bluuebunny/crossref_metadata_2025 huggingface dataset:
On Cracks, Lights and Environments
Authors: Paloniitty Tiina
Published: May 2023

Participatory rights have been the focus of global attention in recent years, as people have fought over access to courts in all things climate-related. Access to information has gained lesser attention, though it is a pivotal first step in holding the decision-makers accountable. In his volume, Sean Whittaker offers the reader a bird’s-eye view on how this aim is conceptualised, understood and executed in three different jurisdictions—England and Wales, the USA and China. Using legal transplant theory prospectively, he explores the potential of each solution to serve as a provider or receiver of transplants. Whittaker’s clear analysis delivers on the calls of contingent and ambitious comparative environmental law scholarship. In carefully examining the substantive and procedural cores of the access to environmental information right, Whittaker explores the details of the right and, by doing so, also presents a fine-tuned understanding of the global normativity of the Aarhus Convention.
Penicillin-resistant gonorrheal endocarditis presenting with large annular abscess and aortic valve destruction
Authors: Kashyap Abhishek K , Abu-Hussain Hamza , GnanaDev Raja , Seliem Ahmed , Salabat Reza
Published: May 2023

Disseminated gonorrheal infections with cardiac involvement are rare, with fewer than 100 cases reported. The increasing prevalence of gonococcal infections and increasing antibiotic resistance represent a concerning challenge to public health. Here we report a case of antibiotic-resistant gonococcal endocarditis presenting with cardiogenic shock and discuss principles of diagnosis and treatment.
Massive hemoptysis as a sole presentation of left atrial myxoma
Authors: Giacaman Narmeen , Marzouqa Nizar , Saeed Elias , Tos Salem M , Emar Ahmad , Nassr Mohammad , Amr Bajis , Marzouqa Bisher
Published: May 2023

Diagnosis of myxoma can be difficult given its variable presentation and while adopting common sense in diagnosing this condition, physicians should also be aware of atypical presentations. Herein we present a 47-year-old heavy smoker presented with massive blood-stained expectoration. He was later diagnosed with cardiac myxoma and managed accordingly.
Lipoma-like hibernoma: a rare soft tissue tumor—a case series and review of the literature
Authors: Gabra Abanoub , Sacasa Astrid , Baig Mirza , Kleinstein Eric
Published: May 2023

Hibernomas are rare and benign lipomatous tumors which consist of lobules of cells derived from brown fat. Less than 200 overall cases are reported in the literature which necessitates the need to report more cases for better understanding of this neoplasm. Hibernomas are slowly progressing, painless tumors most frequently located in the subcutaneous thigh, upper trunk, and neck. Hibernomas generally occur in areas close to the axial skeleton where brown fat used to exist in the fetus and persisted during adulthood. They are rare to be discovered for the first time in older adults, most commonly presenting in adults 20–40 years of age. Core biopsies are simple to perform under local anesthetic and provide pathological differentiation prior to planned excision, hence are recommended, especially with larger lesions. Based on case presentation, watchful waiting for non-growing asymptomatic cases of hibernoma is possible, but more frequent follow-up and/or a core biopsy would be advised to rule out a more aggressive diagnosis. Compared with lipoma-like hibernomas, atypical lipomatous tumor/well-differentiated liposarcoma recurs and might dedifferentiate to a dedifferentiated liposarcoma with the ability to metastasize in up to 5% of the recurrences. Hibernomas, including the lipoma-like variant, are typically treated by surgical excision, rarely recur and do not metastasize. We report two rare presentations of hibernoma and lipoma-like hibernoma. Awareness of this clinicopathological entity is crucial to avoid misdiagnosis as malignant lipomatous neoplasm.
The transcription factor BnaWRKY10 regulates cytokinin dehydrogenase BnaCKX2 to control cytokinin distribution and seed size in Brassica napus
Authors: Yan Guanbo , Li Sijia , Ma Mengya , Quan Chengtao , Tian Xia , Tu Jinxing , Shen Jinxiong , Yi Bin , Fu Tingdong , Ma Chaozhi , Guo Liang , Dai Cheng
Published: May 2023

Cytokinins (CKs) are phytohormones that promote cell division and differentiation. However, the regulation of CK distribution and homeostasis in Brassica napus is poorly understood. Here, the endogenous CKs were first quantified by LC-ESI-MS/MS in rapeseed tissues and visualized by TCSn::GUS reporter lines. Interestingly, the cytokinin oxidase/dehydrogenase BnaCKX2 homologs were mainly expressed in reproductive organs. Subsequently, the quadruple mutants of the four BnaCKX2 homologs were generated. Endogenous CKs were increased in the seeds of the BnaCKX2 quadruple mutants, resulting in a significantly reduced seed size. In contrast, overexpression of BnaA9.CKX2 resulted in larger seeds, probably by delaying endosperm cellularization. Furthermore, the transcription factor BnaC6.WRKY10b, but not BnaC6.WRKY10a, positively regulated BnaA9.CKX2 expression by binding directly to its promoter region. Overexpression of BnaC6.WRKY10b rather than BnaC6.WRKY10a resulted in lower concentration of CKs and larger seeds by activating BnaA9.CKX2 expression, indicating that the functional differentiation of BnaWRKY10 homologs might have occurred during B. napus evolution or domestication. Notably, the haploid types of BnaA9.CKX2 were associated with 1000-seed weight in the natural B. napus population. Overall, the study reveals the distribution of CKs in B. napus tissues, and shows that BnaWRKY10-mediated BnaCKX2 expression is essential for seed size regulation, providing promising targets for oil crop improvement.
PhD-SNPg: updating a webserver and lightweight tool for scoring nucleotide variants
Authors: Capriotti Emidio , Fariselli Piero
Published: May 2023

One of the primary challenges in human genetics is determining the functional impact of single nucleotide variants (SNVs) and insertion and deletions (InDels), whether coding or noncoding. In the past, methods have been created to detect disease-related single amino acid changes, but only some can assess the influence of noncoding variations. CADD is the most commonly used and advanced algorithm for predicting the diverse effects of genome variations. It employs a combination of sequence conservation and functional features derived from the ENCODE project data. To use CADD, a large set of pre-calculated information must be downloaded during the installation process. To streamline the variant annotation process, we developed PhD-SNPg, a machine-learning tool that is easy to install and lightweight, relying solely on sequence-based features. Here we present an updated version, trained on a larger dataset, that can also predict the impact of the InDel variations. Despite its simplicity, PhD-SNPg performs similarly to CADD, making it ideal for rapid genome interpretation and as a benchmark for tool development.
Reducing the inherent auto-inhibitory interaction within the pegRNA enhances prime editing efficiency
Authors: Ponnienselvan Karthikeyan , Liu Pengpeng , Nyalile Thomas , Oikemus Sarah , Maitland Stacy A , Lawson Nathan D , Luban Jeremy , Wolfe Scot A
Published: May 2023

Prime editing systems have enabled the incorporation of precise edits within a genome without introducing double strand breaks. Previous studies defined an optimal primer binding site (PBS) length for the pegRNA of ∼13 nucleotides depending on the sequence composition. However, optimal PBS length characterization has been based on prime editing outcomes using plasmid or lentiviral expression systems. In this study, we demonstrate that for prime editor (PE) ribonucleoprotein complexes, the auto-inhibitory interaction between the PBS and the spacer sequence affects pegRNA binding efficiency and target recognition. Destabilizing this auto-inhibitory interaction by reducing the complementarity between the PBS-spacer region enhances prime editing efficiency in multiple prime editing formats. In the case of end-protected pegRNAs, a shorter PBS length with a PBS-target strand melting temperature near 37°C is optimal in mammalian cells. Additionally, a transient cold shock treatment of the cells post PE-pegRNA delivery further increases prime editing outcomes for pegRNAs with optimized PBS lengths. Finally, we show that prime editor ribonucleoprotein complexes programmed with pegRNAs designed using these refined parameters efficiently correct disease-related genetic mutations in patient-derived fibroblasts and efficiently install precise edits in primary human T cells and zebrafish.
DUF2285 is a novel helix-turn-helix domain variant that orchestrates both activation and antiactivation of conjugative element transfer in proteobacteria
Authors: Jowsey William J , Morris Calum R P , Hall Drew A , Sullivan John T , Fagerlund Robert D , Eto Karina Y , Solomon Paul D , Mackay Joel P , Bond Charles S , Ramsay Joshua P , Ronson Clive W
Published: May 2023

Horizontal gene transfer is tightly regulated in bacteria. Often only a fraction of cells become donors even when regulation of horizontal transfer is coordinated at the cell population level by quorum sensing. Here, we reveal the widespread ‘domain of unknown function’ DUF2285 represents an ‘extended-turn’ variant of the helix-turn-helix domain that participates in both transcriptional activation and antiactivation to initiate or inhibit horizontal gene transfer. Transfer of the integrative and conjugative element ICEMlSymR7A is controlled by the DUF2285-containing transcriptional activator FseA. One side of the DUF2285 domain of FseA has a positively charged surface which is required for DNA binding, while the opposite side makes critical interdomain contacts with the N-terminal FseA DUF6499 domain. The QseM protein is an antiactivator of FseA and is composed of a DUF2285 domain with a negative surface charge. While QseM lacks the DUF6499 domain, it can bind the FseA DUF6499 domain and prevent transcriptional activation by FseA. DUF2285-domain proteins are encoded on mobile elements throughout the proteobacteria, suggesting regulation of gene transfer by DUF2285 domains is a widespread phenomenon. These findings provide a striking example of how antagonistic domain paralogues have evolved to provide robust molecular control over the initiation of horizontal gene transfer.
On the accuracy of the epigenetic copy machine: comprehensive specificity analysis of the DNMT1 DNA methyltransferase
Authors: Adam Sabrina , Klingel Viviane , Radde Nicole E , Bashtrykov Pavel , Jeltsch Albert
Published: May 2023

The specificity of DNMT1 for hemimethylated DNA is a central feature for the inheritance of DNA methylation. We investigated this property in competitive methylation kinetics using hemimethylated (HM), hemihydroxymethylated (OH) and unmethylated (UM) substrates with single CpG sites in a randomized sequence context. DNMT1 shows a strong flanking sequence dependent HM/UM specificity of 80-fold on average, which is slightly enhanced on long hemimethylated DNA substrates. To explain this strong effect of a single methyl group, we propose a novel model in which the presence of the 5mC methyl group changes the conformation of the DNMT1-DNA complex into an active conformation by steric repulsion. The HM/OH preference is flanking sequence dependent and on average only 13-fold, indicating that passive DNA demethylation by 5hmC generation is not efficient in many flanking contexts. The CXXC domain of DNMT1 has a moderate flanking sequence dependent contribution to HM/UM specificity during DNA association to DNMT1, but not if DNMT1 methylates long DNA molecules in processive methylation mode. Comparison of genomic methylation patterns from mouse ES cell lines with various deletions of DNMTs and TETs with our data revealed that the UM specificity profile is most related to cellular methylation patterns, indicating that de novo methylation activity of DNMT1 shapes the DNA methylome in these cells.
Ex vivo to in vivo model of malignant peripheral nerve sheath tumors for precision oncology
Authors: Larsson Alex T , Bhatia Himanshi , Calizo Ana , Pollard Kai , Zhang Xiaochun , Conniff Eric , Tibbitts Justin F , Rono Elizabeth , Cummins Katherine , Osum Sara H , Williams Kyle B , Crampton Alexandra L , Jubenville Tyler , Schefer Daniel , Yang Kuangying , Lyu Yang , Pino James C , Bade Jessica , Gross John M , Lisok Alla , Dehner Carina A , Chrisinger John S A , He Kevin , Gosline Sara J C , Pratilas Christine A , Largaespada David A , Wood David K , Hirbe Angela C
Published: May 2023

Background

Malignant peripheral nerve sheath tumors (MPNST) are aggressive soft tissue sarcomas that often develop in patients with neurofibromatosis type 1 (NF1). To address the critical need for novel therapeutics in MPNST, we aimed to establish an ex vivo 3D platform that accurately captured the genomic diversity of MPNST and could be utilized in a medium-throughput manner for drug screening studies to be validated in vivo using patient-derived xenografts (PDX).

Methods

Genomic analysis was performed on all PDX-tumor pairs. Selected PDX were harvested for assembly into 3D microtissues. Based on prior work in our labs, we evaluated drugs (trabectedin, olaparib, and mirdametinib) ex vivo and in vivo. For 3D microtissue studies, cell viability was the endpoint as assessed by Zeiss Axio Observer. For PDX drug studies, tumor volume was measured twice weekly. Bulk RNA sequencing was performed to identify pathways enriched in cells.

Results

We developed 13 NF1-associated MPNST-PDX and identified mutations or structural abnormalities in NF1 (100%), SUZ12 (85%), EED (15%), TP53 (15%), CDKN2A (85%), and chromosome 8 gain (77%). We successfully assembled PDX into 3D microtissues, categorized as robust (>90% viability at 48 h), good (>50%), or unusable (<50%). We evaluated drug response to “robust” or “good” microtissues, namely MN-2, JH-2-002, JH-2-079-c, and WU-225. Drug response ex vivo predicted drug response in vivo, and enhanced drug effects were observed in select models.

Conclusions

These data support the successful establishment of a novel 3D platform for drug discovery and MPNST biology exploration in a system representative of the human condition.
Missing Americans: Early death in the United States—1933–2021
Authors: Bor Jacob , Stokes Andrew C , Raifman Julia , Venkataramani Atheendar , Bassett Mary T , Himmelstein David , Woolhandler Steffie
Published: May 2023

We assessed how many US deaths would have been averted each year, 1933–2021, if US age-specific mortality rates had equaled the average of 21 other wealthy nations. We refer to these excess US deaths as “missing Americans.” The United States had lower mortality rates than peer countries in the 1930s–1950s and similar mortality in the 1960s and 1970s. Beginning in the 1980s, however, the United States began experiencing a steady increase in the number of missing Americans, reaching 622,534 in 2019 alone. Excess US deaths surged during the COVID-19 pandemic, reaching 1,009,467 in 2020 and 1,090,103 in 2021. Excess US mortality was particularly pronounced for persons under 65 years. In 2020 and 2021, half of all US deaths under 65 years and 90% of the increase in under-65 mortality from 2019 to 2021 would have been avoided if the United States had the mortality rates of its peers. In 2021, there were 26.4 million years of life lost due to excess US mortality relative to peer nations, and 49% of all missing Americans died before age 65. Black and Native Americans made up a disproportionate share of excess US deaths, although the majority of missing Americans were White.
Considering ChatGPT to be the first aid for young adults on mental health issues
Authors: Aminah Siti , Hidayah Nur , Ramli M
Published: May 2023

Providing mental health support and interventions through artificial intelligence can open up more significant space for young people to get good services. ChatGPT’s potential to provide first aid to young adults struggling with mental health issues is noteworthy. Despite its benefits and limitations, today’s technology can be a valuable addition to the mental health support available to young adults.
Low testosterone state inhibits erectile function by downregulating the expression of GIT1 in rat penile corpus cavernosum
Authors: Gu JianBin , Zhu Li-kun , Zhao Xin , Jiang Jun , Jiang Rui
Published: May 2023

Background

The mechanism of erectile dysfunction (ED) caused by a low androgen level is still not clear.

Aim

To explore the influence of the low testosterone state on G protein–coupled receptor kinase interactor 1 (GIT1) and its contact to erectile function.

Methods

Thirty male Sprague-Dawley rats aged 8 weeks were distributed at random into 5 groups: control (sham operated), castration, testosterone supplement after castration, castration + vacant lentiviral transfection, and castration + lentiviral transfection. The testis and epididymis were removed through a scrotal incision to develop castrated rats. Four weeks after castration, a lentivirus carrying the GIT1 gene was injected into the middle of rat penile corpus cavernosum. One week after transfection, maximum intracavernous pressure/mean arterial pressure (ICPmax/MAP), serum testosterone, nitric oxide, GIT1, endothelial nitric oxide synthase (eNOS), phospho-eNOS (p-eNOS), p-eNOS/eNOS, and the interaction between eNOS and GIT1 were assessed in the rats.

Outcomes

The levels of GIT1 in the penile cavernous tissue of castrated rats are significantly lower than that of controls.

Results

GIT1 was expressed in the cytoplasm and cell membrane of vascular endothelial cells and smooth muscle cells in rat penile tissue. In comparison with normal rats, the castrated rats showed lower levels of GIT1 expression, GIT1 and eNOS interaction, p-eNOS/eNOS, nitric oxide, and ICPmax/MAP (P < .01). Overexpression of GIT1 can intensively enhance the expression level of GIT1, the interaction between GIT1 and eNOS, p-eNOS/eNOS, nitric oxide, and ICPmax/MAP in rats (P < .01).

Clinical Translation

Modulating the interaction between eNOS and GIT1 might be a novel method of treating ED caused by a low androgen level.

Strengths and Limitations

The impact of GIT1 phosphorylation on the activity of eNOS and its possible mechanisms affecting erectile function require further study.

Conclusion

A low testosterone state inhibits erectile function in rats by reducing the expression of GIT1 and the protein interaction between GIT1 and eNOS.
Sexual dysfunction is highly prevalent in male survivors of malignant lymphoma
Authors: Micas Pedersen Signe , Nielsen Torsten Holm , Gang Anne Ortved , Poulsen Christian Bjørn , de Nully Brown Peter , Jørgensen Niels , Feltoft Claus Larsen , Pedersen Lars Møller
Published: May 2023

Background

With improved survival in patients with lymphoma, long-term toxicity and quality of life (QoL), including sexual health, have become increasingly important.

Aim

We aimed to (1) determine the prevalence of erectile dysfunction (ED) in adult male lymphoma survivors; (2) determine whether testosterone deficiency, comorbidities, or lifestyle factors were associated; and (3) evaluate their impact on QoL.

Methods

A cross-sectional study including 172 male survivors of Hodgkin lymphoma or diffuse large B cell lymphoma diagnosed in adulthood between 2008 and 2018 was performed. Patients were in complete metabolic remission after first-line treatment and remained in remission at follow-up (3-13 years after diagnosis). Participants completed 3 questionnaires measuring sexual health and general QoL. Serum concentrations of total testosterone were measured and thorough medical history and sociodemographic factors were obtained. The Danish SEXUS Project, European Male Ageing Study, and European Organization of Research and Treatment of Cancer (EORTC) Reference Manual were used as reference values of the general population.

Outcomes

Patient reported outcome measures including the 5-item International Index of Erectile Function, EORTC C30, and EORTC 22-item Sexual Health Questionnaire.

Results

ED was reported by 55.2%, which was higher than in an age-matched Danish population cohort (17.5%). Erectile function score (5-item International Index of Erectile Function) was negatively associated with comorbidity, body mass index, smoking, and age and positively with the number of children conceived before treatment and serum concentration of total testosterone. Overt testosterone deficiency in combination with ED was detected in 10 (5.7%) of 176 survivors, including excluded survivors in hormonal treatment, which is higher than for the general population (0.1%-3.2% for men <70 years of age). Mean EORTC C30 global health score for survivors with ED was lower (67.7) than for survivors without ED (80.1) but was comparable to the general population (71.2). Furthermore, a positive association was seen between sexual function and both sexual and general QoL.

Clinical implications

Sexual health is important for QoL and related to comorbidities. The focus on improving QoL requires that both sexual health and comorbidities are addressed in the follow-up of lymphoma patients.

Strengths and limitations

Despite the relatively high number of included survivors, the cross-sectional design of this study warrants longitudinal studies to clarify the specific underlying causes of sexual dysfunction.

Conclusion

ED was highly prevalent and associated with comorbidity in lymphoma survivors, and more focus on sexual health and treatment related comorbidity is needed to improve sexual and general QoL.
Impact of endometriosis on female sexual function: an updated systematic review and meta-analysis
Authors: Zhu Xiujuan , Wu Yanhui , Jia Jing , Zhao Xinwei , Zhao Xiuping
Published: May 2023

Introduction

Endometriosis can lead to a state of chronic inflammation marked by the presence of scarring and adhesions within the pelvis and/or other parts of the body. Recent estimates suggest that globally this condition affects approximately 10% of women in the reproductive age group.

Aims

In this study we sought updated evidence on the association between endometriosis and sexual function in female patients.

Methods

We used standard assessment tools to conduct a systematic search of the PubMed, EMBASE, and Scopus databases for observational studies that documented the association of endometriosis with female sexual function. A random-effects model was used for the analysis, and effect sizes were reported as the weighted mean difference (WMD) or OR with 95% CIs.

Results

A total of 13 studies were selected for inclusion in our investigation. All of the included studies were cross-sectional in design. The data on sexual function in most of the studies were collected by using the Female Sexual Function Index (FSFI) tool, for which higher scores suggest better sexual function. The risk of sexual dysfunction (based on specific cutoffs for the FSFI score) was higher in women with than in women without endometriosis (OR 1.71; 95% CI, 1.21-2.43). In addition, when we used continuous scores to examine the risk of sexual dysfunction, diagnosis of endometriosis was associated with significantly lower overall FSFI scores (WMD, −3.40; 95% CI, −5.13 to −1.66) and lower scores on all of its 6 domains, ie, desire (WMD, −0.27; 95% CI, −0.53 to −0.02), arousal (WMD, −0.43; 95% CI, −0.79 to −0.07), lubrication (WMD, −0.49; 95% CI, −0.66 to −0.31), orgasm (WMD, −0.65; 95% CI, −1.07 to −0.23), satisfaction (WMD, −0.52; 95% CI, −0.77 to −0.26), and pain (WMD, −1.06; 95% CI, −1.57 to −0.55).

Conclusion

The findings of this study suggest that female patients with endometriosis have suboptimal sexual function compared with healthy female subjects. Patients with endometriosis should be offered sexual counseling and supportive care by a multidisciplinary team of gynecologists, psychologists, and sexual therapists.
0003 Sleep and Circadian Disruption Induced by Simulated Night Shift Work Impair Cardiometabolic Outcomes in Healthy Adults
Authors: Seward Sophie , Higgins Janine , Wright Kenneth , Broussard Josiane
Published: May 2023

Introduction

People who work evening, night or rotating shifts make up 20% of the US workforce and have a heightened risk for Type 2 diabetes (T2D) and cardiovascular disease (CVD) compared to people who work days, possibly due to repeated bouts of sleep and circadian disruption. However, the mechanism(s) by which sleep and circadian disruption impair cardiometabolic health are unclear. We therefore investigated the impact of acute circadian misalignment on factors related to cardiometabolic health including interstitial glucose concentration, glucose tolerance, resting systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR).

Methods

Ten healthy adults (6F, 25.0±4.5y, BMI: 22.0±2.2kg/m2; mean±SD) participated in an ongoing 6d inpatient simulated night shift protocol to induce sleep and circadian disruption. Participants lived at the Sleep and Metabolism Laboratory for 2d to simulate day shift work followed by 2d of simulated night shift work. Continuous interstitial glucose monitoring was conducted throughout the study (FreeStyle Libre Pro; Abbott, Alameda, CA, USA). BP and HR were assessed 30 minutes after waketime (Omron HEM-FL31; Omron Healthcare, Kyoto, JP) followed by an oral glucose tolerance tests (OGTT) conducted 1.5h after waketime in both conditions.

Results

Simulated night shift work was associated with significantly higher interstitial glucose area under the curve (AUC) in response to an identical first meal after waking (18,299±1412 v 14,606±2035 [mg/dl]*min; p=0.009; mean±SEM) as well as plasma glucose AUC during the OGTT (23,239±2574 v 18,776±1832; p=0.002) compared to simulated day work. Furthermore, simulated night shift work was associated with a significant increase in resting DBP (75±3 v 66±2; p=0.017) compared to day work. No changes were observed in resting SBP (p=0.40) or HR (p=0.11) between conditions.

Conclusion

Sleep and circadian disruption induced by simulated night shift work appear to impair markers of cardiometabolic health, even in healthy, lean adults. Night shift work is largely unavoidable in modern society, thus is it critical to identify strategies or countermeasures to prevent the impact of circadian misalignment on T2D and CVD risk in a large portion of the population.

Support (if any)

This project was supported in part by R01DK125653 & F31HL165883.
0007 Later circadian phase and increased sleep variability are associated with overnight blood pressure dipping in chronic shiftworkers
Authors: Shafer Brooke , Christopher Desirae , Shea Steven , Olson Ryan , McHill Andrew
Published: May 2023

Introduction

Nighttime shift work is associated with adverse cardiovascular outcomes, including hypertension. Increased sleep variability and circadian disruption may increase sympathetic activity and waking blood pressure (BP), potentially constraining typical BP reductions during overnight sleep. However, the relationships between circadian phase, sleep variability, and overnight BP reduction (dipping) among day vs. chronic nightshift workers is unknown.

Methods

Ambulatory BP was monitored for up to 48-hours among 10 dayshift (mean□SD; age 33.1□7.0y; BMI 26.3□5.4 kg/m2) and 10 chronic (9.3□6.8y of shiftwork) nightshift workers (age 32.7□9.1y; BMI 27.9□7.6 kg/m2). Overnight BP dipping was calculated from the sleeping/waking systolic BP ratio. Variability in sleep timing was defined as the SD of self-reported daily-diary sleep onset across 1-week. Circadian phase was determined by salivary dim-light melatonin onset (DLMO; 3pg/ml threshold). Differences between groups were assessed using independent t-tests and linear mixed-effects models with group and time-from-sleep-onset or circadian phase as fixed-effects, and participant as a random effect. Relationships between overnight BP dipping and DLMO, sleep onset timing variability, and phase angle of entrainment (DLMO to sleep onset during ambulatory BP) were assessed using Pearson correlation analyses.

Results

Chronic nightshift workers had reduced overnight BP dipping (-9.3□1.5 % vs -13.4□4.4 %; p=0.04), later DLMO timing (21:08□0:53 vs 19:21□1:02; p=0.001), smaller phase angle of entrainment (3.7□1.0 h vs 5.0□0.90 h; p=0.02), and greater variability in sleep onset timing (3.2□1.5 h vs 0.87□0.34 h; p< 0.001). Significant group effects for higher systolic BP across circadian phase and time from sleep onset in the nightshift group were observed compared to dayshift (both p< 0.05). DLMO (r=0.49; p=0.04) and phase angle of entrainment (r=0.53; p=0.02) were associated with overnight BP dip magnitude (later circadian phase correlated with less dipping). Sleep onset timing variability was negatively correlated with magnitude of overnight BP dipping in nightshift (r= -0.76; p=0.04) but not dayshift workers (r=0.26; p=0.61).

Conclusion

Attenuated overnight BP dipping among chronic nightshift workers may be indicative of increased cardiovascular risk. Chronic circadian disruptions (i.e., smaller phase angle of entrainment) as a function of higher sleep variability among nightshift workers may mechanistically contribute to poorer health outcomes compared to dayshift workers.

Support (if any)

R01HL105495, K01HL146992
0022 Characterization and Evaluation of Digital Dim Light Melatonin Onset in a Population-Based Sample
Authors: Reis Daniel , Bahraini Nazanin
Published: May 2023

Introduction

Misalignment between circadian phase (i.e., timing) and the desired sleep window is associated with sleep disturbances including insomnia and circadian rhythm sleep-wake disorders. Identifying circadian-driven sleep disruption requires assessment of circadian phase, the gold-standard of which is dim light melatonin onset (DLMO). While DLMO is traditionally measured in the lab using saliva or blood sampled over multiple hours, novel methods of estimating DLMO using activity and light data obtained from wearable sensors (i.e., digital DLMO) have been recently validated in samples with both healthy and disordered sleep. Such methods could potentially provide pragmatic, low-burden ways of assessing circadian phase, which in turn could be used to aid diagnostic decisions and to adjust circadian-targeted interventions in real time. However, digital DLMO has yet to be characterized within the broader adult population, which is needed to differentiate between normative and clinically salient values. Therefore, this study will examine digital DLMO in a large, population-based sample, as well as explore the potential clinical utility of digital biomarkers related to circadian phase.

Methods

This study will be a secondary analysis of data obtained during the ancillary sleep study of the Multi-Ethnic Study of Atherosclerosis (i.e., MESA Sleep). Seven days of activity and light data, measured via wrist-worn actigraphy, were obtained from 2,237 participants. Digital DLMO will be estimated using the extended Kronauer limit-cycle model of the human circadian pacemaker. Phase angles, or the differences in clock time, between digital DLMO and the following will be calculated: self-reported in-bed time; and sleep onset, midpoint, and offset measured by actigraphy. The associations between these phase angles and sleep outcomes (i.e., sleep onset latency, sleep efficiency, and sleep quality) will be explored using linear regression. Covariates will include the age, gender, and race/ethnicity of the participant. Weekend and weekday averages will be evaluated separately.

Results

Results will include the distribution of digital DLMO in a population-based sample, as well as the associations between digital DLMO phase angles and sleep outcomes.

Conclusion

This study will inform future research into the clinical potential of digital circadian biomarkers.

Support (if any)
0026 Novel home-based circadian phase assessment tool to enhance accessible quality care of circadian rhythm disorders- Circadia Study
Authors: Love Jessica , Lane Jacqueline , Saxena Richa , Scheer Frank , Bormes Gregory
Published: May 2023

Introduction

Circadian rhythm sleep-wake disorders (CRSWDs), shifted timing of physiology with the external environments. CRSWDs, often undiagnosed, can increase risk of cardiometabolic and psychiatric disorders, and may have substantial financial and social impacts. Dim light melatonin onset assay (DLMO), the current gold-standard diagnostic test for CRSWDs is expensive/difficult. We aim to develop a more accessible melatonin assessment.

Methods

Our DLMO assay kit is low-cost, home-based and self-administered. It contains all necessary components for a home-based DLMO assay, including 18 time-stamped saliva samples, a Philips Actiwatch, and a lux meter. We tested the feasibility of the kit across two DLMO assessments, one-week apart, in CRSWDs patients (N=5) and controls (N=5). We concurrently assessed sleep with wrist actigraphy and sleep diary over four weeks through a self-guided study website.

Results

We enrolled participants with delayed (n=4) and advanced (n=1) sleep disorder and 5 control participants (mean 38.2 y.o., SD 11.66 y.o.; biological sex: 9 female, 1 male (control)). Participants were 100% compliant completing questionnaires and sleep logging with little intervention and no in-person guidance. 5/10 participants completed 100% of sleep diary days. Average duration of study involvement, from consent to study completion, was 42.4 days. Participants successfully set up their collection space and collected saliva samples, remaining compliant to objectively-measured study protocols: 86.67% of samples were taken under sufficiently low light, and 82% were taken within five minutes of scheduled collection. We obtained and averaged two DLMO times for 7/10 participants (delayed sleep phase disorder (DPSD): 12:05 AM, controls: 9:56 PM). DLMO times were on average 3:18 hours earlier than self-reported sleep times.

Conclusion

Our pilot study, with both CRSWDs patients (N=5) and controls (N=5), found the at-home kits feasible and reproducible (5/6 participants had their DLMO times within an hour of each other). Using the kit, we confirmed their previous DSPD diagnoses and correctly classified controls through melatonin samples. We used objective measures (Actigraph) to verify the testing procedure was followed. These procedures will be tested on a larger scale in the Circadia Study (circadiastudy.org), set to launch in February 2023.

Support (if any)

This work was supported by the National Institutes of Health [DHHS] (grant number 1R35GM146839-01).
0030 Insomnia is associated with lower serum antioxidants in the Hispanic Community Health Study/Study of Latinos
Authors: Cabrera Jose Santos , Zhang Ying , Huang Tianyi , Chung Joon , Yu Bing , Qi Qibin , Ramos Alberto , Daviglus Martha , Zee Phyllis , Kaplan Robert , González Hector , Isasi Carmen , Redline Susan , Sofer Tamar
Published: May 2023

Introduction

Insomnia is associated with neurodegenerative diseases. Concentration levels of some metabolites are associated with pathophysiologic changes, possibly in a sex-specific manner. Thus, serum metabolite levels are useful for understanding insomnia-related pathophysiological changes and how they potentially relate to neurodegenerative disease.

Methods

The Hispanic Community Health Study/Study of Latinos (HCHS/SOL) is a longitudinal cohort study following self-reported Hispanic/Latino individuals in the U.S. At baseline, insomnia symptoms were captured using the Women Health Initiative Insomnia Rating Scale (WHIIRS), insomnia being defined as WHIIRS≥9. Metabolites were measured using blood serum from the baseline exam in independent discovery and replication datasets. In discovery, metabolite associations with WHIIRS and with insomnia were estimated in linear regression. Significant associations (FDR p-values< 0.05) were tested in the replication dataset and considered replicated if their p-value< 0.05 and the direction of association matched with the discovery analysis. Associations were further studied for gender interaction. All analyses were adjusted for age, self-reported gender, study center, and Hispanic/Latino background.

Results

3,895 individuals participated in the discovery analysis, and 2,121 in replication analysis. Female individuals were aged 46.5 on average (SD 15.1) and 35% had insomnia, while male individuals were aged 44.8 (14.8) and 24% had insomnia. After quality control, 767 metabolites were tested for association. Six metabolites were associated with WHIIRS (FDR< 0.05) in the gender-combined analysis. Three were xenobiotics which were on the benzoate or salicylic acid metabolism pathway. Of the 6 metabolite associations, two replicated (p-value< 0.05): hydrocinnamate and indolepropionate. These two metabolites had spearman correlation of 0.44. Higher levels of both metabolites were associated with lower WHIIRS, showing stronger associations in females (interaction p-value =0.04 for hydrocinnamate and p-value=0.13 for indolepropionate). Associations remained essentially unchanged in a model further adjusting for hypertension and diabetes. When removing 534 individuals who self-reported using sleeping pills at least once a week, the associations attenuated slightly: from an effect estimates of -0.0075 to -0.0069 (hydrocinnamate) and from -0.0082 to -0.0068 (indolepropionate).

Conclusion

Among the HCHS/SOL target population, insomnia symptom severity is associated with a lower concentrations of some plasma metabolite levels which are important antioxidant needed in preventing oxidative stress and neurodegeneration.

Support (if any)

R01HL161012.
0033 Genome-wide association analysis of composite sleep scores in 413,904 individuals
Authors: Goodman Matthew , Nagarajan Pavithra , Lane Jacqueline , Dashti Hassan , Chung Joon , Sofer Tamar , Purcell Shaun , Zhu Xiaofeng , Rutter Martin , Redline Susan , Saxena Richa , Wang Heming
Published: May 2023

Introduction

Recent genome-wide association studies (GWAS) of individual sleep traits have identified hundreds of genetic loci, often implicating genes and biological pathways that are shared across sleep phenotypes. Modeling multiple dimensions of sleep may allow identification of common underlying genetic mechanisms and complement analyses of individual traits. Moreover, construction of novel sleep health scores, accounting for correlations between traits, has the potential to enhance specificity and power for genetic analyses.

Methods

We performed GWAS of composite sleep scores derived from five sleep questions characterizing 413,904 individuals of European ancestry from the UK Biobank. We constructed an additive sleep score (SS-add) as a sum of up to five favorable self-reported sleep behaviors (sleep duration of 7-8 hours, early chronotype, few insomnia symptoms, no snoring, and no excessive daytime sleepiness) as well as five principal component scores (SS-PC1 – SS-PC5) using the underlying sleep traits. SNP-level association studies of the sleep scores were complemented with multiple follow-up analyses, including investigation of pathway, tissue, and cell-type enrichments, as well as comprehensive genetic correlation (381 representative phenotypes) and bi-directional Mendelian randomization (MR) analyses (40 selected phenotypes).

Results

Sleep scores SS-PC1 (interpretable as longer duration sleep without insomnia symptoms) and SS-PC2 (healthy sleep without snoring or sleepiness) showed higher heritability (respectively 11.7% and 9.3%) compared with their primary underlying traits. SS-PC3 (loading strongly on chronotype) showed highest overall heritability (15.3%). Accounting for the six GWAS, we identified 28 significant novel loci (p< 8.3e-9), 31 additional novel loci (p< 5e-8), and 341 loci previously reported (p< 5e-8) by GWAS of individual sleep traits. Associated loci mapped to genes enriched in expression in brain tissues, and in metabolic and neuronal pathways. Numerous neurological, cardiometabolic and other traits were genetically correlated with sleep health (104 with|rho_g|>0.3, p< 2e-5). MR associations (p< 2e-4) pointed to causes (BMI [SS-PC2/PC4], smoking [SS-add/PC1], lower socio-economic status [SS-add/PC5]) and consequences (coronary heart disease [SS-add/PC1], pain [SS-PC1/5]) of poor sleep.

Conclusion

Composite sleep health scores revealed novel genetic mechanisms of related sleep behaviors, helped clarify relationships with neurological and cardiometabolic traits, and deserve further investigation.

Support (if any)

R01HL153814 (to H.W.), NHLBI R35HL135818 (to S.R.), 1R01HL146751 (to R.S.), UKB Application 6818.
0041 Neuronal circuits convergently disrupted in the genetic basis of sleep phenotypes and rare neurodevelopmental syndromes
Authors: Moyses-Oliveira Mariana , Paschalidis Mayara , Souza-Cunha Lais , Guerreiro Pedro , Adami Luana , Kloster Anna , Mosini Amanda , Moreira Gustavo , Doria Sandra , Tempaku Priscila , Pires Gabriel , Andersen Monica , Tufik Sergio
Published: May 2023

Introduction

Sleep-related phenotypes have been frequently reported in early on-set epileptic encephalopathies (EE) and in developmental delay (DD) syndromes, in particular in syndromes related to autism spectrum disorder (ASD). Yet the convergent pathogenetic mechanisms between these comorbidities are largely unknown.

Methods

We first performed a gene enrichment study that identified shared risk genes among rare EE/neurodevelopmental disorders (NDD), rare DD genetic syndromes and sleep disturbances. We then determined cellular and molecular pathways enriched among genes shared between sleep phenotypes and those two early onset mental illnesses, aiming to identify genetic disparities and commonalities among these phenotypic groups.

Results

The sleep gene set was observed as significantly overlapped with the two gene lists associated to rare genetic syndromes (i.e. EE/NDD and DD gene sets), suggesting shared genetic contribution. Similarities across significantly enriched pathways between the two intersect lists comprehended mostly synapse-related pathways, such as retrograde endocannabinoid signaling, serotonergic and GABAergic synapse. Network analysis indicates EE/NDD vs sleep specific clusters and DD vs sleep specific clusters related to synaptic and transcriptional regulation, respectively. Longstanding functional patterns previously described in EE and NDD genetic architecture were recaptured after dissecting the overlap between the genes associated to those developmental phenotypes and sleep disturbances, suggesting that during neurodevelopment different molecular and functional mechanisms are related to alterations on circadian rhythm.

Conclusion

The overlapping gene set and biological pathways highlighted by this study may serve as a primer for new functional investigations of shared molecular mechanisms between sleep disturbances and rare developmental syndromes, with or without epilepsy.

Support (if any)

Our studies are supported by Associação Fundo de Incentivo à Pesquisa (AFIP). S.T. and M.L.A. received CNPq fellowships.
0042 EEP-PROMOTING GABAERGIC NEURONS IN THE PARAFACIAL ZONE PLAY A ROLE IN ANESTHETIC-INDUCED UNCONSCIOUSNESS IN MICE
Authors: Imamura Toshihiro , Wasilczuk Andrzej , Kelz Max , Pack Allan
Published: May 2023

Introduction

Although the precise mechanisms by which anesthetics induce unconsciousness are unknown, there is evidence that shared brain circuits are engaged during sleep and general anesthesia. The parafacial zone (PZ) of the brainstem has recently been identified as a sleep-promoting region. Specifically, GABAergic neurons in the PZ (PZ-GABA) are active during non-REM sleep. The role of sleep-promoting PZ-GABA during anesthetic-induced hypnosis is unknown. We hypothesized that PZ-GABA are active during general anesthesia and contribute to anesthetic-induced hypnosis.

Methods

Adult C57BL6 mice (n = 3) were subjected to two-hour exposures of a hypnotic dose of an inhaled general anesthetic (isoflurane 1.2%) in oxygen, a non-immobilizer, F6, that lacks hypnotic properties (1,2-dicholorohexafluorocyclobutane 3%) in oxygen, or oxygen alone. The brains were harvested immediately following these exposures. The sections of brain were immunostained for c-Fos and GAD. To study the role of PZ-GABA in isoflurane-induced hypnosis, ablation of PZ-GABA was performed in adult bitransgenic Vgat-ires-Cre;Ai6 mice in which GABAergic neurons express a fluorescent reporter (n = 5 per group). Anesthesia sensitivity was evaluated weekly for four weeks after the virus injections by assessing the presence of a righting reflex during the exposures.

Results

In the PZ, a hypnotic dose of Isoflurane selectively activated GAD-positive GABAergic neurons compared to mice treated with F6 and oxygen. (P < 0.01 for both F6 and oxygen vs. Isoflurane, one-way ANOVA) Furthermore, mice lacking PZ-GABA demonstrated resistance to anesthetic induction, decreased sensitivity during the maintenance phase, and consistently increased the propensity to emerge from anesthesia. (For both induction and emergence, the upper 95% CI of EC50 at baseline was lower than the lower 95% CI at all time points after the injections. For maintenance phase, P < 0.05 for baseline vs. 1-week, one-way ANOVA)

Conclusion

PZ-GABA was active during isoflurane exposure. Moreover, the absence of PZ-GABA resulted in resistance to isoflurane-induced hypnosis. These findings suggest that PZ-GABA, a sleep-promoting region in the brainstem, plays an important role in general anesthesia.

Support (if any)

NIH (T32HL7713-27, T32 NS091006), ATS-ASPIRE
0043 Modulation of ultra-slow calcium oscillation in the dentate gyrus during Non-REM sleep
Authors: Turi Gergely , Chen Xinyue , Teng Sasa , Peng Yueqing
Published: May 2023

Introduction

The function of sleep in memory consolidation has been well established. Hippocampal CA1 and CA3 subregions are thought to be particularly important in this process. The current working hypothesis postulates that episodic memory traces captured during waking hours are replayed in the hippocampal CA1-CA3 areas and transferred to the cortex for long-term storage during sleep. Even though the sensory and spatial information from higher-order cortical areas such as the entorhinal cortex primarily enters the hippocampus via the dentate gyrus (DG), this structure has always been considered as the “silent partner” in memory consolidation. Whether and how the captured memory traces are transferred from the DG toward the downstream hippocampal areas is less known.

Methods

Here, we used fiber photometry, two-photon calcium imaging, and EEG recording to examine the activity of DG during sleep.

Results

Strikingly, we found that DG cells are even more active during sleep than wakefulness and the calcium activity in the DG slowly oscillates during non-REM (NREM) sleep epochs. Furthermore, we also found that the cycles of this activity coinciding with microarousals are tightly locked to brief serotonin (5-HT) release during NREM sleep. Pharmacological blockade of 5-HT1a receptors abolished the calcium oscillations in the DG. Furthermore, genetic knockdown of 5-HT1a receptors in the DG leads to memory impairment in spatial and contextual memory tasks.

Conclusion

Together, our results indicate that ultra-slow calcium oscillations in the DG during NREM sleep are driven by serotonin fluctuations and they are required for memory consolidation.

Support (if any)