bluuebunny/crossref_metadata_2025 huggingface dataset:
Similar Outcomes in Treating Major Depressive Disorder With 10 Hz Repetitive Transcranial Magnetic Stimulation (rTMS) Versus Intermittent Theta Burst Stimulation (iTBS): A Naturalistic Observational Study
Authors: Spitz Nathen A. , Ten Eyck Patrick , Nizar Krystal , Boes Aaron D. , Trapp Nicholas T.
Published: March 2022

Background:

Results reported in the existing literature have shown intermittent theta burst stimulation (iTBS) to be noninferior to 10 Hz repetitive transcranial magnetic stimulation (rTMS) in treating major depressive disorder (MDD) when targeted at the left dorsolateral prefrontal cortex. The goal of this naturalistic observational study was to further explore potential differences between these 2 treatment modalities in treating depression in a real-world cohort.

Methods:

The participants were 105 patients, 18 years of age or older with a diagnosis of MDD who received standard clinical 10 Hz rTMS or iTBS treatment between 2016 and 2020. Clinical outcomes of depression treatment were assessed on the basis of changes in scores on the Patient Health Questionnaire-9 and on the Montgomery-Asberg Depression Rating Scale.

Results:

Reduction in depression symptoms was measured with the Patient Health Questionnaire-9 and Montgomery-Asberg Depression Rating Scale from baseline to end of treatment, and no discernible differences in percent change, response, remission, or minimum clinically important difference were found between the 10 Hz rTMS and iTBS treatment groups.

Conclusions:

Findings in an observational, real-world clinical sample showed no significant differences in outcomes between 10 Hz rTMS and iTBS targeted at the left dorsolateral prefrontal cortex in the treatment of MDD. Because of the shorter treatment time involved, the choice of iTBS may reduce hospital exposure and increase savings and the treatment capacity of clinics without sacrificing effectiveness.
Assessment of Sleep and Sleepiness in Patients With Mental Disorders: Applicability of the German Version of the SCOPA-Sleep Questionnaire
Authors: Newe Catarina , Pedersen Anya , Goebel Simone
Published: March 2022

Sleep problems affect the majority of patients with mental disorders and are very clinically relevant. However, they are often not assessed and treated appropriately and, to date, there is no established screening routine for sleep disturbances in this specific patient group. For patients with Parkinson disease, the Scales for Outcomes in Parkinson’s Disease–Sleep (SCOPA-Sleep) questionnaire is widely used. This questionnaire has excellent psychometric properties. It consists of 2 scales for assessing nighttime sleep problems (NS) and daytime sleepiness (DS). As the items in the scales are not disease-specific, the goal of this study was to validate the SCOPA-Sleep for use in patients with mental disorders. A total of 370 German patients with mental disorders completed the SCOPA-Sleep as well as a multitude of other instruments for assessment of validity [eg, Pittsburgh Sleep Quality Index (PSQI), Epworth Sleepiness Scale (ESS)]. For validity estimation, 627 healthy subjects were also included in this study. Internal consistency was good with coefficients of 0.806 and 0.772 for the SCOPA-NS and the SCOPA-DS. Factor analysis revealed 2 factors which could each be assigned to one of the subscales. Convergent validity was high with correlations of 0.672 between the SCOPA-NS and the PSQI and 0.644 between the SCOPA-DS and the ESS. The German version of the SCOPA-Sleep showed good diagnostic accuracy, resulting in an area under the receiver operating curve of 0.959 for the SCOPA-NS and 0.728 for the SCOPA-DS. For patients with mental disorders, the SCOPA-Sleep proved to be an economical, reliable, and valid instrument for assessing NS and DS which is well accepted by patients. It can be recommended for routine assessment of both NS and DS.
Carbamazepine-induced DRESS Syndrome: A Rare Delayed Hypersensitivity Reaction
Authors: Hou Wen-Shun , Tsai Jui-Ping , Chiu Yi-Hang , Lu Mong-Liang
Published: March 2022

Drug rash with eosinophilia and systemic symptoms (DRESS) syndrome is a rare but severe cutaneous drug hypersensitivity reaction. Delays in making a diagnosis of DRESS syndrome and lack of timely treatment may result in morbidity and mortality. However, the presentation can be misinterpreted as other pathologies because of a broad spectrum of clinical presentations, delayed reactions, and prolonged course. Despite the discontinuation of the medication that is the culprit, relapses of drug reactions frequently occur weeks to months later. Several drugs that are associated with DRESS syndrome are prescribed as psychotropic medications. This report describes the case of a patient with DRESS syndrome who was evaluated with the RegiSCAR scoring system as a “definite case,” which was possibly induced by carbamazepine prescribed to treat bipolar I disorder. The young female patient was successfully treated with steroid medication after carbamazepine was discontinued. She was prescribed aripiprazole for mood stabilization without a subsequent recurrence of DRESS syndrome. We recommend that, in cases such as described here, clinicians take DRESS syndrome into consideration and provide proper timely management, particularly for patients receiving psychotropic drugs. A brief review of the literature concerning DRESS syndrome associated with psychotropic drugs and its pathogenesis are outlined and discussed.
A Case Report of Clozapine-induced Pleural Effusion in a 28-Year-Old Chinese Male
Authors: Wolfe Christine L. , Hamilton Joshua , Rogahn Megan , Brooks Stephanie , Ahmed Iqbal
Published: March 2022

Increasing reports have appeared of pleural effusion and peripheral eosinophilia associated with clozapine treatment. These reports describe the onset of pleural effusion from 2 to 5 weeks after initiation of clozapine. Here, we describe a case of a 28-year-old Chinese male who presented with pleural effusion and peripheral eosinophilia ∼17 weeks after initiation of clozapine. We discuss this delayed presentation and examine the potential significance of the patient’s East Asian ethnicity. We recommend clinicians consider ethnicity and other factors that can affect the metabolism of clozapine when choosing a clozapine titration schedule and when monitoring during clozapine treatment.
Color Doppler Imaging of Vascular Abnormalities of the Uterus
Authors: Keller Cody A. , Antil Neha , Jeffrey R. Brooke , Kamaya Aya
Published: March 2022

Many uterine abnormalities present clinically with bleeding encompassing a broad spectrum of patients from postmenopausal spotting to life-threatening hemorrhage. Color and spectral Doppler imaging of the pelvis is often the first crucial investigation used to quickly establish the correct etiology of the uterine bleeding and guide clinical decision making and patient management.
Simultaneous Implant Placement and Sinus Lift Using Leukocyte-Platelet Rich Fibrin: A Retrospective 40-Month Study
Authors: Leighton Yerko , Rosas Eduardo , de Souza Raphael Freitas , Borie Eduardo
Published: March 2022

The aim of this study was to report the 40-month clinical radiographical outcomes of maxillary sinus floor augmentation using leukocyte-platelet rich fibrin (L-PRF) as a single graft in combination with immediately placed implants. Twenty partially edentulous individuals with missing teeth in the region associated to maxillary sinus were included in this retrospective study. A lateral sinus floor elevation was performed considering a complete wear of the bone wall of the maxillary sinus and immediately placing the implant and filled the maxillary sinus with L-PRF clots as a single graft. After 5 months, all the implants received single screwed implant-supported prostheses. Subsequent evaluation was every 4 months assessing the success rate and biological complications. Two maxillary sinus membrane were perforated and covered with L-PRF clots. The mean bone height gain was 62.6%, observing a 100% of success and that in all the cases achieved at least the implant height. In conclusion, the use of platelet-rich fibrin clot as a single graft biomaterial during the maxillary sinus lift procedure combined with immediate implant placement may be a reliable clinical alternative, even to repair the sinus membrane.
Development of an Easily Reproducible Cough Simulator With Droplets and Aerosols for Rapidly Testing Novel Personal Protective Equipment
Authors: Zhou George , Burnett Garrett W. , Shah Ronak S. , Lai Cheuk Yin , Katz Daniel , Fried Eric A.
Published: March 2022

Introduction

The current COVID-19 pandemic has produced numerous innovations in personal protective equipment, barrier devices, and infection mitigation strategies, which have not been validated. During high-risk procedures such as airway manipulation, coughs are common and discrete events that may expose healthcare workers to large amounts of viral particles. A simulated cough under controlled circumstances can rapidly test novel devices and protocols and thus aid in their evaluation and the development of implementation guidelines. Physiologic cough simulators exist but require significant expertise and specialized equipment not available to most clinicians.

Methods

Using components commonly found in healthcare settings, a cough simulator was designed for clinicians to easily assemble and use. Both droplet and aerosol particle generators were incorporated into a bimodal experimental system. High-speed flash photography was used for data collection.

Results

Using a gas flow analyzer, video recordings, and high-speed digital photography, the cough and particle simulators were quantitatively and qualitatively compared with known physiologic cough parameters and in vivo Schlieren imaging of human coughs.

Conclusions

Based on our validation studies, this cough and particle simulator model approximates a physiologic, human cough in the context of testing personal protective equipment, barrier devices, and infection prevention measures.
Sleep Biomarkers Help Predict the Development of Alzheimer Disease
Authors: Grigg-Damberger Madeleine M. , Foldvary-Schaefer Nancy
Published: March 2022

Middle-aged or older adults who self-report sleep–wake disorders are at an increased risk for incident dementia, mild cognitive impairment, and Alzheimer disease. Dementia in people with mild cognitive impairment and Alzheimer disease who complain of sleep–wake disorders progress faster than those without sleep–wake disorders. Removal of amyloid-beta and tau tangles occurs preferentially in non-rapid eye movement 3 sleep and fragmented or insufficient sleep may lead to accumulation of these neurotoxins even in preclinical stages. Selective atrophy in the medial temporal lobe on brain MRI has been shown to predict impaired coupling of slow oscillations and sleep spindles. Impaired slow wave–spindle coupling has been shown to correlate with impaired overnight memory consolidation. Whereas, a decrease in the amplitude of 0.6 to 1 Hz slow wave activity predicts higher cortical Aβ burden on amyloid PET scans. Overexpression of the wake-promoting neurotransmitter orexin may predispose patients with mild cognitive impairment and Alzheimer disease to increased wakefulness, decreasing time they need to clear from the brain the neurotoxic accumulation of amyloid-beta and especially tau. More research exploring these relationships is needed and continuing.
Stroke: What's Sleep Got to Do With It?
Authors: Zafar Atif , Dhruv Parth , Anwar Arsalan , Grigg-Damberger Madeleine M.
Published: March 2022

Ischemic strokes most often occur between 6 am and 12 am after awakening from sleep but up to 30% occur during sleep. Wake-up strokes (WUS) are new focal neurological deficit(s) persisting for ≥ 24 hours attributable to an ischemic event present on patient awakening. Obstructive sleep apnea (OSA) is a major risk factor for WUS because it compounds the instability of the morning environment and increases the likelihood of cardiovascular events, including hypertension, atrial fibrillation, right-to-left shunts, and stroke. Circadian-driven alterations in structural, homeostatic, and serological factors also predispose to WUS. Also, WUS patients are often not considered candidates for time-dependent intravenous thrombolysis therapy because of an uncertain onset time. However, using the tissue clock (positive diffusion weighted imaging-negative fluid-attenuated inversion recovery mismatch) dates the WUS as 3 to 4.5 hours old and permits consideration for intravenous thrombolysis and if needed mechanical thrombectomy. Given the high prevalence of moderate/severe OSA in stroke patients and its impact on stroke outcomes, screening with overnight pulse oximetry and home sleep apnea test is needed. Treating OSA poststroke remains challenging. Polysomnographic changes in sleep architecture following acute/subacute stroke may also impact upon stroke outcome.
Sleep Spindles and K-Complexes Are Favorable Prognostic Biomarkers in Critically Ill Patients
Authors: Grigg-Damberger Madeleine M. , Hussein Omar , Kulik Tobias
Published: March 2022

In this narrative review, we summarize recent research on the prognostic significance of biomarkers of sleep in continuous EEG and polysomnographic recordings in intensive care unit patients. Recent studies show the EEG biosignatures of non-rapid eye movement 2 sleep (sleep spindles and K-complexes) on continuous EEG in critically ill patients better predict functional outcomes and mortality than the ictal–interictal continuum patterns. Emergence of more complex and better organized sleep architecture has been shown to parallel neurocognitive recovery and correlate with functional outcomes in traumatic brain injury and strokes. Particularly interesting are studies which suggest intravenous dexmedetomidine may induce a more biomimetic non-rapid eye movement sleep state than intravenous propofol, potentially providing more restorative sleep and lessening delirium. Protocols to improve intensive care unit sleep and neurophysiological studies evaluating the effect of these on sleep and sleep architecture are here reviewed.
Taxonomy of Rhizobiaceae revisited: proposal of a new framework for genus delimitation
Authors: Kuzmanović Nemanja , Fagorzi Camilla , Mengoni Alessio , Lassalle Florent , diCenzo George C.
Published: March 2022

The alphaproteobacterial family

Rhizobiaceae

is highly diverse, with 168 species with validly published names classified into 17 genera with validly published names. Most named genera in this family are delineated based on genomic relatedness and phylogenetic relationships, but some historically named genera show inconsistent distribution and phylogenetic breadth. The most problematic is

Rhizobium

, which is notorious for being highly paraphyletic, as most newly described species in the family are assigned to this genus without consideration of their proximity to existing genera, or the need to create novel genera. Moreover, many

Rhizobiaceae

genera lack synapomorphic traits that would give them biological and ecological significance. We propose a common framework for genus delimitation within the family

Rhizobiaceae

, wherein genera are defined as monophyletic groups in a core-genome gene phylogeny, that are separated from related species using a pairwise core-proteome average amino acid identity (cpAAI) threshold of approximately 86 %. We further propose that additional genomic or phenotypic evidence can justify division of species into separate genera even if they share greater than 86 % cpAAI. Applying this framework, we propose to reclassify

Rhizobium rhizosphaerae

and

Rhizobium oryzae

into Xaviernesmea gen. nov. Data is also provided to support the formation of Peteryoungia aggregata comb. nov., Endobacterium yantingense comb. nov., Neorhizobium petrolearium comb. nov., Pararhizobium arenae comb. nov., Pseudorhizobium tarimense comb. nov. and Mycoplana azooxidifex comb. nov. Lastly, we present arguments that the unification of the genera

Ensifer

and

Sinorhizobium

in Opinion 84 of the Judicial Commission is no longer justified by current genomic and phenotypic data. Despite pairwise cpAAI values for all

Ensifer

species and all

Sinorhizobium

species being >86 %, additional genomic and phenotypic data suggest that they significantly differ in their biology and ecology. We therefore propose emended descriptions of

Ensifer

and

Sinorhizobium

, which we argue should be considered as separate genera.
Are Dutch General Practitioners willing to prescribe mifepristone and misoprostol?: a mixed-methods study
Authors: Schellekens Julia E A P , Houtvast Claire S E , Leusink Peter , Kleiverda Gunilla , Gomperts Rebecca
Published: March 2022

Background

The World Health Organization (WHO) indicates that General Practitioners (GPs) can effectively and safely provide mifepristone and misoprostol for medical termination of pregnancy (TOP). Dutch GPs are permitted to treat miscarriages with mifepristone and misoprostol, but in practice only guide spontaneous miscarriages. Current Dutch abortion law forbids GPs to prescribe these medications for medical TOP. Medical TOP is limited to the specialized settings of abortion clinics and hospitals. A shift to primary care is debated in the House of Representative, following the example of France and Ireland. It would improve reproductive health care and choices for women. Little is known about GPs’ willingness to provide medical TOP and miscarriage management.

Aim

This study aimed to gain insight into Dutch GPs’ willingness and anticipated obstacles to prescribing mifepristone and misoprostol for medical TOP and miscarriages.

Design and Setting

This is a mixed-method study among Dutch GPs.

Method

A questionnaire provided quantitative data that was analysed using descriptive methods. Thematic analyses were performed on qualitative data collected by in-depth interviews.

Results

The questionnaire was sent to 575 GPs, the response rate was 22.1%. Of the responders, 84.3% were willing to prescribe mifepristone and misoprostol and 58.3% were willing to provide both medical TOP and miscarriage management. 57.5% indicated a need for training. The main barriers influencing GPs’ willingness were lack of experience, knowledge, time and a restrictive abortion law.

Conclusion

Over 80% of the respondents were willing to prescribe mifepristone and misoprostol for medical TOP or miscarriages. Training, (online) education and a revision of the abortion law are recommended.

How this fits in

Medical TOP in the Netherlands can only be provided in abortion clinics and hospitals. GPs may prescribe these same medications for miscarriage management, but in practice only guide spontaneous miscarriages. To improve access to woman-centred care, it is important to allow GPs by law to provide medical TOP. Our study is the first to assess Dutch GPs’ willingness to provide mifepristone and misoprostol and aims to understand enablers and barriers that give insight into the feasibility of a shift in care. Our results illustrate the need to revise laws and to provide training and education in the similar procedure of medical TOP and miscarriage management.
Pegylated-interferon-λ treatment-induced peripheral interferon stimulated genes are associated with SARS-CoV-2 viral load decline despite delayed T cell response in older individuals
Authors: Santer Deanna M , Li Daniel , Ghosheh Yanal , Zahoor Muhammad Atif , Prajapati Dhanvi , Tyrrell D Lorne J , Feld Jordan J , Gehring Adam J
Published: March 2022

Interferons (IFNs) are antiviral cytokines induced very early after SARS-CoV-2 infection and are crucial for viral clearance, shaping immunity, and preventing the development of severe COVID-19. We previously demonstrated that a single injection of peginterferon-lambda1 (PEG-IFN-λ) accelerated viral clearance in COVID-19 patients. To determine if the rapid viral decline was mediated by enhanced immunity, we assessedin vivoresponses to PEG-IFN-λ by single cell RNA sequencing and measured SARS-CoV-2-specific T cell and antibody responses between placebo and PEG-IFN-λ-treated patients. PEG-IFN-λ treatment induced interferon stimulated genes in peripheral immune cells expressingIFNLR1, with plasmacytoid dendritic cells having the greatest response, followed by B cells. PEG-IFN-λ did not significantly affect SARS-CoV-2-specific antibody levels in plasma or the magnitude or functionality of virus-specific T cells. However, we identified a delayed T cell response in older adults, suggesting that PEG-IFN-λ can overcome the delay in adaptive immunity to accelerate viral clearance in patients most at risk for severe disease. Taken together, PEG-IFN-λ offers an early COVID-19 treatment option for outpatients to boost innate antiviral defenses without dampening peripheral SARS-CoV-2 adaptive immunity
Investigating Automatic Speech Emotion Recognition for Children with Autism Spectrum Disorder in interactive intervention sessions with the social robot Kaspar
Authors: Milling Manuel , Bartl-Pokorny Katrin D. , Schuller Björn W.
Published: March 2022

In this contribution, we present the analyses of vocalisation data recorded in the first observation round of the European Commission’s Erasmus Plus project “EMBOA, Affective loop in Socially Assistive Robotics as an intervention tool for children with autism”. In total, the project partners recorded data in 112 robot-supported intervention sessions for children with autism spectrum disorder. Audio data were recorded using the internal and lapel microphone of the H4n Pro Recorder. To analyse the data, we first utilise a child voice activity detection (VAD) system in order to extract child vocalisations from the raw audio data. For each child, session, and microphone, we provide the total time child vocalisations were detected. Next, we compare the results of two different implementations for valence- and arousal-based speech emotion recognition, thereby processing (1) the child vocalisations detected by the VAD and (2) the total recorded audio material. We provide average valence and arousal values for each session and condition. Finally, we discuss challenges and limitations of child voice detection and audio-based emotion recognition in robot-supported intervention settings.
Comprehensive characterization of Candida isolates in a given geographical area for the determination of prevalence and drug sensitivity
Authors: Pamnani Tamanna , Agarwal Saifali , Chourasia Jayati , Sinha Rishika , Choudhary Pankaj , Niraj Ravi Ranjan K , Nagar Ramjas , Shrivastava Sandeep K , Kalra Aakanksha
Published: March 2022

Purpose

An alarming increase in Candidiasis infections has been observed worldwide and in India. This increase is attributed to increase in immune-compromised individuals and an increase in plethora of species causing the disease. The emergence of drug resistance of isolates has further worsened the situation.

Materials & Methods

Candida isolates causing infections were obtained from patients and evaluated for their macroscopic and microscopic characteristics by colony morphology and staining procedures. The patients’ demography was also analyzed to identify any correlation between the isolates. Doubling time was determined to analyze growth characteristics of these isolates. Isolates were also characterized biochemically for their ability to assimilate carbon and nitrogen sources and for fermentative capabilities. Additionally, drug sensitivity profiles of these isolates were analyzed towards Azoles.

Results

Demographic analysis of the isolates suggested that all age groups were affected by Candida via both albicans and non-albican species. The infections were not gender biased but majority isolates were obtained from urine samples suggesting Candida as an important species causing urine-genital infections. Macroscopic and microscopic analysis showed cream colored circular colonies with smooth surface and entire margins with cells in single and budding stage. The doubling time ranged between 40 mins to 180 mins with 81 mins being the average. Biochemical characterization showed sucrose to be the most metabolizable sugar with maximum fermentative capacity with glucose. Surprisingly, very low nitrogen assimilation capacity was observed with all the nitrogen sources tested (nitrate, urea and glycine). Drug sensitivity towards Azoles suggested almost 50% and 90% isolates were resistant to Fluconazole and Itraconazole respectively, the most common Azoles used against fungal infections in the current scenario.

Conclusions

The results obtained from the study suggested differential characteristics of the isolates towards various parameters thereby indicating the relevance of isolate characterization, for appropriate control and prevention of the disease.

Graphical abstract
Discordant population structure inferred from male- and female-type mtDNAs fromMacoma balthica, a bivalve species characterized by doubly uniparental inheritance of mitochondria
Authors: Le Cam Sabrina , Brémaud Julie , Becquet Vanessa , Huet Valérie , Dubillot Emmanuel , Garcia Pascale , Viricel Amélia , Breton Sophie , Pante Eric
Published: March 2022

Doubly Uniparental Inheritance (DUI) of mitochondria is a remarkable exception to the Strictly Maternal Inheritance (SMI) in metazoans. In species characterized by DUI --almost exclusively gonochoric bivalve mollusks--females (F) transmit mitochondria to offspring of both sexes, while males (M) pass on their mitochondria exclusively to their sons. Under DUI, males are heteroplasmic, somatic tissues containing F-transmitted mtDNA and gametic cells containing M-transmitted mtDNAs. The aforementioned transmission routes make M- and F-transmitted mtDNA interesting as sex-specific markers which can differ in their effective population sizes, mutation rates, and selective constraints. For these reasons, looking at both markers can provide significant insights into the genetic structure of populations and investigate its determinants. In this study, we document differences in genetic diversity, divergence, inter-populational differentiation and biogeographic structure between M- and F-typecox1mt genes in the Baltic tellin (Macoma balthica) to test whethercox1mandcox1fgenes bear the marks of similar phylogeographic histories. These markers were sequenced for 302 male individuals sampled from the North Sea to the Gironde Estuary (Southern France) encompassing the intra-subspecificM. b. rubrahybrid zone in the Gulf of Saint Malo. Both genes supported a scenario of cladogenesis ofM.b. rubraclades prior to the last glacial maximum. Nucleotide diversity and net divergence were over twice higher incox1mcompared tocox1f. Genetic differentiation between northern and southern populations was nearly 3 times higher atcox1mcompared tocox1f (global ΦST= 0.345 and 0.126 respectively) and the geographic localization of the strongest genetic break significantly differed between the markers (Finistère Peninsula atcox1f; Cotentin Peninsula atcox1m, ∼250 km apart). A higher mutation rate, relaxed negative selection and differences in effective population sizes (depending on locations) atcox1mcould explain differences in population genetic structure. As both F- and M-type mtDNAs interact with nuclear genes for oxidative phosphorylation and ATP production, geographical discordances in genetic clines in a context of secondary contact could be linked to mito-nuclear genetic incompatibilities.
GraphBio: a shiny web app to easily perform popular visualization analysis for omics data
Authors: Zhao Tian-Xin , Wang Ze-Lin
Published: March 2022

Here, we present GraphBio, a shiny web app to easily perform visualization analysis for omics data. GraphBio provides 15 popular visualization analysis methods, including heatmap, volcano plots, MA plots, network plots, dot plots, chord plots, pie plots, four quadrant diagrams, venn diagrams, cumulative distribution curves, PCA, survival analysis, ROC analysis, correlation analysis and text cluster analysis. It enables experimental biologists without programming skills to easily perform popular visualization analysis and get publication-ready figures.

Availability and implementation

GraphBio, as an online web application, is freely available at http://www.graphbio1.com/en/ (English version) and http://www.graphbio1.com/ (Chinese version).

Contact

databio@163.com
Efficient Gene Knockout in Salivary Gland Epithelial Explant Cultures
Authors: Sekiguchi Rei , Mehlferber Madison M. , Matsumoto Kazue , Wang Shaohe
Published: March 2022

We have developed methods to achieve efficient CRISPR-Cas9 mediated gene knockout in ex vivo mouse embryonic salivary epithelial explants. Salivary epithelial explants provide a valuable model for characterizing cell signaling, differentiation, and epithelial morphogenesis, but research has been limited by a paucity of efficient gene perturbation methods. Here, we demonstrate highly efficient gene perturbation by transient transduction of guide RNA-expressing lentiviruses into Cas9-expressing salivary epithelial buds isolated from Cas9 transgenic mice. We first show that salivary epithelial explants can be cultured in low-concentration, non-solidified Matrigel suspensions in 96-well plates, which greatly increases sample throughput compared to conventional cultures embedded in solidified Matrigel. We further show that salivary epithelial explants can grow and branch with FGF7 alone, while supplementing with ITS (insulin, transferrin, and selenium) enhances growth and branching. We then describe an efficient workflow to produce experiment-ready, high-titer lentiviruses within 1 week after molecular cloning. To track transduced cells, we designed the lentiviral vector to co-express a nuclear fluorescent reporter with the guide RNA. We routinely achieve 80% transduction efficiency when antibiotic selection is used. Importantly, we detected robust loss of targeted protein products when testing 9 guide RNAs for 3 different genes. Moreover, targeting the β1 integrin gene (Itgb1) inhibited branching morphogenesis, which supports the importance of cell-matrix adhesion in driving branching morphogenesis. In summary, we have established a lentivirus-based method that can efficiently perturb genes of interest in salivary epithelial explants, which will greatly facilitate studies of specific gene functions using this system.
Beyond resource selection: emergent spatio-temporal distributions from animal movements and stigmergent interactions
Authors: Potts Jonathan R. , Giunta Valeria , Lewis Mark A.
Published: March 2022

A principal concern of ecological research is to unveil the causes behind observed spatio-temporal distributions of species. A key tactic is to correlate observed locations with environmental features, in the form of resource selection functions or other correlative species distribution models. In reality, however, the distribution of any population both affects and is affected by those surrounding it, creating a complex network of feedbacks causing emergent spatio-temporal features that may not correlate with any particular aspect of the underlying environment. Here, we study the way in which the movements of populations in response to one another can affect the spatio-temporal distributions of ecosystems. We construct a stochastic individual-based modelling (IBM) framework, based on stigmergent interactions (i.e. organisms leave marks which cause others to alter their movements) between and within populations. We show how to gain insight into this IBM via mathematical analysis of a partial differential equation (PDE) system given by a continuum limit. We show how the combination of stochastic simulations of the IBM and mathematical analysis of PDEs can be used to categorise emergent patterns into homogeneous vs. heterogeneous, stationary vs. perpetually-fluctuating, and aggregation vs. segregation. In doing so, we develop techniques for understanding spatial bifurcations in stochastic IBMs, grounded in mathematical analysis. Finally, we demonstrate through a simple example how the interplay between environmental features and between-population stigmergent interactions can give rise to predicted spatial distributions that are quite different to those predicted purely by accounting for environmental covariates.
Thalamic control of sensory enhancement and sleep spindle properties in a biophysical model of thalamoreticular microcircuitry
Authors: Iavarone Elisabetta , Simko Jane , Shi Ying , Bertschy Marine , García-Amado María , Litvak Polina , Kaufmann Anna-Kristin , O’Reilly Christian , Amsalem Oren , Abdellah Marwan , Chevtchenko Grigori , Coste Benoît , Courcol Jean-Denis , Ecker András , Favreau Cyrille , Fleury Adrien Christian , Geit Werner Van , Gevaert Michael , Guerrero Nadir Román , Herttuainen Joni , Ivaska Genrich , Kerrien Samuel , King James G. , Kumbhar Pramod , Lurie Patrycja , Magkanaris Ioannis , Muddapu Vignayanandam Ravindernath , Nair Jayakrishnan , Pereira Fernando L. , Perin Rodrigo , Petitjean Fabien , Ranjan Rajnish , Reimann Michael , Soltuzu Liviu , Sy Mohameth François , Tuncel M. Anıl , Ulbrich Alexander , Wolf Matthias , Clascá Francisco , Markram Henry , Hill Sean L.
Published: March 2022

Thalamoreticular circuitry is known to play a key role in attention, cognition and the generation of sleep spindles, and is implicated in numerous brain disorders, but the cellular and synaptic mechanisms remain intractable. Therefore, we developed the first detailed computational model of mouse thalamus and thalamic reticular nucleus microcircuitry that captures morphological and biophysical properties of ∼14,000 neurons connected via ∼6M synapses, and recreates biological synaptic and gap junction connectivity. Simulations recapitulate multiple independent network-level experimental findings across different brain states, providing a novel unifying cellular and synaptic account of spontaneous and evoked activity in both wakefulness and sleep. Furthermore, we found that: 1.) inhibitory rebound produces frequency-selective enhancement of thalamic responses during wakefulness, in addition to its role in spindle generation; 2.) thalamic interactions generate the characteristic waxing and waning of spindle oscillations; and 3.) changes in thalamic excitability (e.g. due to neuromodulation) control spindle frequency and occurrence. The model is openly available and provides a new tool to interpret spindle oscillations and test hypotheses of thalamoreticular circuit function and dysfunction across different network states in health and disease.
A Single Multipurpose FSH–Blocking Therapeutic for Osteoporosis and Obesity
Authors: Gera Sakshi , Kuo Tan-Chun , Korkmaz Funda , Sant Damini , DeMambro Victoria , Gumerova Anisa , Sudha Kathayani , Padilla Ashley , Prevot Geoffrey , Munitz Jazz , Teunissen Abraham , van Leent Mandy , Post Tomas G.J.M. , Fernandes Jessica C. , Netto Jessica , Sultana Farhath , Shelly Eleanor , Kumar Pushkar , Cullen Liam , Chatterjee Jiya , Miyashita Sari , Kannangara Hasni , Bhongade Megha , Ievleva Kseniia , Muradova Valeriia , Batista Rogerio , Robinson Cemre , Macdonald Anne , Babunovic Susan , Saxena Mansi , Meseck Marcia , Caminis John , Iqbal Jameel , New Maria I. , Ryu Vitaly , Kim Se-Min , Cao Jay J. , Zaidi Neeha , Fayad Zahi , Lizneva Daria , Rosen Clifford J. , Yuen Tony , Zaidi Mone
Published: March 2022

Pharmacological and genetic studies over the past decade have established FSH as an actionable target for diseases affecting millions, notably osteoporosis, obesity and Alzheimer’s disease (AD). Blocking FSH action prevents bone loss, fat gain and AD–like features in mice. We recently developed a first–in–class, humanized, epitope–specific FSH blocking antibody, MS-Hu6, with a KDof 7.52 nM. Using a GLP–compliant platform, we now report the efficacy of MS-Hu6 in preventing obesity and osteoporosis in mice, and parameters of acute safety in monkeys. Biodistribution studies using89Zr–labelled, biotinylated or unconjugated MS-Hu6 in mice and monkeys showed localization to bone, bone marrow and fat depots. MS-Hu6 displayed a β phase t½of 13 days (316 hours) in humanizedTg32mice, and bound endogenous FSH. We tested 215 variations of excipients using the protein thermal shift assay to generate a final formulation that rendered MS-Hu6 stable in solution upon freeze–thaw and at different temperatures, with minimal aggregation, and without self–, cross–, or hydrophobic interactions or appreciable binding to relevant human antigens. MS-Hu6 showed the same level of “humanness” as human IgG1in silico, and was non–immunogenic in ELISPOT assays for IL-2 and IFNγ in human peripheral blood mononuclear cell cultures. We conclude that MS-Hu6 is efficacious, durable and manufacturable, and is therefore poised for future human testing as a multipurpose therapeutic.
Temporal responses of bumblebee gustatory neurons to sugars
Authors: Parkinson Rachel H. , C. Kessler Sébastien , Scott Jennifer , Simpson Alexander , Bu Jennifer , Mahdi Adam , Miriyala Ashwin , Wright Geraldine A.
Published: March 2022

The sense of taste permits the recognition of valuable nutrients and the avoidance of potential toxins. Models of gustatory coding propose that within modalities (e.g. sweet, bitter, etc.), taste ligands are not distinct stimuli. However, these models are based on data from mice or flies that have omnivorous, non-specialist diets. A specialist feeder might, however, be expected to have acuity within modality if stimulus resolution was critical to survival. Previously, we found that bumblebees have a specialized mechanism for sensing sugars whereby two gustatory receptor neurons (GRNs) within the galeal sensilla of the bees’ mouthparts exhibit burst of spikes. Here, we show that the temporal firing patterns of these GRNs separate sugars into four distinct groups that correlate with sugar nutritional value and palatability. We also identified a third GRN that responded to stimulation with relatively high concentrations of fructose, sucrose, and maltose. Sugars that were non-metabolizable or toxic suppressed the responses of bursting GRNs to sucrose. These abilities to encode information about sugar value are a refinement to the bumblebee’s sense of sweet taste that could be an adaptation that enables precise calculations of the nature and nutritional value of floral nectar.
Keratinocyte-tethering modification for biologics enables location-precise treatment in mouse vitiligo
Authors: Hsueh Ying-Chao , Wang Yuzhen , Riding Rebecca L. , Catalano Donna E. , Lu Yu-Jung , Richmond Jillian M. , Siegel Don L. , Rusckowski Mary , Stanley John R. , Harris John E.
Published: March 2022

Despite the central role of IFNγ in vitiligo pathogenesis, systemic IFNγ neutralization is an impractical treatment option due to strong immunosuppression. However, most vitiligo patients present with less than 20% affected body surface area, which provides an opportunity for localized treatments that avoid systemic side effects. After identifying keratinocytes as key cells that amplify IFNγ signaling during vitiligo, we hypothesized that tethering an IFNγ neutralizing antibody to keratinocytes would limit anti-IFNγ effects to the treated skin for the localized treatment. To that end, we developed a bispecific antibody (BsAb) capable of blocking IFNγ signaling while binding to desmoglein expressed by keratinocytes. We characterized the effect of the BsAb in vitro, ex vivo, and in a mouse model of vitiligo. SPECT/CT biodistribution and serum assays after local footpad injection revealed that the BsAb had improved skin retention, faster elimination from the blood, and less systemic IFNγ inhibition than the non-tethered version. Furthermore, the BsAb conferred localized protection almost exclusively to the treated footpad during vitiligo that was not possible by local injection of the non-tethered anti-IFNγ antibody. Thus, keratinocyte-tethering proved effective while significantly diminishing off-tissue effects of IFNγ blockade, offering a new treatment strategy for localized skin diseases, including vitiligo.
Genetic Variation and Disease Severity of Respiratory Syncytial Viruses
Authors: Anderson Christopher S. , Zhang Yun , Corbett Anthony , Chu Chinyi , Grier Alexander , Wang Lu , Qiu Xing , Mccall Mathew , Topham David J. , Walsh Edward E. , Mariani Thomas J. , Scheuermann Richard , Caserta Mary T.
Published: March 2022

Respiratory Syncytial Virus (RSV) disease in newborns ranges from mild symptoms to severe disease requiring hospitalization. RSV is classified into two subtypes (RSVA and RSVB) based on antigenic and genetic differences. The role these genomic variations play in disease severity remains unknown. Genome sequences were obtained using next-generation RNA sequencing on archived frozen nasal swabs of young children (< 8 months-old) infected with RSV in Rochester, NY between 1977-1998. Samples were chosen from both children hospitalized with severe RSV disease (inpatient) and those presenting with mild symptoms (outpatient) during their first cold-season. Both A and B subtypes demonstrated significant differences in the phylogeny and primary-protein structure during this time period. We found a significant association between RSV phylogeny over this time period and disease severity. For both subtypes, the G-protein demonstrated the greatest amino acid substitutions, although the number of amino acid substitutions was higher in the RSVA subtype. We found a significant association between G-protein variation and disease severity for RSVA, but not RSVB. For both subtypes, variation in the M2-2 protein was significantly associated with disease severity. These results suggest that the genetic variability of RSV proteins may contribute to disease severity in humans.

Importance

Each cold-season Respiratory Syncytial Virus (RSV) infects thousands of children in the US. Some will display mild cold symptoms while others develop severe disease, sometimes resulting in lifelong lung problems or fatality. RSV initiates infection and replicates in the nasopharynx. Substitutions in the RSV genome can be found in clinically isolated nasal-swab samples of RSV infected children. Whether these genome variations contribute to severe disease is unknown. Here we found a statistically significant association between RSV phylogeny and disease severity. Furthermore, we found specific RSV proteins (G and M2-2) whose amino acid variation was statistically associated with severe disease, although which protein was associated depended on subtype. Taken together, our results suggest that RSV genotype contributed to disease severity over this time period.
Comparison of the reactogenicity and immunogenicity of a reduced and standard booster dose of the mRNA COVID-19 vaccine in healthy adults after two doses of inactivated vaccine
Authors: Kanokudom Sitthichai , Assawakosri Suvichada , Suntronwong Nungruthai , Chansaenroj Jira , Auphimai Chompoonut , Nilyanimit Pornjarim , Vichaiwattana Preeyaporn , Thongmee Thanunrat , Yorsaeng Ritthideach , Duangchinda Thaneeya , Chantima Warangkana , Pakchotanon Pattarakul , Srimuan Donchida , Thatsanatorn Thaksaporn , Klinfueng Sirapa , Mongkolsapaya Juthathip , Sudhinaraset Natthinee , Wanlapakorn Nasamon , Honsawek Sittisak , Poovorawan Yong
Published: March 2022

The coronavirus disease 2019 (COVID-19) pandemic has been a serious healthcare problem worldwide since December 2019. The third dose of heterologous vaccine was recently approved by World Health Organization. The present study compared the reactogenicity and immunogenicity of the reduced and standard third booster dose of the BNT162b2 and mRNA-1273 vaccine in adults who previously received the two-dose CoronaVac vaccine. Results showed that headache, joint pain, and diarrhea were more frequent in the 15 μg-than the 30 μg-BNT162b2 groups, whereas joint pain and chilling were more frequent in the 100 μg-than the 50 μg-mRNA-1273 groups. No significant differences in immunogenicity were detected. These findings demonstrate that the reduced dose of the mRNA vaccines elicited antibody responses against the SARS-CoV-2 delta and omicron variants that were comparable to the standard dose. The reduced dose could be used to increase vaccine coverage in situations of limited global vaccine supply.

Highlights

The 15 μg- and 30 μg-BNT162b2, and 50 μg- and 100 μg-mRNA-1273 booster doses were compared

Booster vaccination with the mRNA vaccine elicits high Ig and IgG anti-RBD in CoronaVac-vaccinated adults

No differences were observed in antibody responses after the reduced or standard booster dose of the mRNA vaccine in CoronaVac-vaccinated adults

Neutralizing antibodies against the delta and omicron variants were significantly higher after the booster dose

Neutralizing antibody titers were lower against the omicron variant than the delta variant in all vaccinated adults
Direct fluorescent labeling of NF186 and NaV1.6 in living primary neurons using bioorthogonal click chemistry
Authors: Stajković Nevena , Liu Yuanyuan , Arsić Aleksandra , Meng Ning , Lyu Hang , Zhang Nan , Grimm Dirk , Lerche Holger , Nikić-Spiegel Ivana
Published: March 2022

The axon initial segment (AIS) is a highly specialized neuronal compartment that regulates the generation of action potentials and maintenance of neuronal polarity. Despite its importance, live imaging of the AIS is challenging due to the limited number of suitable labeling methods. To overcome this limitation, we established a novel approach for live labeling of the AIS using unnatural amino acids (UAAs) and bioorthogonal click chemistry. The small size of the UAAs and the possibility of introducing them virtually anywhere into the target proteins make this method particularly suitable for live labeling and imaging of complex and spatially restricted proteins. With this approach, we labeled two large AIS components, the 186 kDa isoform of neurofascin (NF186) and the 260 kDa voltage-gated sodium channel (NaV1.6), and performed widefield and confocal microscopy in fixed and living neurons. Moreover, we demonstrated the applicability of this method by studying the localization of two epilepsy-causing NaV1.6 variants with a loss-of-function effect. Finally, to further improve the efficiency of the UAA incorporation, we developed adeno-associated viral (AAV) vectors for click labeling in primary neurons. The use of AAV vectors will facilitate the transfer of UAA-based click labeling technology to more complex biological systems, such as organotypic slice cultures, organoids, and animal models.