bluuebunny/crossref_metadata_2025 huggingface dataset:
Genomic diversity across Candida auris clinical isolates shapes rapid development of antifungal resistance in vitro and in vivo
Authors: Burrack Laura S. , Todd Robert T. , Soisangwan Natthapon , Wiederhold Nathan P. , Selmecki Anna
Published: March 2022

Antifungal drug resistance and tolerance poses a serious threat to global public health. In the human fungal pathogen, Candida auris, resistance to triazole, polyene, and echinocandin antifungals is rising, resulting in multidrug resistant isolates. Here, we use genome analysis and in vitro evolution of seventeen new clinical isolates of C. auris from clades I and IV to determine how quickly resistance mutations arise, the stability of resistance in the absence of drug, and the impact of genetic background on evolutionary trajectories. We evolved each isolate in the absence of drug as well as in low and high concentrations of fluconazole. In just three passages, we observed genomic and phenotypic changes including karyotype alterations, aneuploidy, acquisition of point mutations, and increases in MIC values within the populations. Fluconazole resistance was stable in the absence of drug, indicating little to no fitness cost associated with resistance. Importantly, two isolates substantially increased fluconazole resistance to ≥256µg/ml fluconazole. Multiple evolutionary pathways and mechanisms to increase fluconazole resistance occurred simultaneously within the same population. Strikingly, the sub-telomeric regions of C. auris were highly dynamic as deletion of multiple genes near the sub-telomeres occurred during the three passages in several populations. Finally, we discovered a mutator phenotype in a clinical isolate of C. auris. This isolate had elevated mutation rates compared to other isolates and acquired substantial resistance during evolution in vitro and in vivo supporting that the genetic background of clinical isolates can have a significant effect on evolutionary potential.

Importance

Drug resistant Candida auris infections are recognized by the CDC as an urgent threat. Here, we obtained and characterized a set of clinical isolates of C. auris including multiple isolates from the same patient. To understand how drug resistance arises, we evolved these isolates and found that resistance to fluconazole, the most commonly prescribed antifungal, can occur rapidly and that there are multiple pathways to resistance. During our experiment, resistance was gained, but it was not lost, even in the absence of drug. We also found that some C. auris isolates have higher mutation rates than others and are primed to acquire antifungal resistance mutations. Furthermore, we found that multidrug resistance can evolve within a single patient. Overall, our results highlight the high stability and high rates of acquisition of antifungal resistance of C. auris that allow evolution of pan-resistant, transmissible isolates in the clinic.
Analysis of humoral immunity against emerging SARS-CoV-2 variants: a population-based prevalence study in Yokohama, Japan
Authors: Goto Atsushi , Miyakawa Kei , Nakayama Izumi , Yagome Susumu , Xu Juan , Kaneko Makoto , Ohtake Norihisa , Kato Hideaki , Ryo Akihide
Published: March 2022

Background

Little is known about the population prevalence of antibodies against emerging immune escape variants of SARS-CoV-2.

Methods

A population-based prevalence study was conducted in Yokohama City, the most populous municipality of Japan. Quantitative measurements of immunoglobulin G against SARS-CoV-2 spike protein (SP-IgG) and qualitative measurements of neutralization antibodies against the Omicron BA.1 and BA.2 variants were performed.

Results

Of 6,000 randomly selected residents aged 20–74, 1,277 participated in the study during a period from January 30 to February 28, 2022. Of them, 3% had prior diagnosis of COVID-19, 96% received at least two-doses of SARS-CoV-2 vaccines, and 94% were positive for SP-IgG. The positive rates of neutralizing antibodies were 28% to Omicron BA.1 and BA.2 variants in a random sample of 10% of participants (n=123) and 100% to BA.1 and BA.2 among participants who received the third vaccination at least 7 days before (n=66).

Conclusions

In this population-based prevalence study in Japan, most had SP-IgG antibodies but the overall neutralizing antibody positive rate was 28% against the Omicron BA.1 and BA.2 variants. The population-level insufficient humoral immunity against the Omicron variants may explain the outbreak of COVID-19 during this period in Japan.
Damaging missense variants inIGF1Rimplicate a role for IGF-1 resistance in the aetiology of type 2 diabetes
Authors: Gardner Eugene J. , Kentistou Katherine A. , Stankovic Stasa , Lockhart Samuel , Wheeler Eleanor , Day Felix R. , Kerrison Nicola D. , Wareham Nicholas J. , Langenberg Claudia , O’Rahilly Stephen , Ong Ken K. , Perry John R. B.
Published: March 2022

Type 2 diabetes (T2D) is a chronic metabolic disorder with a significant genetic component. While large-scale population studies have identified hundreds of common genetic variants associated with T2D susceptibility, the role of rare (minor allele frequency < 0.1%) protein coding variation is less clear. To this end, we performed a gene burden analysis of 18,691 genes in 418,436 (n=32,374 T2D cases) individuals sequenced by the UK Biobank (UKBB) study to assess the impact of rare genetic variants on T2D risk. Our analysis identified T2D associations at exome-wide significance (P < 6.9×10-7) with rare, damaging variants within previously identified genes includingGCK, GIGYF1, HNF1A, andTNRC6B. In addition, individuals with rare, damaging missense variants in the genesZEB2(N=31 carriers; OR=5.5 [95% CI=2.5-12.0]; p=6.4×10-7),MLXIPL(N=245; OR=2.3 [1.6-3.2]; p=3.2×10-7), andIGF1R(N=394; OR=2.4 [1.8-3.2]; p=1.3×10-10) have higher risk of T2D. Carriers of damaging missense variants withinIGF1Rwere also shorter (-2.2cm [-1.8-2.7]; p=1.2×10-19) and had higher circulating protein levels of insulin-like growth factor-1 (IGF-1; 2.3 nmol/L [1.7-2.9] p=2.8×10-14), indicating relative IGF-1 resistance. A likely causal role of IGF-1 resistance on T2D was further supported by Mendelian randomisation analyses using common variants. Our results increase our understanding of the genetic architecture of T2D and highlight a potential therapeutic benefit of targeting the Growth Hormone/IGF-1 axis.
Monitoring of the SARS-CoV-2 Omicron BA.1/BA.2 variant transition in the Swedish population reveals higher viral quantity in BA.2 cases
Authors: Lentini Antonio , Pereira Antonio , Winqvist Ola , Reinius Björn
Published: March 2022

Throughout the SARS-CoV-2 pandemic, multiple waves of variants of concern have swept across populations, leading to a chain of new and yet more contagious lineages dominating COVID-19 cases. Here, we tracked the remarkably rapid shift from Omicron BA.1 to BA.2 sub-variant dominance in the Swedish population during January–March 2022. By analysis of 174,933 clinical nasopharyngeal swab samples using a custom variant-typing RT-PCR assay, we uncover nearly two-fold higher levels of viral RNA in cases with Omicron BA.2. Importantly, increased viral load in the upper pharynx upon BA.2 infection may provide part of the explanation why Omicron BA.2 is more transmissible and currently outcompetes the BA.1 variant across populations.
Epigenetic modulators link mitochondrial redox homeostasis to cardiac function
Authors: Elbeck Zaher , Hossain Mohammad Bakhtiar , Siga Humam , Oskolkov Nikolay , Karlsson Fredrik , Lindgren Julia , Walentinsson Anna , Remedios Cristobal Dos , Koppenhöfer Dominique , Jarvis Rebecca , Bürli Roland , Jamier Tanguy , Franssen Elske , Firth Mike , Degasperi Andrea , Bendtsen Claus , Dudek Jan , Kohlhaas Michael , Nickel Alexander G. , Lund Lars H. , Maack Christoph , Végvári Ákos , Betsholtz Christer
Published: March 2022

Excessive production of reactive oxygen species (ROS) is characteristic of numerous diseases, but most studies in this area have not considered the impact of endogenous antioxidative defenses. Here, utilizing multi-omics, we demonstrate that in cardiomyocytes mitochondrial isocitrate dehydrogenase (IDH2) constitutes a major antioxidant defense. In both male and female mice and humans the paradoxical reduction in expression of IDH2 associated with heart failure is compensated for by an increase in the enzyme’s activity. We describe extensive mutual regulation of the antioxidant activities of IDH2 and NRF2 by a network involving 2-oxoglutarate and L2-hydroxyglutarate and mediated in part through unconventional hydroxymethylation of cytosine residues present in introns. Conditional targeting of ROS in a murine model of heart failure improves cardiac function. Together, these insights may explain why previous attempts to treat heart failure with antioxidants have been unsuccessful and open new approaches to personalizing and, thereby, improving such treatment.

Graphical abstract

Highlights

Paradoxical downregulation of mitochondrial isocitrate dehydrogenase (IDH2) in response to oxidative stress leads to the discovery of a robust antioxidative defense in the heart.

An antioxidative loop involving IDH2 coordinates other antioxidative defenses, such as NRF2.

This loop produces epigenetic modifications that link oxidative stress to mitochondrial function.

The conclusion that enhancing antioxidative capacity improves cardiac function only when the endogenous capacity is insufficient opens new approaches to individualized treatment of patients with heart failure.
Phase transitions of a SEC14-like condensate at Arabidopsis plasma membranes regulate root growth
Authors: Liu Chen , Mentzelopoulou Andriani , Papagavriil Fotini , Ramachandran Prashanth , Perraki Artemis , Claus Lucas , Barg Sebastian , Dörmann Peter , Jaillais Yvon , Johnen Philipp , Russinova Eugenia , Gizeli Electra , Schaaf Gabriel , Moschou Panagiotis Nikolaou
Published: March 2022

Protein function can be modulated by phase transitions in their material properties, which can range from liquid-to solid-like; yet the mechanisms that drive these transitions and whether they are important for physiology are still unknown. Using the model plant Arabidopsis, we show that developmental robustness is reinforced by phase transitions of the plasma membrane–bound lipid-binding protein SEC14-like. Using imaging, genetics, andin vitroreconstitution experiments, we show that SEC14-like undergoes liquid-like phase separation in the root stem cells. Outside the stem cell niche, SEC14-like associates with the caspase-like protease separase and conserved microtubule motors at unique polar plasma membrane interfaces. In these interfaces, SEC14-like undergoes abrupt processing by separase, which promotes its liquid-to-solid transition. The SEC14-like liquid-to-solid transition is important for root developmental robustness, as lines expressing an uncleavable SEC14-like variant or mutants of separase, and associated microtubule motors show similar developmental phenotypes. Furthermore, the processed and solidified but not the liquid form of SEC14-like interacts with the polar protein PINFORMED2 at the plasma membrane and perhaps other polar proteins of the PINFORMED family. This work demonstrates that robust development can involve abrupt liquid-to-solid transitions mediated by proteolysis at unique plasma membrane interfaces.
ALS sensitive spinal motor neurons enter a degenerative downward spiral of impaired splicing and proteostasis
Authors: Iannitelli Dylan E. , Tan Albert , Nguyen Emma , Babu Asha , Elorza Setiembre Delfina , Joseph Tyler , Zaaijer Sophie , An Disi , Ward Michael , Mazzoni Esteban O.
Published: March 2022

Despite clear therapeutic potential, the mechanisms that confer differential neuronal sensitivity are not well understood. During Amyotrophic Lateral Sclerosis (ALS), sensitive spinal motor neurons (SpMN) die while a subset of rostral cranial motor neurons (CrMN) survive. In this work, we optimized a protocol to differentiate CrMNs and SpMNs from human induced pluripotent stem cells (iPSCs) by direct programming and positional patterning. Human iCrMNs are more resistant than iSpMNs to proteotoxic stress and rely on the proteasome to maintain proteostasis. iCrMNs better prevent mislocalization of TDP43 from the nucleus, a hallmark of ALS progression. iSpMNs contain more splicing defects than iCrMNs in response to ALS-related stress with genes involved in splicing and proteostasis maintenance. Therefore, iCrMNs resist ALS at two levels, preventing protein accumulation and reducing splicing defects in response to TDP43 nuclear depletion. Thus, ALS-sensitive iSpMNs appear to enter a downward spiral compromising their ability to maintain proteostasis and splicing.
Modulation of RNA binding properties of the RNA helicase UPF1 by its activator UPF2
Authors: Xue Guangpu , Maciej Vincent D. , de Amorim Alexandrina Machado , Pak Melis , Jayachandran Uma , Chakrabarti Sutapa
Published: March 2022

The NMD helicase UPF1 is a prototype of the superfamily 1 (SF1) of RNA helicases that bind RNA with high affinity and translocate on it in an ATP-dependent manner. Previous studies showed that UPF1 has a low basal catalytic activity that is greatly enhanced upon binding of its interaction partner, UPF2. Activation of UPF1 by UPF2 entails a large conformational change that switches the helicase from an RNA-clamping mode to an RNA-unwinding mode. The ability of UPF1 to bind RNA was expected to be unaffected by this activation mechanism. Here we show, using a combination of biochemical and biophysical methods, that binding of UPF2 to UPF1 drastically reduces the affinity of UPF1 for RNA, leading to a release of the bound RNA. Although UPF2 is capable of binding RNA in vitro, our results suggest that dissociation of the UPF1-RNA complex is not a consequence of direct competition in RNA binding but rather an allosteric effect that is likely mediated by the conformational change induced by binding of the helicase to its activator. We discuss these results in light of transient interactions forged during mRNP assembly, particularly in the UPF1-dependent mRNA decay pathways.
Structure of the connexin-43 gap junction channel in a putative closed state
Authors: Qi Chao , Acosta-Gutierrez Silvia , Lavriha Pia , Othman Alaa , Lopez-Pigozzi Diego , Bayraktar Erva , Schuster Dina , Picotti Paola , Zamboni Nicola , Bortolozzi Mario , Gervasio Francesco L. , Korkhov Volodymyr M.
Published: March 2022

Gap junction channels (GJCs) mediate intercellular communication by connecting two neighboring cells and enabling direct exchange of ions and small molecules. Cell coupling via connexin-43 (Cx43) GJCs is important in a wide range of cellular processes in health and disease1-3, yet the structural basis of Cx43 function and regulation has not been determined until now. Here we describe the structure of a human Cx43 GJC solved by cryo-EM and single particle analysis at 2.26 Å resolution. The pore region of Cx43 GJC features several lipid-like densities per Cx43 monomer, located close to a putative lateral access site at the monomer boundary. We found a previously undescribed conformation on the cytosolic side of the pore, formed by the N-terminal domain and the transmembrane helix 2 of Cx43 and stabilized by a small molecule. Structures of the Cx43 GJC and hemichannels in nanodiscs reveal a similar gate arrangement. The features of the Cx43 GJC and hemichannel cryo-EM maps and the channel properties revealed by molecular dynamics simulations suggest that the captured states of Cx43 are consistent with a closed state.
SWEET11 and SWEET12 transporters function in tandem to modulate sugar flux in Arabidopsis: An account of the underlying unique structure–function relationship
Authors: Fatima Urooj , Balasubramaniam D. , Khan Wajahat Ali , Kandpal Manu , Vadassery Jyothilakshmi , Arockiasamy Arulandu , Senthil-Kumar Muthappa
Published: March 2022

Sugar will eventually be exported transporters (SWEETs) have been identified as a unique class of sugar efflux transporters in all biological kingdoms. AtSWEET11 and AtSWEET12 in Arabidopsis act synergistically to perform distinct physiological roles, particularly in apoplasmic phloem loading, seed filling, and sugar level alteration at the site of pathogen infection. Plasma membrane-localized AtSWEET11 and AtSWEET12 transporters exclusively facilitate sucrose transport along the concentration gradient. This article examines the sucrose binding pocket of AtSWEET11 and AtSWEET12 using docking studies, and how they act synergistically in various functions throughout plant development and during abiotic and biotic stresses. Further, we highlight the phylogenetic and the in-silico analyses of AtSWEET11 and AtSWEET12 orthologs from 39 economically important plant species that could provide new platforms for future studies on sugar allocation mechanisms across the different plant families. In-depth understanding of these transporters and their molecular regulatory mechanisms could be harnessed for crop improvement and crop protection.
Dynamic interaction of BcsD subunit in type I bacterial cellulose synthase
Authors: Kondo Tatsuya , Nakamura Yui , Nojima Shingo , Yao Min , Imai Tomoya
Published: March 2022

Cellulose is a promising biological material for supporting sustainable human life. This natural polymer is synthesized by cellulose synthase, a protein complex in the cell membrane. Cellulose synthase in bacteria is a hetero-subunit complex, and its subunit organization varies widely depending on the species. In the type I bacterial cellulose synthase complex, the BcsD (bacterial cellulose synthase D) protein is believed to play an important role in producing cellulose with long slender fiber morphology and high crystallinity, given the phenotype of the bcsD-deficient mutant and the specific existence of the type I operon in bacterial species synthesizing crystalline cellulose microfibrils such as Acetobacter. In this study, we successfully established a heterogeneously expressed Bcs protein in Escherichia coli as an experimental model and conducted biochemical studies for the BcsD protein and the other three major subunits of bacterial cellulose synthase, BcsA, BcsB, and BcsC. It has been shown that the BcsD protein interacts with the functionally required minimal subunits of the BcsAB complex, as well as the BcsC protein. Furthermore, it was shown that BcsD interacts with the BcsAB complex in two modes: direct protein-protein interactions and indirect interactions through the product cellulose. The former and latter modes represent the basal and active states of the type I bacterial cellulose synthase, respectively. This dynamic behavior of the BcsD protein is important for the type I bacterial cellulose synthase complex to regulate the crystallization process of cellulose.
CRISPR-induced exon skipping of β-catenin reveals tumorigenic mutants driving distinct subtypes of liver cancer
Authors: Mou Haiwei , Yue Junjiayu , Jin Ying , Wang Zhikai , Gao Ya , Janowitz Tobias , Meyer Hannah V. , Kucukural Alper , Wilkinson John E , Ozata Deniz M. , Beyaz Semir
Published: March 2022

CRISPR/Cas9-driven cancer modeling studies are based on disruption of tumor suppressor genes (TSGs) by small insertions or deletions (indels) that lead to frame-shift mutations. In addition, CRISPR/Cas9 is widely used to define the significance of cancer oncogenes and genetic dependencies in loss-of function studies. However, how CRISPR/Cas9 influences gain-of-function oncogenic mutations is elusive. Here, we demonstrate that single guide RNA targeting exon 3 of β-catenin results in exon skipping and generates gain-of-function isoforms in vivo. CRISPR/Cas9-mediated exon skipping of β-catenin induces liver tumor formation in synergy with YAPS127A in mice. We define two distinct exon skipping-induced tumor subtypes with different histological and transcriptional features. Notably, ectopic expression of two exon-skipped β-catenin transcript isoforms together with YAPS127A phenocopies the two distinct subtypes of liver cancer. Moreover, we identify similar β-catenin exon skipping events in hepatocellular carcinoma (HCC) patients. Collectively, our findings advance our understanding of β-catenin-related tumorigenesis and reveal that CRISPR/Cas9 can be repurposed, in vivo, to study gain-of-function mutations of oncogenes in cancer.
Solution structure of NPSL2, a regulatory element in the oncomiR-1 RNA
Authors: Liu Yaping , Munsayac Aldrex , Hall Ian , Keane Sarah C.
Published: March 2022

The miR-17~92a polycistron, also known as oncomiR-1, is commonly overexpressed in multiple cancers and has several oncogenic properties. OncomiR-1 encodes six constituent microRNAs (miRs), each enzymatically processed with different efficiencies. However, the structural mechanism that regulates this differential processing remains unclear. Chemical probing of oncomiR-1 revealed that the Drosha cleavage sites of pri-miR-92a are sequestered in a four-way junction. NPSL2, an independent stem loop element, is positioned just upstream of pri-miR-92a and sequesters a crucial part of the sequence that constitutes the basal helix of pri-miR-92a. Disruption of the NPSL2 hairpin structure could promote the formation of a pri-miR-92a structure that is primed for processing by Drosha. Thus, NPSL2 is predicted to function as a structural switch, regulating pri-miR-92a processing. Here, we determined the solution structure of NPSL2 using solution NMR spectroscopy. This is the first high-solution structure of an oncomiR-1 element. NPSL2 adopts a hairpin structure with a large, but highly structured, apical and internal loops. The 10-bp apical loop contains a pH-sensitive A+·C mismatch. Additionally, several adenosines within the apical and internal loops have elevated pKa values. The protonation of these adenosines can stabilize the NPSL2 structure through electrostatic interactions. Our study provides fundamental insights into the secondary and tertiary structure of an important RNA hairpin proposed to regulate miR biogenesis.
Local 5-HT signal bi-directionally regulates the coincidence time window of associative learning
Authors: Zeng Jianzhi , Li Xuelin , Zhangren Zimo , Lv Mingyue , Wang Yipan , Tan Ke , Xia Xiju , Wan Jinxia , Jing Miao , Yang Yang , Li Yan , Li Yulong
Published: March 2022

Temporal coincidence between the conditioned stimulus (CS) and unconditioned stimulus (US) is essential for associative learning across species. Despite its ubiquitous presence, the mechanism that may regulate this time window duration remains unclear yet. Using olfactory associative learning in Drosophila as a model, we find that suppressing or promoting serotonin (5-HT) signal could respectively shorten or prolong the coincidence time window of odor-shock associative learning and synaptic plasticity in mushroom body (MB) Kenyon cells (KCs). Capitalizing on GPCR-activation based (GRAB) sensors for 5-HT and acetylcholine (ACh), we characterized the in vivo 5-HT dynamics in MB lobes during odor and shock stimulations and further dissected this microcircuit. Interestingly, local KC-released ACh activates nicotinic receptors on the dorsal paired medial (DPM) neuron, and in turn the DPM neuron releases 5-HT to inhibit the ACh signal via the 5-HT1a receptor. Finally, we demonstrated that the DPM-mediated serotonergic feedback circuit is sufficient and necessary to regulate the coincidence time window. This work provides a model for studying the temporal contingency of environmental events and their causal relationship.
A note on the correspondence between horizontal and vertical price transmission
Authors: Kinnucan Henry W.
Published: March 2022

Horizontal (vertical) elasticities of price transmission in a competitive market can be positive or negative in sign depending on whether observed changes in market prices are triggered by shifts in the supply or demand for the commodity or by changes in trade (marketing) costs. These facts suggest estimates of horizontal (vertical) elasticities of price transmission obtained from empirical models that exclude trade (marketing) costs will suffer from attenuation bias. The attenuation bias is apt to cause inferences about market efficiency to be more pessimistic than is warranted.
Preparation and physiochemical properties of enzymatically modified octenyl succinate starch
Authors: Chen Xingyu , Hu Zhongshan , Chen Da , Feng Tao
Published: March 2022

The high viscosity and low solubility of octenyl succinate starch (OSAS) in water often lead to low reaction efficiency and uneven distribution of OSA groups, which can be solved by subjecting OSA starch to enzymatic digestion. Waxy corn starch is used to produce OSAS. The molecular weight of enzymolysis modified octenyl succinate starches (E‐OSAS) decreased with the addition of enzyme, and the substitution of enzymatically digested OSAS was larger than that of OSAS. As the addition of pullulanase increased, the substitution of E‐OSAS tended to increase and the apparent viscosity decreased significantly. The smaller its particle size and polydispersity index (PDI), the larger the absolute value of the zeta potential. E40‐OSAS with the smallest particle size and best homogeneity and stability had the smallest weight average molecular weight (1.38 × 106 g/mol), the smallest number average molecular weight (1.37 × 105 g/mol), and the largest degree of substitution of 0.019. The E‐OSAS has not been investigated deeply enough, and the enzymatic treatment by pullulanase is not much studied, so this study is of great importance to provide some basis for the research of new microcapsules.

Practical Application

This work aims to provide wall materials with lower viscosity and better encapsulation properties for the food industry. Waxy corns starch is used to manufacture OSAS. This wall material is a partly biodegradable material that is hydrophilic on the outside and hydrophobic on the inside, and its preparation process is in line with the concept of green chemistry.
Effects of drying temperature on umami taste and aroma profiles of mushrooms (Suillus granulatus)
Authors: Hou Zhenshan , Wei Yunyun , Sun Libin , Xia Rongrong , Xu Heran , Li Yunting , Feng Yao , Fan Wenli , Xin Guang
Published: March 2022

Temperature is one of the most important factors for drying edible mushrooms. To evaluate the effects of different hot‐air drying (HAD) temperatures on the umami taste and aroma profile of Suillus granulatus (S. granulatus) mushrooms, we measured umami substances and volatile compounds of S. granulatus dried at 40°C, 50°C, 60°C, 70°C, and 80°C. Results showed that when dried at 60°C, S. granulatus exhibited significantly higher (p < 0.05) equivalent umami concentration, taste activity values of glutamic acid (Glu) and 5′‐guanosine monophosphate (5′‐GMP), and electronic tongue umami sensory scores. The results identified a total of 71 volatile components of which geranylacetone, benzaldehyde, phenylethyl alcohol, and 3‐methylbutanoic acid were the dominant compounds. Sensory evaluation and relative odor activity values (ROAVs) revealed that 16 volatile compounds were the key volatile organic compounds contributing mushroom‐like and sweet odor to the overall aroma of S. granulatus; these included 1‐octen‐3‐ol (ROAV: 15.11‐62.06) and ethyl phenylacetate (ROAV: 13.62‐79.11). The drying temperature changed the aroma profile of S. granulatus. Furthermore, the mushroom dried at 60°C had a more desirable mushroom‐like and almond odor. It was, therefore, proposed that HAD at 60°C was optimal for retaining a pleasant flavor in S. granulatus. This study provides a theoretical basis for the optimal drying condition selection for the mushroom processing industry.

Practical Application

Hot‐air drying at 60°C can significantly retain the flavor of S. granulatus and is an optimal temperature for mushroom drying.
Identification of zinc‐chelating pumpkin seed (Cucurbita pepo L.) peptides and in vitro transport of peptide–zinc chelates
Authors: Peng Mengyao , Lu Dan , Yu Min , Jiang Bo , Chen Jingjing
Published: March 2022

In this study, pumpkin seeds peptide was purified, characterized, and evaluated for their zinc‐chelating capability, as well as in vitro bioaccessibility and transportation. Raw pumpkin seeds protein hydrolysate (PSPH) was produced by papain hydrolysis. The peptide fractions with the highest zinc‐chelating abilities were purified using immobilized metal affinity chromatography (IMAC) followed by gel filtration chromatography (GF). Eight peptides were identified, two of which with the lowest molecular weights were synthesized (RPKHPLK and RPKHPLSHDL) for determining potential bioaccessibility and bioavailability. Our results showed that the gastrointestinal stability of RPKHPLK‐Zn and RPKHPLSHDL‐Zn was higher than that of inorganic zinc salts in the simulated gastrointestinal tract model. Furthermore, the influence of the peptide zinc chelates on zinc transport was explored in vitro using Caco‐2 cell monolayer model. It was also shown that both RPKHPLK‐Zn and RPKHPLSHDL‐Zn could increase zinc transport rate and may be used to facilitate effective zinc absorption. The result of this study may provide important implications for developing plant protein foods with higher nutritional value.

Practical Application

As a potential alternative protein source, pumpkin seeds may find promising applications in plant‐based foods and drinks to meet the growing market for nonanimal foods. In this study, pumpkin seed protein peptides were prepared and purified, and the zinc‐chelating peptides were identified and evaluated for the abilities to promote the uptake of zinc. This type of mineral peptide chelates could be incorporated into plant‐based foods to increase mineral contents, which is significantly higher in foods originated from animals. The result of our study may provide important information for food industry to increase the nutritional value of plant‐based foods.
2‐Fluorodeschloroketamine has similar abuse potential as ketamine
Authors: Li Feng , Du Han , Wu Bo , Wei Jiayun , Qiao Yanling , Lai Miaojun , Zhou Wenhua , Shen Haowei , Wang Youmei , Xu Peng , Di Bin
Published: March 2022

2‐Fluorodeschloroketamine (2‐FDCK) as a substitute for ketamine has emerged among drug abusers in recent years. However, 2‐FDCK has not been controlled or regulated in many countries, which may be partly related to the lack of evidence on its abuse potential. In this study, we evaluated the abuse potential of 2‐FDCK via the tests of the conditioned place preference (CPP), locomotor sensitization, drug self‐administration and drug discrimination using ketamine as a reference. 2‐FDCK induced significant CPP at a minimum dose of 3 mg/kg in mice, an effect comparable with that of ketamine (3 mg/kg). Acute injections of 2‐FDCK or ketamine at 30 mg/kg enhanced locomotor activity. Repeated treatments with this dose of 2‐FDCK and ketamine induced locomotor sensitization after withdrawal. 2‐FDCK readily induced self‐administration with 0.5 mg/kg/infusion, the same dose for ketamine, and induced the highest seeking response at 1 mg/kg. Drug discrimination test showed that 2‐FDCK dose‐dependently substitute for ketamine with comparable ED50 to ketamine in substitution testing. Taken together, these results strongly suggested that 2‐FDCK has an abuse potential comparable with ketamine.
Sequential class switch recombination to IgE and allergen‐induced accumulation of IgE+ plasmablasts occur in the nasal mucosa of local allergic rhinitis patients
Authors: Testera‐Montes Almudena , Palomares Francisca , Jurado‐Escobar Raquel , Fernandez‐Santamaria Ruben , Ariza Adriana , Verge Jesus , Salas Maria , Campo Paloma , Mayorga Cristobalina , Torres Maria Jose , Rondon Carmen , Eguiluz‐Gracia Ibon
Published: March 2022

Background

The involvement of allergen‐specific (s)IgE in local allergic rhinitis (LAR) has been debated. Here, we investigate the effect of nasal allergen challenge with Dermatophagoides pteronyssinus (NAC‐DP) in mucosal and peripheral B‐cell subpopulations in LAR patients.

Methods

Nine LAR, 5 allergic rhinitis (AR), and 5 non‐atopic healthy control (HC) individuals were subjected to a 3‐day NAC‐DP protocol, and nasal biopsies and blood samples were collected before and after provocation. Nasal biopsies were used for immunohistochemistry and gene expression studies, whereas the frequency of lymphocyte subsets and basophil activation test (BAT) were analyzed in blood samples by flow cytometry. sIgG was measured in sera.

Results

NAC‐DP induced an increase in IgE+CD38+ plasmablasts in the nasal mucosa of LAR patients, but not in AR or HC individuals. Markers of sequential recombination to IgE (εCSR) (from IgG) were observed in 33% of LAR, 20% of AR, and 0% of HC subjects. NAC‐DP increased the proportion of peripheral CD19+CD20+CD38+ plasmablasts in AR and LAR patients, but not in HC. Expression of the mucosal homing receptor CXCR3 in peripheral CD19+CD20+CD38+ plasmablasts from LAR, AR, and HC individuals was 7%, 5%, and 0.5%, respectively. In vitro DP stimulation increased proliferating CD19+CD20+CD38+ plasmablasts in LAR and AR patients, but not in HC. Serum DP‐sIgG was higher in LAR and AR patients as compared to HC. BAT was positive in 33%, 100%, and 0% of LAR, AR, and HC subjects, respectively.

Conclusion

These results suggest that allergen exposure induces the sequential εCSR of IgG+CD19+CD20+CD38+ plasmablasts in the nasal mucosa of LAR patients.
A new person‐fit method based on machine learning in CDM in education
Authors: Zhu Zhemin , Arthur David , Chang Hua‐Hua
Published: March 2022

Cognitive diagnosis models have become popular in educational assessment and are used to provide more individualized feedback about a student's specific strengths and weaknesses than traditional total scores. However, if the testing data are contaminated by certain biases or aberrant response patterns, such predictions may not be accurate. The current research objective is to develop a new person‐fit method that is based on machine learning and improves the functionality of existing person‐fit methods. Various simulations were designed under three aberrant conditions: cheating, sleeping and random guessing. Simulation results showed that the new method was more powerful and effective than previous methods, especially for short‐length tests.
Prescribing and using vitiligo treatments: lessons from a nested process evaluation within the HI-Light vitiligo randomized controlled trial
Authors: Leighton Paul , Chalmers Joanne R. , Batchelor Jonathan M. , Rogers Andy , Akram Perways , Haines Rachel H. , Meakin Garry D. , White Jennifer , Ravenscroft Jane C. , Sach Tracey H. , Santer Miriam , Whitton Maxine E. , Eleftheriadou Viktoria , Thomas Kim S.
Published: March 2022

Background

The HI-Light Trial demonstrated that for active, limited vitiligo, combination treatment with potent topical corticosteroid (TCS) and handheld narrowband ultraviolet B offers a better treatment response than potent TCS alone. However, it is unclear how to implement these findings.

Aim

We sought to answer three questions: (i) Can combination treatment be used safely and effectively by people with vitiligo?; (ii) Should combination treatment be made available as routine clinical care?; and (iii) Can combination treatment be integrated within current healthcare provision?

Methods

This was a mixed-methods process evaluation, including semi-structured interviews with a purposive sample of trial participants, structured interviews with commissioners, and an online survey and focus groups with trial staff. Transcripts were coded by framework analysis, with thematic development by multiple researchers.

Results

Participants found individual treatments easy to use, but the combination treatment was complicated and required nurse support. Both participants and site investigators felt that combination treatment should be made available, although commissioners were less certain. There was support for the development of services offering combination treatment, although this might not be prioritized above treatment for other conditions. A ‘mixed economy’ model was suggested, involving patients purchasing their own devices, although concerns regarding the safe use of treatments mean that training, monitoring and ongoing support are essential. The need for medical physics support may mean that a regional service is more practical.

Conclusion

Combination treatment should be made available for people seeking treatment for vitiligo, but services require partnership with medical physics and ongoing training and support for patients.
DNA image cytometry parameters to identify high‐grade cervical lesions
Authors: Costa Ane Francyne , Pogere Adriane , Pasinato Ana Paula Beltrame Farina , Onofre Alexandre Sherlley Casimiro , Onofre Fabiana Botelho de Miranda
Published: March 2022

Objective

Evaluate the performance of different DNA image cytometry (DNA‐ICM) ploidy parameters in the categorisation of DNA‐ICM results and identification of high‐grade cervical intraepithelial neoplasia or worse (≥ CIN2).

Methods

Cervical samples from 232 women were collected for DNA‐ICM analysis and biopsy confirmation. Five DNA parameters were used to define DNA aneuploidy: number of cells with exceeding events (EE) over 2.5cEE, 4cEE, 5cEE and 9cEE, and aneuploid stemlines. DNA‐ICM results were categorised as normal, suspicious, and abnormal.

Results

For individual DNA ploidy parameters, sensitivity values for 50 cells with 2.5cEE, 45 cells with 4cEE, 1 cell with 9cEE and aneuploid stemline were 72.95%. 54.1%, 69.67% and 54.1%, while specificity values were 80.0%, 90.0%, 89.09% and 95.45%, respectively. For the 5cEE parameter, the sensitivity values for 1, 2, 3, 4 and 5 cells were 93.44%, 85.25%, 81.97%, 77.87% and 75.41%, while specificity values were 46.36%, 63.64%, 74.55%, 76.36% and 80.91%, respectively. For categorised DNA‐ICM results, a suspicious result showed superior sensitivity than an abnormal result (87.70% vs 82.79%, P = 0.031), but lower specificity (54.55% vs 75.45%, P < 0.001). Both types of DNA‐ICM result were statistically significantly different from a normal result (P < 0.05).

Conclusion

For prognostic purposes, 1 cell with 9cEE, 45 cells with 4cEE and aneuploid stemline are the best parameters with which to categorise an abnormal DNA‐ICM result, followed by 50 cells with 2.5cEE and 4 cells with 5cEE. For screening purposes, 10 cells with 2.5cEE, 10 cells with 4cEE, and 2 cells with 5cEE are suitable parameters with which to categorise a suspicious DNA‐ICM result.
The Fracking Frontier in the United States: A Case Study of Foreign Investment, Civil Liberties and Land Ethics in the Shale Industry
Authors: Bratman Eve , Auch Ted , Stinchfield Bryan
Published: March 2022

This article provides a fresh perspective on the unconventional oil and gas (hydraulic fracturing or ‘fracking’) industry in the Appalachian and Midwestern regions of the United States. It presents a case study of this industry in relation to development, democracy and the environment. The article utilizes neocolonialism as a heuristic device and analyses the fracking industry in relation to processes of capital expansion in resource frontiers. It identifies a two‐pronged paradox at the heart of the US energy independence rhetoric. First, the political‐economic reality is one in which the companies involved in promoting energy independence are in fact dependent on considerable amounts of foreign direct investment. Second, despite the purported ‘freedom’ offered through such energy sources, many legal measures associated with the industry ultimately detract from civil liberties, rights to private land and local sovereignty over decision making. Centrally, this article argues that political and economic relationships within this industry may undermine democratic participation and representative government. The case study demonstrates how relationships of coloniality in the fracking industry involve dynamics of land loss, ethical shifts in relation to land, and a lack of local property rights controls that are derivative of natural resource extraction on resource frontiers.
The method of critical phenomenology: Simone de Beauvoir as a phenomenologist
Authors: Oksala Johanna
Published: March 2022

The paper aims to contribute to the ongoing conversation on critical phenomenology with reflections on its method. The key argument is that critical phenomenology should be understood as a form of historico‐transcendental inquiry and therefore it cannot forgo the phenomenological reduction. Rather, this methodological step should be centered in critical phenomenology, and appropriated in problematized and rethought forms. The methodological assessment of critical phenomenology has implications also for how we read its canon. The paper shows that while Simone de Beauvoir did not adopt the phenomenological reduction in its full Husserlian meaning, her analyses of experience did not remain on the level of personal or experiential description either. The contention is that if we want to read her as a critical phenomenologist, we should focus on her seminal modification of the historico‐transcendental method of phenomenology.
“Obvious” indications for Neural antibody testing in Epilepsy or Seizures: The ONES checklist
Authors: Chang Yiu‐Chia , Nouri Maryam Nabavi , Mirsattari Seyed , Burneo Jorge G. , Budhram Adrian
Published: March 2022

Objective

Numerous predictive scores have been developed to help determine which patients with epilepsy or seizures of unknown etiology should undergo neural antibody testing. However, their diagnostic advantage compared to only performing testing in patients with “obvious” indications (e.g., broader features of autoimmune encephalitis, characteristic seizure semiologies) requires further study. We aimed to develop a checklist that identifies patients who have “obvious” indications for neural antibody testing and to compare its diagnostic performance to predictive scores.

Methods

We developed the “Obvious” indications for Neural antibody testing in Epilepsy or Seizures (ONES) checklist through literature review. We then retrospectively reviewed patients who underwent neural antibody testing for epilepsy or seizures at our center between March 2019 and January 2021, to determine and compare the sensitivity and specificity of the ONES checklist to the recently proposed Antibody Prevalence in Epilepsy and Encephalopathy (APE2)/Antibodies Contributing to Focal Epilepsy Signs and Symptoms (ACES) reflex score.

Results

One‐hundred seventy patients who underwent neural antibody testing for epilepsy or seizures were identified. Seventy‐four of 170 (43.5%) with a known etiology were excluded from sensitivity/specificity analyses; none had a true‐positive neural antibody. Of the 96 patients with an unknown etiology, 14 (15%) had a true‐positive neural antibody. The proportion of false‐positives was significantly higher among patients with a known etiology (3/3, 100%) compared to an unknown etiology (2/16, 13%; p = .01). There was no significant difference of the APE2/ACES reflex score compared to the ONES checklist with regard to sensitivity (93% for both, p > .99) or specificity (71% vs. 78%, p = .18) for true‐positive neural antibodies.

Significance

Compared to only performing neural antibody testing in patients with epilepsy or seizures of unknown etiology who have “obvious” indications, predictive scores confer no clear diagnostic advantage. Prespecified definitions of what constitutes a true‐positive neural antibody is required in future studies to avoid false‐positives that can confound results.
Global engagement in the post‐pandemic world: Challenges and responses. Perspective from the UK
Authors: Tsiligkiris Vangelis , Ilieva Janet
Published: March 2022

Higher education (HE), particularly its international mobility element, is one of the sectors that have suffered an immediate and substantial impact by the pandemic. The disruption in international travel and the local restrictions to physical contact has caused disruptions in the academic planning and delivery of higher education worldwide. Many argue that the recent pandemic and the move to online and blended learning delivery will define the future of HE. In this paper, we explore two main themes. Firstly, we explore the pre‐Covid trends in international higher education. Using national and international data from UNESCO, OECD and other sources, we analyse the global developments in international student mobility. Secondly, we focus on the UK to study the position, share and interplay between the international student and transnational education (TNE) markets. We reflect on the findings of our analysis and discuss the pivot to online education and its pedagogical and regulatory implications. The role of sustainability as a central strategic objective for HEIs is examined through the lenses of TNE in generating local impact and promoting the United Nations (UN) Sustainable Development Goals (SDGs).
Preparation and characterization of new tubular kaolino‐illitic ceramic membrane used for dairy wastewater treatment
Authors: Ben Ali Mahassen , Mahmoudi Khaled , Hamdi Noureddine , Srasra Ezzedine , Rodríguez Miguel A.
Published: March 2022

Low cost asymmetric tubular ceramic membrane was developed from kaolino‐illitic clay collected from the region of Medenin (Tunisia). The obtained membrane was designed to be used for ultrafiltration. The effect of pore forming agent on the plasticity, porosity, pore size distribution, mechanical strength, and permeability was evaluated. The support was performed by extrusion using the raw clay and olive pomace (OP) as pore forming agent. The effect of the use of OP was noticeable; it improved the plasticity of the paste. Also it helped to ameliorate both permeability and porosity of the obtained supports. Top layer was prepared using acid activated clay suspension. The obtained membrane had gas permeability of about 13 cm3.s−1.cm−2.bar−1 at a pressure of 1.5 bar and water flux of 1700 L.h−1.m−2 at a pressure of 1 bar. The final membrane was successfully used to eliminate the fat present in a solution containing 10% of milk. Regeneration of the used membrane was carried out by calcination at 550°C. The performances of the used membrane were recovered by 97% after regeneration.
Milk kefir enriched with inulin‐grafted seed extract from white wine pomace: chemical characterisation, antioxidant profile and in vitro gastrointestinal digestion
Authors: Carullo Gabriele , Spizzirri Umile Gianfranco , Montopoli Monica , Cocetta Veronica , Armentano Biagio , Tinazzi Mattia , Sciubba Fabio , Giorgi Gianluca , Enrica Di Cocco Maria , Bohn Torsten , Aiello Francesca , Restuccia Donatella
Published: March 2022

An innovative functional food with symbiotic and functional potential was produced by combining probiotic milk kefir, and white grape‐seed pomace obtained by UAE (ultrasound‐assisted extraction) rich in catechin and glycosylated flavonoids. The milk kefir was fortified by adding, after 24 h of fermentation, a polymer of inulin grafted with seed extract. The inulin/grape extract‐enriched polymer showed a higher antioxidant profile (P < 0.05). Simulated in vitro digestion demonstrated that 50 g of the kefir product would result in the delivery of antioxidants equivalent to 300 mg of ascorbic acid. The integrity of the intestinal barrier functions was not compromised by kefir treatments (at 10 μg mL‐1).