bluuebunny/crossref_metadata_2025 huggingface dataset:
Complexation with Random Methyl-β-Cyclodextrin and (2-Hidroxypropyl)-β-Cyclodextrin Enhances In Vivo Anti-Fibrotic and Anti-Inflammatory Effects of Chrysin via the Inhibition of NF-κB and TGF-β1/Smad Signaling Pathways and Modulation of Hepatic Pro/Anti-Fibrotic miRNA
Authors: Ciceu Alina , Balta Cornel , Herman Hidegard , Gharbia Sami , Ignat Simona-Rebeca , Dinescu Sorina , Váradi Judit , Fenyvesi Ferenc , Gyöngyösi Szilvia , Hermenean Anca , Costache Marieta
Published: February 2021

Chrysin (CHR) is a natural flavonoid with a wide range of pharmacological activities, including hepatoprotection, but poor water solubility. By including water-soluble hydroxypropyl (HPBCD) and randomly methylated (RAMEB) β-cyclodextrin, we aimed to increase its biodisponibility and the effectiveness of the antifibrotic effects of chrysin at oral administration. Liver fibrosis in mice was induced in 7 weeks by CCl4 i.p. administration, and afterwards treated with 50 mg/kg of CHR-HPBCD, CHR-RAMEB, and free chrysin. CCl4 administration increased hepatic inflammation (which was augmented by the upregulation of nuclear factor kappa-light-chain enhancer of activated B cells (NF-kB), tumor necrosis factor (TNF)-α, and interleukin 6 (IL-6) and induced fibrosis, as determined using histopathology and electron microscopy. These results were also confirmed by the upregulation of Collagen I (Col I) and matrix metalloproteinase (MMP) expression, which led to extracellular fibrotic matrix proliferation. Moreover, the immunopositivity of alpha-smooth muscle actin (a-SMA) in the CCl4 group was evidence of hepatic stellate cell (HSC) activation. The main profibrotic pathway was activated, as confirmed by an increase in the transforming growth factor- β1 (TGF-β1) and Smad 2/3 expression, while Smad 7 expression was decreased. Treatment with CHR–HPBCD and CHR–RAMEB considerably reduced liver injury, attenuated inflammation, and decreased extracellular liver collagen deposits. CHR–RAMEB was determined to be the most active antifibrotic complex. We conclude that both nanocomplexes exert anti-inflammatory effects and antifibrotic effects in a considerably stronger manner than for free chrysin administration.
Nrf2 Activation Sensitizes K-Ras Mutant Pancreatic Cancer Cells to Glutaminase Inhibition
Authors: Hamada Shin , Matsumoto Ryotaro , Tanaka Yu , Taguchi Keiko , Yamamoto Masayuki , Masamune Atsushi
Published: February 2021

Pancreatic cancer remains intractable owing to the lack of effective therapy for unresectable cases. Activating mutations of K-ras are frequently found in pancreatic cancers, but these have not yet been targeted by cancer therapies. The Keap1-Nrf2 system plays a crucial role in mediating the oxidative stress response, which also contributes to cancer progression. Nrf2 activation reprograms the metabolic profile to promote the proliferation of cancer cells. A recent report suggested that K-ras- and Nrf2-active lung cancer cells are sensitive to glutamine depletion. This finding led to the recognition of glutaminase inhibitors as novel anticancer agents. In the current study, we used murine pancreatic cancer tissues driven by mutant K-ras and p53 to establish cell lines expressing constitutively activated Nrf2. Genetic or pharmacological Nrf2 activation in cells via Keap1 deletion or Nrf2 activation sensitized cells to glutaminase inhibition. This phenomenon was confirmed to be dependent on K-ras activation in human pancreatic cancer cell lines harboring mutant K-ras, i.e., Panc-1 and MiaPaCa-2 in response to DEM pretreatment. This phenomenon was not observed in BxPC3 cells harboring wildtype K-ras. These results indicate the possibility of employing Nrf2 activation and glutaminase inhibition as novel therapeutic interventions for K-ras mutant pancreatic cancers.
CRISPR/Cas9-Mediated Generation of Pathogen-Resistant Tomato against Tomato Yellow Leaf Curl Virus and Powdery Mildew
Authors: Pramanik Dibyajyoti , Shelake Rahul Mahadev , Park Jiyeon , Kim Mi Jung , Hwang Indeok , Park Younghoon , Kim Jae-Yean
Published: February 2021

Tomato is one of the major vegetable crops consumed worldwide. Tomato yellow leaf curl virus (TYLCV) and fungal Oidium sp. are devastating pathogens causing yellow leaf curl disease and powdery mildew. Such viral and fungal pathogens reduce tomato crop yields and cause substantial economic losses every year. Several commercial tomato varieties include Ty-5 (SlPelo) and Mildew resistance locus o 1 (SlMlo1) locus that carries the susceptibility (S-gene) factors for TYLCV and powdery mildew, respectively. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) is a valuable genome editing tool to develop disease-resistant crop varieties. In this regard, targeting susceptibility factors encoded by the host plant genome instead of the viral genome is a promising approach to achieve pathogen resistance without the need for stable inheritance of CRISPR components. In this study, the CRISPR/Cas9 system was employed to target the SlPelo and SlMlo1 for trait introgression in elite tomato cultivar BN-86 to confer host-mediated immunity against pathogens. SlPelo-knockout lines were successfully generated, carrying the biallelic indel mutations. The pathogen resistance assays in SlPelo mutant lines confirmed the suppressed accumulation of TYLCV and restricted the spread to non-inoculated plant parts. Generated knockout lines for the SlMlo1 showed complete resistance to powdery mildew fungus. Overall, our results demonstrate the efficiency of the CRISPR/Cas9 system to introduce targeted mutagenesis for the rapid development of pathogen-resistant varieties in tomato.
PRKAR1B-AS2 Long Noncoding RNA Promotes Tumorigenesis, Survival, and Chemoresistance via the PI3K/AKT/mTOR Pathway
Authors: Elsayed Abdelrahman M. , Bayraktar Emine , Amero Paola , Salama Salama A. , Abdelaziz Abdelaziz H. , Ismail Raed S. , Zhang Xinna , Ivan Cristina , Sood Anil K. , Lopez-Berestein Gabriel , Rodriguez-Aguayo Cristian
Published: February 2021

Many long noncoding RNAs have been implicated in tumorigenesis and chemoresistance; however, the underlying mechanisms are not well understood. We investigated the role of PRKAR1B-AS2 long noncoding RNA in ovarian cancer (OC) and chemoresistance and identified potential downstream molecular circuitry underlying its action. Analysis of The Cancer Genome Atlas OC dataset, in vitro experiments, proteomic analysis, and a xenograft OC mouse model were implemented. Our findings indicated that overexpression of PRKAR1B-AS2 is negatively correlated with overall survival in OC patients. Furthermore, PRKAR1B-AS2 knockdown-attenuated proliferation, migration, and invasion of OC cells and ameliorated cisplatin and alpelisib resistance in vitro. In proteomic analysis, silencing PRKAR1B-AS2 markedly inhibited protein expression of PI3K-110α and abrogated the phosphorylation of PDK1, AKT, and mTOR, with no significant effect on PTEN. The RNA immunoprecipitation detected a physical interaction between PRKAR1B-AS2 and PI3K-110α. Moreover, PRKAR1B-AS2 knockdown by systemic administration of 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine nanoparticles loaded with PRKAR1B-AS2–specific small interfering RNA enhanced cisplatin sensitivity in a xenograft OC mouse model. In conclusion, PRKAR1B-AS2 promotes tumor growth and confers chemoresistance by modulating the PI3K/AKT/mTOR pathway. Thus, targeting PRKAR1B-AS2 may represent a novel therapeutic approach for the treatment of OC patients.
The In Vivo Selection Method in Breast Cancer Metastasis
Authors: Nakayama Jun , Han Yuxuan , Kuroiwa Yuka , Azuma Kazushi , Yamamoto Yusuke , Semba Kentaro
Published: February 2021

Metastasis is a complex event in cancer progression and causes most deaths from cancer. Repeated transplantation of metastatic cancer cells derived from transplanted murine organs can be used to select the population of highly metastatic cancer cells; this method is called as in vivo selection. The in vivo selection method and highly metastatic cancer cell lines have contributed to reveal the molecular mechanisms of cancer metastasis. Here, we present an overview of the methodology for the in vivo selection method. Recent comparative analysis of the transplantation methods for metastasis have revealed the divergence of metastasis gene signatures. Even cancer cells that metastasize to the same organ show various metastatic cascades and gene expression patterns by changing the transplantation method for the in vivo selection. These findings suggest that the selection of metastasis models for the study of metastasis gene signatures has the potential to influence research results. The study of novel gene signatures that are identified from novel highly metastatic cell lines and patient-derived xenografts (PDXs) will be helpful for understanding the novel mechanisms of metastasis.
Loliolide, a New Therapeutic Option for Neurological Diseases? In Vitro Neuroprotective and Anti-Inflammatory Activities of a Monoterpenoid Lactone Isolated from Codium tomentosum
Authors: Silva Joana , Alves Celso , Martins Alice , Susano Patrícia , Simões Marco , Guedes Miguel , Rehfeldt Stephanie , Pinteus Susete , Gaspar Helena , Rodrigues Américo , Goettert Márcia Ines , Alfonso Amparo , Pedrosa Rui
Published: February 2021

Parkinsons Disease (PD) is the second most common neurodegenerative disease worldwide, and is characterized by a progressive degeneration of dopaminergic neurons. Without an effective treatment, it is crucial to find new therapeutic options to fight the neurodegenerative process, which may arise from marine resources. Accordingly, the goal of the present work was to evaluate the ability of the monoterpenoid lactone Loliolide, isolated from the green seaweed Codium tomentosum, to prevent neurological cell death mediated by the neurotoxin 6-hydroxydopamine (6-OHDA) on SH-SY5Y cells and their anti-inflammatory effects in RAW 264.7 macrophages. Loliolide was obtained from the diethyl ether extract, purified through column chromatography and identified by NMR spectroscopy. The neuroprotective effects were evaluated by the MTT method. Cells’ exposure to 6-OHDA in the presence of Loliolide led to an increase of cells’ viability in 40%, and this effect was mediated by mitochondrial protection, reduction of oxidative stress condition and apoptosis, and inhibition of the NF-kB pathway. Additionally, Loliolide also suppressed nitric oxide production and inhibited the production of TNF-α and IL-6 pro-inflammatory cytokines. The results suggest that Loliolide can inspire the development of new neuroprotective therapeutic agents and thus, more detailed studies should be considered to validate its pharmacological potential.
Zoledronic Acid-Loaded β-TCP Inhibits Tumor Proliferation and Osteoclast Activation: Development of a Functional Bone Substitute for an Efficient Osteosarcoma Treatment
Authors: Kameda Yuka , Aizawa Mamoru , Sato Taira , Honda Michiyo
Published: February 2021

Osteosarcoma has a poor survival rate due to relapse and metastasis. Zoledronic acid (ZOL), an anti-resorptive and anti-tumor agent, is used for treating osteosarcoma. Delivery of ZOL to the target region is difficult due to its high binding affinity to bone minerals. This study developed a novel treatment for osteosarcoma by delivering ZOL to the target region locally and sustainably. In this study, we fabricated a novel bone substitute by loading ZOL on β-tricalcium phosphate (β-TCP). The ZOL-loaded β-TCP (ZOL/β-TCP) would be expected to express the inhibitory effects via both bound-ZOL (bound to β-TCP) and free-ZOL (release from ZOL/β-TCP). To explore the ability to release ZOL from the ZOL/β-TCP, the amount of released ZOL was measured. The released profile indicates that a small amount of ZOL was released, and most of it remained on the β-TCP. Our data showed that ZOL/β-TCP could successfully express the effects of ZOL via both bound-ZOL and free-ZOL. In addition, we examined the biological effects of bound/free-ZOL using osteosarcoma and osteoclasts (target cells). The results showed that two states of ZOL (bound/free) inhibit target cell activities. As a result, ZOL/β-TCP is a promising candidate for application as a novel bone substitute.
Cross-Talk of Toll-Like Receptor 5 and Mu-Opioid Receptor Attenuates Chronic Constriction Injury-Induced Mechanical Hyperalgesia through a Protein Kinase C Alpha-Dependent Signaling
Authors: Chang Ching , Liu Hung-Kai , Yeh Chao-Bin , Yang Ming-Lin , Liao Wen-Chieh , Liu Chiung-Hui , Tseng To-Jung
Published: February 2021

Recently, Toll-like receptors (TLRs), a family of pattern recognition receptors, are reported as potential modulators for neuropathic pain; however, the desired mechanism is still unexplained. Here, we operated on the sciatic nerve to establish a pre-clinical rodent model of chronic constriction injury (CCI) in Sprague-Dawley rats, which were assigned into CCI and Decompression groups randomly. In Decompression group, the rats were performed with nerve decompression at post-operative week 4. Mechanical hyperalgesia and mechanical allodynia were obviously attenuated after a month. Toll-like receptor 5 (TLR5)-immunoreactive (ir) expression increased in dorsal horn, particularly in the inner part of lamina II. Additionally, substance P (SP) and isolectin B4 (IB4)-ir expressions, rather than calcitonin-gene-related peptide (CGRP)-ir expression, increased in their distinct laminae. Double immunofluorescence proved that increased TLR5-ir expression was co-expressed mainly with IB4-ir expression. Through an intrathecal administration with FLA-ST Ultrapure (a TLR5 agonist, purified flagellin from Salmonella Typhimurium, only the CCI-induced mechanical hyperalgesia was attenuated dose-dependently. Moreover, we confirmed that mu-opioid receptor (MOR) and phospho-protein kinase Cα (pPKCα)-ir expressions but not phospho-protein kinase A RII (pPKA RII)-ir expression, increased in lamina II, where they mostly co-expressed with IB4-ir expression. Go 6976, a potent protein kinase C inhibitor, effectively reversed the FLA-ST Ultrapure- or DAMGO-mediated attenuated trend towards mechanical hyperalgesia by an intrathecal administration in CCI rats. In summary, our current findings suggest that nerve decompression improves CCI-induced mechanical hyperalgesia that might be through the cross-talk of TLR5 and MOR in a PKCα-dependent manner, which opens a novel opportunity for the development of analgesic therapeutics in neuropathic pain.
Novel Genetically Modified Mouse Model to Assess Soman-Induced Toxicity and Medical Countermeasure Efficacy: Human Acetylcholinesterase Knock-in Serum Carboxylesterase Knockout Mice
Authors: Marrero-Rosado Brenda M. , Stone Michael F. , de Araujo Furtado Marcio , Schultz Caroline R. , Cadieux C. Linn , Lumley Lucille A.
Published: February 2021

The identification of improved medical countermeasures against exposure to chemical warfare nerve agents (CWNAs), a class of organophosphorus compounds, is dependent on the choice of animal model used in preclinical studies. CWNAs bind to acetylcholinesterase and prevent the catalysis of acetylcholine, causing a plethora of peripheral and central physiologic manifestations, including seizure. Rodents are widely used to elucidate the effects of CWNA-induced seizure, albeit with a caveat: they express carboxylesterase activity in plasma. Carboxylesterase, an enzyme involved in the detoxification of some organophosphorus compounds, plays a scavenging role and decreases CWNA availability, thus exerting a protective effect. Furthermore, species-specific amino acid differences in acetylcholinesterase confound studies that use oximes or other compounds to restore its function after inhibition by CWNA. The creation of a human acetylcholinesterase knock-in/serum carboxylesterase knockout (C57BL/6-Ces1ctm1.1LocAChEtm1.1Loc/J; a.k.a KIKO) mouse may facilitate better modeling of CWNA toxicity in a small rodent species. The current studies characterize the effects of exposure to soman, a highly toxic CWNA, and evaluate the efficacy of anti-seizure drugs in this newly developed KIKO mouse model. Data demonstrate that a combination of midazolam and ketamine reduces seizure duration and severity, eliminates the development of spontaneous recurrent seizures, and protects certain brain regions from neuronal damage in a genetically modified model with human relevance to organophosphorus compound toxicity. This new animal model and the results of this study and future studies using it will enhance medical countermeasures development for both defense and homeland security purposes.
Biofilm Spreading by the Adhesin-Dependent Gliding Motility of Flavobacterium johnsoniae. 1. Internal Structure of the Biofilm
Authors: Sato Keiko , Naya Masami , Hatano Yuri , Kondo Yoshio , Sato Mari , Nagano Keiji , Chen Shicheng , Naito Mariko , Sato Chikara
Published: February 2021

The Gram-negative bacterium Flavobacterium johnsoniae employs gliding motility to move rapidly over solid surfaces. Gliding involves the movement of the adhesin SprB along the cell surface. F. johnsoniae spreads on nutrient-poor 1% agar-PY2, forming a thin film-like colony. We used electron microscopy and time-lapse fluorescence microscopy to investigate the structure of colonies formed by wild-type (WT) F. johnsoniae and by the sprB mutant (ΔsprB). In both cases, the bacteria were buried in the extracellular polymeric matrix (EPM) covering the top of the colony. In the spreading WT colonies, the EPM included a thick fiber framework and vesicles, revealing the formation of a biofilm, which is probably required for the spreading movement. Specific paths that were followed by bacterial clusters were observed at the leading edge of colonies, and abundant vesicle secretion and subsequent matrix formation were suggested. EPM-free channels were formed in upward biofilm protrusions, probably for cell migration. In the nonspreading ΔsprB colonies, cells were tightly packed in layers and the intercellular space was occupied by less matrix, indicating immature biofilm. This result suggests that SprB is not necessary for biofilm formation. We conclude that F. johnsoniae cells use gliding motility to spread and maturate biofilms.
Early-Life Stress Modulates Gut Microbiota and Peripheral and Central Inflammation in a Sex-Dependent Manner
Authors: Park Hae Jeong , Kim Sang A. , Kang Won Sub , Kim Jong Woo
Published: February 2021

Recent studies have reported that changes in gut microbiota composition could induce neuropsychiatric problems. In this study, we investigated alterations in gut microbiota induced by early-life stress (ELS) in rats subjected to maternal separation (MS; 6 h a day, postnatal days (PNDs) 1–21), along with changes in inflammatory cytokines and tryptophan-kynurenine (TRP-KYN) metabolism, and assessed the differences between sexes. High-throughput sequencing of the bacterial 16S rRNA gene showed that the relative abundance of the Bacteroides genus was increased and that of the Lachnospiraceae family was decreased in the feces of MS rats of both sexes (PND 56). By comparison, MS increased the relative abundance of the Streptococcus genus and decreased that of the Staphylococcus genus only in males, whereas the abundance of the Sporobacter genus was enhanced and that of the Mucispirillum genus was reduced by MS only in females. In addition, the levels of proinflammatory cytokines were increased in the colons (IFN-γ and IL-6) and sera (IL-1β) of the male MS rats, together with the elevation of the KYN/TRP ratio in the sera, but not in females. In the hippocampus, MS elevated the level of IL-1β and the KYN/TRP ratio in both male and female rats. These results indicate that MS induces peripheral and central inflammation and TRP-KYN metabolism in a sex-dependent manner, together with sex-specific changes in gut microbes.
Comparing Super-Resolution Microscopy Techniques to Analyze Chromosomes
Authors: Kubalová Ivona , Němečková Alžběta , Weisshart Klaus , Hřibová Eva , Schubert Veit
Published: February 2021

The importance of fluorescence light microscopy for understanding cellular and sub-cellular structures and functions is undeniable. However, the resolution is limited by light diffraction (~200–250 nm laterally, ~500–700 nm axially). Meanwhile, super-resolution microscopy, such as structured illumination microscopy (SIM), is being applied more and more to overcome this restriction. Instead, super-resolution by stimulated emission depletion (STED) microscopy achieving a resolution of ~50 nm laterally and ~130 nm axially has not yet frequently been applied in plant cell research due to the required specific sample preparation and stable dye staining. Single-molecule localization microscopy (SMLM) including photoactivated localization microscopy (PALM) has not yet been widely used, although this nanoscopic technique allows even the detection of single molecules. In this study, we compared protein imaging within metaphase chromosomes of barley via conventional wide-field and confocal microscopy, and the sub-diffraction methods SIM, STED, and SMLM. The chromosomes were labeled by DAPI (4′,6-diamidino-2-phenylindol), a DNA-specific dye, and with antibodies against topoisomerase IIα (Topo II), a protein important for correct chromatin condensation. Compared to the diffraction-limited methods, the combination of the three different super-resolution imaging techniques delivered tremendous additional insights into the plant chromosome architecture through the achieved increased resolution.
Genome-Wide Identification and Characterization of Wheat 14-3-3 Genes Unravels the Role of TaGRF6-A in Salt Stress Tolerance by Binding MYB Transcription Factor
Authors: Shao Wenna , Chen Wang , Zhu Xiaoguo , Zhou Xiaoyi , Jin Yingying , Zhan Chuang , Liu Gensen , Liu Xi , Ma Dongfang , Qiao Yongli
Published: February 2021

14-3-3 proteins are a large multigenic family of general regulatory factors (GRF) ubiquitously found in eukaryotes and play vital roles in the regulation of plant growth, development, and response to stress stimuli. However, so far, no comprehensive investigation has been performed in the hexaploid wheat. In the present study, A total of 17 potential 14-3-3 gene family members were identified from the Chinese Spring whole-genome sequencing database. The phylogenetic comparison with six 14-3-3 families revealed that the majority of wheat 14-3-3 genes might have evolved as an independent branch and grouped into ε and non-ε group using the phylogenetic comparison. Analysis of gene structure and motif indicated that 14-3-3 protein family members have relatively conserved exon/intron arrangement and motif composition. Physical mapping showed that wheat 14-3-3 genes are mainly distributed on chromosomes 2, 3, 4, and 7. Moreover, most 14-3-3 members in wheat exhibited significantly down-regulated expression in response to alkaline stress. VIGS assay and protein-protein interaction analysis further confirmed that TaGRF6-A positively regulated slat stress tolerance by interacting with a MYB transcription factor, TaMYB64. Taken together, our findings provide fundamental information on the involvement of the wheat 14-3-3 family in salt stress and further investigating their molecular mechanism.
Neutralizing Human Antibodies against Severe Acute Respiratory Syndrome Coronavirus 2 Isolated from a Human Synthetic Fab Phage Display Library
Authors: Kim Yu Jung , Lee Min Ho , Lee Se-Ra , Chung Hyo-Young , Kim Kwangmin , Lee Tae Gyu , Kim Dae Young
Published: February 2021

Since it was first reported in Wuhan, China, in 2019, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic outbreak resulting in a tremendous global threat due to its unprecedented rapid spread and an absence of a prophylactic vaccine or therapeutic drugs treating the virus. The receptor-binding domain (RBD) of the SARS-CoV-2 spike protein is a key player in the viral entry into cells through its interaction with the angiotensin-converting enzyme 2 (ACE2) receptor protein, and the RBD has therefore been crucial as a drug target. In this study, we used phage display to develop human monoclonal antibodies (mAbs) that neutralize SARS-CoV-2. A human synthetic Fab phage display library was panned against the RBD of the SARS-CoV-2 spike protein (SARS-2 RBD), yielding ten unique Fabs with moderate apparent affinities (EC50 = 19–663 nM) for the SARS-2 RBD. All of the Fabs showed no cross-reactivity to the MERS-CoV spike protein, while three Fabs cross-reacted with the SARS-CoV spike protein. Five Fabs showed neutralizing activities in in vitro assays based on the Fabs’ activities antagonizing the interaction between the SARS-2 RBD and ACE2. Reformatting the five Fabs into immunoglobulin Gs (IgGs) greatly increased their apparent affinities (KD = 0.08–1.0 nM), presumably due to the effects of avidity, without compromising their non-aggregating properties and thermal stability. Furthermore, two of the mAbs (D12 and C2) significantly showed neutralizing activities on pseudo-typed and authentic SARS-CoV-2. Given their desirable properties and neutralizing activities, we anticipate that these human anti-SARS-CoV-2 mAbs would be suitable reagents to be further developed as antibody therapeutics to treat COVID-19, as well as for diagnostics and research tools.
Sarcopenic Obesity in Liver Cirrhosis: Possible Mechanism and Clinical Impact
Authors: Nishikawa Hiroki , Enomoto Hirayuki , Nishiguchi Shuhei , Iijima Hiroko
Published: February 2021

The picture of chronic liver diseases (CLDs) has changed considerably in recent years. One of them is the increase of non-alcoholic fatty liver disease. More and more CLD patients, even those with liver cirrhosis (LC), tend to be presenting with obesity these days. The annual rate of muscle loss increases with worsening liver reserve, and thus LC patients are more likely to complicate with sarcopenia. LC is also characterized by protein-energy malnutrition (PEM). Since the PEM in LC can be invariable, the patients probably present with sarcopenic obesity (Sa-O), which involves both sarcopenia and obesity. Currently, there is no mention of Sa-O in the guidelines; however, the rapidly increasing prevalence and poorer clinical consequences of Sa-O are recognized as an important public health problem, and the diagnostic value of Sa-O is expected to increase in the future. Sa-O involves a complex interplay of physiological mechanisms, including increased inflammatory cytokines, oxidative stress, insulin resistance, hormonal disorders, and decline of physical activity. The pathogenesis of Sa-O in LC is diverse, with a lot of perturbations in the muscle–liver–adipose tissue axis. Here, we overview the current knowledge of Sa-O, especially focusing on LC.
Stromal CCL5 Promotes Breast Cancer Progression by Interacting with CCR3 in Tumor Cells
Authors: Yamaguchi Mio , Takagi Kiyoshi , Narita Koki , Miki Yasuhiro , Onodera Yoshiaki , Miyashita Minoru , Sasano Hironobu , Suzuki Takashi
Published: February 2021

Chemokines secreted from stromal cells have important roles for interactions with carcinoma cells and regulating tumor progression. C-C motif chemokine ligand (CCL) 5 is expressed in various types of stromal cells and associated with tumor progression, interacting with C-C chemokine receptor (CCR) 1, 3 and 5 expressed in tumor cells. However, the expression on CCL5 and its receptors have so far not been well-examined in human breast carcinoma tissues. We therefore immunolocalized CCL5, as well as CCR1, 3 and 5, in 111 human breast carcinoma tissues and correlated them with clinicopathological characteristics. Stromal CCL5 immunoreactivity was significantly correlated with the aggressive phenotype of breast carcinomas. Importantly, this tendency was observed especially in the CCR3-positive group. Furthermore, the risk of recurrence was significantly higher in the patients with breast carcinomas positive for CCL5 and CCR3 but negative for CCR1 and CCR5, as compared with other patients. In summary, the CCL5-CCR3 axis might contribute to a worse prognosis in breast cancer patients, and these findings will contribute to a better understanding of the significance of the CCL5/CCRs axis in breast carcinoma microenvironment.
Early Covert Appearance of Marginal Zone B Cells in Salivary Glands of Sjögren′s Syndrome-Susceptible Mice: Initiators of Subsequent Overt Clinical Disease
Authors: Peck Ammon B. , Nguyen Cuong Q. , Ambrus Julian
Published: February 2021

The C57BL/6.NOD-Aec1Aec2 mouse model has been extensively studied to define the underlying cellular and molecular bioprocesses critical in the onset of primary Sjögren’s Syndrome (pSS), a human systemic autoimmune disease characterized clinically as the loss of lacrimal and salivary gland functions leading to dry eye and dry mouth pathologies. This mouse model, together with several gene knockout mouse models of SS, has indicated that B lymphocytes, especially marginal zone B (MZB) cells, are necessary for development and onset of clinical manifestations despite the fact that destruction of the lacrimal and salivary gland cells involves a classical T cell-mediated autoimmune response. Because migrations and functions of MZB cells are difficult to study in vivo, we have carried out ex vivo investigations that use temporal global RNA transcriptomic analyses to profile autoimmunity as it develops within the salivary glands of C57BL/6.NOD-Aec1Aec2 mice. Temporal profiles indicate the appearance of Notch2-positive cells within the salivary glands of these SS-susceptible mice concomitant with the early-phase appearance of lymphocytic foci (LF). Data presented here identify cellular bioprocesses occurring during early immune cell migrations into the salivary glands and suggest MZB cells are recruited to the exocrine glands by the upregulated Cxcl13 chemokine where they recognize complement (C’)-decorated antigens via their sphingosine-1-phosphate (S1P) and B cell (BC) receptors. Based on known MZB cell behavior and mobility, we propose that MZB cells activated in the salivary glands migrate to splenic follicular zones to present antigens to follicular macrophages and dendritic cells that, in turn, promote a subsequent systemic cell-mediated and autoantibody-mediated autoimmune T cell response that targets exocrine gland cells and functions. Overall, this study uses the power of transcriptomic analyses to provide greater insight into several molecular events defining cellular bioprocesses underlying SS that can be modelled and more thoroughly studied at the cellular level.
Newborn Screening for CAH—Challenges and Opportunities
Authors: Heather Natasha L. , Nordenstrom Anna
Published: February 2021

Newborn screening for congenital adrenal hyperplasia (CAH) using 17-hydroxyprogesterone (17-OHP) as an indicator of disease was first introduced in the 1970s [...]
An Ambient Intelligence-Based Human Behavior Monitoring Framework for Ubiquitous Environments
Authors: Thakur Nirmalya , Han Chia Y.
Published: February 2021

This framework for human behavior monitoring aims to take a holistic approach to study, track, monitor, and analyze human behavior during activities of daily living (ADLs). The framework consists of two novel functionalities. First, it can perform the semantic analysis of user interactions on the diverse contextual parameters during ADLs to identify a list of distinct behavioral patterns associated with different complex activities. Second, it consists of an intelligent decision-making algorithm that can analyze these behavioral patterns and their relationships with the dynamic contextual and spatial features of the environment to detect any anomalies in user behavior that could constitute an emergency. These functionalities of this interdisciplinary framework were developed by integrating the latest advancements and technologies in human–computer interaction, machine learning, Internet of Things, pattern recognition, and ubiquitous computing. The framework was evaluated on a dataset of ADLs, and the performance accuracies of these two functionalities were found to be 76.71% and 83.87%, respectively. The presented and discussed results uphold the relevance and immense potential of this framework to contribute towards improving the quality of life and assisted living of the aging population in the future of Internet of Things (IoT)-based ubiquitous living environments, e.g., smart homes.
Inverted U-Curve Association between Serum Indoxyl Sulfate Levels and Cardiovascular Events in Patients on Chronic Hemodialysis
Authors: Tsai Ming-Hsien , Chang Chung-Hsin , Liou Hung-Hsiang , Fang Yu-Wei
Published: February 2021

Background: Protein-bound uremic toxins are associated with cardiovascular disease and mortality in patients with chronic kidney disease. We investigated their association with clinical outcomes in patients undergoing chronic hemodialysis (CHD). Methods: A prospective cohort study was conducted on 86 Taiwanese patients undergoing CHD. The predictors were indoxyl sulfate and p-cresyl sulfate concentrations, with each analyzed as three tertiles. Outcomes were cardiovascular events and all-cause mortality. Results: During a 25-month follow up period, there were 23 cardiovascular events and seven all-cause mortality events. In the crude survival analysis, the second indoxyl sulfate tertile was shown to be a powerful predictor of cardiovascular events compared with the third tertile (hazard ratio (HR), 3.14; 95% confidence interval (CI), 1.10–8.94), and the first tertile was shown to have a poor but insignificant cardiovascular outcome (HR, 1.09; 95% CI, 0.30–4.00). Moreover, the predictive power of the second indoxyl sulfate tertile for cardiovascular events remained after adjustment for confounders (HR, 5.42; 95% CI, 1.67–17.60). Conclusions: An inverse U-curve relationship was observed between the total serum indoxyl sulfate level and cardiovascular events in our CHD patients. A large-scale study is needed to confirm this relationship.
Are Quality of Randomized Clinical Trials and ESMO-Magnitude of Clinical Benefit Scale Two Sides of the Same Coin, to Grade Recommendations for Drug Approval?
Authors: Rodriguez Adela , Esposito Francis , Oliveres Helena , Torres Ferran , Maurel Joan
Published: February 2021

The approval of a new drug for cancer treatment by the US Food and Drug Administration (FDA) and the European Medicines Agency (EMA) is based on positive, well-designed randomized phase III clinical trials (RCTs). However, not all of them are analyzed to support the recommendations. For this reason, there are different scales to quantify and evaluate the quality of RCTs and the magnitude of the clinical benefits of new drugs for treating solid tumors. In this review, we discuss the value of the progression-free survival (PFS) as an endpoint in RCTs and the concordance between it and the overall survival (OS) as a measure of the quality of clinical trial designs. We summarize and analyze the different scales to evaluate the clinical benefits of new drugs such as the The American Society of Clinical Oncology value framework (ASCO-VF-NHB16) and European Society for Medical Oncology Magnitude of Clinical Benefit Scale (ESMO-MCBS) and the concordance between them, focusing on metastatic colorectal cancer (mCRC). We propose several definitions that would help to evaluate the quality of RCT, the magnitude of clinical benefit and the appropriate approval of new drugs in oncology.
Skeletal Changes in Growing Cleft Patients with Class III Malocclusion Treated with Bone Anchored Maxillary Protraction—A 3.5-Year Follow-Up
Authors: Steegman Ralph M. , Klein Meulekamp Annemarlien Faye , Dieters Arjan , Jansma Johan , van der Meer Wicher J. , Ren Yijin
Published: February 2021

This prospective controlled trial aimed to evaluate the skeletal effect of 3.5-years bone anchored maxillary protraction (BAMP) in growing cleft subjects with a Class III malocclusion. Subjects and Method: Nineteen cleft patients (11.4 ± 0.7-years) were included from whom cone beam computed tomography (CBCT) scans were taken before the start of BAMP (T0), 1.5-years after (T1) and 3.5 y after (T2). Seventeen age- and malocclusion-matched, untreated cleft subjects with cephalograms available at T0 and T2 served as the control group. Three dimensional skeletal changes were measured qualitatively and quantitatively on CBCT scans. Two dimensional measurements were made on cephalograms. Results: Significant positive effects have been observed on the zygomaticomaxillary complex. Specifically, the A-point showed a displacement of 2.7 mm ± 0.9 mm from T0 to T2 (p < 0.05). A displacement of 3.8 mm ± 1.2 mm was observed in the zygoma regions (p < 0.05). On the cephalograms significant differences at T2 were observed between the BAMP and the control subjects in Wits, gonial angle, and overjet (p < 0.05), all in favor of the treatment of Class III malocclusion. The changes taking place in the two consecutive periods (ΔT1-T0, ΔT2-T1) did not differ, indicating that not only were the positive results from the first 1.5-years maintained, but continuous orthopedic effects were also achieved in the following 2-years. Conclusions: In conclusion, findings from the present prospective study with a 3.5-years follow-up provide the first evidence to support BAMP as an effective and reliable treatment option for growing cleft subjects with mild to moderate Class III malocclusion up to 15-years old.
Optical Coherence Tomography Angiography of the Choriocapillaris in Age-Related Macular Degeneration
Authors: Scharf Jackson , Corradetti Giulia , Corvi Federico , Sadda SriniVas , Sarraf David
Published: February 2021

The advent of optical coherence tomography angiography (OCTA) has allowed for remarkable advancements in our understanding of the role of the choriocapillaris in age-related macular degeneration (AMD). As a relatively new imaging modality, techniques to analyze and quantify choriocapillaris images are still evolving. Quantification of the choriocapillaris requires careful consideration of many factors, including the type of OCTA device, segmentation of the choriocapillaris slab, image processing techniques, and thresholding method. OCTA imaging shows that the choriocapillaris is impaired in intermediate non-neovascular AMD, and the severity of impairment may predict the advancement of disease. In advanced atrophic AMD, the choriocapillaris is severely impaired underneath the area of geographic atrophy, and the level of impairment surrounding the lesion predicts the rate of atrophy enlargement. Macular neovascularization can be readily identified and classified using OCTA, but it is still unclear if neovascularization features with OCTA can predict the lesion’s level of activity. The choriocapillaris surrounding macular neovascularization is impaired while the more peripheral choriocapillaris is spared, implying that choriocapillaris disruption may drive neovascularization growth. With continued innovation in OCTA image acquisition and analysis methods, advancement in clinical applications and pathophysiologic discoveries in AMD are set to follow.
Left Atrial Ejection Fraction Assessed by Prior Cardiac CT Predicts Recurrence of Atrial Fibrillation after Pulmonary Vein Isolation
Authors: Kaufmann Reinhard , Rezar Richard , Strohmer Bernhard , Wernly Bernhard , Lichtenauer Michael , Hitzl Wolfgang , Meissnitzer Matthias , Hergan Klaus , Granitz Marcel
Published: February 2021

Assuming that atrial fibrillation (AF) is associated with left atrial remodeling and dysfunction, we hypothesize that left atrial and left atrial appendage ejection fractions (LAEF and LAAEF) are useful and may be more sensitive outcome predictors of pulmonary vein isolation (PVI). Fifty patients who underwent PVI at our institution with available pre-interventional cardiac computed tomography (CT) for procedure planning were included in this retrospective study. The patients were separated into two groups by recurrence and non-recurrence of AF and subgroups of paroxysmal and persistent AF. Semiautomatic volumetric analysis of the left atrium was used to calculate morphological and functional parameters and optimal cut-offs were calculated using the Youden index. LAEF (accuracy 94%, sensitivity 67%) and LAAEF (accuracy 90%, sensitivity 67%) were significantly reduced in patients with AF recurrence (16% vs. 36%, p = 0.00002; 16% vs. 42%, p = 0.000002), and in the subgroup analysis, the functional parameters were independent from AF type (paroxysmal and persistent). With a cut-off of <23% for both LAEF and LAAEF (area under the curve (AUC) 0.94, 95%CI 0.84–0.99 and AUC 0.96, 95%CI 0.86–0.99, respectively), AF recurrence occurred in 77.8%, within a mean follow-up period of 229 days. In conclusion, left atrial function on prior cardiac CT offers useful parameters for predicting AF recurrence after PVI.
Staphylococcus aureus Keratitis: Incidence, Pathophysiology, Risk Factors and Novel Strategies for Treatment
Authors: Lee Jason W. , Somerville Tobi , Kaye Stephen B. , Romano Vito
Published: February 2021

Bacterial keratitis is a devastating condition that can rapidly progress to serious complications if not treated promptly. Certain causative microorganisms such as Staphylococcus aureus and Pseudomonas aeruginosa are notorious for their resistance to antibiotics. Resistant bacterial keratitis results in poorer outcomes such as scarring and the need for surgical intervention. Thorough understanding of the causative pathogen and its virulence factors is vital for the discovery of novel treatments to avoid further antibiotic resistance. While much has been previously reported on P. aeruginosa, S. aureus has been less extensively studied. This review aims to give a brief overview of S. aureus epidemiology, pathophysiology and clinical characteristics as well as summarise the current evidence for potential novel therapies.
IMPELLA® or Extracorporeal Membrane Oxygenation for Left Ventricular Dominant Refractory Cardiogenic Shock
Authors: Schurtz Guillaume , Rousse Natacha , Saura Ouriel , Balmette Vincent , Vincent Flavien , Lamblin Nicolas , Porouchani Sina , Verdier Basile , Puymirat Etienne , Robin Emmanuel , Van Belle Eric , Vincentelli André , Aissaoui Nadia , Delhaye Cédric , Delmas Clément , Cosenza Alessandro , Bonello Laurent , Juthier Francis , Moussa Mouhamed Djahoum , Lemesle Gilles
Published: February 2021

Mechanical circulatory support (MCS) devices are effective tools in managing refractory cardiogenic shock (CS). Data comparing veno-arterial extracorporeal membrane oxygenation (VA-ECMO) and IMPELLA® are however scarce. We aimed to assess outcomes of patients implanted with these two devices and eligible to both systems. From 2004 to 2020, we retrospectively analyzed 128 patients who underwent VA-ECMO or IMPELLA® in our institution for refractory left ventricle (LV) dominant CS. All patients were eligible to both systems: 97 patients were first implanted with VA-ECMO and 31 with IMPELLA®. The primary endpoint was 30-day all-cause death. VA-ECMO patients were younger (52 vs. 59.4, p = 0.006) and had a higher lactate level at baseline than those in the IMPELLA® group (6.84 vs. 3.03 mmol/L, p < 0.001). Duration of MCS was similar between groups (9.4 days vs. 6 days in the VA-ECMO and IMPELLA® groups respectively, p = 0.077). In unadjusted analysis, no significant difference was observed between groups in 30-day mortality: 43.3% vs. 58.1% in the VA-ECMO and IMPELLA® groups, respectively (p = 0.152). After adjustment, VA-ECMO was associated with a significant reduction in 30-day mortality (HR = 0.25, p = 0.004). A higher rate of MCS escalation was observed in the IMPELLA® group: 32.3% vs. 10.3% (p = 0.003). In patients eligible to either VA-ECMO or IMPELLA® for LV dominant refractory CS, VA-ECMO was associated with improved survival rate and a lower need for escalation.
Accelerating 3D Medical Image Segmentation by Adaptive Small-Scale Target Localization
Authors: Shirokikh Boris , Shevtsov Alexey , Dalechina Alexandra , Krivov Egor , Kostjuchenko Valery , Golanov Andrey , Gombolevskiy Victor , Morozov Sergey , Belyaev Mikhail
Published: February 2021

The prevailing approach for three-dimensional (3D) medical image segmentation is to use convolutional networks. Recently, deep learning methods have achieved human-level performance in several important applied problems, such as volumetry for lung-cancer diagnosis or delineation for radiation therapy planning. However, state-of-the-art architectures, such as U-Net and DeepMedic, are computationally heavy and require workstations accelerated with graphics processing units for fast inference. However, scarce research has been conducted concerning enabling fast central processing unit computations for such networks. Our paper fills this gap. We propose a new segmentation method with a human-like technique to segment a 3D study. First, we analyze the image at a small scale to identify areas of interest and then process only relevant feature-map patches. Our method not only reduces the inference time from 10 min to 15 s but also preserves state-of-the-art segmentation quality, as we illustrate in the set of experiments with two large datasets.
A Model-Based Optimization Framework for Iterative Digital Breast Tomosynthesis Image Reconstruction
Authors: Loli Piccolomini Elena , Morotti Elena
Published: February 2021

Digital Breast Tomosynthesis is an X-ray imaging technique that allows a volumetric reconstruction of the breast, from a small number of low-dose two-dimensional projections. Although it is already used in the clinical setting, enhancing the quality of the recovered images is still a subject of research. The aim of this paper was to propose and compare, in a general optimization framework, three slightly different models and corresponding accurate iterative algorithms for Digital Breast Tomosynthesis image reconstruction, characterized by a convergent behavior. The suggested model-based implementations are specifically aligned to Digital Breast Tomosynthesis clinical requirements and take advantage of a Total Variation regularizer. We also tune a fully-automatic strategy to set a proper regularization parameter. We assess our proposals on real data, acquired from a breast accreditation phantom and a clinical case. The results confirm the effectiveness of the presented framework in reconstructing breast volumes, with particular focus on the masses and microcalcifications, in few iterations and in enhancing the image quality in a prolonged execution.
Manufacturing Concept and Prototype for Train Component Using the FSW Process
Authors: Hoyos Elizabeth , Escobar Santiago , De Backer Jeroen , Martin Jonathan , Palacio Mauricio
Published: February 2021

Friction stir welding (FSW) is a process originally developed for joining light materials, such as aluminum and magnesium, as an answer to their poor weldability by conventional fusion processes. In Colombia, the technique has been studied but its industrial implementation is uncommon, due to the high cost of specialized machinery and the unfamiliarity with the technique of local industries. This article presents an implementation case study of FSW on a 6082-aluminum alloy train component from Metro de Medellín (MdM), aiming to establish the component design changes required to accommodate the FSW process, and conventional machines available in the local area which may be available for welding. Additionally, a simple comparison was made between the cost of this approach versus the manufacturing strategy currently used for the selected component. Initially, welding forces were measured when performing the seam on the selected component using an FSW machine. This data was then used to downselect the local milling machines with these capabilities. A simple but specific tool was designed for the geometry of one of the component features. Finally, a prototype was fabricated, and weld samples were obtained, polished, etched, and examined using a microhardness machine and an optical microscope. Results show a good opportunity for the execution of simple components with uniform geometries, which can be carried out using locally available machinery because they do not surpass their maximum loading capacity, the welds do not present visible discontinuities, and an average hardness of 69.5 HV and mechanical efficiency of 95% can be achieved. Additionally, the manufacturing process is around 30% cheaper compared to traditional methods, making the application viable, economically speaking.