Nervous System Commons^™

Cortisol Production In Female Soccer Players, Katelyn Seagraves

ASPIRE 2024

Cortisol is released from the adrenal glands when there is an apparent stressor. Exercise, while beneficial, is seen as stress to the body and incites increased cortisol release. Cortisol is known to increase with exercise, and higher-intensity exercise tends to raise cortisol more compared to lower intensities. However, less is known about interval training and cortisol concentrations.

This review of literature dives into the background of cortisol: how it's produced, what systems it regulates, and differences in production based on gender, stressors, exercise intensity, and duration. This paper identifies some knowledge gaps and links training intensities in soccer to cortisol …

Zika Virus Microcephaly: Predicting The Symptoms Of Microcephaly So Families Can Prepare, Eleanor J. Wind

Mako: NSU Undergraduate Student Journal

Microcephaly is a rare congenital defect where the circumference of an infants head is smaller than average. The cause is unknown and the risk factors and symptoms of microcephaly are wide ranging. One risk factor is exposure during pregnancy to viruses such as the Zika virus.. A recent study was conducted on infants who were born with Zika virus Microcephaly following the Zika outbreak in Brazil. The researchers conducted MRI scans and developmental testing. Then an algorithm to predict behavioral outcomes of Zika Virus Microcephaly was successfully created. This research will benefit caretakers of infants with Zika virus Microcephaly by …

Adolescent Mental Health In The Twenty-First Century, Arthur Maerlander, Patricia Gail Bray

Center for Brain, Biology, and Behavior: Faculty Publications

Conclusion

Parents do not suddenly relinquish medical decision-making to their children and are a vital part of a process construed within the framework of consent and confidentiality laws. In the transition period from childhood to adulthood, adolescents develop unique healthcare needs and concerns at the very time they are gaining autonomy from their parents. Adolescents progress from being recipients of decisions about their care to gradually becoming active participants in their healthcare and emerge as a primary partner in medical decision-making. Building a strong therapeutic alliance with young people can help them become responsible healthcare decision-makers.

Yet while “healthcare,” including …

Associations Of Sex And Sport Contact‑Level With Recovery Timelines Among Collegiate Athletes With Sport‑Related Concussion, Bernadette A. D'Alonzo, Andrea L. C. Schneider, Ian J. Barnett, Christina L. Master, Abigail C. Bretzin, Douglas J. Wiebe, Ivy League-Big Ten Epidemiology Of Concussion Study Investigators

Center for Brain, Biology, and Behavior: Faculty Publications

Background

Growing interest has motivated recent studies to examine differences in recovery after sportrelated concussion (SRC) by sex. However, heterogeneity in study design, participants, and recovery outcomes has led to mixed findings. Further work is needed to evaluate potential differences by sex and to investigate the role of related characteristics, such as sport contact-level, in recovery timelines. This study aimed to investigate whether concussion recovery trajectories differ by sex, considering a priori clinical and demographic covariates, and accounting for the sequence of recovery outcomes. Our secondary question was whether sport contact-level modifies the relationship between sex and time to outcomes. …

Outpatient Fall Prevention In Ambulatory Adults 65 Years Old And Over, Dorothy L. Osborne-White

Doctor of Nursing Practice (DNP) Scholarly Projects - Archive

Background: In the United States (U.S.), falls are the leading cause of injury among adults 65 and over, resulting in 36 million falls yearly (Moreland et al., 2020). According to the Centers for Disease Control and Prevention (CDC, 2023), one in four older adults experiences a fall each year. Falls are the world's second most prominent cause of accidental deaths (World Health Organization [WHO], 2021). Falls are the leading cause of both fatal and non-fatal injuries among older adults (Moreland et al., 2020).

Methods: A quality improvement project that included a fall bundle was implemented in a primary clinic. A …

An Objective Assessment Of Neuromotor Control Using A Smartphone App After Repeated Subconcussive Blast Exposure, Charlend K. Howard, Masahiro Yamada, Marcia Dovel, Rie Leverett, Alexander Hill, Kenneth A. Manlapaz, David O. Keyser, Rene S. Hernandez, Sheilah S. Rowe, Walter S. Carr, Michael S. Roy, Christopher K. Rhea

Rehabilitation Sciences Faculty Publications

Subconcussive blast exposure has been shown to alter neurological functioning. However, the extent to which neurological dysfunction persists after blast exposure is unknown. This longitudinal study examined the potential short- and long-term effects of repeated subconcussive blast exposure on neuromotor performance from heavy weapons training in military personnel. A total of 214 participants were assessed; 137 were exposed to repeated subconcussive blasts and 77 were not exposed to blasts (controls). Participants completed a short stepping-in-place task while an Android smartphone app placed on their thigh recorded movement kinematics. We showed acute suppression of neuromotor variability 6 h after subconcussive blast …

Central Activation Of Orl-1 Receptors In Heart Failure Models, Jeffrey Angell

Theses & Dissertations

Heart failure is a prevalent and debilitating disease associated with high morbidity and mortality rates worldwide and is the leading cause of death in the United States. While various therapeutic strategies have been developed and approved to manage heart failure, exploration of novel targets and mechanisms are essential for improving patient outcomes. This thesis investigates the therapeutic potential of the central activation of opioid receptor-like 1 (ORL-1) by nociceptin in a rodent heart failure model.

Furthermore, this study aims to elucidate the central mechanisms underlying nociceptin induced cardiovascular and renal effects in heart failure. This involves investigating neuronal pathways and …

Covid‑19‑Related Risk, Resilience, And Mental Health Among Mexican American Mothers Across The First Year Of The Pandemic, Amy L. Non, Elizabeth S. Clausing, Sandraluz Lara‑Cinisomo, Kimberly L. D’Anna Hernandez

Center for Brain, Biology, and Behavior: Faculty Publications

Background Latina mothers have been especially affected by the pandemic and historically exhibit high rates of depression and anxiety. However, few longitudinal studies have assessed the effect of the pandemic on this vulnerable population. We hypothesized that COVID-19-related stressors would associate with psychological distress among Latina mothers across the first year of the pandemic.

Methods We investigated COVID-19-related impact, stigma, and fears across two critical time points and changes in these measures in relation to changes in maternal anxiety and depression among mothers of Mexican descent living in Southern California (n=152). Surveys were administered within 5–16 weeks of …

Brain Microvascular Endothelial Cells Possess A Second Cilium That Arises From The Daughter Centriole, Karthikeyan Thirugnanam, Ankan Gupta, Francisco Nunez, Shubhangi Prabhudesai, Amy Y. Pan, Surya M. Nauli, Ramani Ramchandran

Pharmacy Faculty Articles and Research

Primary cilia from the brain microvascular endothelial cells (ECs) are specialized cell-surface organelles involved in mediating sensory perception, cell signaling, and vascular stability. Immunofluorescence (IF) analysis of human primary brain microvascular ECs reveals two cilia per cell. To confirm the in vitro observation of the two-cilia phenotype in human primary brain ECs, ECs isolated from mouse brain were cultured and stained for cilium. Indeed, brain ECs from a ciliopathic mouse (polycystic kidney disease or Pkd2−/−) also possess more than one cilium. Primary cilium emerges from the mother centriole. Centriole analysis by IF suggests that in brain ECs, markers …

Reduced Grey Matter Volume In Adolescents With Conduct Disorder: A Region‑Of‑Interest Analysis Using Multivariate Generalized Linear Modeling, Ru Zhang, Robert James R. Blair, Karina S. Blair, Matthew Dobbertin, Jaimie Elowsky, Johannah Bashford‑Largo, Ahria J. Dominguez, Melissa Hatch, Sahil Bajaj

Center for Brain, Biology, and Behavior: Faculty Publications

Background

Conduct disorder (CD) involves a group of behavioral and emotional problems that usually begins during childhood or adolescence. Structural brain alterations have been observed in CD, including the amygdala, insula, ventrolateral and medial prefrontal cortex, anterior cingulate cortex, and fusiform gyrus. The current study developed a multivariate generalized linear model (GLM) to differentiate adolescents with CD from typically developing (TD) adolescents in terms of grey matter volume (GMV).

Methods

The whole‐brain structural MRI data were collected from 96 adolescents with CD (mean age = 16.188 ± 1.259 years; mean IQ = 104.292 ± 8.107 ; 63 males) and 90 …

Specialized Late Cingulo-Opercular Network Activation Elucidates The Mechanisms Underlying Decisions About Ambiguity, Jordan E. Pierce, Nathan M. Petro, Elizabeth Clancy, Caterina Gratton, Steven E. Petersen, Maital Neta

Center for Brain, Biology, and Behavior: Faculty Publications

Cortical task control networks, including the cingulo-opercular (CO) network play a key role in decision-making across a variety of functional domains. In particular, the CO network functions in a performance reporting capacity that supports successful task performance, especially in response to errors and ambiguity. In two studies testing the contribution of the CO network to ambiguity processing, we presented a valence bias task in which masked clearly and ambiguously valenced emotional expressions were slowly revealed over several seconds. This slow reveal task design provides a window into the decision-making mechanisms as they unfold over the course of a trial. In …

Assessing Blood Oxygen Level–Dependent Signal Variability As A Biomarker Of Brain Injury In Sport-Related Concussion, Evan D. Anderson, Tanveer Talukdar, Grace Goodwin, Valentina Di Pietro, Kamal M. Yakoub, Christopher E. Zwilling, David Davies, Antonio Belli, Aron K. Barbey

Center for Brain, Biology, and Behavior: Faculty Publications

Mild traumatic brain injury is a complex neurological disorder of significant concern among athletes who play contact sports. Athletes who sustain sport-related concussion typically undergo physical examination and neurocognitive evaluation to determine injury severity and return-to-play status. However, traumatic disruption to neurometabolic processes can occur with minimal detectable anatomic pathology or neurocognitive alteration, increasing the risk that athletes may be cleared for return-to-play during a vulnerable period and receive a repetitive injury. This underscores the need for sensitive functional neuroimaging methods to detect altered cerebral physiology in concussed athletes. The present study compared the efficacy of Immediate Post-concussion Assessment and …

Identification Of Structural Brain Alterations In Adolescents With Depressive Symptomatology, J. Bashford‑Largo, R. James R. Blair, Karina S. Blair, Matthew Dobbertin, Ahria Dominguez, Melissa Hatch, Sahil Bajaj

Center for Brain, Biology, and Behavior: Faculty Publications

Introduction: Depressive symptoms can emerge as early as childhood and may lead to adverse situations in adulthood. Studies have examined structural brain alternations in individuals with depressive symptoms, but findings remain inconclusive. Furthermore, previous studies have focused on adults or used a categorical approach to assess depression. The current study looks to identify grey matter volumes (GMV) that predict depressive symptomatology across a clinically concerning sample of adolescents.

Methods: Structural MRI data were collected from 338 clinically concerning adolescents (mean age = 15.30 SD=2.07; mean IQ = 101.01 SD=12.43; 132 F). Depression symptoms were indexed via the Mood …

Novel Insights Into Oligodendrocyte Biology From Developmental Myelination Studies In Autophagy Deficient Mice And Analysis Of Oligodendrocyte Translatome Response To Contusive Spinal Cord Injury., Michael David Forston

Electronic Theses and Dissertations

Loss of myelin causes severe neurological disorders and functional deficits in white matter injuries (WMI) such as traumatic spinal cord injury (SCI). This dissertation is focused on autophagy in OL development and the OL translatome after SCI. Chapter I describes the history of myelin, OL development, and their involvement in neurodegenerative diseases and SCI. The proteostasis network, in particular autophagy, and its contributions to white matter pathology is discussed. It concludes examining advantages and disadvantages of unbiased omics tools, like RiboTag, to study transcriptional/translational landscapes after SCI. Chapter II focuses on autophagy in OPC/OL differentiation, survival, and proper myelination in …

Specialized Late Cingulo-Opercular Network Activation Elucidates The Mechanisms Underlying Decisions About Ambiguity, Jordan E. Pierce, Nathan M. Petro, Elizabeth Clancy, Caterina Gratton, Steven E. Petersen, Maital Neta

Center for Brain, Biology, and Behavior: Faculty Publications

Cortical task control networks, including the cingulo-opercular (CO) network play a key role in decision-making across a variety of functional domains. In particular, the CO network functions in a performance reporting capacity that supports successful task performance, especially in response to errors and ambiguity. In two studies testing the contribution of the CO network to ambiguity processing, we presented a valence bias task in which masked clearly and ambiguously valenced emotional expressions were slowly revealed over several seconds. This slow reveal task design provides a window into the decision-making mechanisms as they unfold over the course of a trial. In …

Investigating Sexual Dimorphism In Lateral Hypothalamic Melanin-Concentrating Hormone Neuron Subpopulations, Mahitha Juttu

Honors Scholar Theses

The lateral hypothalamic area (LHA) is an area crucial for coordinating key behaviors such as sleep, wakefulness, stress, and feeding. Within this area in the brain, Melanin concentrating hormone (MCH) neurons are exclusive to the LHA and also contribute to these key homeostatic behaviors. These neurons can be divided into two different subpopulations: MCH neurons that express cocaine amphetamine-regulated transcript (CART) tachykinin-3 receptor (NK3R) and MCH neurons that do not express CART and NK3R. These neuronal subpopulations have been implicated in coordinating key behaviors but have not been well studied.

In this paper, our project seeks to analyze how the …

The Last Of Us In Therapy: How Mind-Controlling Fungi And Gut Bacteria Affect Your Mental Health, Anastasia Lyon

Journal of Pharmacology & Nutritional Sciences

The "psilocybiome" represents the mutually beneficial relationship between ourselves, our bacteria, and psychedelic drugs. This short review briefly discusses the benefits and limitations surrounding the potential for psychedelic therapy to synergize with gut bacteria to help regulate and maintain proper balance in the immune system, diet, and stress levels. Psychedelic therapy is a novel treatment strategy that has the potential to improve patient mental health, and, by identifying the types of gut bacteria present in patients, it can aid in personalizing medicine by determining how well their "psilocybiome" may respond.

Immobilization And Its Effects On Functional Recovery After Spinal Cord Injury., William H. Gregory

College of Arts & Sciences Senior Theses

Recovery, or rather the failure to recover, is a perplexing issue of spinal cord injury that is currently being investigated. This issue becomes more puzzling when investigating models in which spinal cord injury (SCI) is studied. There appears to be greater functional recovery experienced by animals within these models after injury as compared to patients in the clinical setting. While many things could contribute to this disparity, one difference that stands out when comparing the basic research model and clinical experience of SCI is the inactivity and limb disuse after the injury. This same level of immobility is rarely accounted …

Supraspinal Reorganization After Pediatric-Onset Spinal Cord Injury., Luis Alvarado

Electronic Theses and Dissertations

Pediatric spinal cord injury (SCI) disrupts the efferent and afferent flow of the developing brain, leading to devastating functional impairments below the injury site, yet our understanding of its impact on the brain remains limited. This study examines supraspinal reorganization in children with SCI using electrophysiology and neuroimaging techniques to understand the relationship between residual spinal transmission and supraspinal reorganization. Chapter 2 discusses the development of a child-centric approach using ‘learn, play, and practice’ to foster a trusting relationship with each child and increase compliance with experimental protocols. Chapter 3 evaluates the residual neural transmission of three spinal pathways in …

Effects Of Diet On Lcn2 Expression And Onset Of Neuroinflammation In An Alzheimer’S Disease Mice Model, Abdirahman A. Hayir

Neuroscience Honors Projects

This project covers the findings regarding the impact of diet on lipocalin 2 (LCN2) and the effects it has on neuroinflammation Alzheimer’s Disease (AD). LCN2 is a protein that is critical to the functionality of mitochondria and inflammatory responses. Evidence has shown that mitochondrial dysfunction is a potential central event in driving AD pathogenesis and contributing to formation of pathological hallmarks such as chronic inflammation. Furthermore, studies have shown that LCN2 can be deficient under metabolic conditions such as high-fat-diet (HFD). This study investigates if HFD induces LCN2 deficiency and increased neuroinflammation in an AD mice model.

Glycemic Challenge Is Associated With The Rapid Cellular Activation Of The Locus Ceruleus And Nucleus Of Solitary Tract: Circumscribed Spatial Analysis Of Phosphorylated Map Kinase Immunoreactivity, Arshad Khan

Selected Works Temporary Series

Rodent studies indicate that impaired glucose utilization or hypoglycemia is associated with the cellular activation of neurons in the medulla (Winslow, 1733) (MY), believed to control feeding behavior and glucose counterregulation. However, such activation has been tracked primarily within hours of the challenge, rather than sooner, and has been poorly mapped within standardized brain atlases. Here, we report that, within 15 min of receiving 2-deoxy-D-glucose (2-DG; 250 mg/kg, i.v.), which can trigger glucoprivic feeding behavior, marked elevations were observed in the numbers of rhombic brain (His, 1893) (RB) neuronal cell profiles immunoreactive for the cellular activation marker(s), phosphorylated p44/42 MAP …

Glycemic Challenge Is Associated With The Rapid Cellular Activation Of The Locus Ceruleus And Nucleus Of Solitary Tract: Circumscribed Spatial Analysis Of Phosphorylated Map Kinase Immunoreactivity, Khan

Departmental Papers (Biology)

Rodent studies indicate that impaired glucose utilization or hypoglycemia is associated with the cellular activation of neurons in the medulla (Winslow, 1733) (MY), believed to control feeding behavior and glucose counterregulation. However, such activation has been tracked primarily within hours of the challenge, rather than sooner, and has been poorly mapped within standardized brain atlases. Here, we report that, within 15 min of receiving 2-deoxy-D-glucose (2-DG; 250 mg/kg, i.v.), which can trigger glucoprivic feeding behavior, marked elevations were observed in the numbers of rhombic brain (His, 1893) (RB) neuronal cell profiles immunoreactive for the cellular activation marker(s), phosphorylated p44/42 MAP …

Higher Sexual Excitation Is Associated With An Increase In Sex-Linked Substance Use In Women With A History Of Unwanted Sexual Contact, Harper R. Jones, Tierney K. Lorenz

Center for Brain, Biology, and Behavior: Faculty Publications

Survivors of unwanted sexual contact have an increased likelihood of using substances in sexual situations, which puts them at heightened risk for intoxication-related harms. Separately, research has indicated that women may intentionally use substances in sexual situations to either enhance pleasure (i.e., increase sexual excitation) and/or reduce sexual anxiety or shame (i.e., reduce sexual inhibition), a phenomenon termed sex-linked substance use (SLSU). A predominant assumption in the literature is that women with unwanted sex histories are more likely to disengage during sex, suggesting greater inhibition-related SLSU; however, there is little prior research directly examining if women who have unwanted sex …

Pre–Post Intervention Exploring Cognitive Function And Relationships With Weight Loss, Intervention Adherence And Dropout, Amanda N. Szabo-Reed, Laura E. Martin, Cary R. Savage, Richard A. Washburn, Joseph E. Donnelley

Center for Brain, Biology, and Behavior: Faculty Publications

Objective: To evaluate the association between baseline cognitive function, intervention dropout, adherence and 3-month weight loss (WL) when controlling for confounding demographic variables.

Methods: 107 (Mage = 40.9 yrs.), BMI in the overweight and obese range (BMI = 35.6 kg/m2), men (N = 17) and women (N = 90) completed a 3-month WL intervention. Participants attended weekly behavioral sessions, comply with a reduced calorie diet, and complete 100 min of physical activity (PA)/wk. Cognitive function tasks at baseline included Flanker (attention), Stroop (executive control) and working memory, demographics, body weight and cardiovascular fitness were assessed at baseline. Session attendance, adherence …

Concussion-Related Disruptions To Hub Connectivity In The Default Mode Network Are Related To Symptoms And Cognition, Heather C. Bouchard, Kate L. Higgins, Grace Amadon, Julia M. Laing-Young, Arthur C. Maerlender, Seima Al-Momani, Maital Neta, Cary R. Savage, Douglas H. Schultz

Center for Brain, Biology, and Behavior: Faculty Publications

Concussions present with a myriad of symptomatic and cognitive concerns; however, the relationship between these functional disruptions and the underlying changes in the brain are not yet well understood. Hubs, or brain regions that are connected to many different functional networks, may be specifically disrupted after concussion. Given the implications in concussion research, we quantified hub disruption within the default mode network (DMN) and between the DMN and other brain networks. We collected resting-state functional magnetic resonance imaging data from collegiate student-athletes (n = 44) at three timepoints: baseline (prior to beginning their athletic season), acute post-injury (approximately 48 hours …

Manydogs Project: A Big Team Science Approach To Investigating Canine Behavior And Cognition, Daniela Alberghina, Emily E. Bray, Daphna Buchsbaum, Sarah- Elizabeth Byosiere, Julia Espinosa, Gitanjali E. Gnanadesikan, C.-N. Alexandrina Guran, Elizabeth Hare, Daniel J. Horschler, Ludwig Huber, Valerie A. Kuhlmeier, Evan L. Maclean, Madeline H. Pelgrim, Bryan Perez, Dana Ravid-Schurr, Liza Rothkoff, Courtney L. Sexton, Zachary A. Silver, Jeffrey R. Stevens

Center for Brain, Biology, and Behavior: Faculty Publications

Dogs have a special place in human history as the first domesticated species and play important roles in many cultures around the world. However, their role in scientific studies has been relatively recent. With a few notable exceptions (e.g., Darwin, Pavlov, Scott, and Fuller), domestic dogs were not commonly the subject of rigorous scientific investigation of behavior until the late 1990s. Although the number of canine science studies has increased dramatically over the last 20 years, most research groups are limited in the inferences they can draw because of the relatively small sample sizes used, along with the exceptional diversity …

The Accuracy Of Recreational Athletes In Self Detecting Ventilation Threshold During A Maximal Exercise Test, Andrew Biegner

Williams Honors College, Honors Research Projects

INTRODUCTION: Ventilatory threshold (VT) is where minute ventilation (VE) increases nonlinearly with increasing exercise intensity. Prior studies have shown that subjects can recognize VT by changes in breathing during exercise. Teaching recreational athletes to use VT as a training method could result in better quality training. PURPOSE: Determine if recreational athletes can accurately perceive changes in ventilation associated with VT during maximal exercise testing. METHODS: Subjects were recruited (n=20, age= 20.5 ± 1.7 yrs.) to participate in the study. Subjects performed a modified maximal treadmill protocol with gas analysis. Subjects indicated when they noticed a considerable change …

The Effect Of Training Program On Autism Spectrum Quotient Scores, Hossein Nazary Sharif, Sarvin Salar, Justin Anthony Haegele

Human Movement Studies & Special Education Faculty Publications

Background: Autism Spectrum Quotient (AQ) functions as an evaluative instrument designed to assess traits associated with autism. Variances exist within the brain of individuals on the autism spectrum. Moreover, certain exercises have the potential to impact specific regions of the brain.

Aim: This study aims to investigate how the program training of this study impact Autism—Spectrum Quotient.

Materials and Methods: Forty-two children aged 7-10 years with ASD were recruited to participate in this study. These children were randomly divided into two groups: the control group (N=10, 7.80±1.35 years) and the experimental group (N=32, age: 8.16±1.16 years). The experimental group underwent …

Explicit And Implicit Emotion Processing In The Cerebellum: A Meta‑Analysis And Systematic Review, Jordan E. Pierce, Marine Thomasson, Philippe Voruz, Garance Selosse, Julie Péron

Center for Brain, Biology, and Behavior: Faculty Publications

The cerebellum’s role in affective processing is increasingly recognized in the literature, but remains poorly understood, despite abundant clinical evidence for affective disruptions following cerebellar damage. To improve the characterization of emotion processing and investigate how attention allocation impacts this processing, we conducted a meta-analysis on task activation foci using GingerALE software. Eighty human neuroimaging studies of emotion including 2761 participants identified through Web of Science and ProQuest databases were analyzed collectively and then divided into two categories based on the focus of attention during the task: explicit or implicit emotion processing. The results examining the explicit emotion tasks identified …

The Role Of Reported Affective Symptoms And Anxiety In Recovery Trajectories After Sport-Related Concussion, Bernadette A. D'Alonzo, Abigail C. Bretzin, Douglas J. Wiebe, Ivy League–Big Ten Epidemiology Of Concussion Study Investigators, Arthur C. Maerlender, Cary R. Savage

Center for Brain, Biology, and Behavior: Faculty Publications

Background: There is growing awareness and clinical interest in athletes with affective symptoms after sport-related concussion (SRC), as these symptoms may contribute to overall symptoms and represent a modifiable risk factor of longer recovery. However, evidence of their effects on the entire return-to-play (RTP) trajectory, particularly among women and men, is limited.

Purpose/Hypothesis: To examine the relationship between affective symptom reporting and RTP progression after SRC among a cohort of Division 1 student-athletes. We hypothesized that those endorsing affective symptoms, specifically nervous-anxious symptoms, spend more time in RTP progression and recovery.

Study Design: Cohort study; Level of evidence, 3.

Methods: …