The Impact of Trauma-Sensitive Yoga on Persons Who Are Incarcerated: Identifying, Understanding, and Controlling Emotions.

BACKGROUND: While persons who are incarcerated have high rates of previous trauma, further traumatization can result from the experience of incarceration. The inability to effectively process trauma can lead to maladaptive behavior, a serious concern for correctional administrators. Acquiring the skills to regulate emotions and mitigate feelings of impulsivity help persons who are incarcerated take responsibility for their actions to make better decisions, simultaneously encouraging prosocial behavior, decreasing institutional misconduct, and reducing behaviors that place one at risk for repeated involvement in the criminal justice system. PURPOSE: Trauma-sensitive yoga (TSY) is one correctional intervention that can effectively address misconduct issues. Yet, the specific impact of TSY on populations who are incarcerated has not been empirically investigated. METHODS: A mixed-methods study, utilizing pretest and posttest interviewing, was conducted with male residents in a New York jail who completed a 4-week TSY program. RESULTS: Data indicated that those who participated in TSY experienced statistically significant improvements in self-esteem, self-control, resilience, institutional conduct, self-efficacy, and emotion regulation. Qualitative data also indicated high levels of connectivity with other TSY class participants. IMPLICATIONS: This research implies that cost-effective interventions like TSY have the ability to produce beneficial outcomes within a short time, which is integral to furthering rehabilitative efforts within the transient nature of the jail setting.

Coping with Depression, Anxiety & Stress: The Healing Effects of a Jail-Based Trauma Sensitive Yoga Program.

Stress, anxiety and depression, often linked to internalizing/externalizing behaviors, are remarkedly high in a jail-based setting when one's future is uncertain. While research demonstrates that carceral yoga programs can provide physiological/psychological benefits, persons who are incarcerated, who have high rates of trauma-related experiences and mental illness, might benefit more from a trauma sensitive approach. Empirical studies examining the specific impact of trauma sensitive yoga (TSY) on populations who are incarcerated appear unavailable, necessitating this exploratory investigation with male residents in a TSY intervention at a New York jail. Through pre and post-test interviewing and a mixed methods data analysis, data indicated that those who participated in TSY experienced statistically significant increases in overall health, in addition to statistically significant reductions in stress, anxiety, depression and institutional misconduct. Qualitative analyses suggested that male participants experienced transcendence of the jail environment, easing the monotony of the correctional setting. TSY also provided them with new opportunities, the ability to regulate emotions/behaviors and initiate personal growth/changes within. Hence, this research implies that TSY, a benign intervention, easily implemented in a correctional setting, has the ability to beget benefits for persons who are incarcerated within a short period of time. This is vital to providing rehabilitative options within the transient nature of the jail setting.

Enrichment analysis of phenotypic data for drug repurposing in rare diseases.

Drug-induced Behavioral Signature Analysis (DBSA), is a machine learning (ML) method for in silico screening of compounds, inspired by analytical methods quantifying gene enrichment in genomic analyses. When applied to behavioral data it can identify drugs that can potentially reverse in vivo behavioral symptoms in animal models of human disease and suggest new hypotheses for drug discovery and repurposing. We present a proof-of-concept study aiming to assess Drug-induced Behavioral Signature Analysis (DBSA) as a systematic approach for drug discovery for rare disorders. We applied Drug-induced Behavioral Signature Analysis to high-content behavioral data obtained with SmartCube®, an automated in vivo phenotyping platform. The therapeutic potential of several dozen approved drugs was assessed for phenotypic reversal of the behavioral profile of a Huntington's Disease (HD) murine model, the Q175 heterozygous knock-in mice. The in silico Drug-induced Behavioral Signature Analysis predictions were enriched for drugs known to be effective in the symptomatic treatment of Huntington's Disease, including bupropion, modafinil, methylphenidate, and several SSRIs, as well as the atypical antidepressant tianeptine. To validate the method, we tested acute and chronic effects of tianeptine (20 mg/kg, i. p.) in vivo, using Q175 mice and wild type controls. In both experiments, tianeptine significantly rescued the behavioral phenotype assessed with the SmartCube® platform. Our target-agnostic method thus showed promise for identification of symptomatic relief treatments for rare disorders, providing an alternative method for hypothesis generation and drug discovery for disorders with huge disease burden and unmet medical needs.

Reversible hypogonadotropic hypogonadism in men with the fertile eunuch/Pasqualini syndrome: A single-center natural history study.

Congenital hypogonadotropic hypogonadism (HH) is a heterogeneous genetic disorder characterized by disrupted puberty and infertility. In most cases, HH is abiding yet 10-15% undergo reversal. Men with HH and absent and partial puberty (i.e., testicular volume <4mL and >4mL respectively) have been well-studied, but the rare fertile eunuch (FE) variant remains poorly characterized. This natural history study of 240 men with HH delineates the clinical presentation, neuroendocrine profile, rate of reversal and genetics of the FE variant. We compared three HH groups: FE (n=38), absent puberty (n=139), and partial puberty (n=63). The FE group had no history of micropenis and 2/38 (5%) had cryptorchidism (p<0.0001 vs. other groups). The FE group exhibited higher rates of detectable gonadotropins, higher mean LH/FSH levels, and higher serum inhibin B levels (all p<0.0001). Neuroendocrine profiling showed pulsatile LH secretion in 30/38 (79%) of FE men (p<0.0001) and 16/36 (44%) FE men underwent spontaneous reversal of HH (p<0.001). The FE group was enriched for protein-truncating variants (PTVs) in GNRHR and FGFR1 and 4/30 (13%) exhibited oligogenic PTVs. Findings suggest men with the FE variant exhibit the mildest neuroendocrine defects of HH men and the FE sub-type represents the first identified phenotypic predictor for reversible HH.

Coupling-dependent metabolic ultradian rhythms in confluent cells.

Ultradian rhythms in metabolism and physiology have been described previously in mammals. However, the underlying mechanisms for these rhythms are still elusive. Here, we report the discovery of temperature-sensitive ultradian rhythms in mammalian fibroblasts that are independent of both the cell cycle and the circadian clock. The period in each culture is stable over time but varies in different cultures (ranging from 3 to 24 h). We show that transient, single-cell metabolic pulses are synchronized into stable ultradian rhythms across contacting cells in culture by gap junction-mediated coupling. Coordinated rhythms are also apparent for other metabolic and physiological measures, including plasma membrane potential (Δψp), intracellular glutamine, α-ketoglutarate, intracellular adenosine triphosphate (ATP), cytosolic pH, and intracellular calcium. Moreover, these ultradian rhythms require extracellular glutamine, several different ion channels, and the suppression of mitochondrial ATP synthase by α-ketoglutarate, which provides a key feedback mechanism. We hypothesize that cellular coupling and metabolic feedback can be used by cells to balance energy demands for survival.

Dialogic Health Education to Reduce COVID-19 Disparities and Increase Health Literacy in Community and Correctional Settings: Protocol for a Two-Pronged Health Education Program.

BACKGROUND: COVID-19 vaccines significantly reduce rates of hospitalization and death for those infected with the SARS-CoV-2 virus. Those facing social oppression, including people of color, experience heightened risk for COVID-19 and comorbidities, but are often mistrustful of governmental agencies and initiatives, contributing to low vaccine uptake and a reluctance to access vital health care services. Dialogue-based health literacy interventions may mitigate mistrust and increase access to health services and information, subsequently increasing rates of vaccination and other behaviors that reduce COVID-19 risk. OBJECTIVE: To improve health literacy and reduce COVID-19 disparities, the Westchester County Department of Health, in partnership with two universities, community- and faith-based organizations, and the Westchester County Department of Correction, co-developed a health education program for community members, correctional officers, and incarcerated jail residents in Westchester, New York. Specific objectives are to increase preventative health behaviors, positive attitudes toward use of public health protocols, full vaccination or intentions to vaccinate, health care information understanding, health provider care access, clear communication with health care providers, and personal health care decision-making. METHODS: Grounded in dialogic learning, the program entails training community-based "trusted messengers" and correctional officers to lead health information sessions in community and correctional settings. During the grant period, the program intends for 80 community-based trusted messengers to receive training from the Department of Health and will be expected to reach a goal of 100 members (N=8000) of their communities. Correctional staff with experience delivering educational programs will be trained to facilitate sessions among 400 correctional facility residents and 600 correctional staff. RESULTS: Pre-post surveys will assess changes in health behaviors, attitudes, and perceptions. The program has been administered in the correctional facility since February 2022, with information sessions expected to cease for correctional staff and residents in June 2022 and November 2022, respectively. An initial cohort of community-based trusted messengers began training in February 2022, and information sessions have been scheduled in various virtual and community settings since March 2022. As of April 2022, the two-pronged health education program has reached 439 correctional officers, 98 jail residents, and 201 community members countywide. Program evaluation findings will be released in future publications after study implementation is complete. CONCLUSIONS: Few studies have evaluated the combined effects of training-of-trainers (ToT) and dialogical learning models on behavior and health literacy. As the first known COVID-19-specific dialogue-based health education program that applies a ToT model in the community-based, correctional, and virtual settings simultaneously, this study fills a gap in current knowledge about health literacy and health behavior in marginalized populations. Thus, this evidence-based framework can remedy COVID-19 disparities while also addressing risks for a host of health-related issues at the community level, potentially serving as a best-practice model for future health programs. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/37713.

Introduction to the Clock System.

Circadian (24-h) rhythms dictate almost everything we do, setting our clocks for specific times of sleeping and eating, as well as optimal times for many other basic functions. The physiological systems that coordinate circadian rhythms are intricate, but at their core, they all can be distilled down to cell-autonomous rhythms that are then synchronized within and among tissues. At first glance, these cell-autonomous rhythms may seem rather straight-forward, but years of research in the field has shown that they are strikingly complex, responding to many different external signals, often with remarkable tissue-specificity. To understand the cellular clock system, it is important to be familiar with the major players, which consist of pairs of proteins in a triad of transcriptional/translational feedback loops. In this chapter, we will go through each of the core protein pairs one-by-one, summarizing the literature as to their regulation and their broader impacts on circadian gene expression. We will conclude by briefly examining the human genetics literature, as well as providing perspectives on the future of the study of the molecular clock.

Do as I Say, Not What I Do: The Practices of HIV Prison Peer Educators.

The rate of new HIV infections has declined over the past several years. Mass education pertaining to preventive measures is often credited for its decline, particularly among incarcerated populations where the infection rates are often higher than in the general community. Interestingly, those tasked with providing the education may not always practice the preventive measures taught. Interviews were conducted with 49 incarcerated women who worked in two HIV prison-based peer programs during their incarceration. Responses indicated that although participants were comfortable discussing their sex/drug histories with partners or potential partners, approximately half of participants did not practice safer sex consistently during/after the time they were working as HIV peer educators.

NPAS4 regulates the transcriptional response of the suprachiasmatic nucleus to light and circadian behavior.

The suprachiasmatic nucleus (SCN) is the master circadian pacemaker in mammals and is entrained by environmental light. However, the molecular basis of the response of the SCN to light is not fully understood. We used RNA/chromatin immunoprecipitation/single-nucleus sequencing with circadian behavioral assays to identify mouse SCN cell types and explore their responses to light. We identified three peptidergic cell types that responded to light in the SCN: arginine vasopressin (AVP), vasoactive intestinal peptide (VIP), and cholecystokinin (CCK). In each cell type, light-responsive subgroups were enriched for expression of neuronal Per-Arnt-Sim (PAS) domain protein 4 (NPAS4) target genes. Further, mice lacking Npas4 had a longer circadian period under constant conditions, a damped phase response curve to light, and reduced light-induced gene expression in the SCN. Our data indicate that NPAS4 is necessary for normal transcriptional responses to light in the SCN and critical for photic phase-shifting of circadian behavior.

Sleeping Sickness Disrupts the Sleep-Regulating Adenosine System.

Patients with sleeping sickness, caused by the parasite Trypanosoma brucei, have disruptions in both sleep timing and sleep architecture. However, the underlying cause of these sleep disturbances is not well understood. Here, we assessed the sleep architecture of male mice infected with T. brucei and found that infected mice had drastically altered sleep patterns. Interestingly, T. brucei-infected mice also had a reduced homeostatic sleep response to sleep deprivation, a response modulated by the adenosine system. We found that infected mice had a reduced electrophysiological response to an adenosine receptor antagonist and increased adenosine receptor gene expression. Although the mechanism by which T. brucei infection causes these changes remains to be determined, our findings suggest that the symptoms of sleeping sickness may be because of alterations in homeostatic adenosine signaling.SIGNIFICANCE STATEMENT Sleeping sickness is a fatal disease that disrupts the circadian clock, causes disordered temperature regulation, and induces sleep disturbance. To examine the neurologic effects of infection in the absence of other symptoms, in this study, we used a mouse model of sleeping sickness in which the acute infection was treated but brain infection remained. Using this model, we evaluated the effects of the sleeping sickness parasite, Trypanosoma brucei, on sleep patterns in mice, under both normal and sleep-deprived conditions. Our findings suggest that signaling of adenosine, a neuromodulator involved in mediating homeostatic sleep drive, may be reduced in infected mice.

Dual-Color Single-Cell Imaging of the Suprachiasmatic Nucleus Reveals a Circadian Role in Network Synchrony.

The suprachiasmatic nucleus (SCN) acts as a master pacemaker driving circadian behavior and physiology. Although the SCN is small, it is composed of many cell types, making it difficult to study the roles of particular cells. Here we develop bioluminescent circadian reporter mice that are Cre dependent, allowing the circadian properties of genetically defined populations of cells to be studied in real time. Using a Color-Switch PER2::LUCIFERASE reporter that switches from red PER2::LUCIFERASE to green PER2::LUCIFERASE upon Cre recombination, we assess circadian rhythms in two of the major classes of peptidergic neurons in the SCN: AVP (arginine vasopressin) and VIP (vasoactive intestinal polypeptide). Surprisingly, we find that circadian function in AVP neurons, not VIP neurons, is essential for autonomous network synchrony of the SCN and stability of circadian rhythmicity.

Mouse model systems of autism spectrum disorder: Replicability and informatics signature.

Phenotyping mouse model systems of human disease has proven to be a difficult task, with frequent poor inter- and intra-laboratory replicability, particularly in behavioral domains such as social and cognitive function. However, establishing robust animal model systems with strong construct validity is of fundamental importance as they are central tools for understanding disease pathophysiology and developing therapeutics. To complete our studies of mouse model systems relevant to autism spectrum disorder (ASD), we present a replication of the main findings from our two published studies of five genetic mouse model systems of ASD. To assess the intra-laboratory robustness of previous results, we chose the two model systems that showed the greatest phenotypic differences, the Shank3/F and Cntnap2, and repeated assessments of general health, activity and social behavior. We additionally explored all five model systems in the same framework, comparing all results obtained in this three-yearlong effort using informatics techniques to assess commonalities and differences. Our results showed high intra-laboratory replicability of results, even for those with effect sizes that were not particularly large, suggesting that discrepancies in the literature may be dependent on subtle but pivotal differences in testing conditions, housing enrichment, or background strains and less so on the variability of the behavioral phenotypes. The overall informatics analysis suggests that in our behavioral assays we can separate the set of tested mouse model system into two main classes that in some aspects lie on opposite ends of the behavioral spectrum, supporting the view that autism is not a unitary concept.

Epigenetic inheritance of circadian period in clonal cells.

Circadian oscillations are generated via transcriptional-translational negative feedback loops. However, individual cells from fibroblast cell lines have heterogeneous rhythms, oscillating independently and with different period lengths. Here we showed that heterogeneity in circadian period is heritable and used a multi-omics approach to investigate underlying mechanisms. By examining large-scale phenotype-associated gene expression profiles in hundreds of mouse clonal cell lines, we identified and validated multiple novel candidate genes involved in circadian period determination in the absence of significant genomic variants. We also discovered differentially co-expressed gene networks that were functionally associated with period length. We further demonstrated that global differential DNA methylation bidirectionally regulated these same gene networks. Interestingly, we found that depletion of DNMT1 and DNMT3A had opposite effects on circadian period, suggesting non-redundant roles in circadian gene regulation. Together, our findings identify novel gene candidates involved in periodicity, and reveal DNA methylation as an important regulator of circadian periodicity.

Noise-driven cellular heterogeneity in circadian periodicity.

Nongenetic cellular heterogeneity is associated with aging and disease. However, the origins of cell-to-cell variability are complex and the individual contributions of different factors to total phenotypic variance are still unclear. Here, we took advantage of clear phenotypic heterogeneity of circadian oscillations in clonal cell populations to investigate the underlying mechanisms of cell-to-cell variability. Using a fully automated tracking and analysis pipeline, we examined circadian period length in thousands of single cells and hundreds of clonal cell lines and found that longer circadian period is associated with increased intercellular heterogeneity. Based on our experimental results, we then estimated the contributions of heritable and nonheritable factors to this variation in circadian period length using a variance partitioning model. We found that nonheritable noise predominantly drives intercellular circadian period variation in clonal cell lines, thereby revealing a previously unrecognized link between circadian oscillations and intercellular heterogeneity. Moreover, administration of a noise-enhancing drug reversibly increased both period length and variance. These findings suggest that circadian period may be used as an indicator of cellular noise and drug screening for noise control.

Prisoner Parenting: Parenting From a Federal Jail.

While mothers are incarcerated, the disruption to the bond with their children places both at risk for emotional and behavioral problems. Parenting skills education can help to restore these bonds. Utilizing a pre- and posttest quasi-experimental design, we evaluated the effects of an evidence-based parenting program on females housed in a federal jail managed by the Federal Bureau of Prisons. Findings indicate statistically significant decreases in depression, parental stress, and anxiety, and increased self-esteem. Participants also had increased contact with their children and reported more confidence in their parenting skills. Results imply that programs developed for women that enhance parenting skills and improve relationships with their children and/or children's caregivers are important jail-based interventions.

Tissue-specific FAH deficiency alters sleep-wake patterns and results in chronic tyrosinemia in mice.

Fumarylacetoacetate hydrolase (FAH) is the last enzyme in tyrosine catabolism, and mutations in the FAH gene are associated with hereditary tyrosinemia type I (HT1 or TYRSN1) in humans. In a behavioral screen of N-ethyl-N-nitrosourea mutagenized mice we identified a mutant line which we named "swingshift" (swst, MGI:3611216) with a nonsynonymous point mutation (N68S) in Fah that caused age-dependent disruption of sleep-wake patterns. Mice homozygous for the mutation had an earlier onset of activity (several hours before lights off) and a reduction in total activity and body weight when compared with wild-type or heterozygous mice. Despite abnormal behavioral entrainment to light-dark cycles, there were no differences in the period or phase of the central clock in mutant mice, indicating a defect downstream of the suprachiasmatic nucleus. Interestingly, these behavioral phenotypes became milder as the mice grew older and were completely rescued by the administration of NTBC [2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione], an inhibitor of 4-hydroxyphenylpyruvate dioxygenase, which is upstream of FAH. Mechanistically, the swst mutation had no effect on the enzymatic activity of FAH, but rather promoted the degradation of the mutant protein. This led to reduced FAH protein levels and enzymatic activity in the liver and kidney (but not the brain or fibroblasts) of homozygous mice. In addition, plasma tyrosine-but not methionine, phenylalanine, or succinylacetone-increased in homozygous mice, suggesting that swst mutants provide a model of mild, chronic HT1.

Circadian clock genes and the transcriptional architecture of the clock mechanism.

The mammalian circadian clock has evolved as an adaptation to the 24-h light/darkness cycle on earth. Maintaining cellular activities in synchrony with the activities of the organism (such as eating and sleeping) helps different tissue and organ systems coordinate and optimize their performance. The full extent of the mechanisms by which cells maintain the clock are still under investigation, but involve a core set of clock genes that regulate large networks of gene transcription both by direct transcriptional activation/repression as well as the recruitment of proteins that modify chromatin states more broadly.

Neuronal Myocyte-Specific Enhancer Factor 2D (MEF2D) Is Required for Normal Circadian and Sleep Behavior in Mice.

The transcription factor, myocyte enhancer factor-2 (MEF2), is required for normal circadian behavior in Drosophila; however, its role in the mammalian circadian system has not been established. Of the four mammalian Mef2 genes, Mef2d is highly expressed in the suprachiasmatic nucleus (SCN) of the hypothalamus, a region critical for coordinating peripheral circadian clocks. Using both conventional and brain-specific Mef2d KO (Mef2d-/-) mouse lines, we demonstrate that MEF2D is essential for maintaining the length of the circadian free-running period of locomotor activity and normal sleep patterns in male mice. Crossing Mef2d-/- with Per2::luc reporter mice, we show that these behavioral changes are achieved without altering the endogenous period of the master circadian oscillator in the SCN. Together, our data suggest that alterations in behavior in Mef2d-/- mice may be the result of an effect on SCN output, rather than an effect on timekeeping within the SCN itself. These findings add to the growing body of evidence that MEF2 proteins play important roles in the brain.SIGNIFICANCE STATEMENT These studies are the first to show a role for MEF2 proteins in the brain outside of the hippocampus, and our findings suggest that these proteins may play diverse roles in the CNS. It is important to continue to build on our understanding of the roles of proteins acting in the SCN because SCN dysfunction underlies jet lag in humans and influences the response to shift work schedules, which are now known as risk factors for the development of cancer. Our work on MEF2D could be the basis for opening new lines of research in the development and regulation of circadian rhythms.

A Hyperkinetic Redox Sensor Drives Flies to Sleep.

Several different potassium channels modulate the activity of sleep-promoting neurons in the Drosophila brain, but the regulation of these channels is not completely understood. A recent study (Kempf et al., Nature, 2019) found that one of the potassium channel subunits, Hyperkinetic, alters the firing properties of sleep-promoting neurons in response to NADPH oxidation. These findings are the first to link cellular redox state and mitochondrial metabolism directly to sleep.

A novel mutation in Slc2a4 as a mouse model of fatigue.

Chronic fatigue is a debilitating disorder with widespread consequences, but effective treatment strategies are lacking. Novel genetic mouse models of fatigue may prove invaluable for studying its underlying physiological mechanisms and for testing treatments and interventions. In a screen of voluntary wheel-running behavior in N-ethyl-N-nitrosourea mutagenized C57BL/6J mice, we discovered two lines with low body weights and aberrant wheel-running patterns suggestive of a fatigue phenotype. Affected progeny from these lines had lower daily activity levels and exhibited low amplitude circadian rhythm alterations. Their aberrant behavior was characterized by frequent interruptions and periods of inactivity throughout the dark phase of the light-dark cycle and increased levels of activity during the rest or light phase. Expression of the behavioral phenotypes in offspring of strategic crosses was consistent with a recessive inheritance pattern. Mapping of phenotypic abnormalities showed linkage with a single locus on chromosome 1, and whole exome sequencing identified a single point mutation in the Slc2a4 gene encoding the GLUT4 insulin-responsive glucose transporter. The single nucleotide change (A-T, which we named "twiggy") was in the distal end of exon 10 and resulted in a premature stop (Y440*). Additional metabolic phenotyping confirmed that these mice recapitulate phenotypes found in GLUT4 knockout mice. However, to the best of our knowledge, this is the first time a mutation in this gene has been shown to result in extensive changes in general behavioral patterns. These findings suggest that GLUT4 may be involved in circadian behavioral abnormalities and could provide insights into fatigue in humans.