bluuebunny/crossref_metadata_2025 huggingface dataset:
Biochemical Characterization of a Novel Redox-Regulated Metacaspase in a Marine Diatom
Authors: Graff van Creveld Shiri , Ben-Dor Shifra , Mizrachi Avia , Alcolombri Uria , Hopes Amanda , Mock Thomas , Rosenwasser Shilo , Vardi Assaf
Published: September 2021

Programmed cell death (PCD) in marine microalgae was suggested to be one of the mechanisms that facilitates bloom demise, yet its molecular components in phytoplankton are unknown. Phytoplankton are completely lacking any of the canonical components of PCD, such as caspases, but possess metacaspases. Metacaspases were shown to regulate PCD in plants and some protists, but their roles in algae and other organisms are still elusive. Here, we identified and biochemically characterized a type III metacaspase from the model diatom Phaeodactylum tricornutum, termed PtMCA-IIIc. Through expression of recombinant PtMCA-IIIc in E. coli, we revealed that PtMCA-IIIc exhibits a calcium-dependent protease activity, including auto-processing and cleavage after arginine. Similar metacaspase activity was detected in P. tricornutum cell extracts. PtMCA-IIIc overexpressing cells exhibited higher metacaspase activity, while CRISPR/Cas9-mediated knockout cells had decreased metacaspase activity compared to WT cells. Site-directed mutagenesis of cysteines that were predicted to form a disulfide bond decreased recombinant PtMCA-IIIc activity, suggesting its enhancement under oxidizing conditions. One of those cysteines was oxidized, detected in redox proteomics, specifically in response to lethal concentrations of hydrogen peroxide and a diatom derived aldehyde. Phylogenetic analysis revealed that this cysteine-pair is unique and widespread among diatom type III metacaspases. The characterization of a cell death associated protein in diatoms provides insights into the evolutionary origins of PCD and its ecological significance in algal bloom dynamics.
Genome-Wide Transcription Factor DNA Binding Sites and Gene Regulatory Networks in Clostridium thermocellum
Authors: Hebdon Skyler D. , Gerritsen Alida T. , Chen Yi-Pei , Marcano Joan G. , Chou Katherine J.
Published: September 2021

Clostridium thermocellum is a thermophilic bacterium recognized for its natural ability to effectively deconstruct cellulosic biomass. While there is a large body of studies on the genetic engineering of this bacterium and its physiology to-date, there is limited knowledge in the transcriptional regulation in this organism and thermophilic bacteria in general. The study herein is the first report of a large-scale application of DNA-affinity purification sequencing (DAP-seq) to transcription factors (TFs) from a bacterium. We applied DAP-seq to > 90 TFs in C. thermocellum and detected genome-wide binding sites for 11 of them. We then compiled and aligned DNA binding sequences from these TFs to deduce the primary DNA-binding sequence motifs for each TF. These binding motifs are further validated with electrophoretic mobility shift assay (EMSA) and are used to identify individual TFs’ regulatory targets in C. thermocellum. Our results led to the discovery of novel, uncharacterized TFs as well as homologues of previously studied TFs including RexA-, LexA-, and LacI-type TFs. We then used these data to reconstruct gene regulatory networks for the 11 TFs individually, which resulted in a global network encompassing the TFs with some interconnections. As gene regulation governs and constrains how bacteria behave, our findings shed light on the roles of TFs delineated by their regulons, and potentially provides a means to enable rational, advanced genetic engineering of C. thermocellum and other organisms alike toward a desired phenotype.
Regulation and Evasion of Host Immune Response by African Swine Fever Virus
Authors: Wu Lei , Yang Bincai , Yuan Xu , Hong Jinxuan , Peng Min , Chen Ji-Long , Song Zhongbao
Published: September 2021

African swine fever (ASF) is an acute lethal hemorrhagic viral disease in domestic pigs and wild boars; is widely epidemic in Africa, Europe, Asia, and Latin America; and poses a huge threat to the pig industry worldwide. ASF is caused by the infection of the ASF virus (ASFV), a cytoplasmic double-stranded DNA virus belonging to the Asfarviridae family. Here, we review how the virus regulates the host immune response and its mechanisms at different levels, including interferon modulation, inflammation, apoptosis, antigen presentation, and cellular immunity.
Comparative Genome Analysis and Spore Heat Resistance Assay Reveal a New Component to Population Structure and Genome Epidemiology Within Clostridium perfringens Enterotoxin-Carrying Isolates
Authors: Jaakkola Kaisa , Virtanen Kira , Lahti Päivi , Keto-Timonen Riikka , Lindström Miia , Korkeala Hannu
Published: September 2021

Clostridium perfringenscauses a variety of human and animal enteric diseases including food poisoning, antibiotic-associated diarrhea, and necrotic enteritis. Yet, the reservoirs of enteropathogenic enterotoxin-producing strains remain unknown. We conducted a genomic comparison of 290 strains and a heat resistance phenotyping of 30C. perfringensstrains to elucidate the population structure and ecology of this pathogen.C. perfringensgenomes shared a conserved genetic backbone with more than half of the genes of an average genome conserved in >95% of strains. The cpe-carrying isolates were found to share genetic context: thecpe-carrying plasmids had different distribution patterns within the genetic lineages and the estimated pan genome ofcpe-carrying isolates had a larger core genome and a smaller accessory genome compared to that of 290 strains. We characterizecpe-negative strains related to chromosomalcpe-carrying strains elucidating the origin of these strains and disclose two distinct groups of chromosomalcpe-carrying strains with different virulence characteristics, spore heat resistance properties, and, presumably, ecological niche. Finally, an antibiotic-associated diarrhea isolate carrying two copies of the enterotoxincpegene and the associated genetic lineage with the potential for the emergence of similar strains are outlined. WithC. perfringensas an example, implications of input genome quality for pan genome analysis are discussed. Our study furthers the understanding of genome epidemiology and population structure of enteropathogenicC. perfringensand brings new insight into this important pathogen and its reservoirs.
Emergence of Multi-Drug Resistance and Its Association With Uncommon Serotypes of Streptococcus agalactiae Isolated From Non-neonatal Patients in Thailand
Authors: Tulyaprawat Orawan , Pharkjaksu Sujiraphong , Shrestha Raj Kumar , Ngamskulrungroj Popchai
Published: September 2021

Group B streptococcus (GBS) or Streptococcus agalactiae is an opportunistic pathogen that causes serious illness in newborns, pregnant women, and adults. However, insufficient detection methods and disease prevention programs have contributed to an increase in the incidence and fatality rates associated with this pathogen in non-neonatal patients. This study aimed to investigate factors of the observed increased incidence by investigation of serotype distribution, virulence factors, and antimicrobial susceptibility patterns from invasive GBS disease among non-neonatal patients in Thailand. During 2017–2018, a total of 109 S. agalactiae isolates were collected from non-pregnant patients. There were 62 males and 47 females, with an average age of 63.5 years (range: 20 – 96). Serotypes were determined by latex agglutination assay and multiplex polymerase chain reaction (PCR)-based assay. Among those isolates, seven virulence genes (rib, bca, pavA, lmb, scpB, cylE, and cfb) were detected by PCR amplification, and were determined for their susceptibility to 20 antimicrobial agents using a SensititreTM Streptococcus species STP6F AST plate. Among the study isolates, serotype III was predominant (52.3%), followed by serotype V and serotype VI (13.8% for each), serotype Ib (11.9%), and other serotypes (8.2%). Of the seven virulence genes, pavA was found in 67.0%. Except for one, there were no significant differences in virulence genes between serotype III and non-serotype III. Study isolates showed an overall rate of non-susceptibility to penicillin, the first-line antibiotic, of only 0.9%, whereas the resistance rates measured in tetracycline, clindamycin, azithromycin, and erythromycin were 41.3, 22.0, 22.0, and 22.0%, respectively. Strains that were resistant to all four of those drugs were significantly associated with non-serotype III (p < 0.001). Using multi-locus sequence typing (MLST), 40.0% of the four-drug-resistant isolates belonged to serotype VI/ST1, followed by serotype Ib/ST1 (35.0%). Cluster analysis with global GBS isolates suggested that the multiple drug-resistant isolates to be strongly associated with the clonal complex (CC) 1 (p < 0.001). Compared to the 2014 study of 210 invasive GBS isolates conducted in 12 tertiary hospitals in Thailand, the proportion of serotype III has dramatically dropped from nearly 90% to about 50%. This suggests that resistances to the second-line antibiotics for GBS might be the selective pressure causing the high prevalence of non-serotype III isolates.
Maturation of Rhodobacter capsulatus Multicopper Oxidase CutO Depends on the CopA Copper Efflux Pathway and Requires the cutF Product
Authors: Öztürk Yavuz , Blaby-Haas Crysten E. , Daum Noel , Andrei Andreea , Rauch Juna , Daldal Fevzi , Koch Hans-Georg
Published: September 2021

Copper (Cu) is an essential cofactor required for redox enzymes in all domains of life. Because of its toxicity, tightly controlled mechanisms ensure Cu delivery for cuproenzyme biogenesis and simultaneously protect cells against toxic Cu. Many Gram-negative bacteria contain extracytoplasmic multicopper oxidases (MCOs), which are involved in periplasmic Cu detoxification. MCOs are unique cuproenzymes because their catalytic center contains multiple Cu atoms, which are required for the oxidation of Cu1+ to the less toxic Cu2+. Hence, Cu is both substrate and essential cofactor of MCOs. Here, we investigated the maturation of Rhodobacter capsulatus MCO CutO and its role in periplasmic Cu detoxification. A survey of CutO activity of R. capsulatus mutants with known defects in Cu homeostasis and in the maturation of the cuproprotein cbb3-type cytochrome oxidase (cbb3-Cox) was performed. This revealed that CutO activity is largely independent of the Cu-delivery pathway for cbb3-Cox biogenesis, except for the cupric reductase CcoG, which is required for full CutO activity. The most pronounced decrease of CutO activity was observed with strains lacking the cytoplasmic Cu chaperone CopZ, or the Cu-exporting ATPase CopA, indicating that CutO maturation is linked to the CopZ-CopA mediated Cu-detoxification pathway. Our data demonstrate that CutO is important for cellular Cu resistance under both aerobic and anaerobic growth conditions. CutO is encoded in the cutFOG operon, but only CutF, and not CutG, is essential for CutO activity. No CutO activity is detectable when cutF or its putative Cu-binding motif are mutated, suggesting that the cutF product serves as a Cu-binding component required for active CutO production. Bioinformatic analyses of CutF-like proteins support their widespread roles as putative Cu-binding proteins for several Cu-relay pathways. Our overall findings show that the cytoplasmic CopZ-CopA dependent Cu detoxification pathway contributes to providing Cu to CutO maturation, a process that strictly relies on cutF.
Effects of Subchronic Copper Poisoning on Cecal Histology and Its Microflora in Chickens
Authors: Huang Cheng , Shi Yan , Zhou Changming , Guo Lianying , Liu Guohui , Zhuang Yu , Li Guyue , Hu Guoliang , Liu Ping , Guo Xiaoquan
Published: September 2021

Copper (Cu) is an important trace element with a two-sided effect on the growth performance of animals, which depends on the timing and dosage of Cu addition, etc. The purpose of this study was to determine the effects of oral copper sulfate (CuSO4, 350 ppm) on growth performance, cecal morphology, and its microflora of chickens (n = 60) after 30, 60, and 90 days. The results showed that after 90 days of copper exposure, the chickens lost weight, the cecum mucosa was detached, and vacuolation and inflammatory infiltration occurred at the base of the lamina propria. In addition, using the 16S rDNA sequencing method, we observed that copper exposure changed the richness and diversity of intestinal microorganisms. At the phylum level, Proteobacteria and Actinobacteria both significantly increased, while Bacteroidetes significantly decreased in the Cu group compared with control check (CK) group. At the genus level, the relative abundance of Rikenellaceae_RC9_gut_group decreased significantly, while Ruminococcaceae_UCG-014, Lachnoclostridium, and [Eubacterium]_coprostanoligenes_group increased significantly after copper exposure, and the change in microflora was most significant at 90 days. Moreover, the relevance of genus-level bacteria was altered. PICRUST analysis revealed potential metabolic changes associated with copper exposure, such as Staphylococcus aureus infection and metabolic disorders of nutrients. To sum up, these data show that subchronic copper exposure not only affects the growth and development of chickens but also causes the imbalance of intestinal microflora, which may further induce metabolic disorders in chickens.
From Merkel Cell Polyomavirus Infection to Merkel Cell Carcinoma Oncogenesis
Authors: Krump Nathan A. , You Jianxin
Published: September 2021

Merkel cell polyomavirus (MCPyV) infection causes near-ubiquitous, asymptomatic infection in the skin, but occasionally leads to an aggressive skin cancer called Merkel cell carcinoma (MCC). Epidemiological evidence suggests that poorly controlled MCPyV infection may be a precursor to MCPyV-associated MCC. Clearer understanding of host responses that normally control MCPyV infection could inform prophylactic measures in at-risk groups. Similarly, the presence of MCPyV in most MCCs could imbue them with vulnerabilities that-if better characterized-could yield targeted intervention solutions for metastatic MCC cases. In this review, we discuss recent developments in elucidating the interplay between host cells and MCPyV within the context of viral infection and MCC oncogenesis. We also propose a model in which insufficient restriction of MCPyV infection in aging and chronically UV-damaged skin causes unbridled viral replication that licenses MCC tumorigenesis.
Bioinspired Smart Materials With Externally-Stimulated Switchable Adhesion
Authors: Wang Jie , Wan Yiyang , Wang Xiaowei , Xia Zhenhai
Published: September 2021

Living organisms have evolved, over billions of years, to develop specialized biostructures with switchable adhesion for various purposes including climbing, perching, preying, sensing, and protecting. According to adhesion mechanisms, switchable adhesives can be divided into four categories: mechanically-based adhesion, liquid-mediated adhesion, physically-actuated adhesion and chemically-enhanced adhesion. Mimicking these biostructures could create smart materials with switchable adhesion, appealing for many engineering applications in robotics, sensors, advanced drug-delivery, protein separation, etc. Progress has been made in developing bioinspired materials with switchable adhesion modulated by external stimuli such as electrical signal, magnetic field, light, temperature, pH value, etc. This review will be focused on new advance in biomimetic design and synthesis of the materials and devices with switchable adhesion. The underlying mechanisms, design principles, and future directions are discussed for the development of high-performance smart surfaces with switchable adhesion.
Spatial Navigation Is Impaired in Elderly Patients With Cerebral Small Vessel Disease
Authors: Zhào Hóngyi , Chi Liyi , Zhang Yanhai , Huang Yonghua , Tian Hongyan
Published: September 2021

Cerebral small vessel disease (SVD) refers to a heterogeneous group of pathological processes that result from damage to the small penetrating vessels in the brain. Spatial navigation, one of the most fundamental behaviors, has lately attracted considerable clinical interest. This study aimed to determine whether spatial navigation performance is impaired in elderly SVD patients. In total, 18 elderly patients with severe SVD, 40 elderly patients with non-severe SVD, and 41 age-matched healthy volunteers were classified according to the Fazekas scale. Spatial navigation was evaluated by Amunet (a computer-based analogy of Morris water maze software), and a mini-mental scale evaluation (MMSE), animal category verbal fluency test (VFT), clock drawing test (CDT), and trail making test (TMT) -B were also applied. Compared to healthy controls, severe SVD, rather than non-severe SVD patients, exhibited significantly worse performance on “allocentric + egocentric” (41.74 ± 29.10 vs. 31.50 ± 16.47 vs. 29.21 ± 19.03; p = 0.031). Furthermore, the different abilities of spatial navigation among groups reached a statistical level on allocentric subtests (46.93 ± 31.27 vs. 43.69 ± 23.95 vs. 28.56 ± 16.38; p = 0.003), but not on egocentric subtest (56.16 ± 39.85 vs. 56.00 ± 28.81 vs. 43.06 ± 25.07; p = 0.105). The linear regression analysis revealed that allocentric navigation deficit was significantly correlated with TMT-B (p = 0.000, standardized β = 0.342) and VFT (p = 0.016, standardized β = −0.873) performance in elderly SVD patients. These results elucidated that spatial navigation ability could be a manifestation of cognitive deficits in elderly patients with SVD.
Impacts of NF1 Gene Mutations and Genetic Modifiers in Neurofibromatosis Type 1
Authors: Wang Wei , Wei Cheng-Jiang , Cui Xi-Wei , Li Yue-Hua , Gu Yi-Hui , Gu Bin , Li Qing-Feng , Wang Zhi-Chao
Published: September 2021

Neurofibromatosis type 1 (NF1) is a tumor predisposition genetic disorder that directly affects more than 1 in 3,000 individuals worldwide. It results from mutations of the NF1 gene and shows almost complete penetrance. NF1 patients show high phenotypic variabilities, including cafe-au-lait macules, freckling, or other neoplastic or non-neoplastic features. Understanding the underlying mechanisms of the diversities of clinical symptoms might contribute to the development of personalized healthcare for NF1 patients. Currently, studies have shown that the different types of mutations in the NF1 gene might correlate with this phenomenon. In addition, genetic modifiers are responsible for the different clinical features. In this review, we summarize different genetic mutations of the NF1 gene and related genetic modifiers. More importantly, we focus on the genotype–phenotype correlation. This review suggests a novel aspect to explain the underlying mechanisms of phenotypic heterogeneity of NF1 and provides suggestions for possible novel therapeutic targets to prevent or delay the onset and development of different manifestations of NF1.
Altered Coupling Between Resting-State Cerebral Blood Flow and Functional Connectivity Strength in Cervical Spondylotic Myelopathy Patients
Authors: Wei Wuzeng , Wang Tao , Abulizi Tuersong , Li Bing , Liu Jun
Published: September 2021

Background: Changes in regional neural activity and functional connectivity in cervical spondylotic myelopathy (CSM) patients have been reported. However, resting-state cerebral blood flow (CBF) changes and coupling between CBF and functional connectivity in CSM patients are largely unknown.

Methods: Twenty-seven CSM patients and 24 sex/age-matched healthy participants underwent resting-state functional MRI and arterial spin labeling imaging to compare functional connectivity strength (FCS) and CBF between the two groups. The CBF–FCS coupling of the whole gray matter and specific regions of interest was also compared between the groups.

Results: Compared with healthy individuals, CBF–FCS coupling was significantly lower in CSM patients. The decrease in CBF–FCS coupling in CSM patients was observed in the superior frontal gyrus, bilateral thalamus, and right calcarine cortex, whereas the increase in CBF–FCS coupling was observed in the middle frontal gyrus. Moreover, low CBF and high FCS were observed in sensorimotor cortices and visual cortices, respectively.

Conclusion: In general, neurovascular decoupling at cortical level may be a potential neuropathological mechanism of CSM.
The Common Neurological Presentations and Clinical Outcomes of Coronavirus Disease 2019 in Saudi Arabia
Authors: Alkeridy Walid A. , Alanazy Mohammed H. , Alamri Nada , Alqahtani Awyshah , Alhazzani Adel , Muayqil Taim
Published: September 2021

Background: Neurological manifestations have increasingly become recognized in COVID-19. People from different ethnic backgrounds are experiencing different outcomes related to SARS-CoV-2 infection. Several cohort studies reported the common neurological manifestations and complications associated with COVID-19 disease around the world however, the prevalence of neurological complications associated with SARS-CoV-2 infection in the Arab countries and Saudi Arabia is still unknown.

Objective: To study the prevalence, risk factors, and characteristics of the neurological complications associated with COVID-19 and their relationship with clinical outcomes.

Methods: We conducted a prospective, single-center, observational, cohort study of consecutive hospitalized adults COVID-19 patients with and without neurological manifestation admitted between March 2020 until the end of December 2020. Data was collected prospectively using electronic medical records; Cases and controls were observed until they either get discharged from the hospital or died. The primary outcomes were death, survival, and survival with sequalae.

Results: Among 497 patients with COVID-19, 118 patients (23.7%) had neurological complications, 94 patients (18.9%) had encephalopathy, and 16 patients (3.2%) had cerebrovascular accidents (CVA). Patients with COVID-19-related neurological complications were older and more likely to have a pre-existing neurological disease. The most common neurological syndrome associated with COVID-19 were encephalopathy (18.9%) and headache (13.7%). Pre-existing neurological disease and an elevated neutrophil count were the strongest predictors of developing any neurological complications. Death form COVID-19 was associated with age (OR 1.06, 95% CI 1.02–1.10, P = 0.001), invasive ventilation (OR 37.12, 95% CI 13.36–103.14), COVID-19-related-neurological complications (OR 3.24, 95% CI 1.28–8.21, P = 0.01), and elevated CRP level (OR 1.01, 95% CI 1.00–1.01, P = 0.01).

Conclusions: COVID-19 is associated with a wide range of neurological manifestations in people living in Saudi Arabia, with older individuals and those with underlying neurological disorders being most at risk. The presence of neurological complications was associated with increased mortality and poor outcomes.
Preliminary Evaluations of [11C]Verubulin: Implications for Microtubule Imaging With PET
Authors: Lindberg Anton , Mossine Andrew V. , Aliaga Arturo , Hopewell Robert , Massarweh Gassan , Rosa-Neto Pedro , Shao Xia , Bernard-Gauthier Vadim , Scott Peter J. H. , Vasdev Neil
Published: September 2021

[11C]Verubulin (a.k.a.[11C]MCP-6827), [11C]HD-800 and [11C]colchicine have been developed for imaging microtubules (MTs) with positron emission tomography (PET). The objective of this work was to conduct an in vivo comparison of [11C]verubulin for MT imaging in mouse and rat brain, as well as an in vitro study with this radiotracer in rodent and human Alzheimer’s Disease tissue. Our preliminary PET imaging studies of [11C]verubulin in rodents revealed contradictory results between mouse and rat brain uptake under pretreatment conditions. In vitro autoradiography with [11C]verubulin showed an unexpected higher uptake in AD patient tissue compared with healthy controls. We also conducted the first comparative in vivo PET imaging study with [11C]verubulin, [11C]HD-800 and [11C]colchicine in a non-human primate. [11C]Verubulin and [11C]HD-800 require pharmacokinetic modeling and quantification studies to understand the role of how these radiotracers bind to MTs before translation to human use.
A Compartmentalized Neuronal Cell-Culture Platform Compatible With Cryo-Fixation by High-Pressure Freezing for Ultrastructural Imaging
Authors: Tran Hung Tri , Lucas Miriam S. , Ishikawa Takashi , Shahmoradian Sarah H. , Padeste Celestino
Published: September 2021

The human brain contains a wide array of billions of neurons and interconnections, which are often simplified for analysis in vitro using compartmentalized microfluidic devices for neuronal cell culturing, to better understand neuronal development and disease. However, such devices are traditionally incompatible for high-pressure freezing and high-resolution nanoscale imaging and analysis of their sub-cellular processes by methods including electron microscopy. Here we develop a novel compartmentalized neuronal co-culture platform allowing reconstruction of neuronal networks with high variable spatial control, which is uniquely compatible for high-pressure freezing. This cryo-fixation method is well-established to enable high-fidelity preservation of the reconstructed neuronal networks and their sub-cellular processes in a near-native vitreous state without requiring chemical fixatives. To direct the outgrowth of neurites originating from two distinct groups of neurons growing in the two different compartments, polymer microstructures akin to microchannels are fabricated atop of sapphire disks. Two populations of neurons expressing either enhanced green fluorescent protein (EGFP) or mCherry were grown in either compartment, facilitating the analysis of the specific interactions between the two separate groups of cells. Neuronally differentiated PC12 cells, murine hippocampal and striatal neurons were successfully used in this context. The design of this device permits direct observation of entire neuritic processes within microchannels by optical microscopy with high spatial and temporal resolution, prior to processing for high-pressure freezing and electron microscopy. Following freeze substitution, we demonstrate that it is possible to process the neuronal networks for ultrastructural imaging by electron microscopy. Several key features of the embedded neuronal networks, including mitochondria, synaptic vesicles, axonal terminals, microtubules, with well-preserved ultrastructures were observed at high resolution using focused ion beam – scanning electron microscopy (FIB-SEM) and serial sectioning – transmission electron microscopy (TEM). These results demonstrate the compatibility of the platform with optical microscopy, high-pressure freezing and electron microscopy. The platform can be extended to neuronal models of brain disease or development in future studies, enabling the investigation of subcellular processes at the nanoscale within two distinct groups of neurons in a functional neuronal pathway, as well as pharmacological testing and drug screening.
Eating and Lifestyle Habits in Youth With Down Syndrome Attending a Care Program: An Exploratory Lesson for Future Improvements
Authors: Roccatello Giulia , Cocchi Guido , Dimastromatteo Rosa Tullia , Cavallo Alessandra , Biserni Giovanni Battista , Selicati Mariella , Forchielli Maria Luisa
Published: September 2021

Introduction:Children with Down Syndrome (DS) have nutritional problems with unknown implications besides increased potential for obesity. Their food habits are unknown. We aim to delineate eating and lifestyle habits of DS children attending a multispecialist program to identify the challenges they face and the potential improvements.

Patients and Methods:We interacted with 34 DS children (22 males, 12 females, 2–16 years old) and their families. Food habits, medical conditions and treatments, degrees of development and physical activity, anthropometric and laboratory data were recorded over 6 months and analyzed. A 3-day food diary and a 24-h recall food frequency questionnaire were administered.

Results:Twenty-nine (85%) children completed meals, only 11 (32%) received alternative food such as milk. Weaning regularly started in 25 (73%) children. Preschool children introduced adequate calories and nutrients. School children and adolescents did not reach recommendations. All age groups, as the general pediatric population, excessively ate protein and saturated fat, and preferred bread, pasta, fruit juices, meat and cold cuts. Peculiarly, pulses and fish were adequately assumed by preschool and school children, respectively. Five children (15%) were overweight/obese.

Conclusions:Dietary excesses commonly found in the general pediatric population are also present in this DS group, proving a narrowing gap between the two. DS group performed better nutritionally in the early years and overweight/obesity occurrence seems contained. DS children may benefit from a practical yet professional care-program in which nutrition education may improve their growth, development and transition into adulthood.
Wheat–Fusarium graminearum Interactions Under Sitobion avenae Influence: From Nutrients and Hormone Signals
Authors: Luo Kun , Ouellet Thérèse , Zhao Huiyan , Wang Xiukang , Kang Zhensheng
Published: September 2021

The English grain aphid Sitobion avenae and phytopathogen Fusarium graminearum are wheat spike colonizers. “Synergistic” effects of the coexistence of S. avenae and F. graminearum on the wheat spikes have been shown in agroecosystems. To develop genetic resistance in diverse wheat cultivars, an important question is how to discover wheat–F. graminearum interactions under S. avenae influence. In recent decades, extensive studies have typically focused on the unraveling of more details on the relationship between wheat-aphids and wheat-pathogens that has greatly contributed to the understanding of these tripartite interactions at the ecological level. Based on the scientific production available, the working hypotheses were synthesized from the aspects of environmental nutrients, auxin production, hormone signals, and their potential roles related to the tripartite interaction S. avenae–wheat–F. graminearum. In addition, this review highlights the relevance of preexposure to the herbivore S. avenae to trigger the accumulation of mycotoxins, which stimulates the infection process of F. graminearum and epidemic of Fusarium head blight (FHB) in the agroecosystems.
Apigenin Inhibits the Histamine-Induced Proliferation of Ovarian Cancer Cells by Downregulating ERα/ERβ Expression
Authors: Liu Manman , Zhang Yani , Xu Qiqi , Liu Guirong , Sun Na , Che Huilian , He Tao
Published: September 2021

Background

Apigenin (APG), a natural flavonoid, can affect the development of a variety of tumors, but its role in ovarian cancer remains unclear. There has been an increasing amount of evidence supporting the vital role played by mast cells and the bioactive mediators they release, as components of the tumor microenvironment, in the progression of ovarian cancer (OC); however, the mechanism warrants further exploration.

Methods and Results

In this study, a combination of transcriptomics analysis and application of TCGA database was performed, and we found that the expression of genes related to mast cell degranulation in ovarian cancer tissues changed remarkably. We then explored whether histamine, a major constituent of mast cell degranulation, could affect the development of ovarian cancer through immunohistochemistry analysis and cell proliferation assays. The results showed that a certain concentration of histamine promoted the proliferation of ovarian cancer cells by upregulating the expression of estrogen receptor α (ERα)/estrogen receptor β (ERβ). Additionally, we found that the inhibition of ERα or the activation of ERβ could inhibit the proliferation of ovarian cancer cells induced by histamine through real-time PCR and western blot assays. Finally, we demonstrated the attenuation effect imparted by apigenin in histamine-mediated ovarian cancer via the PI3K/AKT/mTOR signaling pathway.

Conclusion

Our research revealed that apigenin decelerated ovarian cancer development by downregulating ER-mediated PI3K/AKT/mTOR expression, thus providing evidence of its applicability as a potentially effective therapeutic agent for ovarian cancer treatment.
A Real-World, Population-Based Retrospective Analysis of Therapeutic Survival for Recurrent Localized Renal Cell Carcinoma After Nephrectomy
Authors: Kim Sung Han , Choi Min Gee , Shin Ji Hye , Kim Young-Ae , Chung Jinsoo
Published: September 2021

We retrospectively analyzed therapeutic strategies and risk factors for overall survival (OS) in disease recurrence following curative nephrectomy for localized renal cell carcinoma (loRCC) using the Korean National Cancer Registry Database. We selected 1295 recurrent loRCC patients who underwent either partial or radical nephrectomy from 2007–2013. Patients were excluded for age <19 years, secondary RCC, multiple primary tumors, other SEER stages except for a localized or regional stage, postoperative recurrence within 3-month, and non-nephrectomized cases. Four therapeutic groups were statistically analyzed for OS and risk factors: surgery (OP, 12.0%), other systemic therapy (OST, 59.5%), radiotherapy (RT, 2.8%), and targeted therapy (TT, 25.8%). The overall mortality rate for recurrent loRCC was 32.5%, including 82.4% for RCC-related deaths. The baseline comparison among groups showed statistical differences for the diagnostic age of cancer and the SEER stage (p<0.05). Multivariate analysis of OS showed significance for the TT (hazard ratio [HR]: 6.27), OST (HR: 7.05), and RT (HR: 7.47) groups compared with the OP group, along with significance for the sex, SEER stage, and the time from nephrectomy to treatment for disease recurrence (p<0.05). The median OS curve showed a significantly better OS in the OP group (54.9 months) compared with the TT, OST, and RT groups (41.7, 42.9, and 38.0 months, respectively; p<0.001). In conclusion, the surgery-treated group had the best OS among the different therapeutic strategies for recurrent loRCC after nephrectomy, and the importance of the time from nephrectomy to secondary treatment was a significant prognostic factor.
A Phase I Trial of Dasatinib and Osimertinib in TKI Naïve Patients With Advanced EGFR-Mutant Non-Small-Cell Lung Cancer
Authors: Kim Chul , Liu Stephen V. , Crawford Jennifer , Torres Tisdrey , Chen Vincent , Thompson Jillian , Tan Ming , Esposito Giuseppe , Subramaniam Deepa S. , Giaccone Giuseppe
Published: September 2021

Background

Osimertinib is an effective first-line therapy option for EGFR-mutant NSCLC, but virtually all patients develop resistance. CRIPTO, through Src activation, has been implicated in resistance to EGFR tyrosine kinase inhibitor (EGFR-TKI) therapy. Dasatinib, a Src inhibitor, has shown preclinical synergy with EGFR-TKI therapy.

Method

This is a single-arm phase I/II trial of osimertinib and dasatinib in TKI-naïve advanced EGFR-mutant NSCLC (NCT02954523). A 3 + 3 design was used in the phase I to establish the recommended phase II dose (RP2D). Osimertinib 80 mg QD was combined with dasatinib 70 mg BID (DL2), 50 mg BID (DL1), 70 mg QD (DL-1), and 50 mg QD (DL-2).

Results

Ten patients (DL2: 3, DL1: 6, DL -1: 1) were enrolled. 3 (50%) of 6 patients at DL1 experienced a DLT (grade 3 headaches/body pain, neutropenia, rash, one each). Common treatment-related adverse events included pleural effusion (n=10), diarrhea (n=8), rash (n=7), transaminitis (n=7), thrombocytopenia (n=7), and neutropenia (n=7). While the MTD was not determined by protocol-defined DLT criteria, DL-2 was chosen as the RP2D, considering overall tolerability. Nine (90%) patients had a PR, including 1 unconfirmed PR. Median PFS was 19.4 months and median OS 36.1 months. The trial was closed to accrual prematurely due to slow accrual after the approval of osimertinib as first-line therapy.

Conclusions

The combination of dasatinib and osimertinib demonstrated anticancer activity. The treatment was limited by chronic toxicities mainly attributed to dasatinib. To improve the safety and tolerability of Src and EGFR co-inhibition, Src inhibitors with a more favorable safety profile should be utilized in future studies.

Clinical Trial Registration

https://clinicaltrials.gov/ct2/show/NCT02954523
Prolonged Slow Expiration Technique and Gastroesophageal Reflux in Infants Under the Age of 1 Year
Authors: Lievens Laure , Vandenplas Yvan , Vanlaethem Sylvie , Van Ginderdeuren Filip
Published: September 2021

Background:The Prolonged Slow Expiration Technique (PSE) is an airway clearance technique (ACT) carried out in newborn children with bronchial obstruction and hypersecretion to clear away the mucus from the respiratory tract. Evidence about the effect of PSE on gastroesophageal reflux (GER) is currently lacking in the literature. This study aimed to evaluate the influence of PSE on GER in infants under the age of 1 year.

Methods:Infants were observed using multichannel intraluminal impedance-pH monitoring (MII-pH) over 24 h. During monitoring, the participants were treated with one 20 min intervention of PSE in supine position, 2 h after feeding. In this controlled trial with intra-subject design, the number of reflux episodes (REs) during PSE were compared to 20 min before and after PSE.

Results:Fifty infants younger than 1 year were screened of whom 22 had a pathological GER. For the entire group, no significant difference was seen in the total number of REs between before, during, or after the PSE treatment (P= 0.76). No significant difference in total REs was found between the three measuring points (P= 0.59) in the group of infants with an abnormal MII-pH (n= 22).

Conclusion:PSE does not cause a significant difference in REs in infants younger than 1 year.

Registration number:NCT03341585.
Triptolide Downregulates the Expression of NRF2 Target Genes by Increasing Cytoplasmic Localization of NRF2 in A549 Cells
Authors: Nam Le Ba , Choi Won Jun , Keum Young-Sam
Published: September 2021

We have identified triptolide as a novel NRF2 inhibitor, which significantly attenuates ARE-luciferase activity at nanomolar concentrations. Triptolide did not affect the level of NRF2, but significantly inhibited the expression of NRF2 target genes in A549 cells. We found that NRF2 possesses a previously unrecognized NES in the Neh2 domain, and that triptolide promotes an interaction between NRF2 and CRM1. Triptolide also decreased nuclear accumulation of NRF2, suggesting that it promotes nuclear export of NRF2. In addition, we show that triptolide decreased the expression of NRF2 target genes and increased intracellular oxidative stress, suppressing invasion and promoting cisplatin-induced apoptosis in A549 cells. Finally, oral administration of triptolide suppressed the growth of A549 xenografts in athymic mice by decreasing the expression of NRF2 target genes and promoting oxidative damages via the nuclear export of NRF2 and CRM1 in vivo. To the best of our knowledge, triptolide is the first type of compound to inhibit NRF2 by increasing cytoplasmic localization of NRF2.
A High Performance Terahertz Photoconductive Antenna Array Detector With High Synthesis Efficiency
Authors: Shi Wei , Wang Zhiquan , Hou Lei , Wang Haiqing , Wu Meilin , Li Chaofan
Published: September 2021

A 2 × 2 terahertz photoconductive antenna (PCA) array detector with high efficiency synthesis characteristic that improves the signal-to-noise ratio (SNR) of the detected signals has been reported in this paper. By processing the substrate material through a special micromachining process, the current signal generated by the adjacent antenna elements as opposed to that generated by the antenna gap is eliminated. Experiments show that the amplitude of the current signal output by the PCA array detector is consistent with the amplitude of the synchronous superposition of the current signals output by antenna elements, and the synthesis efficiency of the device achieves 93.7%. At the same time, the antenna array detector has low current noise, and its highest SNR is 62 dB under the excitation of different light energy, which is related to the number of antenna array elements.
Theoretical Study on Characteristics of Glow Discharged Neon Gas and Its Interaction With Terahertz Waves
Authors: Hou Lei , Wang Yaodong , Wang Junnan , Yang Lei , Shi Wei
Published: September 2021

Discharge gases have been used to detect terahertz (THz) waves, however, there are few relevant theoretical studies. The neon glow discharge model is established by COMSOL Multiphysics software, the characteristics of glow discharged neon and the interaction of the discharged gas with THz waves were investigated. The results show that with the increase of THz wave’s frequency, the transmittance increases, the change of plasma discharge characteristics caused by THz wave can be used for THz wave detection. The results provide a theoretical basis for the development of cheap, room temperature THz wave detector with fast response speed, and high sensitivity.
Phosphorylation at Serines 157 and 161 Is Necessary for Preserving Cardiac Expression Level and Functions of Sarcomeric Z-Disc Protein Telethonin
Authors: Lewis Hannah R. , Eminaga Seda , Gautel Mathias , Avkiran Metin
Published: September 2021

Aims:In cardiac myocytes, the sarcomeric Z-disc protein telethonin is constitutively bis-phosphorylated at C-terminal residues S157 and S161; however, the functional significance of this phosphorylation is not known. We sought to assess the significance of telethonin phosphorylationin vivo, using a novel knock-in (KI) mouse model generated to express non-phosphorylatable telethonin (TcapS157/161A).

Methods and Results:TcapS157/161Aand wild-type (WT) littermates were characterized by echocardiography at baseline and after sustained β-adrenergic stimulationviaisoprenaline infusion. Heart tissues were collected for gravimetric, biochemical, and histological analyses. At baseline,TcapS157/161Amice did not show any variances in cardiac structure or function compared with WT littermates and mutant telethonin remained localized to the Z-disc. Ablation of telethonin phosphorylation sites resulted in a gene-dosage dependent decrease in the cardiac telethonin protein expression level in mice carrying the S157/161A alleles, without any alteration in telethonin mRNA levels. The proteasome inhibitor MG132 significantly increased the expression level of S157/161A telethonin protein in myocytes fromTcapS157/161Amice, but not telethonin protein in myocytes from WT mice, indicating a role for the ubiquitin–proteasome system in the regulation of telethonin protein expression level.TcapS157/161Amice challenged with sustained β-adrenergic stimulationviaisoprenaline infusion developed cardiac hypertrophy accompanied by mild systolic dysfunction. Furthermore, the telethonin protein expression level was significantly increased in WT mice following isoprenaline stimulation but this response was blunted inTcapS157/161Amice.

Conclusion:Overall, these data reveal that telethonin protein turnoverin vivois regulated in a novel phosphorylation-dependent manner and suggest that C-terminal phosphorylation may protect telethonin against proteasomal degradation and preserve cardiac function during hemodynamic stress. Given that human telethonin C-terminal mutations have been associated with cardiac and skeletal myopathies, further research on their potential impact on phosphorylation-dependent regulation of telethonin protein expression could provide valuable mechanistic insight into those myopathies.
Genome Assembly and Sex-Determining Region of Male and Female Populus × sibirica
Authors: Melnikova Nataliya V. , Pushkova Elena N. , Dvorianinova Ekaterina M. , Beniaminov Artemy D. , Novakovskiy Roman O. , Povkhova Liubov V. , Bolsheva Nadezhda L. , Snezhkina Anastasiya V. , Kudryavtseva Anna V. , Krasnov George S. , Dmitriev Alexey A.
Published: September 2021

The genus Populus is presented by dioecious species, and it became a promising object to study the genetics of sex in plants. In this work, genomes of male and female Populus × sibirica individuals were sequenced for the first time. To achieve high-quality genome assemblies, we used Oxford Nanopore Technologies and Illumina platforms. A protocol for the isolation of long and pure DNA from young poplar leaves was developed, which enabled us to obtain 31 Gb (N50 = 21 kb) for the male poplar and 23 Gb (N50 = 24 kb) for the female one using the MinION sequencer. Genome assembly was performed with different tools, and Canu provided the most complete and accurate assemblies with a length of 818 Mb (N50 = 1.5 Mb) for the male poplar and 816 Mb (N50 = 0.5 Mb) for the female one. After polishing with Racon and Medaka (Nanopore reads) and then with POLCA (Illumina reads), assembly completeness was 98.45% (87.48% duplicated) for the male and 98.20% (76.77% duplicated) for the female according to BUSCO (benchmarking universal single-copy orthologs). A high proportion of duplicated BUSCO and the increased genome size (about 300 Mb above the expected) pointed at the separation of haplotypes in a large part of male and female genomes of P. × sibirica. Due to this, we were able to identify two haplotypes of the sex-determining region (SDR) in both assemblies; and one of these four SDR haplotypes, in the male genome, contained partial repeats of the ARR17 gene (Y haplotype), while the rest three did not (X haplotypes). The analysis of the male P. × sibirica SDR suggested that the Y haplotype originated from P. nigra, while the X haplotype is close to P. trichocarpa and P. balsamifera species. Moreover, we revealed a Populus-specific repeat that could be involved in translocation of the ARR17 gene or its part to the SDR of P. × sibirica and other Populus species. The obtained results expand our knowledge on SDR features in the genus Populus and poplar phylogeny.
Cotton CC-NBS-LRR Gene GbCNL130 Confers Resistance to Verticillium Wilt Across Different Species
Authors: Li Tinggang , Zhang Qianqian , Jiang Xilong , Li Ran , Dhar Nikhilesh
Published: September 2021

Verticillium wilt (VW) is a destructive disease in cotton caused by Verticillium dahliae and has a significant impact on yield and quality. In the absence of safe and effective chemical control, VW is difficult to manage. Thus, at present, developing resistant varieties is the most economical and effective method of controlling Verticillium wilt of cotton. The CC-NBS-LRR (CNL) gene family is an important class of plant genes involved in disease resistance. This study identified 141 GbCNLs in Gossypium barbadense genome, with 37.5% (53 genes) GbCNLs enriched in 12 gene clusters (GC01–GC12) based on gene distribution in the chromosomes. Especially, seven GbCNLs from two largest clusters (GC11 and GC12) were significantly upregulated in the resistant cultivar (Hai No. 7124) and the susceptible (Giza No. 57). Virus-induced gene silencing of GbCNL130 in G. barbadense, one typical gene in the gene cluster 12 (GC12), significantly altered the response to VW, compromising plant resistance to V. dahliae. In contrast, GbCNL130 overexpression significantly increased the resistance to VW in the wild-type Arabidopsis thaliana. Based on our research findings presented here, we conclude that GbCNL130 promotes resistance to VW by activating the salicylic acid (SA)-dependent defense response pathway resulting in strong accumulation of reactive oxygen species and upregulation of pathogenesis-related (PR) genes. In conclusion, our study resulted in the discovery of a new CNL resistance gene in cotton, GbCNL130, that confers resistance to VW across different hosts.