bluuebunny/crossref_metadata_2025 huggingface dataset:
Proinsulin C-peptide is a major source of HLA-DQ8 restricted HIPs recognized by human Islet-Infiltrating CD4+T cells
Authors: Bhattacharjee Pushpak , Pakusch Miha , Lacorcia Matthew , Tresoldi Eleonora , Rubin Alan F. , Foster Abby , King Laura S. , Chiu Chris , Kay Thomas W.H. , Karas John A. , Cameron Fergus J. , Mannering Stuart I.
Published: May 2024

Type 1 diabetes (T1D) is an autoimmune disease that develops when T cells destroy the pancreatic insulin-producing beta cells that reside in the pancreatic islets. Immune cells, including T cells infiltrate the islets and gradually destroy the beta cells. Human islet-infiltrating CD4+T cells recognize peptide epitopes derived from proinsulin, particularly C-peptide. Hybrid Insulin peptides (HIPs) are neoepitopes formed by the fusion of two peptides derived from beta-cell granule proteins and are known to be the targets of pathogenic CD4+T cells in the NOD mouse and human islet-infiltrating CD4+T cells. Proinsulin is widely recognized as a central antigen in T1D, but its role in forming HIPs is unclear. We developed a method to functionally screen TCRs derived from human islet-infiltrating CD4+T cells and applied this to the identification of new proinsulin-derived HIPs. We generated a library of 4,488 candidate HIPs formed by fusion of proinsulin fragments and predicted to bind to HLA-DQ8. This library was screened against 109 islet-infiltrating CD4+T-cell TCRs isolated from four organ donors who had T1D. We identified 13 unique HIPs recognized by 9 different TCRs from two organ donors. HIP specific T-cell avatars responded specifically to a peptide extract from human islets. These new HIPs predominantly stimulated CD4+T-cell proliferation in PBMCs from people with T1D in contrast to HLA-matched controls. This is the first unbiased functional, islet-infiltrating T-cell based, screen to identify proinsulin derived HIPs. It has revealed many new HIPs and a central role of proinsulin C-peptide in their formation.

Type 1 diabetes is an autoimmune disease caused by T cells destroying the pancreatic insulin-producing beta cells. The antigens/epitopes seen by disease promoting CD4+T cells are poorly understood. Hybrid insulin peptides (HIPs) are a new class of CD4+antigen recognized by pathogenic NOD mouse CD4+T cells. In humans very few HIPs recognized by human islet-infiltrating CD4+T cells are known. We show that proinsulin HIPs are recognized by human islet-infiltrating CD4+T cells from T1D donors and describe 13 new HIPs formed by fusion of proinsulin peptides. This work shows that proinsulin, particularly C-peptide, is a major contributor to the pool HIPs recognized by human islet-infiltrating CD4+T cells and are therefore central to autoimmunity in T1D.
Impairments in proprioceptively-referenced limb and eye movements in chronic stroke
Authors: Tulimieri Duncan T. , Decarie Amelia , Singh Tarkeshwar , Semrau Jennifer A.
Published: May 2024

Background

Upper limb proprioceptive impairments are common after stroke and can affect daily function. Recent work has shown that stroke survivors have difficulty using visual information to improve proprioception. It is unclear how eye movements are impacted to guide action of the arm after stroke. Here, we aimed to understand how upper limb proprioceptive impairments impact eye movements in individuals with stroke.

Methods

Control (N=20) and stroke participants (N=20) performed a proprioceptive matching task with upper limb and eye movements. A KINARM exoskeleton with eye tracking was used to assess limb and eye kinematics. The upper limb was passively moved by the robot and participants matched the location with either an arm or eye movement. Accuracy was measured as the difference between passive robot movement location and the active limb matching (Hand-End Point Error) or the active eye movement matching (Eye-End Point Error).

Results

We found that individuals with stroke had significantly larger Hand and Eye-End Point Errors compared to controls. Further, we found that stroke participants had proprioceptive errors of the hand and eye were highly correlated [r=0.67, p=0.001], a relationship not observed for controls.

Conclusions

Eye movement accuracy declined as a function of proprioceptive impairment of the more-affected limb, which was used as a proprioceptive reference. The inability to use proprioceptive information of the arm to coordinate eye movements suggests that disordered proprioception impacts integration of sensory information across different modalities. These results have important implications for how vision is used to actively guide limb movement during rehabilitation.
High Throughput Screening with a Primary Human Mucociliary Airway Model Identifies a Small Molecule with Anti-SARS-CoV-2 Activity
Authors: Sen Chandani , Rickabaugh Tammy M. , Jeyachandran Arjit Vijey , Yuen Constance , Ghannam Maisam , Durra Abdo , Aziz Adam , Castillo Kristen , Garcia Gustavo , Purkayastha Arunima , Han Brandon , Boulton Felix W. , Chekler Eugene , Garces Robert , Wolff Karen C. , Riva Laura , Kirkpatrick Melanie G. , Gebara-Lamb Amal , McNamara Case W. , Betz Ulrich A.K. , Arumugaswami Vaithilingaraja , Damoiseaux Robert , Gomperts Brigitte N.
Published: May 2024

Respiratory viruses (e.g. influenza, RSV, SARS etc.) attack the proximal airway and cause a wide spectrum of diseases for which we have limited therapies. To date, a few primary human stem cell-based models of the proximal airway have been reported for drug discovery but scaling them up to a higher throughput platform remains a significant challenge. Here we present a microscale, primary human stem cell-based proximal airway model of SARS-CoV-2 infection, which is amenable to moderate-to-high throughput drug screening. The model recapitulates the heterogeneity of infection seen among different patients and with different SARS-CoV-2 variants. We applied this model to screen 2100 compounds from targeted drug libraries using an image-based quantification method. While there were heterogeneous responses across variants for host factor targeting compounds, the direct acting antivirals showed a consistent response and we characterized a new antiviral drug that is effective against both the parental strain and the Omicron variant.
Amyloid fibril polymorphism in the heart of an ATTR amyloidosis patient with polyneuropathy attributed to the V122Δ variant
Authors: Ahmed Yasmin , Nguyen Binh An , Afrin Shumaila , Singh Virender , Evers Bret , Singh Preeti , Pedretti Rose , Wang Lanie , Bassett Parker , Fernandez-Ramirez Maria del Carmen , Pekala Maja , Kluve-Beckerman Barbara , Saelices Lorena
Published: May 2024

ATTR amyloidosis is a phenotypically heterogeneous disease characterized by the pathological deposition of transthyretin in the form of amyloid fibrils into various organs. ATTR amyloidosis may stem from mutations in variant (ATTRv) amyloidosis, or aging in wild-type (ATTRwt) amyloidosis. ATTRwt generally manifests as a cardiomyopathy phenotype, whereas ATTRv may present as polyneuropathy, cardiomyopathy, or mixed, in combination with many other symptoms deriving from secondary organ involvement. Over 130 different mutational variants of transthyretin have been identified, many of them being linked to specific disease symptoms. Yet, the role of these mutations in the differential disease manifestation remains elusive. Using cryo-electron microscopy, here we structurally characterized fibrils from the heart of an ATTRv patient carrying the V122Δ mutation, predominantly associated with polyneuropathy. Our results show that these fibrils are polymorphic, presenting as both single and double filaments. Our study alludes to a structural connection contributing to phenotypic variation in ATTR amyloidosis, as polymorphism in ATTR fibrils may manifest in patients with predominantly polyneuropathic phenotypes.
Normal male fertility in a mouse model of KPNA2 deficiency
Authors: Rother Franziska , Hweidi Dalia Abu , Hartmann Enno , Bader Michael
Published: May 2024

The nuclear transport of proteins is mediated by karyopherins and has been implicated to be crucial for germ cell and embryonic development. Deletion of distinct members of the karyopherin alpha family has been shown to cause male and female infertility in mice. Using a genetrap approach, we established mice deficient for KPNA2 (KPNA2 KO) and investigated the role of this protein in male germ cell development and fertility. Breeding of male KPNA2 KO mice leads to healthy offsprings in all cases albeit the absence of KPNA2 resulted in a reduction in sperm number by 60%. Analyses of the KPNA2 expression in wild-type mice revealed a strong KPNA2 presence in meiotic germ cells of all stages while a rapid decline is found in round spermatids. The high KPNA2 expression throughout all meiotic stages of sperm development suggests a possible function of KPNA2 during this phase, hence in its absence the spermatogenesis is not completely blocked. In KPNA2 KO mice, a higher portion of sperms presented with morphological abnormalities in the head and neck region, but a severe spermiogenesis defect was not found. Thus, we conclude that the function of KPNA2 in round spermatids is dispensable, as our mice do not show any signs of infertility. Our data provide evidence that KPNA2 is not crucial for male germ cell development and fertility.
Neuropeptide Y co-opts neuronal ensembles for memory lability and stability
Authors: Wu Yan-Jiao , Gu Xue , Kong Yalei , Yang Shuo , Wang Huan , Xu Miao , Wang Qi , Yi Xin , Lin Ze-Jie , Jiao Zhi-Han , Cheung Hoiyin , Zhao Xin-Yu , Bian Xin , Jiang Qin , Li Ying , Zhu Michael X. , Wang Lu-Yang , Li Yulong , Huang Ju , Li Qian , Li Wei-Guang , Xu Tian-Le
Published: May 2024

Memory engrams are formed by activity-dependent recruitment of distinct subsets of excitatory principal neurons (or neuronal ensembles) whereas inhibitory neurons pivot memory lability and stability1–5. However, the molecular logic for memory engrams to preferentially recruit specific type of interneurons over other subtypes remains enigmatic. Using activity-dependent single-cell transcriptomic profiling6–8in mice with training of cued fear memory and extinction, we discovered that neuropeptide Y (NPY)-expressing (NPY+) GABAergic interneurons in the ventral hippocampal CA1 (vCA1) region exert fast GABAergic inhibition to facilitate the acquisition of memory, but bifurcate NPY-mediated slow peptidergic inhibition onto distinct sub-ensembles underlying the extinction of single memory trace. Genetically encoded calcium and NPY sensors revealed that both calcium dynamics of NPY+neurons and their NPY release in vCA1 ramp up as extinction learning progresses while behavioral state switches from “fear-on” to “fear-off”. Bidirectional manipulations of NPY+neurons or NPY itself demonstrated NPY is both necessary and sufficient to control the rate and degree of memory extinction by acting on two physically non-overlapping sub-ensembles composed of NPY1R- and NPY2R-expressing neurons. CRISPR/Cas9-mediated knockout of NPY2R or NPY1R further unravels that NPY co-opts its actions on these two sub-ensembles to gate early fast and late slow stages of extinction. These findings exemplify the intricate spatiotemporal orchestration of slow peptidergic inhibitions from single subtype of GABAergic interneurons to fine-tune engram lability verse stability of memory.
Advancing species identification: A non-invasive molecular approach through spider silk proteome analysis
Authors: Yamamoto Phillip K , Takasuka Keizo , Mori Masaru , Masuda Takeshi , Kono Nobuaki
Published: May 2024

Species identification is crucial in various scientific disciplines such as biology, ecology, medicine, and agriculture. While traditional methods rely on morphological characteristics, DNA barcoding has gained popularity due to its molecular biology approach. Nonetheless, DNA barcoding can be problematic for small animals such as insects, as it requires damaging their bodies for DNA extraction, impacting subsequent breeding and experiments. In this paper, we propose a non-invasive molecular method for species identification that examines the protein composition of animal produced biomaterials. We chose spider silk, with species-specific protein sequences, as our subject of analysis. First, we established a universal silk-dissolving method that applies to silks from various species. We constructed a bioinformatics pipeline employing metrics of significant difference through proteomic analysis to identify spider species by analyzing peptide sequences present in silk proteins. As a result, we achieved a species identification accuracy of 86% across15 species. An appropriate reference dataset was successfully created, in addition, we also discovered some species are difficult to distinguish due to sequence similarities. This technology has been confirmed to be applicable to spider webs taken from the field. This non-invasive approach can complement DNA barcoding, especially in situations where it is infeasible, such as in studies involving spider-parasitoid wasps that eat spiders. Furthermore, it can be applied to other organisms that release biological substances, such as silkworm pupae, termite digestive enzymes, and tick saliva, aiding in species identification and pest control efforts.
Identifiability investigation of within-host models of acute virus infection
Authors: Liyanage Yuganthi R. , Heitzman-Breen Nora , Tuncer Necibe , Ciupe Stanca M.
Published: May 2024

Uncertainty in parameter estimates from fitting within-host models to empirical data limits the model’s ability to uncover mechanisms of infection, disease progression, and to guide pharmaceutical interventions. Understanding the effect of model structure and data availability on model predictions is important for informing model development and experimental design. To address sources of uncertainty in parameter estimation, we use four mathematical models of influenza A infection with increased degrees of biological realism. We test the ability of each model to reveal its parameters in the presence of unlimited data by performing structural identifiability analyses. We then refine the results by predicting practical identifiability of parameters under daily influenza A virus titers alone or together with daily adaptive immune cell data. Using these approaches, we present insight into the sources of uncertainty in parameter estimation and provide guidelines for the types of model assumptions, optimal experimental design, and biological information needed for improved predictions.

Author summary

Within-host models of virus infections fitted to data have improved our understanding of mechanisms of infection, disease progression, and allowed us to provide guidelines for pharmaceutical interventions. Given their predictive power, it is essential that we properly uncover and address uncertainty in model predictions and parameter estimation. Here, we focus on the effect of model structure and data availability on our ability to uncover unknown parameters. To address these questions, we use four mathematical models of influenza A infection with increased degrees of biological realism. We test the ability of each model to reveal its parameters in the presence of unlimited data by performing structural identifiability analysis. We then refine the results by predicting practical identifiability of parameters under daily influenza A virus titers alone or together with daily adaptive immune cell data. Using these approaches, we present insights into the sources of uncertainty in parameter estimations and provide guidelines for the types of model assumptions, optimal experimental design, and biological information needed for improved predictions.
The Implantable System That Restores Hemodynamic Stability After Spinal Cord Injury
Authors: Phillips Aaron A. , Gandhi Aasta P. , Hankov Nicolas , Hernandez-Charpak Sergio D. , Rimok Julien , Incognito Anthony , Nijland Anouk E.J. , D'Ercole Marina , Watrin Anne , Berney Maxime , Damianaki Aikaterini , Dumont Gregory , Macellari Nicolo , De Herde Laura , Intering Nadine , Smith Donovan , Miller Ryan , Smith Meagan N. , Lee Jordan , Baaklini Edeny , Ledoux Jean-Baptist , Ordonnez Javier G. , Newton Taylor , Meliado Ettore F. , Duguet Lea , Jacquet Charlotte , Bole-Feysot Lea , Rieger Markus , Gelenitis Kristen , Dumeny Yoann , Caban Miroslav , Ganty Damien , Paoles Edoardo , Baumgartner Thomas ,   Clinical Study Team,   Onward Team, Harte Cathal , Sasportes Charles David , Romo Paul , Vouga Tristan , Fasola Jemina , Ravier Jimmy , Gautier Matthieu , Merlos Frederic , Buschman Rik , Milekovic Tomislav , Rowald Andreas , Mandija Stefano , van den Berg Cornelis A.T. , Kuster Niels , Neufeld Esra , Pralong Etienne , Hirt Lorenz , Carda Stefano , Becce Fabio , Aleton Etienne , Rogan Kyle , Schoettker Patrick , Wuerzner Gregoire , Langerak Nelleke , Keijsers Noel L.W. , Kwon Brian K. , Guest James D. , Ross Erika , Murphy John , Kurt Erkan , Casha Steve , Girgis Fady , van Nes Ilse , Larkin-Kaiser Kelly A. , Demesmaeker Robin , Asboth Leonie , Squair Jordan W. , Bloch Jocelyne , Courtine Gregoire
Published: May 2024

A spinal cord injury (SCI) causes immediate and sustained hemodynamic instability that threatens neurological recovery and impacts quality of life. Here, we establish the clinical burden of chronic hypotensive complications due to SCI in 1,479 participants, and expose the ineffective treatment of these complications with conservative measures. To address this clinical burden, we developed a purpose-built implantable system based on biomimetic epidural electrical stimulation (EES) of the spinal cord that immediately triggered robust pressor responses. The system durably reduced the severity of hypotensive complications in people with SCI, removed the necessity for conservative treatments, improved quality of life, and enabled engagement in activities of daily living. Central to the development of this therapy was the head-to-head demonstration in the same participants that EES must target the last three thoracic segments, and not the lumbosacral segments, to achieve the safe and effective regulation of blood pressure in people with SCI. These findings in 14 participants establish a path for a pivotal device trial that evaluates the safety and efficacy of EES to treat the underappreciated, treatment-resistant hypotensive complications due to SCI.
Myomectomies during cesarean section and outcomes
Authors: Ruhliada Nikolai N.
Published: May 2024

The problem of myomectomy during cesarean section is relevant and disputable. Currently, there are no clear guidelines available for conditions and recommendations for performing this procedure during cesarean deliveries. Many obstetricians are hesitant to expand the scope of surgery due to the potential complications, such as increased intraoperative bleeding, postoperative hematomas, the risk of postoperative sepsis, venous thromboembolism, and abdominal adhesions. Additionally, long-term complications can arise from scarring on the uterus that can affect future fertility. This article presents an analysis of case series of cesarean deliveries with and without myomectomies performed at the Department of Obstetrics and Gynecology at St. Petersburg State Medical University in 2022.
Can drinking water, sanitation, handwashing, and nutritional interventions reduce antibiotic use in young children?
Authors: Ercumen Ayse , Mertens Andrew N. , Butzin-Dozier Zachary , Jung Da Kyung , Ali Shahjahan , Achando Beryl S. , Rao Gouthami , Hemlock Caitlin , Pickering Amy J. , Stewart Christine P. , Tan Sophia T. , Grembi Jessica A. , Benjamin-Chung Jade , Wolfe Marlene , Ho Gene G. , Rahman Md. Ziaur , Arnold Charles D. , Dentz Holly N. , Njenga Sammy M , Meerkerk Dorie , Chen Belinda , Nadimpalli Maya , Islam Mohammad Aminul , Hubbard Alan E. , Null Clair , Unicomb Leanne , Rahman Mahbubur , Colford John M. , Luby Stephen P. , Arnold Benjamin F. , Lin Audrie
Published: May 2024

Frequent antibiotic use in areas with high infection burdens can lead to antimicrobial resistance and microbiome alterations. Reducing pathogen exposure and child undernutrition can reduce infections and subsequent antibiotic use. We assessed effects of water, sanitation, handwashing (WSH) and nutrition interventions on pediatric antibiotic use in Bangladesh and Kenya, using longitudinal data from birth cohorts (at ages 3-28 months) enrolled in the WASH Benefits cluster-randomized trials. Over 50% of children used antibiotics at least once in the last 90 days. In Bangladesh, the prevalence of using antibiotics at least once was 10-14% lower in groups receiving WSH (prevalence ratio [PR]=0.90 (0.82-0.99)), nutrition (PR=0.86 (0.78-0.94)), and nutrition+WSH (PR=0.86 (0.79-0.93)) interventions. The prevalence of using antibiotics multiple times was 24-35% lower in intervention arms. Intervention effects were strongest when the birth cohort was younger. In Kenya, interventions did not affect antibiotic use. Improving WSH and nutrition can reduce antibiotic use in some low- and middle-income settings. Studies should assess whether such reductions translate to reduced carriage of antimicrobial resistance.
Incubation period, serial interval, generation time and reproduction number of mpox clade I
Authors: Marziano Valentina , Guzzetta Giorgio , Longini Ira , Merler Stefano
Published: May 2024

We estimate that the generation time of mpox clade I is distinctly longer than clade IIb and may depend on the transmission route (mean 17.5 days in households vs. 11.4 in hospitals). We estimate a mean reproduction number of 1.22-1.33 in the Democratic Republic of the Congo between August 2023-March 2024.
Characterising the contribution of rare protein-coding germline variants to prostate cancer risk and severity in 37,184 cases
Authors: Mitchell Jonathan , Camacho Niedzica , Shea Patrick , Stopsack Konrad H. , Joseph Vijai , Burren Oliver , Dhindsa Ryan , Nag Abhishek , Berchuck Jacob E. , O’Neill Amanda , Abbasi Ali , Zoghbi Anthony W. , Alegre-Díaz Jesus , Kuri-Morales Pablo , Berumen Jaime , Tapia-Conyer Roberto , Emberson Jonathan , Torres Jason M. , Collins Rory , Wang Quanli , Goldstein David , Matakidou Athena , Haefliger Carolina , Anderson-Dring Lauren , March Ruth , Jobanputra Vaidehi , Dougherty Brian , Carss Keren , Petrovski Slavé , Kantoff Philip W. , Offit Kenneth , Mucci Lorelei A. , Pomerantz Mark , Fabre Margarete A.
Published: May 2024

The etiology of prostate cancer, the second most common cancer in men globally, has a strong heritable component. While rare coding germline variants in several genes have been identified as risk factors from candidate gene and linkage studies, the exome-wide spectrum of causal rare variants remains to be fully explored. To more comprehensively address their contribution, we analysed data from 37,184 prostate cancer cases and 331,329 male controls from five cohorts with germline exome/genome sequencing and one cohort with imputed array data from a population enriched in low-frequency deleterious variants. Our gene-level collapsing analysis revealed that rare damaging variants inSAMHD1as well as genes in the DNA damage response pathway (BRCA2,ATMandCHEK2) are associated with the risk of overall prostate cancer. We also found that rare damaging variants inAOX1andBRCA2were associated with increased severity of prostate cancer in a case-only analysis of aggressive versus non-aggressive prostate cancer. At the single-variant level, we found rare non-synonymous variants in three genes (HOXB13,CHEK2,BIK) significantly associated with increased risk of overall prostate cancer and in four genes (ANO7,SPDL1,AR,TERT) with decreased risk. Altogether, this study provides deeper insights into the genetic architecture and biological basis of prostate cancer risk and severity.
Cross-platform Clinical Proteomics using the Charité Open Standard for Plasma Proteomics (OSPP)
Authors: Wang Ziyue , Farztdinov Vadim , Sinn Ludwig Roman , Tober-Lau Pinkus , Ludwig Daniela , Freiwald Anja , Amari Fatma , Textoris-Taube Kathrin , Niewienda Agathe , Welter Anna Sophie , Wei Alan An Jung , Luckau Luise , Kurth Florian , Selbach Matthias , Hartl Johannes , Mülleder Michael , Ralser Markus
Published: May 2024

The role of plasma and serum proteomics in characterizing human disease, identifying biomarkers, and advancing diagnostic technologies is rapidly increasing. However, there is an ongoing need to improve proteomic workflows in terms of accuracy, reproducibility, platform transferability, and cost-effectiveness. Here, we present the CharitéOpen PeptideStandard forPlasmaProteomics(OSPP), a panel of 211 extensively pre-selected, stable-isotope-labeled peptides combined in an open, versatile, and cost-effective internal standard for targeted and untargeted proteomic studies. The selected peptides are well suited for chemical synthesis, and distribute well over the captured analytical dynamic range and chromatographic gradients, and show consistent quantification properties across platforms, in serum, as well as in EDTA-, citrate, and heparin plasma. Quantifying proteins that function in a wide range of biological processes, including several that are routinely used in clinical tests or are targets of FDA-approved drugs, the OSPP quantifies proteins that are important for human disease. On an acute COVID-19 in-patient cohort, we demonstrate the application of the OSPP to i) achieve patient classification and biomarker identification ii) generate comparable quantitative proteomics data with both targeted and untargeted approaches, and iii) estimate peptide quantities for successful cross-platform alignment of proteomic data. The OSPP adds low costs per proteome sample, thus making the use of an internal standard accessible. In addition to the standards, corresponding spectral libraries and optimized acquisition methods for several platforms are made openly available.
Cognitive Behavioural Therapy in Adults with Psychogenic Non-Epileptic Seizures: A Systematic Review and Meta-Analysis of Randomised Controlled Trials
Authors: Moro Pierludovico , Lattanzi Simona , Beier Christoph P , Bonaventura Carlo Di , Irelli Emanuele Cerulli
Published: May 2024

Background

Randomised controlled clinical trials (RCTs) investigating cognitive-behavioural therapy (CBT) among adults with psychogenic non-epileptic seizures (PNES) has become increasingly available, prompting the opportunity to critically appraise the efficacy and safety of CBT in this population.

Methods

We conducted a systematic review and meta-analysis including RCTs comparing CBT in addition to standardised medical treatment (SMT) versus SMT alone for adults with PNES. The primary outcome was seizure freedom at the end of treatment. Secondary outcomes included measures of quality of life, anxiety and depression assessed via standardised clinical questionnaires.

Results

Three RCTs were included comprising 228 participants treated with CBT and 222 with SMT. The intervention was significantly associated with seizure freedom (Odds Ratio [OR] 1.98; 95% confidence interval [CI] 1.14, 3.46; p = 0.02; I2 = 0%), reductions in anxiety (standardised mean difference [SMD] −0.21; 95% CI −0.41, −0.003; p = 0.047; I2 = 0%) and improvements in quality of life (SMD 0.34; 95% CI 0.12, 0.57; p = 0.003; I2 = 0%) at the end of treatment. Conversely, no significant differences between groups were observed regarding depression symptoms (SMD −0.19; 95% CI −0.39, 0.02; p = 0.08; I2= 0%). There was no statistically significant increase in the risk of suicidal ideation and self-harm with CBT (OR 2.11; 95% CI 0.81, 5.48; p = 0.13; I2= 0%) nor were there differences in terms of discontinuation rates during follow-up (OR 0.92; 95% CI 0.49, 1.72; p = 0.79; I2= 7%).

Conclusions

There is high-quality evidence supporting the efficacy and safety of CBT in treating PNES. Future research should investigate whether combining CBT with other therapeutic methods could potentially enhance treatment efficacy.
Molecular Myelin Dysfunction in the Most Common Inherited Peripheral Neuropathies – CMT1A and HNPP
Authors: Moss Kathryn R. , Gutierrez Dave , Mi Ruifa , Cetinkaya-Fisgin Aysel , Höke Ahmet
Published: May 2024

Increased and decreased dosage of thePeripheral Myelin Protein 22(PMP22) gene cause dysmyelinating peripheral neuropathy. Charcot-Marie-Tooth Disease Type 1A (CMT1A,PMP22duplication) and Hereditary Neuropathy with Liability to Pressure Palsies (HNPP,PMP22deletion) are the most common inherited peripheral neuropathies, yet gaps remain about their pathophysiology and pathomechanims. Our previous results with CMT1A model mice demonstrate that muscle atrophy occurs without evidence of secondary axon degeneration suggesting that primary myelin dysfunction may contribute to functional deficits in CMT1A and motivating investigation of myelin dysfunction. Here we used CMT1A and HNPP model mice and confocal immunofluorescence imaging of teased tibial nerve fibers to determine how altered PMP22 expression disrupts myelin integrity and reveal CMT1A and HNPP pathomechanisms. We identified dramatic changes to molecular machinery at Schmidt-Lanterman incisures (SLIs) and Nodes of Ranvier that led us to propose two potential pathomechanisms for CMT1A and HNPP: impaired metabolic support and axonal ion disequilibrium. We also developed a working model for these pathomechanims that is driven by PMP22-mediated regulation of adherens junctions. Ongoing studies are aimed at testing these proposed pathomechanisms and determining how altered PMP22 and adherens junctions cause these defects. This work will provide insight into CMT1A and HNPP pathogenesis and may reveal novel approaches for developing therapeutics.
Key role of Pro230 in the hinge region on the IgG architecture and function
Authors: Koseki Yuuki , Yamaguchi Yuki , Aoyama Michihiko , Tada Minoru , Senoo Akinobu , Ishii-Watabe Akiko , Uchihashi Takayuki , Uchiyama Susumu , Kato Koichi , Yanaka Saeko , Caaveiro Jose M.M.
Published: May 2024

Immunoglobulin G (IgG) is a molecule that plays an important role in biological defense; IgG molecules have been applied as drugs due to their high specificity for antigens and their ability to activate immunity via effector molecules on immune cells. On the other hand, the flexibility of the hinge region makes it difficult to apply conventional structural biology approaches due to its dynamic conformational changes, and the mechanism of action of the molecule as a whole has not been elucidated. Here, we introduced a deletion amino acid mutation in the hinge region to elucidate the role of the hinge region and its effect on the structure and function of the IgG molecule. Deletion of Pro230 resulted in the formation of a half-molecular in which the interaction between heavy chains was lost. We elucidated the mechanism of half-IgG formation by structural analysis using nuclear magnetic resonance (NMR) measurements and by disulfide quantification using peptide mapping using LC-MS/MS. For this purpose, a new NMR stable isotope labeling method was introduced. Finally, cell assay revealed that the IgG half-molecules have specific FcγRI-mediated activity. This report provides new insights into the higher-order structure formation of IgG molecules and is expected to contribute to the elucidation of the molecular basis of the Fcγ receptor-mediated activation mechanism of the immune system.
A regulatory gene network that couples floral transition to shoot apical meristem morphology in Arabidopsis
Authors: de Olalla Enric Bertran Garcia , Rodríguez-Maroto Gabriel , Cerise Martina , Vayssières Alice , Severing Edouard , Sampere Yaiza López , Wang Kang , Schäfer Sabine , Formosa-Jordan Pau , Coupland George
Published: May 2024

Plants flower in response to environmental signals. These signals change the shape and developmental identity of the shoot apical meristem (SAM), causing it to form flowers and inflorescences rather than leaves. How the changes in SAM shape and identity are coordinated is poorly understood. Using genetics, confocal microscopy and RNA sequencing, we show that reciprocal repression of the APETALA2 (AP2) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) transcription factors in the SAM is crucial in coupling these processes. During vegetative development, AP2 repressesSOC1transcription to delay floral transition. Later,SOC1is activated during floral transition in response to environmental cues and repressesAP2transcription. However, during floral transition and prior to its stable repression by SOC1, AP2 rapidly promotes an increase in the size, and changes in the morphology, of the SAM. Thus, direct reciprocal repression of AP2 and SOC1 couples floral transition and changes in SAM morphology, which initiate inflorescence development.
Unveiling Gene Perturbation Effects through Gene Regulatory Networks Inference from single-cell transcriptomic data
Authors: Corridori Clelia , Romeike Merrit , Nicoletti Giorgio , Buecker Christa , Suweis Samir , Azaele Sandro , Martello Graziano
Published: May 2024

1

Physiological and pathological processes are governed by a network of genes called gene regulatory networks (GRNs). By reconstructing GRNs, we can accurately model how cells behave in their natural state and predict how genetic changes will affect them. Transcriptomic data of single cells are now available for a wide range of cellular processes in multiple species. Thus, a method building predictive GRNs from single-cell RNA sequencing (scRNA-seq) data, without any additional prior knowledge, could have a great impact on our understanding of biological processes and the genes playing a key role in them. To this aim, we developed IGNITE (Inference of Gene Networks using Inverse kinetic Theory and Experiments), an unsupervised machine learning framework designed to infer directed, weighted, and signed GRNs directly from unperturbed single-cell RNA sequencing data. IGNITE uses the GRNs to generate gene expression data upon single and multiple genetic perturbations. IGNITE is based on the inverse problem for a kinetic Ising model, a model from statistical physics that has been successfully applied to biological networks. We tested IGNITE on murine pluripotent stem cells (PSCs) transitioning from the naïve to formative states. Using as input only scRNA-seq data of unperturbed PSCs, IGNITE simulated single and triple gene knockouts. Comparison with experimental data revealed high accuracy, up to 74%, outperforming currently available methods. In sum, IGNITE identifies predictive GRNs from scRNA-seq data without additional prior knowledge and faithfully simulates single and multiple gene perturbations. Applications of IGNITE range from studying cell differentiation to identifying genes specifically active under pathological conditions.
Switching patterns of cortical-subcortical interaction in the human brain
Authors: Nazzi Alessandro , Favaretto Chiara , Vallesi Antonino , Corbetta Maurizio , Allegra Michele
Published: May 2024

Resting-state fMRI studies show that functional connectivity (FC) undergoes rapid fluctuations. Although the underlying neural mechanisms are poorly understood, a recent contribution analyzing stroke patients suggested that FC fluctuations involve a dynamic reconfiguration of cortico-subcortical interactions. Here, we analyze cortical-subcortical dynamic FC in a large cohort of healthy subjects (Human Connectome Project data base). Our analysis confirms that FC shifts are synchronized in cortex and subcortex. Two core subcortical 'clusters' comprising, respectively, limbic regions (hippocampus and amygdala) and subcortical nuclei (thalamus and basal ganglia) change their connectivity pattern with cortical regions. We consistently identify two recurring FC patterns (states). In state 1, limbic regions couple with the default mode network, in state 2 with sensorimotor networks. An opposite pattern is observed for thalamus/basal ganglia. Our findings hint at a general relevance of cortico-subcortical interactions in the generation of whole-brain spontaneous FC patterns.
Seasonal recurrence and modular assembly of an Arctic pelagic marine microbiome
Authors: Priest Taylor , Oldenburg Ellen , Popa Ovidiu , Dede Bledina , Metfies Katja , von Appen Wilken-Jon , Torres-Valdés Sinhué , Bienhold Christina , Fuchs Bernhard M. , Amann Rudolf , Boetius Antje , Wietz Matthias
Published: May 2024

Deciphering how microbial communities are shaped by environmental variability is fundamental for understanding the structure and function of ocean ecosystems. Thus far, we know little about the structuring of community functionality and the coupling between taxonomy and function over seasonal environmental gradients. To address this, we employed autonomous sampling devices andin situsensors to investigate the taxonomic and functional dynamics of a pelagic Arctic Ocean microbiome over a four-year period. We demonstrate that the dominant prokaryotic and microeukaryotic populations exhibit recurrent, unimodal fluctuations each year, with community gene content following the same trend. The recurrent dynamics within the prokaryotic microbiome are structured into five temporal modules that represent distinct ecological states, characterised by unique taxonomic and metabolic signatures and connections to specific microeukaryotic populations and oceanographic conditions. For instance,Cand. Nitrosopumilus and the machinery to oxidise ammonia and reduce nitrite are signatures of early polar night, along with Radiolarians. In contrast, late summer is characterised byAmylibacter, sulfur compound metabolism and diverse Haptophyta lineages. Exploring the composition of modules further along with their degree of functional redundancy and the structuring of genetic diversity within functions over time revealed seasonal heterogeneity in environmental selection processes. In particular, we observe strong selection pressure on a functional level in spring while late polar night features weaker selection pressure that likely acts on an organismal level. By integrating taxonomic, functional, and environmental information, our study provides fundamental insights into how microbiomes are structured under pronounced environmental variability in understudied, yet rapidly changing polar marine ecosystems.
A whole cell luminescence-based screen for inhibitors of the bacterial Sec machinery
Authors: Salter Tia , Collinson Ian , Allen William J.
Published: May 2024

There is a pressing need for new antibiotics to combat rising resistance to those already in use. The bacterial general secretion (Sec) system has long been considered a good target for novel antimicrobials thanks to its irreplacable role in maintaining cell envelope integrity; yet the lack of a robust, high-throughput method to screen for Sec inhibition has so far hampered efforts to realise this potential. Here, we have adapted our recently-developedin vitroassay for Sec activity – based on the split NanoLuc luciferase – to work at scale and in living cells. A simple counterscreen allows compounds that specifically target Sec to be distinguished from those with other effects on cellular function. As proof of principle we have applied this assay to a library of 5000 compounds and identified a handful of moderately effectivein vivoinhibitors of Sec. We therefore anticipate that the methods presented here will be scalable to larger compound libraries, in the ultimate quest for Sec inhibitors with clinically relevant properties.
Striatal output regulates the postnatal maturation of cortical circuits
Authors: Deemyad Tara , Janeček Michael , Shih Yi-Chun , Valle Vicente , D’Agostino Andrew , Perez Megan S. , Ketchesin Kyle D. , da Silva Susana , Peixoto Rui T.
Published: May 2024

The dorsomedial prefrontal cortex (dmPFC) is interconnected with the basal ganglia (BG) through large-scale circuit loops that regulate critical motor and cognitive functions. In mice, these circuits undergo extensive postnatal maturation with marked changes in neural activity and expansion of synaptic connectivity. While cortical activity is known to regulate the development of downstream striatal circuits, the role of the basal ganglia in cortical maturation remains unknown. Here, we used mesoscale two-photon microscopy and whole-cell electrophysiology to examine whether striatal output during early postnatal development impacts the maturation of upstream dmPFC circuits. We found that ablating spiny projection neurons of the direct or indirect pathways of the striatum during the first two postnatal weeks causes bidirectional changes in dmPFC neural activity, similar to what is observed in mature circuits. In addition, these manipulations alter the maturation of synaptic connectivity of dmPFC layer 2/3 pyramidal neurons, shifting the balance of excitation and inhibition of cortical circuits. These findings demonstrate that striatal output regulates the activity of cortical circuits during early postnatal development and suggest a regulatory role of the basal ganglia in the establishment of cortical circuits.
Cannabinoid receptor 1 positive allosteric modulator ZCZ011 shows differential effects on behavior and the endocannabinoid system in HIV-1 Tat transgenic female and male mice
Authors: Yadav-Samudrala Barkha J , Dodson Hailey , Ramineni Shreya , Kim Elizabeth , Poklis Justin L , Lu Dai , Ignatowska-Jankowska Bogna M , Lichtman Aron H , Fitting Sylvia
Published: May 2024

The cannabinoid receptor type 1 (CB1R) is a promising therapeutic target for various neurodegenerative diseases, including HIV-1-associated neurocognitive disorder (HAND). However, the therapeutic potential of CB1R by direct activation is limited due to its psychoactive side effects. Therefore, research has focused on indirectly activating the CB1R by utilizing positive allosteric modulators (PAMs). Studies have shown that CB1R PAMs (ZCZ011 and GAT211) are effective in mouse models of Huntington’s disease and neuropathic pain, and hence, we assess the therapeutic potential of ZCZ011 in a well-established mouse model of neuroHIV. The current study investigates the effect of chronic ZCZ011 treatment (14 days) on various behavioral paradigms and the endocannabinoid system in HIV-1 Tat transgenic female and male mice. Chronic ZCZ011 treatment (10 mg/kg) did not alter body mass, locomotor activity, or anxiety-like behavior regardless of sex or genotype. However, differential effects were noted in hot plate latency, motor coordination, and recognition memory in female mice only, with ZCZ011 treatment increasing hot plate latency and improving motor coordination and recognition memory. Only minor effects or no alterations were seen in the endocannabinoid system and related lipids except in the cerebellum, where the effect of ZCZ011 was more pronounced in female mice. Moreover, AEA and PEA levels in the cerebellum were positively correlated with improved motor coordination in female mice. In summary, these findings indicate that chronic ZCZ011 treatment has differential effects on antinociception, motor coordination, and memory, based on sex and HIV-1 Tat expression, making CB1R PAMs potential treatment options for HAND without the psychoactive side effects.
“What” and “when” predictions jointly modulate speech processing
Authors: Auksztulewicz Ryszard , Ödül Ozan Bahattin , Helbling Saskia , Böke Ana , Cappotto Drew , Luo Dan , Schnupp Jan , Melloni Lucía
Published: May 2024

Adaptive behavior rests on forming predictions based on previous statistical regularities encountered in the environment. Such regularities pertain not only to the contents of the stimuli (“what”) but also their timing (“when”), and both interactively modulate sensory processing. In speech streams, predictions can be formed at multiple hierarchical levels, both in terms of contents (e.g. single syllables vs. words) and timing (e.g., faster vs. slower time scales). Whether and how these hierarchies map onto each other in terms of integrating “what” and “when” predictions remains unknown. Under one hypothesis neural hierarchies may link “what” and “when” predictions within sensory processing areas: with lower cortical regions mediating interactions for smaller units e.g., syllables, and higher cortical areas mediating interactions for larger units e.g., words. Alternatively, interactions between “what” and “when” predictions might rest on a generic, sensory-independent mechanism, mediated by common attention-related (e.g., frontoparietal) networks. To address those questions, we manipulated “what” and “when” predictions at two levels – single syllables and disyllabic pseudowords – while recording neural activity using magnetoencephalography (MEG) in healthy volunteers (N=22). We studied how syllable and/or word deviants are modulated by “when” predictability, both analyzing event-related fields and using source reconstruction and dynamic causal modeling to explain the observed effects in terms of the underlying effective connectivity. “When” predictions modulated “what” mismatch responses in a specific way with regards to speech hierarchy, such that mismatch responses to deviant words (vs. syllables) were amplified by temporal predictions at a slower (vs. faster) time scale. However, these modulations were source-localized to a shared network of cortical regions, including frontal and parietal sources. Effective connectivity analysis showed that, while mismatch responses to violations of “what” predictions modulated connectivity between regions, the integration of “what” and “when” predictions selectively modulated connectivity within regions, consistent with gain effects. These results suggest that the brain integrates “what” and “when” predictions that are congruent with respect to their hierarchical level, but this integration is mediated by a shared and distributed cortical network. This contrasts with recent studies indicating separable networks for different levels of hierarchical speech processing.
Two-photon voltage imaging with rhodopsin-based sensors
Authors: Grimm Christiane , Sims Ruth R. , Tanese Dimitrii , Mohamed Lafirdeen Aysha S. , Chan Chung Yuen , Faini Giulia , Putti Elena , Del Bene Filippo , Papagiakoumou Eirini , Emiliani Valentina
Published: May 2024

The recent advances in sophisticated optical techniques, coupled with two-photon sensitive genetic voltage indicators (GEVIs), have enabled in-depth voltage imagingin vivoat single spike and single-cell resolution. To date, these results have been only achieved using ASAP-type sensors, as the complex photocycle of rhodopsin-based voltage indicators posed challenges for their two-photon use, restricting their application to one-photon approaches. In this work, we demonstrate that rhodopsin-based GEVIs (FRET-opsin) can be used under two-photon illumination when their peculiar light intensity dependence of kinetics and sensitivity are considered. We rationally engineer a fully genetically-encoded, rhodopsin-based voltage indicator with the brightest known fluorophore AaFP1, Jarvis, and demonstrate its utility under both one- and two-photon illumination. We also showed two-photon usability of the similar FRET-opsin sensor pAce. Our comparison of 2P scanless with fast 2P scanning illumination revealed that the latter approach is less suitable for this class of indicators and, on the contrary, both sensors responded well when scanless approaches were used. Furthermore, utilising Jarvis, we demonstrated high-fidelity, high-SNR action potential detection at kilohertz-imaging rates both in mouse hippocampal slices and in zebrafish larvae. To the best of our knowledge, this study represents the first report of a fully genetically-encoded rhodopsin-based voltage indicator for high contrast action potential detection under two-photon illuminationin vitroandin vivo.