bluuebunny/crossref_metadata_2025 huggingface dataset:
Dismantling cognitive-behavioral therapy for chronic insomnia: a protocol for a systematic review and component network meta-analysis
Authors: Furukawa Yuki , Sakata Masatsugu , Funada Satoshi , Kikuchi Shino , Furukawa Toshi A. , Ostinelli Edoardo G. , Efthimiou Orestis , Perlis Michael
Published: June 2022

Introduction

Insomnia is highly prevalent and disabling. Clinical practice guidelines recommend cognitive-behavioral therapy for insomnia (CBTI) as the first-line treatment. However, CBTI includes various combinations of many different components and its clinical benefits have been shown as a package, whereas the effect of each component remains unclear. In this study, we will explore the effect of each component of CBTI with the use of component network meta-analysis.

Methods and analysis

We will include all randomized controlled trials that compared any form of CBTI against another form of CBTI or a control condition in the treatment of adults with chronic insomnia. Concomitant treatments will be allowed as long as they are equally distributed among the arms. We will include both primary and secondary insomnia. The primary outcome of interest in this study is (1) treatment efficacy (remission defined as reaching a satisfactory state at endpoint measured by any validated self-reported scale) at four weeks post-treatment or at its closest time point. Secondary outcomes are (2) acceptability, (3) sleep diary measures and (4) efficacy at long-term follow-up. We will systematically search in PubMed, CENTRAL, PsycINFO and WHO International Clinical Trials Registry Platform. We will assess risk of bias using Cochrane Risk of Bias 2.0 tool. We will conduct component network meta-analysis with the netmeta package in R.

Ethics and dissemination

This study will use published data and does not require ethical approval. Findings will be disseminated in a peer-reviewed journal.

PROSPERO

CRD42022324233.
Comparative analysis of capture methods for genomic profiling of circulating tumor cells in colorectal cancer
Authors: Alves Joao M. , Estévez-Gómez Nuria , Valecha Monica , Prado-López Sonia , Tomás Laura , Alvariño Pilar , Piñeiro Roberto , Muinelo-Romay Laura , Mondelo-Macía Patricia , Salgado Mercedes , Iglesias-Gómez Agueda , Codesido-Prada Laura , Cubiella Joaquin , Posada David
Published: June 2022

The genomic profiling of circulating tumor cells (CTCs) in the bloodstream should provide clinically relevant information on therapeutic efficacy and help predict cancer survival. However, the molecular characterization of CTCs has so far proven extremely difficult. A variety of technologies have been developed for CTC isolation, but so far the impact on the genomic assessment of CTCs has not been fully evaluated. To fill this gap, here we contrasted the genomic profiles of CTC pools recovered from blood samples obtained from four metastatic colorectal cancer (mCRC) patients using three different enrichment strategies (CellSearch, Parsortix, and FACS). Our results suggest clear differences in the mutational burden of CTC pools depending on the enrichment method used, with all evaluated methods returning a somewhat limited representation of the mutational spectrum of individual tumors, potentially due to allelic dropout during whole-genome amplification. Nevertheless, the CTC pools from Parsortix, and in part, CellSearch, showed diversity estimates, mutational signatures and drug-suitability scores remarkably close to the ones found in matching primary tumor samples. In contrast, FACS CTC pools were substantially enriched in apparent sequencing artifacts, which led to much higher estimates of genomic diversity. Although CTC genomics still faces technical challenges, our results suggest that CTC-derived metrics can reflect the diversity scores seen in primary tumor lesions thus highlighting the utility of CTCs to assess the heterogeneity status of individual tumors, and to help clinicians prioritize drugs in mCRC.
Delays in COVID-19 Diagnosis and Hospitalization and Outcomes — New York City, New York, USA, October 2020–November 2021
Authors: Graf Laura E. , Peterson Eric R. , Baumgartner Jennifer , Fine Anne , Thompson Corinne N. , Blaney Kathleen , Greene Sharon K.
Published: June 2022

COVID-19 patients diagnosed ≥3 days after symptom onset had increased odds of hospitalization. The 75th percentile for diagnosis delay was 5 days for residents of low-privilege areas and Black and Hispanic people diagnosed before SARS-CoV-2 Delta predominance, compared with 4 days for other patients, indicating inequities in prompt diagnosis.
Glucagon/GLP-1 receptor co-agonist NNC9204-1177 reduced body weight in adults with overweight or obesity but was associated with safety issues
Authors: Friedrichsen Martin , Endahl Lars , Kreiner Frederik Flindt , Goldwater Ronald , Kankam Martin , Toubro Søren , Nygård Sune Boris
Published: June 2022

Glucagon/glucagon-like peptide-1 (GLP-1) receptor co-agonists may provide greater weight loss than agonists targeting the GLP-1 receptor alone. We report results from three phase 1 trials investigating the glucagon/GLP-1 receptor co-agonist NNC9204-1177 (NN1177) for once-weekly subcutaneous use in adults with overweight or obesity.

Our focus was a 12–week multiple ascending dose (MAD), placebo-controlled, double-blind trial in which adults (N=99) received NN1177 (dose-escalated to treatment doses of 200, 600, 1,300, 1,900, 2,800, 4,200, and 6,000 μg) or placebo. Two other trials also contribute to the findings in this report: a first human dose (FHD) / single ascending dose (SAD), placebo-controlled, double-blind trial in which adults (N=49) received NN1177 (treatment doses of 10, 40, 120, 350, 700 and 1,100 μg) or placebo, and a drug–drug interaction (DDI), open-label, single-sequence trial in which adults (N=45) received a 4,200 μg dose of NN1177. Pharmacokinetic, safety and tolerability, and pharmacodynamic endpoints were assessed.

For the MAD and FHD/SAD trials, baseline characteristics were generally balanced across groups. The half-life of NN1177 was estimated at between 77.3 and 111 hours. NN1177 appeared tolerable across trials; however, a number of safety concerns were observed, including an increase in heart rate (range 5–22 beats per minute) and decrease in reticulocyte count, which were both dose dependent, and increased markers of inflammation (fibrinogen and C-reactive protein), hepatic disturbances (increased aspartate aminotransferase and alanine aminotransferase), impaired glucose tolerance (dose groups 2,800–6,000 ug) and reduced blood levels of some amino acids. Clinically relevant weight loss was achieved (up to 12.6% at week 12; 4,200 ug in the MAD trial), but this was not accompanied by cardiometabolic improvements.

In conclusion, although treatment with NN1177 was associated with dose-dependent and clinically relevant weight loss, unacceptable safety concerns precluded further clinical development.
Outcomes Associated with Rural Emergency Department Provider-to-Provider Telehealth for Sepsis Care: A Multicenter Cohort Study
Authors: Mohr Nicholas M. , Okoro Uche , Harland Karisa K. , Fuller Brian M. , Campbell Kalyn , Swanson Morgan B. , Wymore Cole , Faine Brett , Zepeski Anne , Parker Edith A. , Mack Luke , Bell Amanda , DeJong Katie , Mueller Keith , Chrischilles Elizabeth , Carpenter Christopher R. , Wallace Kelli , Jones Michael P. , Ward Marcia M.
Published: June 2022

Objective

To test the hypothesis that provider-to-provider tele-emergency department (tele-ED) care is associated with more 28-day hospital-free days and improved Surviving Sepsis Campaign (SSC) guideline adherence in rural emergency departments (EDs).

Methods

Multicenter (n=23), propensity-matched, cohort study using medical records of sepsis patients from rural hospitals in a well-established, on-demand, rural video tele-ED network in the upper Midwest between August 2016 and June 2019. The primary outcome was 28-day hospital-free days, with secondary outcomes of 28-day in-hospital mortality and SSC guideline adherence.

Results

A total of 1,191 patients were included in the analysis, with tele-ED used for 326 (27%). Tele-ED cases were more likely to be transferred to another hospital (88% vs. 8%, difference 79%, 95% CI 75–83%). After matching and regression adjustment, tele-ED cases did not have more 28-day hospital-free days (difference 0.80 days longer for non-tele-ED, 95% confidence interval [CI] [-0.87]–2.47) or 28-day in-hospital mortality (adjusted odds ratio [aOR] 1.61, 95% CI 0.74-3.57). Adherence with both the SSC 3-hour bundle (aOR 0.80, 95% CI 0.24– 2.70) and complete bundle (aOR 0.81, 95% CI 0.15–4.41) were similar. An a priori–defined subgroup analysis of patients treated by advanced practice providers suggested that the mortality was lower in the cohort with tele-ED use (aOR 0.19, 95% CI 0.04–0.90) despite no significant difference in complete SSC bundle adherence (aOR 2.48, 95% CI 0.45–13.76).

Conclusions

Rural ED patients treated with provider-to-provider tele-ED care in a mature network appear to have similar clinical outcomes to those treated without.
Data Source Concordance for Infectious Disease Epidemiology
Authors: Majumder Maimuna , Cusick Marika Mae , Rose Sherri
Published: June 2022

Background

As highlighted by the COVID-19 pandemic, researchers are eager to make use of a wide variety of data sources, both government-sponsored and alternative, to characterize the epidemiology of infectious diseases. To date, few studies have investigated the strengths and limitations of sources currently being used for such research. These are critical for policy makers to understand when interpreting study findings.

Methods

To fill this gap in the literature, we compared infectious disease reporting for three diseases (measles, mumps, and varicella) across four different data sources: Optum (health insurance billing claims data), HealthMap (online news surveillance data), Morbidity and Mortality Weekly Reports (official government reports), and National Notifiable Disease Surveillance System (government case surveillance data). We reported the yearly number of national- and state-level disease-specific case counts and disease clusters according to each of our sources during a five-year study period (2013–2017).

Findings

Our study demonstrated drastic differences in reported infectious disease incidence across data sources. When compared against the other three sources of interest, Optum data showed substantially higher, implausible standardized case counts for all three diseases. Although there was some concordance in identified state-level case counts and disease clusters, all four sources identified variations in state-level reporting.

Interpretation

Researchers should consider data source limitations when attempting to characterize the epidemiology of infectious diseases. Some data sources, such as billing claims data, may be unsuitable for epidemiological research within the infectious disease context.
Gendered time, financial & nutritional benefits from access to pay-as-you-go LPG for cooking in an informal settlement in Nairobi, Kenya
Authors: Shupler Matthew , Karl Jonathan , O’Keefe Mark , Osiolo Helen Hoka , Perros Tash , Simiyu Willah Nabukwangwa , Gohole Arthur , Lorenzetti Federico , Puzzolo Elisa , Mwitari James , Pope Daniel , Nix Emily
Published: June 2022

Introduction

Few studies have examined gendered benefits of transitioning from polluting cooking fuels (e.g. charcoal, kerosene) to cleaner fuels (e.g. liquefied petroleum gas (LPG)). This study investigates pathways between adoption of pay-as-you-go (PAYG) LPG and women’s empowerment in Nairobi, Kenya.

Methods

Female (N=304) and male (N=44) primary cooks in an informal settlement in Nairobi were surveyed from December 2021-January 2022. The majority (84%; N=293) were customers of PayGo Energy, a company offering PAYG LPG. Other individuals (16%; N=55) cooking with full cylinder LPG or polluting fuels were randomly sampled from the community. The 45-minute telephonic survey examined how access to PAYG LPG affected the livelihoods of PayGo Energy’s customers.

Results

PayGo Energy customers were 50% more likely to cook exclusively with LPG (60%) than those using full cylinder LPG (40%). Due to reduced cooking times (average reduction: 42 min/day among previous polluting fuel users) from the adoption of PAYG LPG, the majority (58%; N=70) of female household heads took on additional employment compared with 36% (N=55) of females living in male-headed households. A greater proportion of married female household heads used their monetary savings from cooking with PAYG LPG for investment (41%) or savings (35%), compared with married women that were not household heads (3% and 21%, respectively). Increased dietary diversity and consumption of protein-rich foods (legumes, meat, fish) from cooking with PAYG LPG was reported by 15% of female household heads.

Conclusion

Female household heads were more likely than non-household heads to experience economic and nutritional gains when adopting PAYG LPG, illustrating how the agency of women influences their social co-benefits when undergoing clean energy transitions.
Less is more in language production: Shorter sentences contain more informative words
Authors: Rezaii Neguine , Ren Boyu , Quimby Megan , Hochberg Daisy , Dickerson Bradford
Published: June 2022

Agrammatism is characterized by short sentences, the omission of function words, a higher ratio of heavy to light verbs, and a decreased use of verbs relative to nouns. Despite the observation of these phenomena more than two centuries ago, there has been no unifying theory to explain all features of agrammatism. Here, by first examining the language of patients with primary progressive aphasia, we show that the seemingly heterogeneous features of agrammatism can be explained by a process that selects lower frequency words over their higher frequency alternatives in the context of a limitation in sentence production, likely to increase the informational content of sentences. We further show that when healthy speakers are constrained to produce short sentences, features of agrammatism emerge in their language. Finally, we show that these findings instantiate a general property in healthy language production in which shorter sentences are constructed by selecting lower frequency words.
Association between mild traumatic brain injury, brain structure, and mental health outcomes in the Adolescent Brain Cognitive Development Study
Authors: Lopez Daniel A. , Christensen Zachary P. , Foxe John J. , Ziemer Laura R. , Nicklas Paige R. , Freedman Edward G.
Published: June 2022

Background

Children that experience a mild traumatic brain injury (mTBI) are at an increased risk of neural alterations that can deteriorate mental health. We test the hypothesis that mTBI is associated with behavioral and emotional problems and that structural brain metrics (e.g., volume, area) meaningfully mediate the relation in an adolescent population.

Methods

We analyzed behavioral and brain MRI data from 11,876 children who participated in the Adolescent Brain Cognitive Development (ABCD) Study. Mixed-effects models were used to examine the longitudinal association between mTBI and mental health outcomes. Bayesian methods were used to investigate brain regions that are intermediate between mTBI and symptoms of poor mental health.

Results

There were 199 children with mTBI and 527 with possible mTBI across the three ABCD Study visits. There was a 7% (IRR = 1.07, 95% CI: 1.01, 1.13) and 15% (IRR = 1.16, 95% CI: 1.05, 1.26) increased risk of emotional or behavioral problems in children that experienced possible mTBI or mTBI, respectively. Possible mTBI was associated with a 17% (IRR: 1.17, 95% CI: 0.99, 1.40) increased risk of experiencing distress following a psychotic-like experience. We did not find any brain regions that meaningfully mediated the relationship between mTBI and mental health outcomes. Analysis of volumetric measures found that 3 to 5% of the total effect of mTBI on mental health outcomes operated through total cortical volume. Image intensity measure analyses determined that 2 to 5% of the total effect was mediated through the left-hemisphere of the dorsolateral prefrontal cortex.

Conclusion

Results indicate an increased risk of emotional and behavioral problems in children that experienced possible mTBI or mTBI. Mediation analyses did not elucidate the mechanisms underlying the association between mTBI and mental health outcomes.
A recurrent de novo splice site variant involving DNM1 exon 10a causes developmental and epileptic encephalopathy through a dominant-negative mechanism
Authors: Parthasarathy Shridhar , Ruggiero Sarah M , Gelot Antoinette , Soardi Fernanda C , Ribeiro Bethânia F R , Pires Douglas E V , Ascher David B , Schmitt Alain , Rambaud Caroline , Xie Hongbo M , Lusk Laina , Wilmarth Olivia , McDonnell Pamela Pojomovsky , Juarez Olivia A , Grace Alexandra N , Buratti Julien , Mignot Cyril , Gras Domitille , Nava Caroline , Pierce Samuel R , Keren Boris , Kennedy Benjamin C , Pena Sergio D J , Helbig Ingo , Cuddapah Vishnu Anand
Published: June 2022

Heterozygous pathogenic variants in DNM1 cause developmental and epileptic encephalopathy (DEE) due to a dominant-negative mechanism impeding vesicular fission. Thus far, pathogenic variants in DNM1 have been studied using a canonical transcript that includes the alternatively spliced exon 10b. However, after performing RNA sequencing in thirty-nine pediatric brain samples, we find the primary transcript expressed in the brain includes the downstream exon 10a instead. Using this information, we evaluated genotype-phenotype correlations of variants affecting exon 10a and identified a cohort of eleven previously unreported individuals. Eight individuals harbor a recurrent de novo splice site variant, NG_029726.1(NM_001288739.1):c.1197-8G>A, which affects exon 10a and leads to DEE consistent with the classical DNM1 phenotype. We find this splice site variant leads to disease through an unexpected dominant-negative mechanism. Functional testing reveals an in-frame upstream splice acceptor causing insertion of two amino acids predicted to impair oligomerization-dependent activity. This is supported by neuropathological samples showing accumulation of synaptic vesicles adherent to the plasma membrane consistent with impaired vesicular fission. Two additional individuals with missense variants affecting exon 10a, p.(Arg399Trp) and p.(Gly401Asp), had a similar DEE phenotype. In contrast, a single individual with a missense variant affecting exon 10b, p.(Pro405Leu), which is less expressed in the brain, had a correspondingly less severe presentation. Thus, we implicate variants affecting exon 10a as causing the severe DEE typically associated with DNM1-related disorders. We highlight the importance of considering relevant isoforms for disease-causing variants, as well as the possibility of splice site variants acting through a dominant-negative mechanism.
Transcriptional activity is shaped by the chromatin landscapes in Arabidopsis
Authors: Jamge Bhagyshree , Lorković Zdravko J. , Axelsson Elin , Yelagandula Ramesh , Akimcheva Svetlana , Berger Frédéric
Published: June 2022

Summary

How histone variants and histone modifications shape nucleosome-mediated transcriptional repression, and how transcriptional activity shapes the enrichment of histone modifications and variants remain unclear. To clarify these relationships, we examined chromatin organization in the Arabidopsis thaliana genome, identifying a limited number of chromatin landscapes that distinguish transposon families and distinct groups of genes based on their transcriptional regulation. Unexpectedly, H2A variants are strong determinants of the landscape architecture. Combinations of H2A.W and four histone modifications define six domains that are occupied by specific transposon families and organized concentrically around the centromere. Moreover, H2A.Z defines transcriptional gene repression in specific domains. Expressed genes occupy four chromatin landscapes with specific RNA Polymerase II profiles. Although the composition of each chromatin landscape is invariant, they cover genes with a wide range of expression levels. Therefore, chromatin landscapes control the range of transcriptional activity, but transcriptional activity has little effect on chromatin composition.

One Sentence Summary

Histone variants and histone modifications build a limited number of distinct chromatin landscapes that instruct the transcriptional regulation of genes and transposons in Arabidopsis.
Research Productivity and Training Support for Doctoral Students in the Biological and Biomedical Sciences
Authors: Schaller Michael D.
Published: June 2022

Training of doctoral students as part of the next generation of the biomedical workforce is essential for sustaining the scientific enterprise in the US. Training of doctoral students primarily occurs at institutions of higher education and these trainees comprise an important part of the workforce at these institutions. Federal investment in the support of doctoral students in the biological and biomedical sciences is distributed differently than the distribution of students across different types of institutions, e.g. Public vs Private. Institutions in states that historically receive less federal support for research also receive less support for doctoral student training. Analysis of F31 awardees at different types of institution reveal little difference in research productivity and subsequent receipt of additional NIH awards. Analysis of a large cohort of doctoral students at different types of institutions also reveal little difference in research productivity, with the exception of citations. Thus, training outcomes, which are related to the quality of the student and training environment, are similar across different institutions. Factors that correlate with F31 funding include R01 funding levels and program size. The findings suggest strategies for institutions to increase success at securing F31s and modification of policy to promote more equitable distribution of F31s across institutions.
A Genetically Encoded Far-Red Fluorescent Indicator for Imaging Synaptically-Released Zn2+
Authors: Wu Tianchen , Kumar Manoj , Zhao Shengyu , Drobizhev Mikhail , Tian Xiaodong , Tzounopoulos Thanos , Ai Hui-wang
Published: June 2022

Synaptic Zn2+ has emerged as a key neuromodulator in the brain. However, the lack of research tools for directly tracking synaptic Zn2+ in the brain in live animals hinders our rigorous understanding of the physiological and pathological roles of synaptic Zn2+. In this study, we developed a genetically encoded far-red fluorescent indicator for monitoring synaptic Zn2+ dynamics in the nervous system. Our engineered far-red fluorescent indicator for synaptic Zn2+ (FRISZ) displayed a substantial Zn2+-specific turn-on response and low micromolar affinity. We genetically anchored FRISZ to the mammalian extracellular membrane via a transmembrane α-helix. We further successfully used membrane-tethered FRISZ (FRISZ-TM) to image synaptic Zn2+ dynamics in response to sound in the primary auditory cortex (A1) in awake mice. This study thus establishes a new technology for studying the roles of synaptic Zn2+ in the nervous system.
“The anti-apoptotic effect ofLonomia obliquahemolymph is associated with the mitochondria pathway”
Authors: Mendonça Ronaldo Z. , Martins Luciana Moreira
Published: June 2022

The apoptosis death is a very important factors in production processes that limited the industrial production of some proteins of economic interest. However, one of the forms to increase the cellular productivity would be inhibit or attenuate the cellular death. Recently we have demonstrated the presence of a potent anti-apoptotic protein inLonomia obliquahemolymph which extends the cell culture viability through apoptosis prevention. By the other side, has been reported that mitochondria have one important action in the apoptosis control process, being that mitochondria membrane permeabilization (MMP) can be an important stage in this process. MMP associated or not with the loss of the electrochemical potential of the mitochondria and alteration of the matrix is responsible for the inter membranes protein release (e.g. cytochrome c, the AIF, etc) of cytosol. The result obtained showed that the addition of a protein fromLonomia obliquahemolymph in the culture lead to a prolongation of the cellular life (3-4 days) and the cells leading a high electrochemical potential of the mitochondria. This protein can has its action in mitochondria membrane, avoiding the loss of the membrane permeability and the Cytochrome-C release. As positive control, apoptosis death in these cultures was induced by 50 μm of t-BHP or 600 μm of H202. The presence of apoptosis was characterized by flow citometry, microscopy electronic and agarose gel electrophoresis. The potential electrochemical of the mitochondria was determined by JC-1, Hoechst 33324 and DIOC6. Cytochrome C was identified in cytosol by an anti-cytochrome antibody.
Chemical screening of food-related chemicals for human fatty liver risk: Combining high content imaging of cellular responses with in vitro to in vivo extrapolation
Authors: Müller Fabrice A. , Stamou Marianna , Englert Felix , Frenzel Ole , Diedrich Sabine , Wambaugh John F. , Sturla Shana J.
Published: June 2022

1

Nonalcoholic fatty liver disease (NAFLD) is an increasingly prevalent human disease with accumulating evidence linking its pathophysiology and co-morbidities to chemical exposures. The complex pathophysiology of NAFLD has limited the elucidation of potential chemical etiologies. In this study we generated a high-content imaging analysis method for the simultaneous quantification of sentinel steatosis cellular markers in chemically exposed human liver cells in vitro combined with a computational model for the extrapolation of human oral equivalent doses (OED). First, the in vitro test method was generated using 14 reference chemicals with known capacities to induce cellular alterations in nuclear morphology, lipid accumulation, mitochondrial membrane potential and oxidative stress. These effects were quantified on a single cell- and population-level, and then, using physiologically based pharmacokinetic modelling and reverse dosimetry, OEDs were extrapolated from these in vitro data. The extrapolated OEDs were confirmed to be within biologically relevant exposure ranges for the reference chemicals. Next, we tested 14 chemicals found in food, selected from thousands of putative chemicals on the basis of structure-based prediction for nuclear receptor activation. Amongst these, orotic acid had an extrapolated OED overlapping with realistic exposure ranges. By the strategy developed in this study, we were able to characterize known NAFLD-inducing chemicals and translate to data scarce food-related chemicals, amongst which we identified orotic acid to induce steatosis. This strategy addresses needs of next generation risk assessment, and can be used as a first chemical prioritization hazard screening step in a tiered approach to identify chemical risk factors for NAFLD.
Calcium-Mediated Modulation of Blood-Brain Barrier Permeability by Laser Stimulation of Endothelial-Targeted Nanoparticles
Authors: Li Xiaoqing , Cai Qi , Wilson Blake A. , Fan Hanwen , Giannotta Monica , Bachoo Robert , Qin Zhenpeng
Published: June 2022

The blood-brain barrier (BBB) maintains an optimal environment for brain homeostasis but excludes most therapeutics from entering the brain. Strategies that can reversibly increase BBB permeability will be essential for treating brain diseases and is the focus of significant preclinical and translational interest. Recently, we reported that picosecond laser excitation of molecular-targeted gold nanoparticles (AuNPs) induced a graded and reversible increase in BBB permeabilityin vivo(OptoBBB). Here we investigate how to increase the targeting efficiency and how picosecond lase stimulation of AuNP leads to an increase in endothelial paracellular permeability. Our results suggest that targeting brain endothelial glycoproteins leads to >20-fold higher targeting efficiency compared with tight junction targeting. We report that OptoBBB is associated with a transient elevation of Ca2+that propagates to adjacent endothelial cells after laser excitation and extends the region of BBB opening. The Ca2+response involves both internal Ca2+depletion and Ca2+influx. Furthermore, we demonstrate that the involvement of actin polymerization and Ca2+-dependent phosphorylation of ERK1/2 lead to cytoskeletal activation, increasing paracellular permeability. Taken together, we provide mechanistic insight into how excitation of endothelial targeted AuNPs leads to an increase in BBB permeability. These insights will be critical for guiding the future developments of this technology for brain disease treatment.
The developmental progression of eight opsin spectral signals recorded from the zebrafish retinal cone layer is altered by the timing and cell type expression of thyroxin receptor β2 (trβ2) gain-of-function transgenes
Authors: Nelson Ralph F. , Balraj Annika , Suresh Tara , Elias Leah J. , Yoshimatsu Takeshi , Patterson Sara S.
Published: June 2022

Zebrafish retinal cone signals shift in spectral shape through larval, juvenile, and adult development as expression patterns of eight cone-opsin genes change. An algorithm extracting signal amplitudes for the component cone spectral types is developed and tested on two thyroxin receptor β2 (trβ2) gain-of-function lines crx:mYFP-2A-trβ2 and gnat2:mYFP-2A-trβ2, allowing correlation between opsin signaling and opsin immunoreactivity in lines with different developmental timing and cell-type expression of this red-opsin-promoting transgene. Both adult transgenics became complete, or nearly complete, ‘red-cone dichromats’, with disproportionately large LWS1 opsin amplitudes as compared to controls, where LWS1 and LWS2 amplitudes were about equal, and significant signals from SWS1, SWS2, and Rh2 opsins were detected. But in transgenic larvae and juveniles of both lines it was LWS2 amplitudes that increased, with LWS1 cone signals rarely encountered. In gnat2:mYFP-2A-trβ2 embryos at 5 days post fertilization (dpf), red-opsin immunoreactive cone density doubled, but red-opsin amplitudes (LWS2) increased < 10%, and green, blue and UV opsin signals were unchanged, despite co-expressed red opsins, and the finding that an sws1 UV-opsin reporter gene was shut down by the gnat2:mYFP-2A-trβ2 transgene. By contrast both LWS2 red-cone amplitudes and the density of red-cone immunoreactivity more than doubled in 5 dpf crx:mYFP-2A-trß2 embryos, while UV-cone amplitudes were reduced 90%. Embryonic cones with trβ2 gain-of-function transgenes were morphologically distinct from control red, blue or UV cones, with wider inner segments and shorter axons than red cones, suggesting cone spectral specification, opsin immunoreactivity and shape are influenced by the abundance and developmental timing of trβ2 expression.

Significance Statement

As different combinations of eight cone-opsin mRNAs are successively expressed during zebrafish development and maturation, the composite cone-ERG spectral signal shifts. Amplitudes of each of the eight resulting cone signals are inferred computationally from the composite signal, both in controls and in two thyroxin-receptor β2 (trβ2) gain-of-function transgenics, crx:mYFP-2A-trβ2 and gnat2:mYFP-2A-trβ2, trβ2 being a transcription factor required for expression of the red-cone opsins LWS1 and LWS2. Adult transgenics become red cone dichromats with excess LWS1 amplitudes, but larvae and juveniles evoke excess LWS2 amplitudes. Controls retain 5 to 6 cone signals of changing composition throughout development. The progression of transgene-induced amplitude alterations is slower in gnat2:mYFP-2A-trβ2, with supernormal red-opsin antigenicity not immediately correlating with red-cone signaling.
Stable and ancient endocytic structures navigate the complex pellicle of apicomplexan parasites
Authors: Koreny Ludek , Mercado-Saavedra Brandon N. , Klinger Christen M. , Barylyuk Konstantin , Butterworth Simon , Hirst Jennifer , Rivera-Cuevas Yolanda , Zaccai Nathan R. , Holzer Victoria J. C. , Klingl Andreas , Dacks Joel B. , Carruthers Vern B. , Robinson Margaret S. , Gras Simon , Waller Ross F.
Published: June 2022

Apicomplexan parasites have an immense impact on humanity, but their basic cellular processes are often poorly understood. The sites of endocytosis, the conservation of this process with other eukaryotes, and its functions across Apicomplexa are major unanswered questions. Yet endocytosis inPlasmodiumis implicated in antimalarial drug failure. Using the apicomplexan modelToxoplasma, we identified the molecular composition and behavior of unusual, fixed endocytic structures. Here, stable complexes of endocytic proteins differ markedly from the dynamic assembly/disassembly of these machineries in other eukaryotes. Moreover, conserved molecular adaptation of this structure is seen in Apicomplexa, including the kelch-domain protein K13 central to malarial drug-resistance. We determine that an essential function of endocytosis inToxoplasmais plasma membrane homeostasis, rather than parasite nutrition, and that these specialized endocytic structures originated early in infrakingdom Alveolata, likely in response to the complex cell pellicle that defines this medically and ecologically important ancient eukaryotic lineage.
Topical Delivery of Elastic Liposomal Vesicles for Treatment of Middle and Inner Ear Disease
Authors: Sadabad Raana Kashfi , Xia Anping , Benkafadar Nesrine , Faniku Chrysovalantou , Preciado Diego , Yang Stella , Valdez Tulio A.
Published: June 2022

We present a topical drug delivery mechanism through the ear canal to the middle and inner ear using liposomal nanoparticles without disrupting the integrity of the tympanic membrane. The current delivery method provides a non-invasive,and safer alternative to trans-tympanic membrane injections, ear tubes followed by ear drops administration and systemic drug formulations. We investigate the capability of liposomal NPs, particularly transfersomes (TLipo), used as drug delivery vesicles to penetrate the tympanic membrane (TM) and round window membrane (RWM) with high affinity, specificity, and retention time. The TLipo is applied to the ear canal and found to pass through tympanic membrane quickly in 3 hours post drug administration. They are identified in the middle ear cavity 6 hours and in the inner ear 24 hours after drug administration. We performed cytotoxicity in vitro and ototoxicity in vivo studies. Cell viability shows no significant difference between the applied TLipo concentration and control. Furthermore, auditory brainstem response (ABR) reveals no hearing loss in 1 week and 1 month post administration. Immunohistochemistry results demonstrates no evidence of hair cell loss in the cochlea at 1 month following TLipo administration. Together, the data suggested that TLipo can be used as a vehicle for topical drug delivery to the middle ear and inner ear.
Novel insights into the fundamentals of palatal shelf elevation dynamics in normal mouse embryos
Authors: Goering Jeremy P. , Moedritzer Michael , Stetsiv Marta , Isai Dona Greta , Hufft-Martinez Brittany M. , Umar Zaid , Rickabaugh Madison K. , Keselman Paul , Chauhan Munish , Brooks William M. , Fischer Kenneth J. , Czirok Andras , Saadi Irfan
Published: June 2022

Embryonic palate development involves bilateral vertical growth of palatal shelves – extensions from the maxillary processes – next to the tongue until embryonic day (E) 13.5. Following vertical growth, palatal shelves elevate and adhere above the tongue by E14.5. Current models indicate that this process of elevation involves a complex vertical to horizontal reorienting of the palatal shelves. While earlier studies have implied that this is a rapid process, the precise timing has not been resolved. To understand the dynamics of palatal shelf elevation, we employed time-restricted pregnancies with a one-hour resolution and magnetic resonance imaging of intermediate stages. Our data showed that in almost all C57BL/6J embryos, palatal shelves have not yet elevated by E14.0. However, six hours later at E14.25, palatal shelves have completed elevation in 80% of embryos. Interestingly, all E14.25 embryos with unelevated palatal shelves (20%) were female, suggesting a delay in female embryos. In FVB/NJ embryos, the elevation window started earlier (E13.875-E14.25) without any noticeable sex differences. We frequently captured an intermediate stage with unilateral elevation of either right or left palatal shelf. Magnetic resonance imaging of various stages showed that palatal shelf elevation began with the formation of bilateral bulges in the posterior. These bulges progressed laterally and anteriorly over time. During elevation, we observed increased cell proliferation in the lingual region of the palatal shelf. Within the bulge, cell orientation was acutely tilted towards the tongue and actomyosin activity was increased, which together may participate in the projection of the bulge in the horizontal direction. Thus, our data reveal novel insights into the rapid dynamic changes in palatal shelf elevation that lay the foundation for future studies of normal and abnormal palatogenesis.
Transcriptome analysis of human preimplantation embryo reveals expressed waves associated with blastulation failure based on embryonic grade and age
Authors: Yuan Ping , Liu Ying , Zhao Haijing , Ma Guangwei , Zheng Lingyan , Zhang Qingxue , Chen Hui , Wang Wenjun , Guo Yabin
Published: June 2022

In thein vitrofertilization and embryo transfer (IVF-ET) treatments, blastocyst culture is the method of choice for the generation of the embryos. Blastocysts can present different growth, quality, availability, and morphological characteristics that can be used to evaluate them. Although extreme blastocyst formation failures have been associated with the alteration of a single gene, the molecular factors responsible for arrested embryos remain unknown. RNA-sequencing (RNA-seq) is a promising tool for facilitating transcriptomic studies in early human embryos, thus allowing the investigation of gene expression discrepancies associated with different morphological criteria. Herein, we performed transcriptome analyses of the different stages of arrested human embryos. We identified candidate genes and related cell signaling pathways potentially associated with either arrested or developed embryos. Specifically, the three genes (MOV10L1, DDX4, andFKBP6) related to both DNA methylation and piRNA metabolic pathway might be involved in embryo development. Additionally, the transcriptome of arrested early blastocysts was significantly different from developed late blastocysts. Although the gene expression profiles identified were not significantly different between low- and high-quality late blastocysts, a significant difference in the profiles of day 5 and day 6 available late blastocysts was observed, which may be related to the clinical pregnancy rate associated with IVF-ET. Furthermore, we show that some chimeric RNAs may be functional in blastocyst development. Our findings uncovered new molecular markers that can be used for embryonic development detection, which might act as a tool for blastocyst selection for subsequent transfer.
Characterizing the role of Ca2+ fluxes in defining fast and slow components of spontaneous Ca2+ local transients in OPCs using computational modeling
Authors: Desjardins Nicolas , Jiang Xiaoyu , Oprea Laurentiu , Sareen Kushagra , Zheng James Q. , Khadra Anmar
Published: June 2022

1

Spontaneous Ca2+ local transients (SCaLTs) in isolated oligodendrocyte precursor cells (OPCs) are largely regulated by the following fluxes: store-operated Ca2+ entry (SOCE), Na+/Ca2+ exchange (NCX), Ca2+ pumping through Ca2+-ATPases, and Ca2+-induced Ca2+-release through Ryanodine receptors (RyR) and inositoltriphosphate receptors (IP3R). However, the relative contributions of these fluxes in mediating fast spiking and slow baseline oscillations seen in SCaLTs remain incompletely understood. Here, we developed a stochastic spatiotemporal computational model to simulate SCaLTs in a homogeneous medium with ion flow between the extracellular, cytoplasmic and endoplasmic-reticulum compartments. By simulating the model and plotting both the histograms of SCaLTs obtained experimentally and from the model as well as the standard deviation of interspike intervals (ISI) against ISI averages of multiple model and experimental realizations we revealed that: SCaLTs exhibit very similar characteristics between the two datasets, they are mostly random, they encode information in their frequency, and the slow baseline oscillations could be due to the stochastic slow clustering of IP3R (modeled as an Ornstein-Uhlenbeck noise process). Bifurcation analysis of a deterministic temporal version of the model shows that the contribution of fluxes to SCaLTs depends on the parameter regime and that the combination of excitability, stochasticity, and mixed-mode oscillations are responsible for irregular spiking and doublets in SCaLTs. Additionally, our results demonstrate that blocking each flux reduces SCaLTs frequency and that the reverse (forward) mode of NCX decreases (increases) SCaLTs. Taken together, these results provide a quantitative framework for SCaLT formation in OPCs.
Inertial Focusing of Particles and Cells in the Microfluidic Labyrinth Device: Role of Sharp Turns
Authors: Gangadhar Anirudh , Vanapalli Siva A.
Published: June 2022

Inertial, size-based focusing was investigated in the microfluidic labyrinth device consisting of several U-shaped turns along with circular loops. Turns are associated with tight curvature, and therefore induce strong Dean forces for separating particles, however, systematic studies exploring this possibility do not exist. We characterized the focusing dynamics of different-sized rigid particles, cancer cells and white blood cells over a range of fluid Reynolds numbers Ref. Streak widths of the focused particle streams at all the turns showed intermittent fluctuations which were substantial for smaller particles and at higher Ref. In contrast, cell streaks were less prone to fluctuations. Computational fluid dynamics simulations revealed the existence of strong turn-induced Dean vortices which help explain the intermittent fluctuations seen in particle focusing. Next, we developed a measure of pairwise separability to evaluate the quality of separation between focused streams of two different particle sizes. Using this, we assessed the impact of a single sharp turn on separation. In general, the separability was found to vary significantly as particles traversed the tight-curvature U-turn. Comparing the separability at the entry and exit sections, we found that turns either improved or reduced separation between different-sized particles depending on Ref. Finally, we evaluated the separability at the downstream expansion section to quantify the performance of the labyrinth device in terms of achieving size-based enrichment of particles and cells. Overall, our results show that turns are better for cell focusing and separation given that they are more immune to curvature-driven fluctuations in comparison to rigid particles.
Gene expression patterns associated with fin shape differ between two lamprologine cichlids
Authors: Ahi Ehsan Pashay , Richter Florian , Sefc Kristina M.
Published: June 2022

Comparing gene regulatory patterns between seemingly similar phenotypic traits can provide important insights on the molecular mechanisms underlying the evolution of those traits. In this study, we investigate the molecular basis of the formation of a spade-shaped caudal fin, which is a rare phenotype among teleost fish characterized by an elongated medial region of the fin. We examined the expression patterns of candidate fin-shape genes in the spade-shaped caudal fin of the related speciesLamprologus tigripictilis, an East African cichlid in the tribe Lamprologini. The candidate gene set consisted of a previously identified gene regulatory network (GRN) associated with the elongation of fin regions in another Lamprologini cichlid species and further genes selected on the basis of co-expression data and transcription factor prediction. Unexpectedly, the anatomical features of elongated fin rays differed and gene expression patterns associated with fin elongation were only weakly conserved between the two related species. We report 20 genes and transcription factors (includingangptl5, cd63, csrp1a, cx43, esco2, gbf1andrbpj), whose expression levels differed between the elongated and the short caudal fin regions ofL. tigripictilis, and which are therefore candidates for the regulation of the spade-like fin shape.
Understanding spatiotemporal effects of food supplementation on host-parasite interactions using community-based science
Authors: Knutie Sarah A. , Bahouth Rachel , Bertone Matthew A. , Webb Caroline , Mehta Mahima , Nahom Mia , Barta Rachael M. , Ghai Sharan , Love Ashley C. , Horan Sydney , Soldo Alexandria , Cochrane Elizabeth , Bartholomew Jenna , Cowan Emily , Bjerke Heather , Balenger Susan L. , Butler Michael W. , Cornell Allison , Kennedy Ashley C. , Rolland Virginie , Schultz Elizabeth M. , Stanback Mark , Taff Conor C. , Albery Gregory F.
Published: June 2022

Supplemental feeding can increase the overall health of animals but also can have variable effects on how animals defend themselves against parasites. However, the spatiotemporal effects of food supplementation on host-parasite interactions remain poorly understood, likely because large-scale, coordinated efforts are difficult.

Here, we introduce the Nest Parasite Community Science Project, which is a community-based science project that coordinates studies with bird nest box “stewards” from the public and scientific community. This project was established to understand broad ecological patterns between hosts and their parasites.

The goal of this study was to determine the effect of food supplementation on eastern bluebirds (Sialia sialis) and their nest parasite community across the geographic range of the bluebirds from 2018–2021. We received 646 nests from 68 stewards in 26 states in the eastern United States. Nest box stewards reported whether or not they fed their bluebirds mealworms or suet, then followed the nesting success of the birds (number of eggs laid and hatched, percent hatched, number and percent fledged). We then identified and quantified parasites in the nests.

We found that food supplementation increased fledgling numbers and proportional fledging success. The main nest parasite taxa were parasitic blow flies (Protocalliphora sialia), but a few nests contained fleas (Ceratophyllus idius, C. gallinae, Orchopeas leucopus) and mites (Dermanyssusspp. andOrnithonyssusspp.). Blow flies were primarily found at northern latitudes, where food supplementation affected blow fly abundance. However, the direction of this effect varied substantially in direction and magnitude across years. More stewards fed bluebirds at southern latitudes than at northern latitudes, which contradicted the findings of other community-based science projects.

Overall, food supplementation of birds was associated with increased host fitness but did not appear to play a consistent role in defense against these parasites across all years. Our study demonstrates the importance of coordinated studies across years and locations to understand the effects of environmental heterogeneity, including human-based food supplementation, on host-parasite dynamics. Studies during a single year or considering only a single population might not provide the necessary data to develop management strategies for species.