bluuebunny/crossref_metadata_2025 huggingface dataset:
I Don’t Want to Go Back
Authors: Liu Zihan , Van Egdom Drake , Flin Rhona , Spitzmueller Christiane , Adepoju Omolola , Krishnamoorti Ramanan
Published: August 2020

Objective:

We study employee perspectives on return to physical workspaces to ultimately inform employers’ and policy makers’ decision making around the return to work during COVID-19.

Methods:

We tested the three-component conceptual model using survey data collected in the United States in May 2020 from samples of energy workers (N = 333).

Results:

Women, non-Caucasians, and employees living in multi-generational households were less willing to return. Concerns about childcare were negatively related to willingness to return, whereas organizational strategies for mitigating COVID-19 transmission at work were positively related to willingness to return. COVID-19 infections in an employees’ network were also negatively related to employees’ willingness to return.

Conclusions:

Blanket policies may miss the nuanced needs of different employee groups. Employers and policy makers should adopt flexible approaches to ensure a return to workspaces that addresses employee concerns and needs.
A Literature Review of Nursing Extern Program Outcomes
Authors: Rugs Deborah , Nedd Nicole , Deitrick Lynn , Hall Kimberly S.
Published: August 2020

This literature review gathers in one place recent published literature on extern programs. Fourteen extern articles published between 2009 and 2019 were identified, and program information was extracted. Student benefits were increased in confidence, clinical skills, and learning the nursing environment. Organizational benefits were reduced orientation, reduced recruitment costs, and hiring nurses that were a good fit with the organization. For a healthcare organization, extern programs may ease the shortage of available registered nurses.
One-step Multiplex Real-time RT-PCR for Molecular Detection and Typing of Dengue Virus Infection From Paraffin-embedded Tissues During the Brazilian 2019 Outbreak
Authors: Mariotti Guerra Juliana , Santos da Silva Ferreira Camila , Rosa Fernandes Beraldo Karolina , Midori Kimura Lidia , Possatto Fernandes Takahashi Juliana , Salas-Gómez David , Sequetin Cunha Mariana , Fletcher Buss Lewis , Silva Nogueira Juliana , Yurika Maeda Adriana , José Tadeu de Araújo Leonardo
Published: August 2020

Formalin-fixed paraffin-embedded (FFPE) tissues are an important source for investigation of dengue virus (DENV) infection, particularly when blood or fresh frozen (FF) samples are unavailable. Histopathologic features and immunohistochemistry may have poor sensitivity and serotype determination is not always possible. Viral RNA genome detection tests are faster and considered the most sensitive technique for this kind of analysis, however, the use of molecular methods applied to FFPE tissues is still limited. The authors applied a single-step multiplex reverse transcription-quantitative polymerase chain reaction (RT-qPCR) for the investigation of DENV infection and typing to FFPE samples of 32 fatal cases received during the 2019 outbreak that occurred in São Paulo state, Brazil. The authors compared the results with those obtained using FF tissues. Of the 24 cases with both FF and FFPE samples, 22 (91.67%) of the FF and 19 (76.20%) of the FFPE specimens were positive. Two cases (8.33%) tested negative in both types of samples. All 8 cases with only FFPE samples available were positive. The accuracy (87.5%) of the RT-qPCR for DENV in FFPE samples were satisfactory. Although the cycle quantification (Cq) values were significantly higher in these materials (P<0.0001, Wilcoxon signed-rank test) when compared with FF tissues, Spearman’s rank coefficient indicated a good correlation between the Cq values from both sample types (P=0.0063; rho=0.576). RT-qPCR applied to FFPE samples improved detection of DENV in fatal cases and represents a useful tool for diagnosis and epidemiologic studies.
Urothelial Carcinoma In Situ of the Bladder: Correlation of CK20 Expression With Adaptive Immune Resistance, Response to BCG Therapy, and Clinical Outcome
Authors: Lombardo Kara A. , Murati Amador Belkiss , Parimi Vamsi , Hoffman-Censits Jean , Choi Woonyoung , Hahn Noah M. , Kates Max , Bivalacqua Trinity J. , McConkey David , Hoque Mohammad O. , Matoso Andres
Published: August 2020

Immunohistochemical stains have been suggested to aid in diagnostically challenging cases of urothelial carcinoma in-situ (CIS). Although full thickness immunostaining for CK20 is supportive of CIS, a subset of CIS cases is CK20(−), the clinical significance of which was unknown. This study included 43 patients with primary diagnosis of bladder CIS including 32 with only CIS, 5 with CIS and separate noninvasive high-grade papillary urothelial carcinoma, and 6 with CIS and separate high-grade urothelial carcinoma with lamina propria invasion. Digital morphometric image analysis showed that the average nuclear areas of enlarged nuclei were similar in CK20(+) and CK20(−) CIS (26.9 vs. 24.5 µM2; P=0.31). Average Ki67 index for CK20(+) CIS was higher than CK20(−) CIS (31.1% vs. 18.3%; P=0.03). Patients with CK20(+) CIS [28 (65%)] and patients with CK20(−) CIS [15 (35%)] had the same rates of Bacillus Calmete-Guerin (BCG) failure but patients with CK20(−) CIS had higher stage progression [3 CK20(+) (11%) vs. 6 CK20(−) (40%); P=0.02]. Given recent approval of immune checkpoint inhibitors in patients with CIS refractory to BCG, programmed death ligand-1 expression and colocalization with CD8(+) lymphocytes was investigated as signature of adaptive immune response and was seen in 8 patients regardless of CK20 status and exclusively among patients who failed BCG. Our results confirm that negative CK20 IHC does not exclude CIS and that those patients have similar clinical outcomes as patients with CK20(+) CIS. Programmed death ligand-1 and CD8 colocalization seen among patients who failed BCG therapy is an easy assay to perform to identify patients who could potentially benefit from combined BCG therapy and immune checkpoint inhibition.
Virtual Rehabilitation in Parkinson Disease
Authors: Toldo Juliana M. P. , Arjona Michelly , Campos Neto Guilherme C. , Vitor Taise , Nogueira Solange Amorim , Amaro Edson , Saba Roberta A. , Silva Sonia M. C. A. , Ferraz Henrique B. , Felício André C.
Published: August 2020

Objective

The aim of the study was to verify the effect of a virtual rehabilitation protocol for patients with Parkinson disease, primarily assessing striatal dopamine transporters and secondarily motor symptoms and quality of life.

Design

Nineteen patients with Parkinson disease underwent an 8-wk virtual rehabilitation protocol using XBOX 360S. Evaluation of dopamine transporters was performed by single-photon emission computed tomography using TRODAT-1 as the radioligand. Participants were clinically assessed using the Unified Parkinson Disease Rating Scale to quantify motor symptoms. Moreover, the Parkinson Disease Questionnaire and Short-Form Health Status Survey were used to assess quality of life and the Berg Balance Scale to assess balance.

Results

Regarding our primary outcome, dopamine transporter was significantly increased in the putamen contralateral to the clinically most affected body side (P = 0.034) considering preintervention and postintervention measurements. Furthermore, we observed significant improvement in Unified Parkinson Disease Rating Scale (10-point reduction, P = 0.001), Parkinson Disease Questionnaire (11.3-point reduction, P = 0.001), Short-Form Health Status Survey (“Functional capacity,” P = 0.001; “Pain,” P = 0.006; and “Mental Health” domains, P < 0.001), and Berg Balance Scale (5-point increase, P = 0.015).

Conclusions

In our group of Parkinson disease patients, this virtual rehabilitation protocol enabled a dopamine transporter increase in the region of the putamen contralateral to the clinically most affected body side. Moreover, motor signs and quality of life were significantly improved.
Updates in technology-based interventions for attention deficit hyperactivity disorder
Authors: Guan Lim Choon , Lim-Ashworth Nikki S.J. , Fung Daniel S.S.
Published: August 2020

Purpose of review

Technological advancement has led to the development of novel treatment approaches for attention deficit hyperactivity disorder (ADHD). This review aims to review recent studies which employ the use of technology to treat ADHD, with particular focus on studies published during a 1-year period from February 2019 to February 2020.

Recent findings

Most recent studies involved children aged 12 years and below. Interventions included cognitive training through games, neurofeedback and a combination of several approaches. More novel approaches included trigeminal nerve stimulation and brain–computer interface, and studies had utilized technology such as X-box Kinect and eye tracker. There was a shift towards delivering intervention at home and in school, enabled by technology. The study outcomes were variable and mainly included executive functioning measures and clinical ratings. These interventions were generally safe with few reported adverse events.

Technology has enabled interventions to be delivered outside of the clinic setting and presented an opportunity for increased access to care and early intervention. Better quality studies are needed to inform on the efficacy of these interventions.
Emotion-focused parenting interventions for prevention and treatment of child and adolescent mental health problems: a review of recent literature
Authors: Havighurst Sophie S. , Radovini Alessandra , Hao Brandon , Kehoe Christiane E.
Published: August 2020

Purpose of review

To review recent studies on emotion-focused parenting interventions to provide clinicians with knowledge about how these approaches might be used in prevention and treatment of mental health difficulties for children, adolescents and their families.

Recent findings

A number of emotion-focused parent interventions are reported in the literature, including emotion coaching/communication parenting programs, emotion-focused family therapy, attachment-focused parenting interventions (including those that address parental reflective functioning/mentalization), mindfulness parenting programs and behavioral programs with added emotion components. All target emotions or emotional communication to assist parents and children understand and work through emotional experiences so they are less likely to impede healthy functioning. These interventions target four main domains: exploring family of origin or early attachment/relational experiences with emotion, targeting parents’ own emotion awareness and regulation, shifting parents responses to or communication with their children when emotions occur, and promoting parents’ skills for assisting children to regulate emotions and behavior. This review from the last 18 months found 50 studies that evaluated programs addressing these domains.

Whilst the dominant approach in evidence-based parenting programs has been teaching behavioral strategies, it has been recognized that a focus on emotion-related processes is important. This is especially when working to improve the attachment relationship or when parents and children experience emotion dysregulation. This review demonstrates extensive evidence to support emotion-focused parenting interventions.
Eating disorders in adolescents with type 1 diabetes mellitus
Authors: Wagner Gudrun , Karwautz Andreas
Published: August 2020

Purpose of review

Eating disorders represent one of the most common psychiatric disorders in adolescents with type 1 diabetes mellitus (T1DM), with a prevalence twice as high as in individuals without diabetes. The increased risk for acute and long-term medical complications and a three-fold mortality rate with this dual condition makes it extremely important to screen adolescents for eating disorders as soon as possible.

Recent findings

A number of 13 surveys in the last two years focussed on prevalence rates of disordered eating and associations with sociodemographic, eating disorder-specific and diabetes-related factors in this population. Variations in study design, sample size, age range, sex, instruments to assess eating disorders and outcome make comparisons difficult. Healthcare professionals report challenges in detecting and treating adolescents with this dual condition because of a lack of validated screening tools, guidelines, and integration across specialist care. Studies on psychological treatments for the comorbidity of eating disorders and T1DM are scarce, especially for adolescents.

The development and evaluation of psychological treatments for adolescents with eating disorders and T1DM are urgently needed.
The Coronavirus Health and Impact Survey (CRISIS) reveals reproducible correlates of pandemic-related mood states across the Atlantic
Authors: Nikolaidis Aki , Paksarian Diana , Alexander Lindsay , Derosa Jacob , Dunn Julia , Nielson Dylan M. , Droney Irene , Kang Minji , Douka Ioanna , Bromet Evelyn , Milham Michael , Stringaris Argyris , Merikangas Kathleen R.
Published: August 2020

The COVID-19 pandemic and its social and economic consequences have had adverse impacts on physical and mental health worldwide and exposed all segments of the population to protracted uncertainty and daily disruptions. The CoRonavIruS health and Impact Survey (CRISIS) was developed for use as an easy to implement and robust questionnaire covering key domains relevant to mental distress and resilience during the pandemic. In the current work, we demonstrate the feasibility, psychometric structure and construct validity of this survey. We then show that pre-existing mood states, perceived COVID risk, and lifestyle changes are strongly associated with negative mood states during the pandemic in population samples of adults and in parents reporting on their children in the US and UK. Ongoing studies using CRISIS include international studies of COVID-related ill health conducted during different phases of the pandemic and follow-up studies of cohorts characterized before the COVID pandemic.
On stability of Canonical Correlation Analysis and Partial Least Squares with application to brain-behavior associations
Authors: Helmer Markus , Warrington Shaun , Mohammadi-Nejad Ali-Reza , Ji Jie Lisa , Howell Amber , Rosand Benjamin , Anticevic Alan , Sotiropoulos Stamatios N. , Murray John D.
Published: August 2020

Associations between datasets can be discovered through multivariate methods like Canonical Correlation Analysis (CCA) or Partial Least Squares (PLS). A requisite property for interpretability and generalizability of CCA/PLS solutions is stability of feature patterns driving an association. However, stability of CCA/PLS in high-dimensional datasets is questionable, as found in empirical characterizations. To study these issues in a systematic manner, we developed a generative modeling framework to simulate synthetic datasets, parameterized by dimensionality, variance structure, and association strength. We found that when sample size is relatively small, but comparable to typical studies, CCA/PLS associations are highly unstable and inaccurate; both in their magnitude and importantly in the latent pattern underlying the discovered association. We confirmed these trends across two neuroimaging modalities, functional and diffusion MRI, and in independent datasets, Human Connectome Project (n≈1000) and UK Biobank (n≈20000) and found that only the latter comprised sufficient samples for stable mappings between imaging-derived and behavioral features. We further developed a power calculator to provide sample sizes required for stability and reliability of multivariate analyses for future studies.
The sources of sex differences in aging in annual fishes
Authors: Reichard Martin , Blažek Radim , Žák Jakub , Kačer Petr , Tomášek Oldřich , Albrecht Tomáš , Cellerino Alessandro , Polačik Matej
Published: August 2020

Sex differences in lifespan and aging are widespread among animals, with males usually the shorter-lived sex. Despite extensive research interest, it is unclear how lifespan differences between the sexes are modulated by genetic, environmental and social factors. We combined comparative data from natural populations of annual killifishes with experimental results on replicated captive populations, showing that females consistently outlived males in the wild. This sex-specific survival difference persisted in social environment only in two most aggressive species, and ceased completely when social and physical contacts were prevented. Demographically, neither an earlier start nor faster rate of aging accounted for shorter male lifespans, but increased baseline mortality and the lack of mortality deceleration in the oldest age shortened male lifespan. The sexes did not differ in any measure of functional aging we recorded. Overall, we demonstrate that sex differences in lifespan and aging may be ameliorated by modulating social and environmental conditions.
Integrating barcoded neuroanatomy with spatial transcriptional profiling reveals cadherin correlates of projections shared across the cortex
Authors: Sun Yu-Chi , Chen Xiaoyin , Fischer Stephan , Lu Shaina , Gillis Jesse , Zador Anthony M.
Published: August 2020

Functional circuits consist of neurons with diverse axonal projections and gene expression. Understanding the molecular signature of projections requires high-throughput interrogation of both gene expression and projections to multiple targets in the same cells at cellular resolution, which is difficult to achieve using current technology. Here, we introduce BARseq2, a technique that simultaneously maps projections and detects multiplexed gene expression byin situsequencing. We determined the expression of cadherins and cell-type markers in 29,933 cells, and the projections of 3,164 cells in both the mouse motor cortex and auditory cortex. Associating gene expression and projections in 1,349 neurons revealed shared cadherin signatures of homologous projections across the two cortical areas. These cadherins were enriched across multiple branches of the transcriptomic taxonomy. By correlating multi-gene expression and projections to many targets in single neurons with high throughput, BARseq2 provides a path to uncovering the molecular logic underlying neuronal circuits.
Local and substrate-specific S-palmitoylation determines subcellular localization of Gαo
Authors: Solis Gonzalo P. , Kazemzadeh Arghavan , Abrami Laurence , Valnohova Jana , Alvarez Cecilia , van der Goot F. Gisou , Katanaev Vladimir L.
Published: August 2020

Peripheral membrane proteins (PMPs) associate with cellular membranes through post-translational modifications like S-palmitoylation. The Golgi apparatus is generally viewed as the transitory station where palmitoyl acyltransferases (PATs) modify PMPs, which are then transported to their ultimate destinations such as the plasma membrane (PM). However, little substrate specificity among the many PATs has been determined. Here we describe the inherent partitioning of Gαo – α-subunit of heterotrimeric Go proteins – to PM and Golgi, independent from Golgi-to-PM transport. A minimal code within Gαo N-terminus governs its compartmentalization and re-coding produces G protein versions with shifted localization. We establish the S-palmitoylation at the outer nuclear membrane assay (“SwissKASH”) to probe substrate specificity of PATs in intact cells. With this assay, we show that PATs localizing to different membrane compartments display remarkable substrate selectivity, which is the basis for PMP compartmentalization. Our findings uncover a mechanism governing protein localization and establish the basis for innovative drug discovery.
Which actin genes are necessary for zebrafish heart development and function?
Authors: Prill Kendal , Ojehomon Matiyo , Sandhu Love , Dutta Suchandrima , Dawson John F.
Published: August 2020

Heart failure is the number one cause of mortality in the world, contributed to by cardiovascular disease. Many diseases of the heart muscle are caused by mutations in genes encoding contractile proteins, including cardiac actin mutations. Zebrafish are an advantageous system for modeling cardiac disease since embryos can develop without a functional heart. However, genome duplication in the teleost lineage creates a unique obstacle by increasing the number of genes involved in heart development. Four actin genes are expressed in the zebrafish heart:acta1b; actc1c;and duplicates ofactc1aon chromosome 19 and 20. Here, we characterize the actin genes involved in early zebrafish heart development usingin situhybridization and CRISPR targeting to determine which gene is best to model changes seen in human patients with heart disease. Theactc1aandacta1bgenes are predominant during embryonic heart development, resulting in severe cardiac phenotypes when targeted with CRISPRs. Targeting these two cardiac genes with CRISPRs simultaneously results in a more severe phenotype than their individual counterparts, with the results suggesting compensation for lost actin genes by other actin paralogues. Given the duplication of theactc1agene, we recommendacta1bas the best gene for targeted cardiac actin research.
The SARS-CoV-2 Spike mutation D614G increases entry fitness across a range of ACE2 levels, directly outcompetes the wild type, and is preferentially incorporated into trimers
Authors: Michaud William A. , Boland Genevieve M. , Rabi S. Alireza
Published: August 2020

Early in the current pandemic, the D614G mutation arose in the Spike protein of SARS-CoV-2 and quickly became the dominant variant globally. Mounting evidence suggests D614G enhances viral entry. Here we use a direct competition assay with single-cycle viruses to show that D614G outcompetes the wildtype. We developed a cell line with inducible ACE2 expression to confirm that D614G more efficiently enters cells with ACE2 levels spanning the different primary cells targeted by SARS-CoV-2. Using a new assay for crosslinking and directly extracting Spike trimers from the pseudovirus surface, we found an increase in trimerization efficiency and viral incorporation of D614G protomers. Our findings suggest that D614G increases infection of cells expressing a wide range of ACE2, and informs the mechanism underlying enhanced entry. The tools developed here can be broadly applied to study other Spike variants and SARS-CoV-2 entry, to inform functional studies of viral evolution and vaccine development.
Dual RNAseq highlights the kinetics of skin microbiome and fish host responsiveness to bacterial infection
Authors: Le Luyer J. , Schull Q. , Auffret P. , lopez P. , Crusot M. , Belliard C. , Basset C. , Carradec Q. , Poulain J. , Planes S. , Saulnier D.
Published: August 2020

a)

Background

Tenacibaculum maritimumis a worldwide-distributed fish pathogen known for causing dramatic damages on a broad range of wild and farmed marine fish populations. Recently sequenced genome ofT. maritimumstrain NCIMB 2154Tprovided unprecedented information on the possible molecular mechanisms involved in virulence for this species. However, little is known on the dynamic on the infectionin vivo, and information are lacking on both the intrinsic host response (gene expression) and its associated microbiome community. Here, we applied complementary omic approaches, including dual RNAseq and 16S rRNA gene metabarcoding sequencing using Nanopore and short-reads Illumina technologies to unravel the host-pathogens interplay in experimental infection system using the tropical fishPlatax orbicularisas model.

b)

Results

We show thatT. maritimumtranscriptomic landscape during infection is characterized by an enhancement of antibiotic catalytic and glucan catalytic functions while decreasing specific sulphate assimilation process, compared toin vitrocultures. Simultaneously, fish host display a large palette of immune effectors, notably involving innate response and triggering acute inflammatory response. In addition, results suggest that fish activate adaptive immune response visible through stimulation of T-helper cells, Th17, with congruent reduction of Th2 and T-regulatory cells. Fish were however largely sensitive to infection, and less than 25% of them survived after 96hpi. These surviving fish showed no evidence of stress (cortisol levels) as well as no significant difference in microbiome diversity compared to control at the same sampling time. The presence ofTenacibaculumin resistant fish skin and the total absence of any skin lesion suggest that these fish did not escape contact with the pathogen but rather prevent the pathogen entry. In these individuals we detected the up-regulation of specific immune-related genes differentiating resistant from control at 96hpi, which suggests a possible genomic basis of resistance while no genetic variations in coding regions was reported.

c)

Conclusion

Here we refine the interplay between common fish pathogens and host immune response during experimental infection. We further highlight key actors of defense response, pathogenicity and possible genomic bases of resistance toT. maritimum.
Natural resistance to worms exacerbates bovine tuberculosis severity independently of worm coinfection
Authors: Ezenwa Vanessa O. , Budischak Sarah A. , Buss Peter , Seguel Mauricio , Luikart Gordon , Jolles Anna E. , Sakamoto Kaori
Published: August 2020

Pathogen interactions arising during coinfection can exacerbate disease severity, for example, when the immune response mounted against one pathogen negatively affects defense of another. It is also possible that host immune responses to a pathogen shaped by historical evolutionary interactions between host and pathogen, may modify host immune defenses in ways that have repercussions for other pathogens. In this case, negative interactions between two pathogens could emerge even in the absence of concurrent infection. Parasitic worms and tuberculosis (TB) are involved in one of the most geographically extensive of pathogen interactions, and during coinfection, worms can exacerbate TB disease outcomes. Here, we show that in a wild mammal, natural resistance to worms affects bovine tuberculosis (BTB) severity independently of active worm infection. We found that worm-resistant individuals were more likely to die of BTB than were non-resistant individuals, and their disease progressed more quickly. Anthelmintic treatment moderated, but did not eliminate, the resistance effect, and the effects of resistance and treatment were additive with untreated, resistant individuals experiencing the highest mortality. Interestingly, resistance and anthelmintic treatment had non-overlapping effects on BTB pathology. The effects of resistance manifested in the lungs (the primary site of BTB infection), while the effects of treatment manifested almost entirely in the lymph nodes (the site of disseminated disease), suggesting that resistance and active worm infection affect BTB progression via distinct mechanisms. Our findings reveal that interactions between pathogens can occur as a consequence of processes arising on very different timescales.
Heterologous expression of cryptomaldamide in a cyanobacterial host
Authors: Taton Arnaud , Ecker Andrew , Diaz Brienna , Moss Nathan A. , Anderson Brooke , Reher Raphael , Leão Tiago F. , Simkovsky Ryan , Dorrestein Pieter C. , Gerwick Lena , Gerwick William H. , Golden James W.
Published: August 2020

Filamentous marine cyanobacteria make a variety of bioactive molecules that are produced by polyketide synthases, non-ribosomal peptide synthetases, and hybrid pathways that are encoded by large biosynthetic gene clusters. These cyanobacterial natural products represent potential drugs leads; however, thorough pharmacological investigations have been impeded by the limited quantity of compound that is typically available from the native organisms. Additionally, investigations of the biosynthetic gene clusters and enzymatic pathways have been difficult due to the inability to conduct genetic manipulations in the native producers. Here we report a set of genetic tools for the heterologous expression of biosynthetic gene clusters in the cyanobacteriaSynechococcus elongatusPCC 7942 andAnabaena(Nostoc) PCC 7120. To facilitate the transfer of gene clusters in both strains, we engineered a strain ofAnabaenathat containsS. elongatushomologous sequences for chromosomal recombination at a neutral site and devised a CRISPR-based strategy to efficiently obtain segregated double recombinant clones ofAnabaena. These genetic tools were used to express the large 28.7 kb cryptomaldamide biosynthetic gene cluster from the marine cyanobacteriumMoorena(Moorea)producensJHB in both model strains.S. elongatusdid not produce cryptomaldamide, however high-titer production of cryptomaldamide was obtained inAnabaena. The methods developed in this study will facilitate the heterologous expression of biosynthetic gene clusters isolated from marine cyanobacteria and complex metagenomic samples.

Abstract Figure
Aberrant DJ-1 expression underlies L-type calcium channel hypoactivity in tuberous sclerosis complex and Alzheimer’s disease
Authors: Niere Farr , Uneri Ayse , McArdle Colin J. , Cacheaux Luisa P. , Egido-Betancourt Hailey X. , Namjoshi Sanjeev , Reynoldson Cameron , Penaranda Juan , Wang Xin , Taylor William C. , Craft Suzanne , Keene Christopher Dirk , Ma Tao , Raab-Graham Kimberly F.
Published: August 2020

L-type voltage-dependent Ca2+channels (L-VDCC) integrate synaptic signals to facilitate a plethora of cellular mechanisms. L-VDCC dysfunction is implicated in several neurological and psychiatric diseases. Despite their importance, signals upstream of L-VDCC activity that regulate their channel density, however, are poorly defined. In disease models with overactive mammalian target of rapamycin complex 1 (mTORC1) signaling (or mTORopathies), including tuberous sclerosis (TS) and Alzheimer’s disease (AD), we report a novel mechanism downstream of mTORC1 signaling that results in a deficit in dendritic L-VDCC activity. Deficits in L-VDCC activity are associated with increased expression of the mTORC1-regulated RNA-binding protein DJ-1. DJ-1 binds the mRNA coding the auxiliary Ca2+channel subunit α2δ2 responsible for shuttling L-VDCC to the membrane and represses its expression. Moreover, this novel DJ-1/α2δ2/L-VDCC pathway is disrupted in human AD and preclinical models of AD and TS. Our discovery that DJ-1 directs L-VDCC activity and L-VDCC-associated protein α2δ2 at the synapse suggests that DJ-1/α2δ2/L-VDCC is a common, fundamental pathway disrupted in TS and AD that can be targeted in clinical mTORopathies.

Significance Statement

Many neurological disorders share symptoms, despite disparity among diseases. Treatments are prescribed based on diagnosis rather than individual symptoms. While only treating symptoms may obscure the disease, mechanism-based drug development allows the two approaches to converge. Hub proteins, those that coordinate the expression of proteins that mediate specific cellular functions, may be dysregulated across a broad range of disorders. Herein, we show that the RNA-binding protein DJ-1 controls the activity of L-type voltage-dependent calcium channels (L-VDCC), via the expression of its auxiliary subunit alpha2delta2 (α2δ2). Importantly, we demonstrate that this novel DJ-1/α2δ2/L-VDCC pathway is commonly disrupted among neurological disorders, namely Alzheimer’s disease (AD) and Tuberous Sclerosis (TS). Collectively, these data rationalize mechanism-based drug therapy to treat disease.
Specific decorations of 17-hydroxygeranyllinalool diterpene glycosides solve the autotoxicity problem of chemical defense inNicotiana attenuata
Authors: Heiling Sven , Llorca Lucas Cortes , Li Jiancai , Gase Klaus , Schmidt Axel , Schäfer Martin , Schneider Bernd , Halitschke Rayko , Gaquerel Emmanuel , Baldwin Ian Thomas
Published: August 2020

17-hydroxygeranyllinalool diterpene glycosides (HGL-DTGs) are abundant and potent anti-herbivore defense metabolites inNicotiana attenuatawhose glycosylation and malonylation biosynthetic steps are regulated by jasmonate signaling. To characterize the biosynthetic pathway of HGL-DTGs, we conducted a genome-wide analysis of uridine diphosphate glycosyltransferases (UGTs) and identified 107 members of family-1 UGTs. Tissue-specific time-course transcriptional profiling revealed that the transcripts of three UGTs were highly correlated with two HGL-DTG key biosynthetic genes: geranylgeranyl diphosphate synthase (NaGGPPS)and geranyllinalool synthase (NaGLS). NaGLS’s role in HGL-DTG biosynthesis was confirmed by virus-induced gene-silencing. Silencing the UDP-rhamnosyltransferase,UGT91T1,indicated its role in the rhamnosylation of HGL-DTGs.In vitroenzyme assays revealed that UGT74P3 and UGT74P4 use UDP-glucose for the glucosylation of 17-hydroxygeranyllinalool (17-HGL) to lyciumoside I.UGT74P3andUGT74P5 stablysilenced plants were severely developmentally deformed, suggesting a phytotoxic effect of 17-HGL. Applications of synthetic 17-HGL and silencing of these UGTs in HGL-DTG-free plants confirmed the phytotoxic effect of 17-HGL. Feeding assays withManduca sextalarvae revealed the defensive functions of the glucosylation and rhamnosylation steps in HGL-DTG biosynthesis. Glucosylation is a critical step that contributes to the metabolites’ defensive function and solves the autotoxicity problem of this potent chemical defense.
Low sperm to egg ratio required for successful in vitro fertilisation in a pair-spawning teleost, Senegalese sole (Solea senegalensis)
Authors: Ramos-Júdez Sandra , González-López Wendy Ángela , Ostos Jhons Huayanay , Mamani Noemí Cota , Alemán Carlos Marrero , Beirão José , Duncan Neil
Published: August 2020

Cultured Senegalese sole (Solea senegalensis) breeders fail to spawn fertilised eggs and this bottleneck could be solved with the implementation of large-scalein vitrofertilisation protocols. However, low production of poor-quality sperm has frustrated the development ofin vitrofertilisation protocols. Cultured females were induced to ovulate with a 5 µg kg-1single injection of gonadotropin releasing hormone agonist (GnRHa) and good quality eggs (82.6 ± 9.2% fertilisation) were stripped 41:57 ± 1:46 h after the injection. Sperm was collected from cultured males, diluted in modified Leibovitz and used fresh to fertilise the eggs. A non-linear regression, an exponential rise to a maximum (R = 0.93, P < 0.0001) described the number of motile spermatozoa required to fertilise a viable egg and 1617 motile spermatozoa were sufficient to fertilise 99 ± 12% (± 95% CI) of viable eggs. Similar, spermatozoa egg-1ratios of 592 ± 611 motile spermatozoa egg-1were used in large-scalein vitrofertilisations with 190,512 ± 38,471 eggs. The sperm from a single male (145 ± 50 µL or 8.0 ± 6.8 × 108spermatozoa) was used to fertilise the eggs. The mean hatching rate of the large-scalein vitrofertilisations was 70 ± 14 % to provide 131,540 ± 34,448 larvae per fertilisation. When unfertilised eggs were stored at room temperature the percentage of viable eggs decreased gradually and indicated the sooner eggs were fertilised after stripping the higher the viability of the eggs. The collection of sperm directly into a syringe containing modified Leibovitz significantly increased the percentage of motile spermatozoa (33.4 ± 12.2 %) compared to dilution in modified Leibovitz immediately after collection (6.6 ± 4.9 %). Senegalese sole have a pair-spawning reproductive behaviour characterised by external gamete fertilisation in close proximity with no sperm competition. The low spermatozoa egg-1ratio required for maximum fertilisation was consistent with this reproductive behaviour and strategy. The provision of a large-scalein vitrofertilisation protocol (200 µL of sperm per 100 mL of eggs) will enable the industry to operate sustainably and implement breeding programs to improve production.
Tetanus neurotoxin sensitive SNARE-mediated glial signaling limits motoneuronal excitability
Authors: Böhme Mathias A. , McCarthy Anthony W. , Berezeckaja Monika , Ponimaskin Kristina , Walter Alexander M.
Published: August 2020

Peripheral nerves contain motoneuron axons coated by glial cells, which essentially contribute to function but cellular reactions remain poorly understood. We here identify non-neuronal Synaptobrevin (Syb) as the essential vesicular SNARE in glia to insulate and metabolically supplyDrosophilamotoneurons. Interfering with Syb-functionality by glial knockdown, or glial expression of tetanus neurotoxin light chain (TeNT-LC) caused motonerve disintegration, blocked axonal transport, induced tetanic muscle hyperactivity and caused lethal paralysis. Surprisingly, not the established TeNT-LC-target, neuronal Synaptobrevin (nSyb), is the relevant SNARE, but non-neuronal Synaptobrevin (Syb): Knockdown of Syb- (but not nSyb-) phenocopied glial TeNT-LC expression whose effects were reverted by a TeNT-LC-insensitive Syb mutant. We link Syb-necessity to two distinct glia: to establish nerve insulating septate junctions in subperineurial glia and to integrate monocarboxylate transporters along the nerve in wrapping glia for motoneuronal metabolic supply. Our study identifies crucial roles of Syb in glial subtypes for nerve function and pathology, animal motility and survival.
Evidence of hidden hunger in Darwin's finches as a result of non-native species invasion of the Galapagos cloud forest
Authors: Hood-Nowotny Rebecca , Rabitsch Ingrid , Cimadom Arno , Suarez-Rubio Marcela , Watzinger Andrea , Schmidt Yáñez Paul , Schulze Christian , Zechmeister-Boltenstern Sophie , Jäger Heinke , Tebbich Sabine
Published: August 2020

Invasive species pose a major threat to forest biodiversity, particularly on islands, such as the Galapagos. Here, invasive plants are threatening the remnants of the unique cloud forest and its iconic Darwin's finches. We posit that food web disturbances caused by invasive Rubus niveus (blackberry), but also the management measures used to control it, could contribute to the rapid decline of the insectivourous warbler finch (Certhidae olivacea). We compared changes in long-term management, short-term management and unmanaged areas. We measured C:N ratios, δ15N-nitrogen and δ13C-carbon signatures in bird blood and arthropods, as indicators of resource use change, in addition to mass abundance and diversity of arthropods. We reconstructed the bird's diets using isotope mixing models. The results revealed that finches in (Rubus-invaded) unmanaged areas foraged on abundant yet low quality arthropods and had shorter tarsi. Is this the first evidence of hidden hunger in degraded terrestrial ecosystems in Galapagos?
Antibody-based delivery of Interleukin-9 to neovascular structures: therapeutic evaluation in cancer and arthritis
Authors: Gouyou Baptiste , Ongaro Tiziano , Cazzamalli Samuele , Luca Roberto De , Kerschenmeyer Anne , Valet Philippe , Villa Alessandra , Neri Dario , Matasci Mattia
Published: August 2020

Interleukin-9 (IL9) is a cytokine with multiple functions, including the ability to activate group 2 innate lymphoid cells (ILC2s), which has been postulated to be therapeutically active in mouse models of arthritis. Similarly, IL9 has been suggested to play an important role in tumor immunity. Here, we describe the cloning, expression and characterization of three fusion proteins based on murine IL9 and the F8 antibody, specific to the alternatively-spliced EDA domain of fibronectin. EDA is strongly expressed in cancer and in various arthritic conditions, while being undetectable in the majority of healthy organs. IL9-based fusion proteins with an irrelevant antibody specific to hen egg lysozyme served as negative control in our study. The fusion proteins were characterized by quantitative biodistribution analysis in tumor-bearing mice using radioiodinated protein preparations. The highest tumor uptake and best tumor:organ ratios were observed for a format, in which the IL9 moiety was flanked by two units of the F8 antibody in single-chain Fv format. Biological activity of IL9 was retained when the payload was fused to antibodies. However, the targeted delivery of IL9 to the disease site resulted in a modest anti-tumor activity in three different murine models of cancer (K1735M2, CT26 and F9), while no therapeutic benefit was observed in a collagen induced model of arthritis. Collectively, these results confirm the possibility to deliver IL9 to the site of disease but cast doubts about the alleged therapeutic activity of this cytokine in cancer and arthritis, which has been postulated in previous publications.
Prenatal Stress Dysregulates Resting-State Functional Connectivity and Sensory Motifs
Authors: Rezaei Zahra , Jafari Zahra , Afrashteh Navvab , Torabi Reza , Singh Surjeet , Kolb Bryan E. , Davidsen Jörn , Mohajerani Majid H.
Published: August 2020

Prenatal stress (PS) can impact fetal brain structure and function and contribute to higher vulnerability to neurodevelopmental and neuropsychiatric disorders. To understand how PS alters evoked and spontaneous neocortical activity and intrinsic brain functional connectivity, mesoscale voltage imaging was performed in adult C57BL/6NJ mice that had been exposed to auditory stress on gestational days 12-16, the age at which neocortex is developing. PS mice had a four-fold higher basal corticosterone level and reduced amplitude of cortical sensory-evoked responses to visual, auditory, whisker, forelimb, and hindlimb stimuli. Relative to control animals, PS also led to a general reduction of resting-state functional connectivity, as well as reduced inter-modular connectivity, enhanced intra-modular connectivity, and altered frequency of auditory and forelimb spontaneous sensory motifs. These resting-state changes resulted in a cortical connectivity pattern featuring disjoint but tight modules and a decline in network efficiency. The findings demonstrate that cortical connectivity is sensitive to PS and exposed offspring may be at risk for adult stress-related neuropsychiatric disorders.
Engineering murine GITRL for antibody-mediated delivery to tumor-associated blood vessels
Authors: Mock Jacqueline , Rascon Itzel Astiazaran , Stringhini Marco , Catalano Marco , Neri Dario
Published: August 2020

Preclinical evidence has suggested that the Glucocorticoid-Induced TNFR-related protein (GITR) may be valuable a target for the development of anticancer therapeutics, but clinical studies with GITR ligand (GITRL) have been disappointing. Here, we report the development of a fusion protein featuring GITR ligand (GITRL) fused to the F8 antibody which targets the alternatively-spliced EDA domain of fibronectin, a tumor-associated antigen often found around the tumor neovasculature. Five different formats for F8-GITRL fusion proteins were cloned and characterized, but quantitative biodistribution studies failed to evidence a preferential accumulation at the tumor site. Thein vivotumor targeting properties of F8-GITRL could be substantially improved by enzymatic deglycosylation or site-directed mutagenesis of theN-glycosylation consensus sequence. However, therapy studies in a murine model of cancer with the glycoengineered F8-GITRL N74S and N157T variant failed to elicit a durable anti-tumor response, both in monotherapy and in combination with PD-1 blockade.

HIGHLIGHTS

Different formats of fusion proteins featuring Glucocorticoid-induced TNFR-related protein ligand (GITRL) fused to a tumor-targeting antibody were produced.

The tumor uptake of the fusion proteins could be increased by enzymatic deglycosylation of the fusion protein or by site-directed mutagenesis of theN-glycosylation consensus sequences.

The fusion protein developed in this study failed to show any anti-tumor activity either alone or in combination with PD-1 inhibition.
Large-Scale Transcriptional Profiling of Molecular Perturbations Reveals Cell Type Specific Responses and Implications for Environmental Screening
Authors: Zhang Kun , Zhao Yanbin
Published: August 2020

Cell-based assays represent nearly half of all high-throughput screens currently conducted for risk assessment of environmental chemicals. However, the sensitivity and heterogeneity among cell lines has long been concerned but explored only in a limited manner. Here, we address this question by conducting a large scale transcriptomic analysis of the responses of discrete cell lines to specific small molecules. Our results illustrate heterogeneity of the extent and timing of responses among cell lines. Interestingly, high sensitivity and/or heterogeneity was found to be cell type-specific or universal depending on the different mechanism of actions of the compounds. Our data provide a novel insight into the understanding of cell-small molecule interactions and have substantial implications for the design, execution and interpretation of high-throughput screening assays.
Corticosteroids and cellulose purification improve respectively the in vivo translation and vaccination efficacy of self-amplifying mRNAs
Authors: Zhong Zifu , Cafferty Séan Mc , Opsomer Lisa , Wang Haixiu , Huysmans Hanne , De Temmerman Joyca , Lienenklaus Stefan , Portela Catani João Paulo , Combes Francis , Sanders Niek N.
Published: August 2020

Synthetic mRNAs are an appealing therapeutic platform with multiple biomedical applications ranging from protein replacement therapy to vaccination. In comparison to conventional mRNA, synthetic self-amplifying mRNAs (sa-mRNAs) are gaining increased interest due to their higher and longer-lasting expression. However, sa-mRNAs also elicit an innate immune response, which may complicate the clinical translation of this platform. Approaches to reduce the innate immunity of sa-mRNAs have not been studied in detail. In this work we investigated the effect of several innate immune inhibitors and a novel cellulose-based mRNA purification approach on the type I interferon (IFN) response, translation and vaccination efficacy of our formerly developed sa-mRNA vaccine against Zika virus. Among the investigated inhibitors, we found that topical application of clobetasol at the sa-mRNA injection site was the most efficient in suppressing the type I IFN response and increasing the translation of sa-mRNA. However, clobetasol prevented the formation of antibodies against sa-mRNA encoded antigens and should therefore be avoided in a vaccination context. Residual dsRNA by-products of the in vitro transcription reaction are known inducers of immediate type I IFN responses. We additionally demonstrate drastic reduction of these dsRNA by-products upon cellulose-based purification, consequently reducing the innate immune response and improving sa-mRNA vaccination efficacy.