. G.
Published: November 2021

Cement hydration is a very complex set of processes. The evolution of the crystalline phases during hydration can be accurately followed by X-ray powder diffraction data evaluated by the Rietveld method. However, accurate measurements of some microstructural features, including porosity and amorphous content developments, are more challenging. Here, we combine laboratory X-ray powder diffraction and computed microtomography (μCT) to better understand the results of the μCT analyses. Two pastes with different water–cement ratios, 0.45 and 0.65, filled within capillaries of two sizes, ϕ = 0.5 and 1.0 mm, were analysed at 50 days of hydration. It was shown that within the spatial resolution of the measured μCTs, ~2 μm, the water capillary porosity was segmented within the hydrated component fraction. The unhydrated part could be accurately quantified within 2 vol% error. This work is a first step to accurately determining selected hydration features like the hydration degree of amorphous phases of supplementary cementitious materials within cement blends.