bluuebunny/crossref_metadata_2025_split huggingface dataset:
0029 Bimodality in Undergraduate Students Is Associated with Negative Health and Sleep Outcomes
Authors: Vallim Julia , Tsuji Heloisa , Lima Gabriela , Xylaras Beatriz , Louzada Fernando , Tufik Sergio , D'Almeida Vania 
Published: April 2024

Abstract

Introduction
The bimodal chronotype is a fourth circadian phenotype proposed by re-scoring the Morningness-Eveningness Questionnaire as first described by Martynhak and colleagues in 2010. Bimodal subjects are classified as “intermediate”, but they answer some of the questions as a morning person and others as an evening person. The present work aimed to describe the prevalence of the bimodal chronotype in a sample of undergraduate students in the city of Sao Paulo and to characterize the bimodal type in terms of their health and sleep-related outcomes.

Methods
A web-based cross-sectional study conducted between September 2018 and March 2021, using a convenience sampling method. The sample was composed of undergraduate students from two higher education institutions in Sao Paulo, who completed an electronic form that included the Morningness and Eveningness Questionnaire, the Pittsburgh Sleep Quality Index, the Self-Compassion Scale, the Epworth Sleepiness Scale, the Hospital Anxiety and Depression Scale, and the World Health Organization Subjective Well-Being Index. The final sample consisted of 615 students (82% female, mean age: 23.4±6.5 years).

Results
Of the 615 students, 108 (18%) had positive bimodality indexes. Bimodal subjects comprised 48 students (79% female, 19% male, 2% other gender), 8% of the total sample. Bimodal subjects had poorer subjective sleep quality (F(3,598)=20.0 p&lt; 0.001), more daytime sleepiness (F(3,611)=7.1 p&lt; 0.001), lower subjective well-being (F(3,611)=14.0 p&lt; 0.001), greater anxiety (F(3,611)=9.6 p&lt; 0.001) and depression symptoms (F(3,611)=11.8 p&lt; 0.001), and lower self-compassion (F(3,611)=10.1 p&lt; 0.001) than morning and/or intermediate types. However, they did not differ from evening types.

Conclusion
We found a bimodality prevalence of 8% and show that this profile is associated with negative health and sleep outcomes. The description of bimodality prevalence in a sample of college students is interesting because they are often an at-risk population for abnormal sleep and circadian disruption, which may be of interest for the design of academic public policies more consistent with the circadian reality of students.

Support (if any)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Associação Fundo de Incentivo à Pesquisa (AFIP).
0034 Sleep Is Associated with Telomere Shortening: A Population-Based Longitudinal Study
Authors: Tempaku Priscila , D'Almeida Vania , Andersen Monica , Tufik Sergio 
Published: April 2024

Abstract

Introduction
Evidence has shown that as the chronological age of an organism increases, there is a decrease in the telomere length (TL) via oxidative stress and inflammation pathways. Epidemiological studies have measured this parameter as the mean leukocyte TL and several associations between TL and age-related diseases have been described. Since the major pathophysiological factors related to inadequate sleep (including sleep complaints and sleep disorders) contribute to the exacerbation of inflammation and oxidative stress, the association of sleep and TL has been proposed in many cross-sectional studies. Thus, the aim of this study was to evaluate the association between sleep-related variables with TL in a population-based longitudinal framework.

Methods
We used data derived from the São Paulo Epidemiologic Sleep Study cohort, which was followed over 8 years. All individuals answered sleep-related questionnaires, underwent a full-night polysomnography (PSG) and had their blood collected for DNA extraction. TL was measured through quantitative real time polymerase chain reaction. For analysis in a single point in time, the Generalized Linear Model was used. The Generalized Estimating Equations test was applied for the comparison between baseline and follow-up, considering age, sex and body mass index (BMI) as covariables; and Spearman correlations were applied to compare the delta values between baseline and follow-up.

Results
Of the 1,042 individuals in the EPISONO cohort, 68.3% accepted to participate in the follow-up study (n=712). We have found that baseline SpO2 (ß=0.007, p=0.013), medium SpO2 (ß=0.009, p=0.004) and total sleep time &lt; 90% (ß=-0.142, p=0.037) had an effect on TL from the follow-up. The 8-year TL attrition was inversely associated with sleep quality, sleepiness, total sleep time, sleep efficiency, sleep architecture variables, wake after sleep onset, arousal index, oxygen-related variables baseline and the presence of obstructive sleep apnea (OSA).

Conclusion
Individuals with worse sleep quality, alterations in sleep architecture and OSA had greater TL attrition over the 8 years. Using a longitudinal approach, these findings confirm previous cross-sectional evidence linking sleep with accelerated biological aging.

Support (if any)
This work was supported by grants from AFIP, CNPq and FAPESP.
0035 Characterization of Metabolites in a Randomized, Controlled Trial of Clarithromycin for Pathological Sleepiness
Authors: Saini Prabhjyot , Keating Glenda , Rye David , Bliwise Donald , Ortlund Eric , Trotti Lynn Marie 
Published: April 2024

Abstract

Introduction
Pathologic sleepiness in CDH is associated with the presence of an endogenous, cerebrospinal fluid peptide that can positively, allosterically modulate GABA-A receptors. The macrolide antibiotic clarithromycin has been shown to benefit patients with hypersomnolence that is refractory to standard of care agents and is a negative allosteric modulator of GABA-A receptors. One explanation of its wake-promoting effects is the alteration of gastrointestinal microbiota composition. Here we characterize the metabolomics of serum and stool samples collected as part of a randomized, controlled trial of clarithromycin.

Methods
Ten patients and 10 Healthy controls had serum and stool samples taken on baseline study day and after a 2 week trial of clarithromycin dosed at 500mg bid, A total of 60 serum and stool samples were analyzed using Mass Spectrometry with a metabolomics panel. A total of 214 metabolites were detected, 122 in serum and 148 in stool with 56 overlapping metabolites.

Results
Samples were normalized by QC samples, log transformed and scaled separately for serum and stool. When comparing the 3 groups at the metabolite level, univariate analyses revealed 7 in serum and 4 in stool that had different levels between at nominal significance (p&lt; 0.05). The 4 in stool were 2-Deoxyadenosine, 3-Hydroxybenzaldehyde, Imidazole acetic acid and N-Acetyl aspartate. The first 2 were driven by lower values in the Treatment group vs Healthy Controls, and the latter 2 were the opposite. The 7 in serum were (R)-2,3-Dihydroxy-isovalerate, Homoserine, L-Allothreonine, L-Threonine, O-Acetyl-L-Serine, Palmitoyl carnitine, Riboflavin. Paired log2 fold-change analysis in serum between treatment and baseline revealed nominally significant decrease in the expression of Theobromine (P=0.4). In stool, the analysis revealed changes in 3-Hydroxybutyric acid, 2-Deoxyadenosine,3-2-Hydroxyethylindole,3-Hydroxy-2-Methylpyridine, Glycerol 2-phosphate, N-Acetylaspartate, Nicotinic Acid, Mononucleotide, Riboflavin, and Theobromine. PCA loadings of serum and stool samples, overall and in each group, revealed the first 2 PC’s consisting of 3-Hydroxybutyric acid and (R)-2,3-Dihydroxy-isovalerate

Conclusion
Despite similar profiles in serum and stool in patients, before and after treatment and healthy controls; subtle differences in particular metabolites may need to be further studied. Alternative pathways not currently captured in metabolomics in this study may be better suited to capture evidence of alternative mechanisms of clarithromycin efficacy.

Support (if any)
NS111280
0036 Exploring Common Genetic Factors Between Obstructive Sleep Apnea and Asthma: Insights into Inflammatory Pathways
Authors: Souza Maingredy , Moysés-Oliveira Mariana , Porcacchia Allan , Rosa Daniela , Tufik Sergio , Andersen Monica 
Published: April 2024

Abstract

Introduction
Obstructive sleep apnea (OSA) and asthma are globally prevalent respiratory disorders that share risk factors, symptoms and pathophysiological mechanisms. Emerging evidence suggests a bidirectional relationship, with each condition having a negative impact on the clinical course of the other. Although both diseases have a strong hereditary component, research assessing the genetic basis of their co-occurrence remains limited. Thus, the aim of this study was to analyze the interaction of risk genes and common biological pathways between OSA and asthma.

Methods
Two sets of genes associated with OSA (2,159 genes) and asthma (786 genes) were manually curated from significant single nucleotide polymorphisms (SNP) revealed in genome-wide association studies (GWAS). These lists were subsequently compared to identify intersecting genes, and the statistical significance of the overlap was assessed using Fisher's Exact Test. Pathway enrichment analysis was conducted utilizing the Benjamini-Hochberg test with a significance threshold set at an adjusted p-value&lt; 0.05.

Results
There were 187 overlapping genes between OSA and asthma gene sets, indicating a significantly higher occurrence than expected by chance (p&lt; 1.07E–29; odds ratio=2.3). The pathway overrepresentation analysis of these intersecting genes identified processes associated with immune system functions, encompassing human leucocyte antigen (HLA), antigen presentation, T cell differentiation, cell signaling, and positive regulation of inflammatory mediators.

Conclusion
The shared genetic basis between OSA and asthma suggests dysregulation of the immune response and pro-inflammatory processes as underlying pathophysiological mechanisms contributing to the susceptibility and manifestation of these comorbid conditions.

Support (if any)
Associação Fundo de Incentivo à Pesquisa (AFIP); Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).
0037 Investigating Common Genetic Factors Between Obstructive Sleep Apnea and Prostate Cancer
Authors: Porcacchia Allan , Moysés-Oliveira Mariana , Pires Gabriel , Andersen Monica , Tufik Sergio 
Published: April 2024

Abstract

Introduction
Cancer is one of the leading causes of death in the world. Studies have highlighted the possible association between obstructive sleep apnea (OSA) and an increased risk of incidence or prevalence of some types of cancer, especially prostate cancer. However, the extent to which genes that are related to both conditions contribute to their comorbidity is still unclear. The aim of this study was to evaluate the intersection of risk genes found in both OSA and prostate cancer.

Methods
Two sets of genes associated respectively with OSA and prostate cancer were manually curated and then compared to obtain an intersection gene list. Fisher’s exact test was employed to evaluate the statistical significance of the overlap, with a significance threshold of p-value&lt; 0.05. The identification of enriched pathways among the intersection gene list was performed using the Benjamini–Hochberg test, with a significance threshold of adjusted p-value&lt; 0.05. A functional network was generated from the intersect gene list which was used as an input in a protein–protein interaction analysis conducted using the String database.

Results
Sixty-eight genes overlapped between prostate cancer risk genes and OSA-associated genes, indicating more overlap than expected by chance (odds ratio=2.665; p=2.606E–10). This intersect list was associated mainly with hypoxia, apoptosis, oxidative stress, and cell cycle, proliferation, or cell damage. The hypoxia-inducible factor (HIF)-1 signaling pathway stands out from the list of enriched pathways reported in both conditions (adjusted p-value&lt; 0.0006). A 17-node network was retrieved from the overlapping gene list and included key proteins from the aforementioned enriched pathways, which comprised TNF, STAT3, BCL-2, TGF, and ATM.

Conclusion
The genetic basis of prostate cancer and OSA was significantly shared, indicating common molecular etiology in respect of this comorbidity. Relevant proliferation, apoptosis, and hypoxia pathways were significantly enriched among the genes associated with both conditions, suggesting their role in pathophysiological processes.

Support (if any)
Associação Fundo de Incentivo à Pesquisa (AFIP); Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).
0038 Kinesin Binding as a Shared Pathway Underlying the Genetic Basis of Male Infertility and Insomnia
Authors: Adami Luana , Moyses-Oliveira Mariana , Tufik Sergio , Andersen Monica 
Published: April 2024

Abstract

Introduction
Fertility problems are becoming increasingly common, with issues relating to men accounting for half of the cases. In addition to all the known causes of male infertility, some complaints are of idiopathic origin, and lifestyle, including sleep habits, might be an important underlying factor. Sleep disorders such as insomnia, which are prevalent in the population, might interfere in male fertility. Therefore, we aimed to perform an in silico study to test whether male infertility and insomnia share genetic risk variants, and to identify any molecular, cellular, and biological interactions between them.

Methods
Two lists of gene sets were manually curated through a literature review using PUBMED, 1 associated with male infertility (454 genes) and the other with insomnia (921 genes). The statistical significance of the gene list overlap was performed by the Fisher Exact test using Nematode Bioinformatics and Molbiotools online tools. This analysis considered a total of 21,196 genes in the human genome with a statistical threshold of P-value&lt; 0.05. The enriched terms over-represented in the intersect gene list were acquired from the Benjamini-Hochberg and the protein-protein interaction (PPI) analysis was performed via String database.

Results
Among these genes, 28 were common for both lists, representing more overlap than expected by chance (Fisher's exact test, p-value=0.041). The biological function of the genes contained in the intersection gene list suggested that pathways related to kinesin binding were commonly found in both conditions (p-value=0.038). A protein-protein interaction (PPI) analysis using the intersection list as input retrieved 25 nodes and indicated that 2 of them were kinesin-related proteins (PLEKHM2 and KCL1).

Conclusion
One of the major physiological mechanisms supporting the crosstalk between the sperm and the neuron through kinesis-related processes is mitochondria recycling. The intersecting gene list and the enriched pathways retrieved from our analysis might reflect how the rhythm control of neural circuits may affect spermatogenesis and male fertility. These results might serve as preliminary findings for future clinical trials focused on the identification of pharmacological interventions tailored to this comorbidity.

Support (if any)
AFIP, FAPESP, CNPq.
0041 Drug Screening and CRISPR/Cas9 Screening of HCN Channels
Authors: Smieszek Sandra , Doldur-Balli Fusun , Keenan Brendan , Zimmerman Amber , Veatch Olivia , Polymeropoulos Christos , Birznieks Gunther , Polymeropoulos Mihael 
Published: April 2024

Abstract

Introduction
A growing body of evidence suggests a role for HCN (hyperpolarization-activated cyclic nucleotide-gated) channels in regulation of sleep. We first tested effects of three HCN channel blockers (Corlanor, Zatebradine Hydrochloride and ZD7288) on sleep/wake behavior in wild type zebrafish compared to DMSO control. Then, to more definitively investigate roles of HCN channels, we knocked out genes encoding for HCN channel subunits in zebrafish by utilizing CRISPR/Cas9 technique and we assessed sleep/wake behavior in the HCN channel mutants.

Methods
The compounds were tested at six concentrations varying between 0.1-30 ◻M (.i.e., 0.1 μM, 0.3 μM, 1.0 μM, 4.5 μM, 10 μM and 30 μM) in zebrafish larvae in our drug screening study. Control groups were administered DMSO. In CRISPR screening, single cell-stage zebrafish embryos were injected with preformed ribonucleoprotein complexes containing Cas9 protein and two crRNAs targeting each homologous gene of human HCN1, HCN2, HCN3, and HCN4. Control groups were negative control crRNA injected embryos. Sleep phenotyping was performed by placing individual larva into the wells of 96 well plates and by recording their activity via commercially available video monitoring equipment.

Results
Drug screening study showed shorter latency to sleep at 0.1 μM dose of Ivabradine, moderate reductions in average activity at 30 μM dose of Zatebradine Hydrochloride, and increased sleep at 4.5 μM dose of ZD7288. Our CRISPR/Cas9 screening resulted in decreased activity in hcn1 crispants, fewer sleep bouts in daytime in hcn2 crispants, and no difference in sleep parameters in hcn3 crispants. hcn4 crispants had shorter sleep in both day and nighttime, hyperactivity and shorter sleep bouts at night and fewer sleep bouts in daytime.

Conclusion
Blocking HCN channels decreased wakefulness in our drug screening study. Genetically targeting hcn1 and hcn2 genes resulted in the same direction with the results of drug screening study and knocking out hcn3 didn’t change sleep parameters. Change in sleep parameters were in the opposite direction in hcn4 crispants. HCN channel blockers might be selecting HCN1 and HCN2 however further work is required to confirm this suggestion.

Support (if any)
0043 The Role of Median Preoptic Nucleus Astrocytes in Mediating E2’s Effects on Sleep-Wake Behavior
Authors: Onwukwe Chimdiya , Viechweg Shaun , Byrd Carissa , Mong Jessica 
Published: April 2024

Abstract

Introduction
Although 50-70 million Americans suffer from sleep disorders, women are 2x as likely as men to experience sleep disruptions. This discrepancy emerges at puberty and is strongly associated with fluctuations in estrogen, suggesting a role for estrogens in sleep disorders. Our previous work has identified the median preoptic nucleus (MnPO) as a key site regulating estradiol (E2) effects on sleep-wake behavior. We have also shown that E2 increases extracellular adenosine in the MnPO. Astrocytes are a major source of adenosine in the central nervous system, and cortical astrocytes have been shown to regulate sleep homeostasis. Thus, we hypothesize that MnPO astrocytes play a key role in the estrogenic mechanisms regulating sleep-wake behaviors.

Methods
To clarify the role of astrocytes in E2- modulation of sleep, we expressed Gi- or Gq-DREADDs in MnPO astrocytes of ovariectomized rats. Rats were treated with subcutaneous Oil injections (baseline) followed by injections of low (5ug) and high (10ug) dose E2, 24 hours apart. Rats also received Vehicle or Clozapine-N-oxide (CNO; 1.7mg/kg) at the time of their Oil/E2 injections. EEG/EMG recordings were acquired throughout treatment. To inhibit Ca2+signaling, and, ultimately, gliotransmission in astrocytes, we used a viral approach to express Pleckstrin Homology domain of Phospholipase C (PLC)-like protein p130 (p130PH) in MnPO astrocytes. p130PH is an IP3 buffer that prevents IP3 from initiating intracellular Ca2+ release and was previously shown to reduce astrocytic Ca2+signaling and gliotransmission (PMID: 20736051). Experimental (p130PH) and Control (tdTomato) animals were then treated with the Oil/E2 paradigm described above.

Results
In Oil-treated animals, CNO activation of Gq-DREADD increased wake and decreased sleep, mimicking E2’s effects on Sleep-Wake behavior. Unexpectedly, CNO activation of Gi-DREADD produced similar effects, increasing wake and decreasing sleep. These observations may support literature findings that Gi-DREADDs, while inhibitory in neurons, may enhance activity in astrocytes. Thus, Gi-DREADDs may not be an ideal candidate for inhibiting astrocytic activity. Animals expressing p130PH (n=3) exhibited less wake and more NREM sleep than animals injected with Control virus (n=3).

Conclusion
These preliminary findings suggest that E2 may increase wake by enhancing activity of MnPO astrocytes and highlight a need for further investigation.

Support (if any)
0044 Sleep Need-dependent Plasticity of a Thalamic Circuit Generates Persistence of Sleep Drive
Authors: Lee Sang Soo , Liu Qiang , Kim Thomas , Palmer Isabelle , Park Kristen , Muenzberg Heike , Blackshaw Seth , Wu Mark 
Published: April 2024

Abstract

Introduction
Prolonged wakefulness leads to persistent, deep recovery sleep. However, the neuronal circuits mediating this process remain elusive. Here, starting from a large circuit screen, we identify and characterize a group of thalamic nucleus reuniens (RE) neurons activated by sleep pressure and required for sleep homeostasis.

Methods
To identify upstream excitatory neuronal populations inducing NREM sleep, we performed rabies virus-mediated screening in mice. Sleep/wake states were assayed using tethered EEG/EMG recordings. Activation or inhibition of neuronal activity was performed using chemogenetic and/or optogenetic techniques. Behavior during optogenetic RE activation was assessed using video recordings. To genetically access the sleep pressure-activated RE neurons, Targeted Recombination in Active Populations (TRAP2) mice were used. Viral expression of caspase3 was used for selective ablation of cells. For projection analyses, virally expressed Synaptophysin was used as a presynaptic marker. To visually examine synaptic connectivity, we performed enhanced Green fluorescent protein Reconstruction Across Synaptic Partners (eGRASP) analysis. Functional connectivity between RE to downstream neurons was tested by slice patch-clamp recordings.

Results
From a circuit screen in mice, we identified a group of excitatory thalamic RE neurons activated by sleep pressure and required for sleep homeostasis. Optogenetic activation of these neurons leads to an unusual phenotype: sleep-preparatory behaviors (e.g., nesting) followed by prolonged, intense sleep resembling recovery sleep. The activity of RE neurons (assessed by Fos) is increased by sleep deprivation (SD), and ablation of these Fos+ RE cells markedly impairs recovery sleep following SD. Moreover, inhibiting RE activity during SD impairs subsequent recovery sleep, suggesting that these neurons signal sleep need. RE neurons act upstream of sleep-promoting zona incerta (ZI) cells, and, remarkably, SD triggers plasticity of this circuit to strengthen its connectivity, as assessed by eGRASP analysis and patch-clamp recordings. Finally, our results indicate that CaMKII signaling is required for morphological plasticity of the RE-ZI circuit and persistence of homeostatic NREM sleep.

Conclusion
A subset of RE neurons encode sleep pressure and generate persistent NREM sleep, via a projection to sleep-promoting ZI neurons. Increased sleep need triggers plasticity of the RE-ZI projection which enhances functional connectivity facilitating the transmission of homeostatic sleep drive.

Support (if any)
NIH grants: R01NS094571, R35NS122181, S10OD023548, NS05027