bluuebunny/crossref_metadata_2025 huggingface dataset:
Demographic and psychosocial correlates of measurement error and reactivity bias in a 4-d image-based mobile food record among adults with overweight and obesity
Authors: Whitton Clare , Healy Janelle D. , Dhaliwal Satvinder S. , Shoneye Charlene , Harray Amelia J. , Mullan Barbara A. , McVeigh Joanne A. , Boushey Carol J. , Kerr Deborah A.
Published: May 2022

Improving dietary reporting among people living with obesity is challenging as many factors influence reporting accuracy. Reactive Reporting may occur in response to dietary recording, but little is known about how image-based methods influence this process. Using a 4-d image-based mobile food record (mFRTM), this study aimed to identify demographic and psychosocial correlates of measurement error and reactivity bias, among adults with BMI 25–40 kg/m2. Participants (n 155, aged 18–65 years) completed psychosocial questionnaires and kept a 4-d mFRTM. Energy expenditure (EE) was estimated using ≥ 4 d of hip-worn accelerometer data, and energy intake (EI) was measured using mFRTM. EI:EE ratios were calculated, and participants in the highest tertile were considered to have Plausible Intakes. Negative changes in EI according to regression slopes indicated Reactive Reporting. Mean EI was 72 % (sd = 21) of estimated EE. Among participants with Plausible Intakes, mean EI was 96 % (sd = 13) of estimated EE. Higher BMI (OR 0·81, 95 % CI 0·72, 0·92) and greater need for social approval (OR 0·31, 95 % CI 0·10, 0·96) were associated with lower likelihood of Plausible Intakes. Estimated EI decreased by 3 % per d of recording (interquartile range − 14 %,6 %) among all participants. The EI of Reactive Reporters (n 52) decreased by 17 %/d (interquartile range − 23 %,–13 %). A history of weight loss (> 10 kg) (OR 3·4, 95 % CI 1·5, 7·8) and higher percentage of daily energy from protein (OR 1·1, 95 % CI 1·0, 1·2) were associated with greater odds of Reactive Reporting. Identification of reactivity to measurement, as well as Plausible Intakes, is recommended in community-dwelling studies to highlight and address sources of bias.
Italian Field Hospital Experience in Mozambique: Report of Ordinary Activities in an Extraordinary Context
Authors: Sacchetto Daniela , Raviolo Mario , Lovesio Silvia , Salio Flavio , Hubloue Ives , Ragazzoni Luca
Published: May 2022

On March 15, 2019, Cyclone Idai made landfall near the port city of Beira in central Mozambique causing significant casualties and serious damage to infrastructure. The Emergency Medical Team Type 2 – Italy Regione Piemonte (EMT2-ITA) was deployed approximately two weeks after the disaster to support the country in need, providing essential medical and surgical care.

The EMT2-ITA staff was composed of 77 team members including two rotations and integrating local staff. A total of 1,121 patients (1,183 triage admissions) were treated during the 27 days of field hospital activity; among all the admissions, only few cases (17; 1%) were directly or indirectly attributed to the disaster event. Only three cases of cholera were confirmed and transferred to one of the treatment centers set up in Beira. The EMT2-ITA performed a total of 62 surgical operations (orthopedic, gynecological, general, and plastic surgery), of which more than one-half were elective procedures.

The objective of this manuscript is to report the mission of the EMT2-ITA in Mozambique, raising interesting points of discussion regarding the impact of timing on the mission outcomes, the operational and clinical activities in the field hospital, and the great importance to integrate local staff into the team.
Confirmation of glufosinate-resistant Palmer amaranth and response to other herbicides
Authors: Priess Grant L , Norsworthy Jason K , Godara Navdeep , Mauromoustakos Andy , Butts Thomas R , Roberts Trenton L , Barber Tom
Published: May 2022

The ability of weed populations to evolve resistance to herbicides affects management strategies and the profitability of crop production. The objective of this research was to screen Palmer amaranth accessions from Arkansas for glufosinate resistance. Additional efforts focused on the effectiveness of various herbicides, across multiple sites of action (SOAs), on each putative-resistant accession. The three putative accessions were selected from 60 Palmer amaranth accessions collected in 2019 and 2020 and screened with to 0.5× and 1× rates of glufosinate. A dose-response experiment was conducted for glufosinate on accessions A2019, A2020, and B2020. The effectiveness of various preemergence- and postemergence-applied herbicides were evaluated on each accession. Resistance ratios of A2019, A2020, and B2020 to glufosinate ranged from 5.1 to 27.4 when comparing LD50 values to two susceptible accessions, thus all three accessions were resistant to glufosinate. All three accessions (A2019, A2020, and B2020) were found to have a reduction equal to or greater than 20 percentage points in mortality to at least one herbicide from five different SOAs equal to or greater than five sites of action. Herbicides from nine different SOAs controlled A2019 at least 20 percentage points less than the susceptible accessions, which points to a need for additional research to characterize the response of this accession.
Testing of the Seismic Gap Hypothesis in a Model With Realistic Earthquake Statistics
Authors: Petrillo G. , Rosso A. , Lippiello E.
Published: May 2022

The seismic gap hypothesis states that fault regions where no large earthquake has recently occurred, are more prone than others to host the next large earthquake. It can lead to the idea of immunity after local disaster which, notwithstanding it sounds reasonable, it has been frequently rejected by objective testing. More generally, the estimate of the occurrence probability of the next big shock on the basis of the time delay from the last earthquake still represents a big challenge. The problem is that this issue cannot be addressed only on the basis of historical catalogs which contain too few well documented big shocks, and decades of future observations appear necessary. On the other hand, recent results have shown that important insights can be obtained from the spatial organization of aftershocks and its relationship to the mainshock slip profile. Here, we address this issue by monitoring the stress evolution together with the occurrence of big shocks and their aftershocks in a model where the fault is described as an elastic interface embedded in a ductile medium. The model reproduces all relevant statistical features of earthquake occurrence and allows us to perform accurate testing of the seismic gap hypothesis and its consequences, particularly on the side of aftershock spatial patterns. We show that large earthquakes do not regularly repeat in time, but it is possible to achieve insights on the time until the next big shock from the percentage of aftershocks occurring inside the mainshock slip contour.
Towards Raman imaging of centimeter scale tissue areas for real-time opto-molecular visualization of tissue boundaries for clinical applications
Authors: Ilchenko Oleksii , Pilhun Yurii , Kutsyk Andrii
Published: May 2022

Raman spectroscopy combined with augmented reality and mixed reality to reconstruct molecular information of tissue surface.
A novel polymer-based nitrocellulose platform for implementing a multiplexed microfluidic paper-based enzyme-linked immunosorbent assay
Authors: Lin Dong , Li Bowei , Fu Longwen , Qi Ji , Xia Chunlei , Zhang Yi , Chen Jiadong , Choo Jaebum , Chen Lingxin
Published: May 2022

Nitrocellulose (NC) membranes, as porous paper-like substrates with high protein-binding capabilities, are very popular in the field of point-of-care immunoassays. However, generating robust hydrophobic structures in NC membranes to fabricate microfluidic paper-based analytical devices (μPADs) remains a great challenge. At present, the main method relies on an expensive wax printer. In addition, NC membranes very easy to adhere during the printing process due to electrostatic adsorption. Herein, we developed a facile, fast and low-cost strategy to fabricate μPADs in NC membranes by screen-printing polyurethane acrylate (PUA) as a barrier material for defining flow channels and reaction zones. Moreover, hydrophobic barriers based on UV-curable PUA can resist various surfactant solutions and organic solvents that are generally used in immunoassays and biochemical reactions. To validate the feasibility of this PUA-based NC membrane for immunoassays in point-of-care testing (POCT), we further designed and assembled a rotational paper-based analytical device for implementing a multiplexed enzyme-linked immunosorbent assay (ELISA) in a simple manner. Using the proposed device under the optimal conditions, alpha fetoprotein (AFP) and carcinoembryonic antigen (CEA) could be identified, with limits of detection of 136 pg/mL and 174 pg/mL, respectively, which are below the threshold values of these two cancer biomarkers for clinical diagnosis. We believe that this reliable device provides a promising platform for the diagnosis of disease based on ELISA or other related bioassays in limited settings or remote regions.
Heme cross-feeding can augment Staphylococcus aureus and Enterococcus faecalis dual species biofilms
Authors: Ch’ng Jun-Hong , Muthu Mugil , Chong Kelvin K L , Wong Jun Jie , Tan Casandra A Z , Koh Zachary J S , Lopez Daniel , Matysik Artur , Nair Zeus J , Barkham Timothy , Wang Yulan , Kline Kimberly A
Published: May 2022

The contribution of biofilms to virulence and as a barrier to treatment is well-established for Staphylococcus aureus and Enterococcus faecalis, both nosocomial pathogens frequently isolated from biofilm-associated infections. Despite frequent co-isolation, their interactions in biofilms have not been well-characterized. We report that in combination, these two species can give rise to augmented biofilms biomass that is dependent on the activation of E. faecalis aerobic respiration. In E. faecalis, respiration requires both exogenous heme to activate the cydAB-encoded heme-dependent cytochrome bd, and the availability of O2. We determined that the ABC transporter encoded by cydDC contributes to heme import. In dual species biofilms, S. aureus provides the heme to activate E. faecalis respiration. S. aureus mutants deficient in heme biosynthesis were unable to augment biofilms whereas heme alone is sufficient to augment E. faecalis mono-species biofilms. Our results demonstrate that S. aureus-derived heme, likely in the form of released hemoproteins, promotes E. faecalis biofilm formation, and that E. faecalis gelatinase activity facilitates heme extraction from hemoproteins. This interspecies interaction and metabolic cross-feeding may explain the frequent co-occurrence of these microbes in biofilm-associated infections.
The aspartyl protease DDI2 drives adaptation to proteasome inhibition in multiple myeloma
Authors: Op Mélanie , Ribeiro Sérgio T. , Chavarria Claire , De Gassart Aude , Zaffalon Léa , Martinon Fabio
Published: May 2022

Proteasome inhibitors, such as bortezomib, are first-line therapy against multiple myeloma (MM). Unfortunately, patients frequently become refractory to this treatment. The transcription factor NRF1 has been proposed to initiate an adaptation program that regulates proteasome levels. In the context of proteasome inhibition, the cytosolic protease DDI2 cleaves NRF1 to release an active fragment that translocates to the nucleus to promote the transcription of new proteasome subunits. However, the contribution of the DDI2-NRF1 pathway to bortezomib resistance is poorly understood. Here we show that upon prolonged bortezomib treatment, MM cells become resistant to proteasome inhibition by increasing the expression of DDI2 and consequently activation of NRF1. Furthermore, we found that many MM cells became more sensitive to proteasome impairment in the context of DDI2 deficiency. Mechanistically, we demonstrate that both the protease and the HDD domains of DDI2 are required to activate NRF1. Finally, we show that partial inhibition of the DDI2-protease domain with the antiviral drug nelfinavir increased bortezomib susceptibility in treated MM cells. Altogether, these findings define the DDI2-NRF1 pathway as an essential program contributing to proteasome inhibition responses and identifying DDI2 domains that could be targets of interest in bortezomib-treated MM patients.
NF-κB-inducing kinase (NIK) is activated in pancreatic β-cells but does not contribute to the development of diabetes
Authors: Xiao Peng , Takiishi Tatiana , Violato Natalia Moretti , Licata Giada , Dotta Francesco , Sebastiani Guido , Marselli Lorella , Singh Sumeet Pal , Sze Mozes , Van Loo Geert , Dejardin Emmanuel , Gurzov Esteban Nicolas , Cardozo Alessandra Kupper
Published: May 2022

The transcription factor nuclear factor-κB (NF-κB) has a key role in the pathogenesis of diabetes and its complications. Although activation of the canonical NF-κB pathway in β-cells is generally deleterious, little is known about the role of the non-canonical NF-κB signalling and its main regulator, the NF-κB-inducing kinase (NIK), on pancreatic β-cell survival and function. Previous studies based on models of NIK overexpression in pancreatic islet cells showed that NIK induced either spontaneous β-cell death due to islet inflammation or glucose intolerance during diet-induced obesity (DIO) in mice. Therefore, NIK has been proposed as a potential target for diabetes therapy. However, no clear studies showed whether inhibition of NIK improves diabetes development. Here we show that genetic silencing of NIK in pancreatic β-cells neither modifies diabetes incidence nor inflammatory responses in a mouse model of immune-mediated diabetes. Moreover, NIK silencing in DIO mice did not influence body weight gain, nor glucose metabolism. In vitro studies corroborated the in vivo findings in terms of β-cell survival, function, and downstream gene regulation. Taken together, our data suggest that NIK activation is dispensable for the development of diabetes.
Oxidative stress induces meiotic defects of oocytes in a mouse psoriasis model
Authors: Zhang ZhiQin , Tang ShouBin , Jiang YuYing , Long FangYuan , He Fang , Liu Jian , Gu ShouYong , Lu Yan , Yin ZhiQiang
Published: May 2022

Psoriasis, an immune-mediated inflammatory disease, is associated with poor pregnancy outcomes. Emerging evidence indicates that these defects are likely attributed to compromised oocyte competence. Nevertheless, little is known about the underlying associated mechanisms between psoriasis and poor oocyte quality. In this study, we construct an imiquimod-induced chronic psoriasis-like mouse model to review the effects of psoriasis on oocyte quality. We discover that oocytes from psoriasis-like mice display spindle/chromosome disorganization, kinetochore-microtubule mis-attachment, and aneuploidy. Importantly, our results show that melatonin supplement in vitro and in vivo not only increases the rate of matured oocytes but also significantly attenuates oxidative stress and meiotic defects by restoring mitochondrial function in oocytes from psoriasis-like mice. Altogether, our data uncover the adverse effects of psoriasis symptoms on oocytes, and melatonin supplement ameliorates oxidative stress and meiotic defects of oocytes from psoriatic mice.
Structural transition and re-emergence of iron's total electron spin in (Mg,Fe)O at ultrahigh pressure
Authors: Hsu Han , Umemoto Koichiro
Published: May 2022

Fe-bearing MgO [(Mg1−xFex)O] is considered a major constituent of terrestrial exoplanets. Crystallizing in the B1 structure in the Earth’s lower mantle, (Mg1−xFex)O undergoes a high-spin (S = 2) to low-spin (S = 0) transition at ∼45 GPa, accompanied by anomalous changes of this mineral’s physical properties, while the intermediate-spin (S = 1) state has not been observed. In this work, we investigate (Mg1−xFex)O (x ≤ 0.25) up to 1.8 TPa via first-principles calculations. Our calculations indicate that (Mg1−xFex)O undergoes a simultaneous structural and spin transition at ∼0.6 TPa, from the B1 phase low-spin state to the B2 phase intermediate-spin state, with Fe’s total electron spin S re-emerging from 0 to 1 at ultrahigh pressure. Upon further compression, an intermediate-to-low spin transition occurs in the B2 phase. Depending on the Fe concentration (x), metal–insulator transition and rhombohedral distortions can also occur in the B2 phase. These results suggest that Fe and spin transition may affect planetary interiors over a vast pressure range.
SARS-CoV-2 transmission and impacts of unvaccinated-only screening in populations of mixed vaccination status
Authors: Bubar Kate M. , Middleton Casey E. , Bjorkman Kristen K. , Parker Roy , Larremore Daniel B.
Published: May 2022

Screening programs that test only the unvaccinated population have been proposed and implemented to mitigate SARS-CoV-2 spread, implicitly assuming that the unvaccinated population drives transmission. To evaluate this premise and quantify the impact of unvaccinated-only screening programs, we introduce a model for SARS-CoV-2 transmission through which we explore a range of transmission rates, vaccine effectiveness scenarios, rates of prior infection, and screening programs. We find that, as vaccination rates increase, the proportion of transmission driven by the unvaccinated population decreases, such that most community spread is driven by vaccine-breakthrough infections once vaccine coverage exceeds 55% (omicron) or 80% (delta), points which shift lower as vaccine effectiveness wanes. Thus, we show that as vaccination rates increase, the transmission reductions associated with unvaccinated-only screening decline, identifying three distinct categories of impact on infections and hospitalizations. More broadly, these results demonstrate that effective unvaccinated-only screening depends on population immunity, vaccination rates, and variant.
Mitochondrial calcium uniporter stabilization preserves energetic homeostasis during Complex I impairment
Authors: Balderas Enrique , Eberhardt David R. , Lee Sandra , Pleinis John M. , Sommakia Salah , Balynas Anthony M. , Yin Xue , Parker Mitchell C. , Maguire Colin T. , Cho Scott , Szulik Marta W. , Bakhtina Anna , Bia Ryan D. , Friederich Marisa W. , Locke Timothy M. , Van Hove Johan L. K. , Drakos Stavros G. , Sancak Yasemin , Tristani-Firouzi Martin , Franklin Sarah , Rodan Aylin R. , Chaudhuri Dipayan
Published: May 2022

Calcium entering mitochondria potently stimulates ATP synthesis. Increases in calcium preserve energy synthesis in cardiomyopathies caused by mitochondrial dysfunction, and occur due to enhanced activity of the mitochondrial calcium uniporter channel. The signaling mechanism that mediates this compensatory increase remains unknown. Here, we find that increases in the uniporter are due to impairment in Complex I of the electron transport chain. In normal physiology, Complex I promotes uniporter degradation via an interaction with the uniporter pore-forming subunit, a process we term Complex I-induced protein turnover. When Complex I dysfunction ensues, contact with the uniporter is inhibited, preventing degradation, and leading to a build-up in functional channels. Preventing uniporter activity leads to early demise in Complex I-deficient animals. Conversely, enhancing uniporter stability rescues survival and function in Complex I deficiency. Taken together, our data identify a fundamental pathway producing compensatory increases in calcium influx during Complex I impairment.
A truncated anti-CRISPR protein prevents spacer acquisition but not interference
Authors: Philippe Cécile , Morency Carlee , Plante Pier-Luc , Zufferey Edwige , Achigar Rodrigo , Tremblay Denise M. , Rousseau Geneviève M. , Goulet Adeline , Moineau Sylvain
Published: May 2022

CRISPR-Cas systems in prokaryotic cells provide an adaptive immunity against invading nucleic acids. For example, phage infection leads to addition of new immunity (spacer acquisition) and DNA cleavage (interference) in the bacterial model species Streptococcus thermophilus, which primarily relies on Cas9-containing CRISPR-Cas systems. Phages can counteract this defense system through mutations in the targeted protospacers or by encoding anti-CRISPR proteins (ACRs) that block Cas9 interference activity. Here, we show that S. thermophilus can block ACR-containing phages when the CRISPR immunity specifically targets the acr gene. This in turn selects for phage mutants carrying a deletion within the acr gene. Remarkably, a truncated acrIIA allele, found in a wild-type virulent streptococcal phage, does not block the interference activity of Cas9 but still prevents the acquisition of new immunities, thereby providing an example of an ACR specifically inhibiting spacer acquisition.
Modulation of biological motion perception in humans by gravity
Authors: Wang Ying , Zhang Xue , Wang Chunhui , Huang Weifen , Xu Qian , Liu Dong , Zhou Wen , Chen Shanguang , Jiang Yi
Published: May 2022

The human visual perceptual system is highly sensitive to biological motion (BM) but less sensitive to its inverted counterpart. This perceptual inversion effect may stem from our selective sensitivity to gravity-constrained life motion signals and confer an adaptive advantage to creatures living on Earth. However, to what extent and how such selective sensitivity is shaped by the Earth’s gravitational field is heretofore unexplored. Taking advantage of a spaceflight experiment and its ground-based analog via 6° head-down tilt bed rest (HDTBR), we show that prolonged microgravity/HDTBR reduces the inversion effect in BM perception. No such change occurs for face perception, highlighting the particular role of gravity in regulating kinematic motion analysis. Moreover, the reduced BM inversion effect is associated with attenuated orientation-dependent neural responses to BM rather than general motion cues and correlated with strengthened functional connectivity between cortical regions dedicated to visual BM processing (i.e., pSTS) and vestibular gravity estimation (i.e., insula). These findings suggest that the neural computation of gravity may act as an embodied constraint, presumably implemented through visuo-vestibular interaction, to sustain the human brain’s selective tuning to life motion signals.
Poly(GR) and poly(GA) in cerebrospinal fluid as potential biomarkers for C9ORF72-ALS/FTD
Authors: Krishnan Gopinath , Raitcheva Denitza , Bartlett Daniel , Prudencio Mercedes , McKenna-Yasek Diane M. , Douthwright Catherine , Oskarsson Björn E. , Ladha Shafeeq , King Oliver D. , Barmada Sami J. , Miller Timothy M. , Bowser Robert , Watts Jonathan K. , Petrucelli Leonard , Brown Robert H. , Kankel Mark W. , Gao Fen-Biao
Published: May 2022

GGGGCC repeat expansion inC9ORF72, which can be translated in both sense and antisense directions into five dipeptide repeat (DPR) proteins, including poly(GP), poly(GR), and poly(GA), is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Here we developed sensitive assays that can detect poly(GA) and poly(GR) in the cerebrospinal fluid (CSF) of patients withC9ORF72mutations. CSF poly(GA) and poly(GR) levels did not correlate with age at disease onset, disease duration, or rate of decline of ALS Functional Rating Scale, and the average levels of these DPR proteins were similar in symptomatic and pre-symptomatic patients withC9ORF72mutations. However, in a patient withC9ORF72-ALS who was treated with antisense oligonucleotide (ASO) targeting the aberrantC9ORF72transcript, CSF poly(GA) and poly(GR) levels decreased approximately 50% within 6 weeks, indicating they may serve as sensitive fluid-based biomarkers in studies directed against the production of GGGGCC repeat RNAs or DPR proteins.
Shielding of actin by the endoplasmic reticulum impacts nuclear positioning
Authors: Janota Cátia Silva , Pinto Andreia , Pezzarossa Anna , Machado Pedro , Costa Judite , Campinho Pedro , Franco Cláudio A. , Gomes Edgar R.
Published: May 2022

Nuclear position is central to cell polarization, and its disruption is associated with various pathologies. The nucleus is moved away from the leading edge of migrating cells through its connection to moving dorsal actin cables, and the absence of connections to immobile ventral stress fibers. It is unclear how these asymmetric nucleo-cytoskeleton connections are established. Here, using an in vitro wound assay, we find that remodeling of endoplasmic reticulum (ER) impacts nuclear positioning through the formation of a barrier that shields immobile ventral stress fibers. The remodeling of ER and perinuclear ER accumulation is mediated by the ER shaping protein Climp-63. Furthermore, ectopic recruitment of the ER to stress fibers restores nuclear positioning in the absence of Climp-63. Our findings suggest that the ER mediates asymmetric nucleo-cytoskeleton connections to position the nucleus.
TUBB4A interacts with MYH9 to protect the nucleus during cell migration and promotes prostate cancer via GSK3β/β-catenin signalling
Authors: Gao Song , Wang Shuaibin , Zhao Zhiying , Zhang Chao , Liu Zhicao , Ye Ping , Xu Zhifang , Yi Baozhu , Jiao Kai , Naik Gurudatta A. , Wei Shi , Rais-Bahrami Soroush , Bae Sejong , Yang Wei-Hsiung , Sonpavde Guru , Liu Runhua , Wang Lizhong
Published: May 2022

Human tubulin beta class IVa (TUBB4A) is a member of the β-tubulin family. In most normal tissues, expression of TUBB4A is little to none, but it is highly expressed in human prostate cancer. Here we show that high expression levels of TUBB4A are associated with aggressive prostate cancers and poor patient survival, especially for African-American men. Additionally, in prostate cancer cells, TUBB4A knockout (KO) reduces cell growth and migration but induces DNA damage through increased γH2AX and 53BP1. Furthermore, during constricted cell migration, TUBB4A interacts with MYH9 to protect the nucleus, but either TUBB4A KO or MYH9 knockdown leads to severe DNA damage and reduces the NF-κB signaling response. Also, TUBB4A KO retards tumor growth and metastasis. Functional analysis reveals that TUBB4A/GSK3β binds to the N-terminal of MYH9, and that TUBB4A KO reduces MYH9-mediated GSK3β ubiquitination and degradation, leading to decreased activation of β-catenin signaling and its relevant epithelial-mesenchymal transition. Likewise, prostate-specific deletion of Tubb4a reduces spontaneous tumor growth and metastasis via inhibition of NF-κB, cyclin D1, and c-MYC signaling activation. Our results suggest an oncogenic role of TUBB4A and provide a potentially actionable therapeutic target for prostate cancers with TUBB4A overexpression.
The phosphatase PTEN links platelets with immune regulatory functions of mouse T follicular helper cells
Authors: Chen Xue , Xu Yanyan , Chen Qidi , Zhang Heng , Zeng Yu , Geng Yan , Shen Lei , Li Fubin , Chen Lei , Chen Guo-Qiang , Huang Chuanxin , Liu Junling
Published: May 2022

Beyond a function in hemostasis and thrombosis, platelets can regulate innate and adaptive immune responses. Hyperactive platelets are frequently associated with multiple human autoimmune diseases, yet their pathogenic functions in these diseases have not been fully established. Emerging studies show an essential function of the phosphatase and tensin homolog (PTEN) in maintenance of immune homeostasis. Here, we show that mice with platelet-specific deletion of Pten, develop age-related lymphoproliferative diseases and humoral autoimmunity not seen in wildtype animals. Platelet-specific Pten-deficient mice have aberrant T cell activation, excessive T follicular helper (Tfh) cell responses and accumulation of platelet aggregates in lymph nodes. Transferred Pten-deficient platelets are able to infiltrate into the peripheral lymphoid tissues and form more aggregates. Moreover, Pten-deficient platelets are hyperactive and overproduce multiple Tfh-promoting cytokines via activation of the PDK1/mTORC2-AKT-SNAP23 pathway. Pten-deficient platelets show enhanced interaction with CD4+ T cells and promote conversion of CD4+ T cells into Tfh cells. Our results implicate PTEN in platelet-mediated immune homeostasis, and provide evidence that hyperactive platelets function as an important mediator in autoimmune diseases using mouse models.
C. elegans enteric motor neurons fire synchronized action potentials underlying the defecation motor program
Authors: Jiang Jingyuan , Su Yifan , Zhang Ruilin , Li Haiwen , Tao Louis , Liu Qiang
Published: May 2022

C. elegansneurons were thought to be non-spiking until our recent discovery of action potentials in the sensory neuron AWA; however, the extent to which theC. elegansnervous system relies on analog or digital coding is unclear. Here we show that the enteric motor neurons AVL and DVB fire synchronous all-or-none calcium-mediated action potentials following the intestinal pacemaker during the rhythmicC. elegansdefecation behavior. AVL fires unusual compound action potentials with each depolarizing calcium spike mediated by UNC-2 followed by a hyperpolarizing potassium spike mediated by a repolarization-activated potassium channel EXP-2. Simultaneous behavior tracking and imaging in free-moving animals suggest that action potentials initiated in AVL propagate along its axon to activate precisely timed DVB action potentials through the INX-1 gap junction. This work identifies a novel circuit of spiking neurons inC. elegansthat uses digital coding for long-distance communication and temporal synchronization underlying reliable behavioral rhythm.
Neuronal hyperexcitability drives central and peripheral nervous system tumor progression in models of neurofibromatosis-1
Authors: Anastasaki Corina , Mo Juan , Chen Ji-Kang , Chatterjee Jit , Pan Yuan , Scheaffer Suzanne M. , Cobb Olivia , Monje Michelle , Le Lu Q. , Gutmann David H.
Published: May 2022

Neuronal activity is emerging as a driver of central and peripheral nervous system cancers. Here, we examined neuronal physiology in mouse models of the tumor predisposition syndrome Neurofibromatosis-1 (NF1), with different propensities to develop nervous system cancers. We show that central and peripheral nervous system neurons from mice with tumor-causing Nf1 gene mutations exhibit hyperexcitability and increased secretion of activity-dependent tumor-promoting paracrine factors. We discovered a neurofibroma mitogen (COL1A2) produced by peripheral neurons in an activity-regulated manner, which increases NF1-deficient Schwann cell proliferation, establishing that neurofibromas are regulated by neuronal activity. In contrast, mice with the Arg1809Cys Nf1 mutation, found in NF1 patients lacking neurofibromas or optic gliomas, do not exhibit neuronal hyperexcitability or develop these NF1-associated tumors. The hyperexcitability of tumor-prone Nf1-mutant neurons results from reduced NF1-regulated hyperpolarization-activated cyclic nucleotide-gated (HCN) channel function, such that neuronal excitability, activity-regulated paracrine factor production, and tumor progression are attenuated by HCN channel activation. Collectively, these findings reveal that NF1 mutations act at the level of neurons to modify tumor predisposition by increasing neuronal excitability and activity-regulated paracrine factor production.
A bio-functional polymer that prevents retinal scarring through modulation of NRF2 signalling pathway
Authors: Parikh Bhav Harshad , Liu Zengping , Blakeley Paul , Lin Qianyu , Singh Malay , Ong Jun Yi , Ho Kim Han , Lai Joel Weijia , Bogireddi Hanumakumar , Tran Kim Chi , Lim Jason Y. C. , Xue Kun , Al-Mubaarak Abdurrahmaan , Yang Binxia , R Sowmiya , Regha Kakkad , Wong Daniel Soo Lin , Tan Queenie Shu Woon , Zhang Zhongxing , Jeyasekharan Anand D. , Barathi Veluchamy Amutha , Yu Weimiao , Cheong Kang Hao , Blenkinsop Timothy A. , Hunziker Walter , Lingam Gopal , Loh Xian Jun , Su Xinyi
Published: May 2022

One common cause of vision loss after retinal detachment surgery is the formation of proliferative and contractile fibrocellular membranes. This aberrant wound healing process is mediated by epithelial-mesenchymal transition (EMT) and hyper-proliferation of retinal pigment epithelial (RPE) cells. Current treatment relies primarily on surgical removal of these membranes. Here, we demonstrate that a bio-functional polymer by itself is able to prevent retinal scarring in an experimental rabbit model of proliferative vitreoretinopathy. This is mediated primarily via clathrin-dependent internalisation of polymeric micelles, downstream suppression of canonical EMT transcription factors, reduction of RPE cell hyper-proliferation and migration. Nuclear factor erythroid 2–related factor 2 signalling pathway was identified in a genome-wide transcriptomic profiling as a key sensor and effector. This study highlights the potential of using synthetic bio-functional polymer to modulate RPE cellular behaviour and offers a potential therapy for retinal scarring prevention.
Topography-induced large-scale antiparallel collective migration in vascular endothelium
Authors: Leclech Claire , Gonzalez-Rodriguez David , Villedieu Aurélien , Lok Thévy , Déplanche Anne-Marie , Barakat Abdul I.
Published: May 2022

Collective migration of vascular endothelial cells is central for embryonic development, angiogenesis, and wound closure. Although physical confinement of cell assemblies has been shown to elicit specific patterns of collective movement in various cell types, endothelial migration in vivo often occurs without confinement. Here we show that unconfined endothelial cell monolayers on microgroove substrates that mimic the anisotropic organization of the extracellular matrix exhibit a specific type of collective movement that takes the form of a periodic pattern of antiparallel cell streams. We further establish that the development of these streams requires intact cell-cell junctions and that stream sizes are particularly sensitive to groove depth. Finally, we show that modeling the endothelial cell sheet as an active fluid with the microgrooves acting as constraints on cell orientation predicts the occurrence of the periodic antiparallel cell streams as well as their lengths and widths. We posit that in unconfined cell assemblies, physical factors that constrain or bias cellular orientation such as anisotropic extracellular matrix cues or directed flow-derived shear forces dictate the pattern of collective cell movement.
Multiplex base- and prime-editing with drive-and-process CRISPR arrays
Authors: Yuan Qichen , Gao Xue
Published: May 2022

Current base- and prime-editing technologies lack efficient strategies to edit multiple genomic loci simultaneously, limiting their applications in complex genomics and polygenic diseases. Here, we describe drive-and-process (DAP) CRISPR array architectures for multiplex base-editing (MBE) and multiplex prime-editing (MPE) in human cells. We leverage tRNA as the RNA polymerase III promoter to drive the expression of tandemly assembled tRNA-guide RNA (gRNA) arrays, of which the individual gRNAs are released by the cellular endogenous tRNA processing machinery. We engineer a 75-nt human cysteine tRNA (hCtRNA) for the DAP array, achieving up to 31-loci MBE and up to 3-loci MPE. By applying MBE or MPE elements for deliveries via adeno-associated virus (AAV) and lentivirus, we demonstrate simultaneous editing of multiple disease-relevant genomic loci. Our work streamlines the expression and processing of gRNAs on a single array and establishes efficient MBE and MPE strategies for biomedical research and therapeutic applications.
Structural mechanism of TRPV3 channel inhibition by the anesthetic dyclonine
Authors: Neuberger Arthur , Nadezhdin Kirill D. , Sobolevsky Alexander I.
Published: May 2022

Skin diseases are common human illnesses that occur in all cultures, at all ages, and affect between 30% and 70% of individuals globally. TRPV3 is a cation-permeable TRP channel predominantly expressed in skin keratinocytes, implicated in cutaneous sensation and associated with numerous skin diseases. TRPV3 is inhibited by the local anesthetic dyclonine, traditionally used for topical applications to relieve pain and itch. However, the structural basis of TRPV3 inhibition by dyclonine has remained elusive. Here we present a cryo-EM structure of a TRPV3-dyclonine complex that reveals binding of the inhibitor in the portals which connect the membrane environment surrounding the channel to the central cavity of the channel pore. We propose a mechanism of TRPV3 inhibition in which dyclonine molecules stick out into the channel pore, creating a barrier for ion conductance. The allosteric binding site of dyclonine can serve as a template for the design of new TRPV3-targeting drugs.
Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro
Authors: Zeng Leiping , Liu Yanxia , Nguyenla Xammy Huu , Abbott Timothy R. , Han Mengting , Zhu Yanyu , Chemparathy Augustine , Lin Xueqiu , Chen Xinyi , Wang Haifeng , Rane Draven A. , Spatz Jordan M. , Jain Saket , Rustagi Arjun , Pinsky Benjamin , Zepeda Adrianna E. , Kadina Anastasia P. , Walker John A. , Holden Kevin , Temperton Nigel , Cochran Jennifer R. , Barron Annelise E. , Connolly Michael D. , Blish Catherine A. , Lewis David B. , Stanley Sarah A. , La Russa Marie F. , Qi Lei S.
Published: May 2022

A major challenge in coronavirus vaccination and treatment is to counteract rapid viral evolution and mutations. Here we demonstrate that CRISPR-Cas13d offers a broad-spectrum antiviral (BSA) to inhibit many SARS-CoV-2 variants and diverse human coronavirus strains with >99% reduction of the viral titer. We show that Cas13d-mediated coronavirus inhibition is dependent on the crRNA cellular spatial colocalization with Cas13d and target viral RNA. Cas13d can significantly enhance the therapeutic effects of diverse small molecule drugs against coronaviruses for prophylaxis or treatment purposes, and the best combination reduced viral titer by over four orders of magnitude. Using lipid nanoparticle-mediated RNA delivery, we demonstrate that the Cas13d system can effectively treat infection from multiple variants of coronavirus, including Omicron SARS-CoV-2, in human primary airway epithelium air-liquid interface (ALI) cultures. Our study establishes CRISPR-Cas13 as a BSA which is highly complementary to existing vaccination and antiviral treatment strategies.
Multimodal confined water dynamics in reverse osmosis polyamide membranes
Authors: Foglia Fabrizia , Frick Bernhard , Nania Manuela , Livingston Andrew G. , Cabral João T.
Published: May 2022

While polyamide (PA) membranes are widespread in water purification and desalination by reverse osmosis, a molecular-level understanding of the dynamics of both confined water and polymer matrix remains elusive. Despite the dense hierarchical structure of PA membranes formed by interfacial polymerization, previous studies suggest that water diffusion remains largely unchanged with respect to bulk water. Here, we employ neutron spectroscopy to investigate PA membranes under precise hydration conditions, and a series of isotopic contrasts, to elucidate water transport and polymer relaxation, spanning ps-ns timescales, and Å-nm lengthscales. We experimentally resolve, for the first time, the multimodal diffusive nature of water in PA membranes: in addition to (slowed down) translational jump-diffusion, we observe a long-range and a localized mode, whose geometry and timescales we quantify. The PA matrix is also found to exhibit rotational relaxations commensurate with the nanoscale confinement observed in water diffusion. This comprehensive ‘diffusion map’ can anchor molecular and nanoscale simulations, and enable the predictive design of PA membranes with tuneable performance.
Multidimensional analysis of adult patients’ care trajectories before a first diagnosis of schizophrenia
Authors: Vanasse Alain , Courteau Josiane , Courteau Mireille , Roy Marc-André , Stip Emmanuel , Fleury Marie-Josée , Lesage Alain , Brodeur Sébastien
Published: May 2022

For patients at high-risk for developing schizophrenia, a delayed diagnosis could be affected, among many reasons, by their patterns of healthcare use. This study aims to describe and generate a typology of patients’ care trajectories (CTs) in the 2 years preceding a first diagnosis of schizophrenia, over a medico-administrative database of 3712 adults with a first diagnosis between April 2014 and March 2015 in Quebec, Canada. This study applied a multidimensional approach of State Sequence Analysis, considering together sequences of patients’ diagnoses, care settings and care providers. Five types of distinct CTs have emerged from this data-driven analysis: The type 1, shared by 77.6% of patients, predominantly younger men, shows that this group sought little healthcare, among which 17.5% had no healthcare contact for mental disorders. These individuals might benefit from improved promotion and prevention of mental healthcare at the community level. The types 2, 3 and 4, with higher occurrence of mental disorder diagnoses, represent together 19.5% of the study cohort, mostly middle-aged and women. These CTs, although displaying roughly similar profiles of mental disorders, revealed very dissimilar sequences and levels of care providers encounters, primary and specialized care use, and hospitalizations. Surprisingly, patients of these CTs had few consultations with general practitioners. An increased attentiveness for middle-aged patients and women with high healthcare use for mental disorders could help to reduce delayed diagnosis of schizophrenia. This calls for further consideration of healthcare services for severe mental illness beyond those offered to young adults.
Proton-gated anion transport governs macropinosome shrinkage
Authors: Zeziulia Mariia , Blin Sandy , Schmitt Franziska W. , Lehmann Martin , Jentsch Thomas J.
Published: May 2022

Intracellular organelles change their size during trafficking and maturation. This requires the transport of ions and water across their membranes. Macropinocytosis, a ubiquitous form of endocytosis of particular importance for immune and cancer cells, generates large vacuoles that can be followed optically. Shrinkage of macrophage macropinosomes depends on TPC-mediated Na+efflux and Cl−exit through unknown channels. Relieving osmotic pressure facilitates vesicle budding, positioning osmotic shrinkage upstream of vesicular sorting and trafficking. Here we identify the missing macrophage Cl−channel as the proton-activated Cl−channel ASOR/TMEM206. ASOR activation requires Na+-mediated depolarization and luminal acidification by redundant transporters including H+-ATPases and CLC 2Cl−/H+exchangers. As corroborated by mathematical modelling, feedback loops requiring the steep voltage and pH dependencies of ASOR and CLCs render vacuole resolution resilient towards transporter copy numbers.TMEM206disruption increased albumin-dependent survival of cancer cells. Our work suggests a function for the voltage and pH dependence of ASOR and CLCs, provides a comprehensive model for ion-transport-dependent vacuole maturation and reveals biological roles of ASOR.
Rituximab versus tocilizumab in rheumatoid arthritis: synovial biopsy-based biomarker analysis of the phase 4 R4RA randomized trial
Authors: Rivellese Felice , Surace Anna E. A. , Goldmann Katriona , Sciacca Elisabetta , Çubuk Cankut , Giorli Giovanni , John Christopher R. , Nerviani Alessandra , Fossati-Jimack Liliane , Thorborn Georgina , Ahmed Manzoor , Prediletto Edoardo , Church Sarah E. , Hudson Briana M. , Warren Sarah E. , McKeigue Paul M. , Humby Frances , Bombardieri Michele , Barnes Michael R. , Lewis Myles J. , Pitzalis Costantino ,   the R4RA collaborative group, Rivellese Felice , Giorli Giovanni , Nerviani Alessandra , Fossati-Jimack Liliane , Thorborn Georgina , Humby Frances , Bombardieri Michele , Lewis Myles J. , Durez Patrick , Buch Maya H. , Rizvi Hasan , Mahto Arti , Montecucco Carlomaurizio , Lauwerys Bernard , Ng Nora , Ho Pauline , Romão Vasco C. , da Fonseca João Eurico Cabral , Verschueren Patrick , Kelly Stephen , Sainaghi Pier Paolo , Gendi Nagui , Dasgupta Bhaskar , Cauli Alberto , Reynolds Piero , Cañete Juan D. , Ramirez Julio , Celis Raquel , Moots Robert , Taylor Peter C. , Edwards Chris J. , Isaacs John , Sasieni Peter , Choy Ernest , Thompson Charlotte , Bugatti Serena , Bellan Mattia , Congia Mattia , Holroyd Christopher , Pratt Arthur , White Laura , Warren Louise , Peel Joanna , Hands Rebecca , Hadfield Gaye , Pitzalis Costantino
Published: May 2022

Patients with rheumatoid arthritis (RA) receive highly targeted biologic therapies without previous knowledge of target expression levels in the diseased tissue. Approximately 40% of patients do not respond to individual biologic therapies and 5–20% are refractory to all. In a biopsy-based, precision-medicine, randomized clinical trial in RA (R4RA; n = 164), patients with low/absent synovial B cell molecular signature had a lower response to rituximab (anti-CD20 monoclonal antibody) compared with that to tocilizumab (anti-IL6R monoclonal antibody) although the exact mechanisms of response/nonresponse remain to be established. Here, in-depth histological/molecular analyses of R4RA synovial biopsies identify humoral immune response gene signatures associated with response to rituximab and tocilizumab, and a stromal/fibroblast signature in patients refractory to all medications. Post-treatment changes in synovial gene expression and cell infiltration highlighted divergent effects of rituximab and tocilizumab relating to differing response/nonresponse mechanisms. Using ten-by-tenfold nested cross-validation, we developed machine learning algorithms predictive of response to rituximab (area under the curve (AUC) = 0.74), tocilizumab (AUC = 0.68) and, notably, multidrug resistance (AUC = 0.69). This study supports the notion that disease endotypes, driven by diverse molecular pathology pathways in the diseased tissue, determine diverse clinical and treatment–response phenotypes. It also highlights the importance of integration of molecular pathology signatures into clinical algorithms to optimize the future use of existing medications and inform the development of new drugs for refractory patients.
Exploring body consciousness of dancers, athletes, and lightly physically active adults
Authors: Virtanen Niia , Tiippana Kaisa , Tervaniemi Mari , Poikonen Hanna , Anttila Eeva , Kaseva Kaisa
Published: May 2022

Body consciousness is associated with kinetic skills and various aspects of wellbeing. Physical activities have been shown to contribute to the development of body consciousness. Methodological studies are needed in improving the assessment of body consciousness in adults with distinct physical activity backgrounds. This study (1) examined whether dancers, athletes, and lightly physically active individuals differed regarding the level of their body consciousness, and (2) evaluated the usability of different methods in assessing body consciousness. Fifty-seven healthy adults (aged 20–37) were included in the study. Three experimental methods (aperture task, endpoint matching, and posture copying) and two self-report questionnaires (the Private Body Consciousness Scale, PBCS, and the Body Awareness Questionnaire, BAQ) were used in assessing body consciousness. Athletes outperformed the lightly physically active participants in the posture copying task with the aid of vision when copying leg postures. Dancers performed better than the athletes without the aid of vision when their back and upper body were involved, and better than the lightly active participants when copying leg postures. Dancers and athletes had higher self-reported cognitive and perceptual knowledge of their body than lightly physically active participants. To examine the role of different physical activities in developing body consciousness, experimental methods involving the use of the whole body might be most suitable. Subjective measures may provide complementary evidence for experimental testing.