Manipulation Protocols Research Articles

Short-term effectiveness of the flexion-distraction technique in comparison with high-velocity vertebral manipulation in patients suffering from low-back pain

ObjectivesTo determine the short-term effects of a modified Flexion-Distraction (FD) technique in comparison with a high-velocity low-back spinal manipulation (HVLA-SM) protocol on patients suffering from chronic low-back pain (CLBP). Design and methodsA randomized controlled trial. The sample was composed of 150 patients suffering from CLBP, who were randomly assigned to either a FD (n = 75) or a HVLA-SM (n = 75) group. The variables used to study pain were the scores of the Visual Analogue Scale (VAS) and the Pressure Pain Threshold (PPT) on trigger points (TrPs) of the quadratus lumborum. In addition, the Oswestry Disability Index (ODI) was used to measure disability, and Schober’s test and the Finger Floor Distance test (FFDT) to measure changes in low-back spine motion. An Analysis of Covariance (ANCOVA) was used to measure group effect, and Number Needed to Treat (NNT) for effect size. ResultsGreater improvements occurred in the FD group, with a statistically significant group effect (p < 0.001) for all outcome variables. The ETA2 value was larger than 0.100 in the Schober’s and FDD tests, larger than 0.200 in the case of ODI and PPT, and larger than 0.300 for VAS. OR = 0.07 [IC 95% = 0.03 to 0.18] and NNT = 2.08 [IC 95% = 1.64–2.84) yielded improved values for the FD group. ConclusionFor patients suffering from CLBP, greater improvements in pain and function were observed in the group receiving the modified FD treatment than in the HVLA-SM group.

Manipulation of oral medication for children by parents and nurses occurs frequently and is often not supported by instructions.

Due to a lack of age-appropriate formulations, administration of drugs to children remains a challenge. This study aimed to identify the problems experienced in both the outpatient setting and the clinical setting. Between June 2017 and January 2018, we performed a cross-sectional, prospective study at the Sophia Children's Hospital, The Netherlands. The study comprised of a structured interview on drug manipulations with parents visiting the outpatient clinic, and an observational study of drug manipulations by nurses at the wards. A total of 201 questionnaires were collected, accounting for 571 drugs and 169 manipulations (30%). Drug substances that were most often mentioned as manipulated were macrogol (n=23), esomeprazole (n=15), paracetamol (n=8), methylphenidate (n=7) and melatonin (n=7). Of all manipulated medicines, 93/169 (55%) were manipulated according to the instructions or recommendations of the Summary of Product Characteristics (SmPC) or patient information leaflet. During the observational study, manipulation was performed by 21/35 of observed nurses (60%), of whom 11 deviated from the hospital protocol for manipulation or SmPC (52%). Manipulation was a widely used method to administer drugs to children. Validated information regarding manipulation of drugs for both parents and nursing staff is needed.

Laboratory Growth and Genetic Manipulation of Eimeria tenella

Eimeria is a genus of apicomplexan parasites that contains a large number of species, most of which are absolutely host-specific. Seven species have been recognized to infect chickens. Infection of susceptible chickens results in an intestinal disease called coccidiosis, characterized by mucoid or hemorrhagic enteritis, which is associated with impaired feed conversion or mortality in severe cases. Intensive farming practices have increased the significance of coccidiosis since parasite transmission is favored by high-density housing of large numbers of susceptible chickens. Routine chemoprophylaxis and/or vaccination with live parasite vaccines provides effective control of Eimeria, although the emergence of drug resistance and the relative cost and production capacity of current vaccine lines can prove limiting. As pressure to reduce drug use in livestock production intensifies, novel vaccination strategies are needed. Development of effective protocols supporting genetic complementation of Eimeria species has until recently been hampered by their inability to replicate efficiently in vitro. Now, the availability of such protocols has raised the prospect of generating transgenic parasite lines that function as vaccine vectors to express and deliver heterologous antigens. For example, this technology has the potential to streamline the production of live anticoccidial vaccines through the generation of parasite lines that co-express immunoprotective antigens derived from multiple Eimeria species. In this paper we describe detailed protocols for genetic manipulation, laboratory growth, and in vivo propagation of Eimeria tenella parasites, which will encourage future work from other researchers to expand biological understanding of Eimeria through reverse genetics. © 2019 by John Wiley & Sons, Inc.

Analysis of Neuro-Neuronal Synapses Using Embryonic Chick Ciliary Ganglion via Single-Axon Tracing, Electrophysiology, and Optogenetic Techniques.

The calyx-type synapse is a giant synaptic structure in which a presynaptic terminal wraps around a postsynaptic neuron in a one-to-one manner. It has been used for decades as an experimental model system of the synapse due to its simplicity and high accessibility in physiological recording methods. In particular, the calyx of the embryonic chick ciliary ganglion (CG) has enormous potential for synapse science because more flexible genetic manipulations are available compared with other synapses. Here, we describe methods to study presynaptic morphology, physiology, and development using CGs and cutting-edge molecular tools. We outline step-by-step protocols for presynaptic gene manipulation using in ovo electroporation, preparation of isolated CGs, 3-D imaging for single-axon tracing in transparent CGs, electrophysiology of the presynaptic terminal, and an all-optical approach using optogenetic molecular reagents. These methods will facilitate studies of the synapse and neuronal circuits in the future. © 2019 by John Wiley & Sons, Inc.

Tamoxifen Improves Glucose Tolerance in a Delivery-, Sex-, and Strain-Dependent Manner in Mice.

Tamoxifen, a selective estrogen-receptor modulator, is widely used in mouse models to temporally control gene expression but is also known to affect body composition. We report that tamoxifen has significant and sustained effects on glucose tolerance, independent of effects on insulin sensitivity, in mice. IP, but not oral, tamoxifen delivery improved glucose tolerance in three inbred mouse strains. The extent and persistence of tamoxifen-induced effects were sex and strain dependent. These findings highlight the need to revise commonly used tamoxifen-based protocols for gene manipulation in mice by including longer chase periods after injection, oral delivery, and the use of tamoxifen-treated littermate controls.

Stable transfection of Eimeria necatrix through nucleofection of second generation merozoites

Eimeria spp., the causative agents of coccidiosis, are the most common protozoan pathogens of chickens. Infection with these parasites can result in poor development or death of animals leading to a devastating economic impact on poultry production. The establishment of transfection protocols for genetic manipulation of Eimeria species and stable expression of genes would help advance the biology of these parasites as well as establish these organisms as novel vaccine delivery vehicles. Here, we report the selection of the first stable transgenic E. necatrix population, EnHA1, consitutively expressing the EYFP reporter following transfection of the 2nd generation merozoites with a linear DNA fragment harboring the EYFP reporter gene, the HA1 gene from the avian influenza virus H9N2 and the TgDHFR-TS selectable marker, which confers resistance to pyrimethamine. Transfected merozoites were inoculated into chickens via the cloacal route, and feces from 18 h to 72 h post inoculation were collected and subjected to subsequent serial passages, FACS sorting and pyrimethamine selection. A gradual increase in the number of EYFP-expressing sporulated oocysts was noticed with more than 90% EYFP + oocysts obtained after five passages. Immunofluorescence assay confirmed successful expression of the HA1 antigen in the EnHA1 population. The ability to genetically manipulate E. necatrix merozoites and express heterologous genes in this parasite will pave the way for possible use of this organism as a vaccine-delivery vehicle.

Regulatory-sequence mechanical biosensor: A versatile platform for investigation of G-quadruplex/label-free protein interactions and tunable protein detection

Mechanical biosensors can be used to quantitatively explore DNA-protein binding mechanisms by detecting targets at low concentrations or measuring force in single-molecule force spectroscopy. However, restrictions in single-molecule manipulation and labelling protocols have hindered the application for bulk analysis of label-free protein detection. Here, we present the integration of molecular force measurement and finely tunable detection of label-free proteins into one mechanical sensor. Regulatory-sequence force spectroscopy was obtained to investigate the binding force of DNA G-quadruplexes (GQ) and label-free protein. The dual control of regulatory sequences and mechanical forces induces the structure switching from DNA duplex to GQ/protein complex. It exhibits a synergistic effect, enabling the rational fine-tuning of the dynamic range for biosensing protein concentrations over eight orders of magnitude. Furthermore, this method was exploited to estimate the stability of the human telomeric DNA GQ by Ku protein and ligand methylpyridostatin. The results revealed that human telomeric GQ has two different binding sites for Ku protein and ligand. Force spectroscopy integrating label-free force measurement and tunable target detection holds great promise for use in biosensing, drug screening, targeted therapies, DNA nanotechnology, and fields in which GQ are of rapidly increasing importance.

Generation and Coherent Control of Pulsed Quantum Frequency Combs.

We present a method for the generation and coherent manipulation of pulsed quantum frequency combs. Until now, methods of preparing high-dimensional states on-chip in a practical way have remained elusive due to the increasing complexity of the quantum circuitry needed to prepare and process such states. Here, we outline how high-dimensional, frequency-bin entangled, two-photon states can be generated at a stable, high generation rate by using a nested-cavity, actively mode-locked excitation of a nonlinear micro-cavity. This technique is used to produce pulsed quantum frequency combs. Moreover, we present how the quantum states can be coherently manipulated using standard telecommunications components such as programmable filters and electro-optic modulators. In particular, we show in detail how to accomplish state characterization measurements such as density matrix reconstruction, coincidence detection, and single photon spectrum determination. The presented methods form an accessible, reconfigurable, and scalable foundation for complex high-dimensional state preparation and manipulation protocols in the frequency domain.

Effects of subcutaneous melatonin implants and short-term intravaginal progestagen treatments on estrus induction and fertility of Kivircik ewes on seasonal anestrus.

The aim of this study was to compare the efficacy of estrus induction and fertility by using subcutaneous melatonin (MEL, T1) and short-term intravaginal medroxyprogesteronacetate plus pregnant mare serum gonadotropin treatments (sMAP+eCG, T2) in ewes on seasonaly anestrus. In this study, 105 mature clinically healthy Kivircik ewes in anestrus season and 4 rams were used. After synchronization applications, ewes exhibiting estrus signs were hand-mated with rams known to be fertile. Blood samples were collected at different times in order to determine progesterone (P4) concentrations. Results showed that estrus manipulation protocols induced significant improvement in pregnancy rate. All the fertility results obtained with the sMAP+eCG or MEL groups were similar, in seasonal anestrus. The efficacy duration of P4 in the MEL group was longer than that in short-term progestagen group. Plasma P4 concentrations was significantly different between the first (I) and last (III) measurement days (p0.01). Increase in P4 concentration in T2 group was faster than that in T1 group, and blood P4 concentrations at higher levels could successfully be achieved by using any of the protocols in this study during the seasonal anestrus. In conclusion, according to the results obtained, the hormone application groups received very high estrus response. In addition, the twin ratio was found to be higher in T1 group compared to those determined in the other groups (T2 and Control group). Furthermore, plasma P4 concentrations and high birth rates were obtained in ewes in T1 and T2 groups. These procedures can be considered a good alternative to traditional procedures due to its flexibility under field conditions.

Safety of Appetite Manipulation in Children with Feeding Disorders Admitted to an Inpatient Feeding Program.

Appetite manipulation can be effective in weaning children off gastrostomy tube feeding dependence but can cause dehydration, hypoglycaemia, and ketone body production, which is anorexigenic. As the safety of this approach has not been described, our aim was to describe adverse events observed when weaning children from G-tube dependence using our appetite manipulation protocol. This was a retrospective study of prospectively collected data of patients who completed our inpatient tube-weaning protocol. Daily safety parameters included twice-daily urine specific gravities and urine ketones and fasting capillary blood glucose. Graded clinical interventions to manage adverse events were collected. A total of 143 children with a mean age of 4.8 ± 2.4 years were seen in the inpatient feeding program of which 74 (51.7%) were male. The children were hospitalized 10.1 ± 2.5 days with the vast majority being discharged between days 11 and 14. Overall, 78.2% of patients experienced at least 1 adverse event: urine specific gravity >1.020 was seen in 60.5%, ketonuria in 48.9%, and hypoglycemia (≤60 mg/dL) in 13.4%. Only 2 children had blood glucose levels <40 mg/dL and these were corrected with oral supplementation. Graded clinical interventions to manage adverse events included oral rehydration in 89.9% of children and supplemental tube feeding in 25.2%. Adverse effects are common when appetite manipulation is used to wean children off G-tube dependence. Anticipating, monitoring, and having a clear intervention plan in a closely monitored setting are necessary to safely use this method.

A single-step transconjugation system for gene deletion in Aggregatibacter actinomycetemcomitans

Aggregatibacter (A.) actinomycetemcomitans is a periodontopathogenic bacterium causing aggressive periodontitis. Here we describe a single-step transconjugation system as novel and easily applicable protocol for site-specific genetic manipulation of A. actinomycetemcomitans. Deletion of PgaC, which is involved in the synthesis of biofilm matrix, led to a reduced biofilm formation.

Autonomous quantum rotator

We consider a minimal model of a quantum rotator composed of a single particle confined in a harmonic potential and driven by two temperature-biased heat reservoirs. In the case the particle potential is rendered asymmetric and rotated an angle, a finite angular momentum develops, corresponding to a directed rotary motion. At variance with the classical case, the thermal fluctuations in the baths give rise to a non-vanishing average torque contribution; this is a genuine quantum effect akin to the Casimir effect. In the steady state the heat current flowing between the two baths is systematically converted into particle rotation. We derive exact expressions for the work rate and heat currents in the case in which the system is driven by an external time-periodic mechanical force. We show, in agreement with previous works on classical systems, that for this choice of external manipulation protocol, the rotator cannot work either as a heat pump or as a heat engine. We finally use our exact results to extend an ab initio quantum simulation algorithm to the out-of-equilibrium regime.

Inducing more sleep on school nights reduces sedentary behavior without affecting physical activity in short-sleeping adolescents

Objectives/BackgroundShort sleep duration during adolescence is associated with increased dietary intake and greater risk for overweight/obesity. However, findings are mixed on the relationship between sleep and physical activity (PA) during the school year, when short sleep duration is most common. Furthermore, there is concern that increasing sleep duration may interfere with opportunities for PA, yet this has not been directly tested. This study examined the impact of an at-home experimental sleep extension protocol on PA during the school year among short-sleeping adolescents. Participants/MethodsParticipants included 18 adolescents (67% female, 78% white) who reported regularly sleeping between 5–7 h on school nights. Adolescents completed a five-week, at-home sleep manipulation protocol with an initial baseline week followed in a randomized, counterbalanced order by two experimental conditions, each lasting two weeks. During prescribed habitual sleep (HAB), bedtimes and rise times were set to match the baseline sleep pattern, and during sleep extension (EXT), adolescents were instructed to increase time in bed on school nights by 1.5 h per night relative to baseline. Wrist-mounted actigraphy was employed to monitor sleep and waist-mounted accelerometers were used to measure daytime PA. ResultsAdolescents slept for an average duration of 71 min longer on school nights during EXT than during HAB (p < 0.001). During HAB, adolescents spent more time in sedentary behavior (p = 0.002) than during EXT, but there were no cross-condition differences in light activity (p = 0.184) or moderate-to-vigorous PA (p = 0.102). ConclusionsExtending sleep duration on school nights in short-sleeping adolescents reduces time spent in sedentary behavior, without having a negative impact on health-promoting moderate-to-vigorous PA.

Euploidy rate is indpendently associated with the number of ova retrieved in a fresh donor cycles

Studies have demonstrated that > 50% of human embryos are aneuploid following ovarian stimulation and IVF. Embryonic aneuploidy is correlated with maternal age. The reported aneuploidy rates range from 30% in women <30 years of age to >70% in women >40 years old (Munne et al., 2007). Experimental evidence also suggests that other treatment-related factors, including culture conditions, gamete manipulation, gamete aging and ovarian stimulation protocols may influence aneuploidy rates in human embryos. A recent study also reported that euploid blastocyst rates in cycles using donor eggs varied greatly between infertility clinics ranging from 39.5% to 82.5% (Munne et al., 2017). We examined the impact of the length of the ovarian stimulation, the number of ova retrieved, and the influence of the treating physician in fresh donor egg cycles from a single fertility center on: 1) the percentage of euploid blastocysts and 2) the frequency of blastocyst formation. A total of 161 consecutive fresh donor oocyte cycles performed between 2015 and 2016 were included in the study. The donors had a mean age of 25.1 +/- 3.5 years. A total of 1352 blastocysts were biopsied. The biopsies were analyzed in a single PGS laboratory using either high density oligonucleotide aCGH or NGS. The euploid and overall blastocyst rates were examined as a function of a) the length of the ovarian stimulation (stratified into three groups: < 8 =days (n=23), 9-11 days (n=117) and >12 days (n=21)); b) number of ova retrieved (stratified into three groups: < 20 eggs (n=35), 21-40 eggs (n=100), >41 eggs (n=26)); and c) between the four treating physicians. The euploid rate was calculated by dividing the number of euploid embryos by the number of biopsied blastocysts. Embryos with no diagnosis and/or degraded DNA (<2%) were excluded from this analysis. The percentage data were transformed by arcsine transformation and then analyzed using ANOVA and Tukey test. Differences with p<0.05 were considered significant in all comparisons. The euploid rates were 78.7%, 78.4% and 80.0% respectively when comparing by the length of ovarian stimulation, with no observed statistical differences (p=0.94). When analysed by the number of eggs retrieved, the euploid rates were: 73.2%, 79.9%, and 82.1%, respectively. A significant higher euploidy rate was observed in the group retrieved > 40 eggs (82.1%) compared to the group had < 20 eggs retrieved (p= 0.039). Four physicians managed the egg donor cycles with similar euploid rates of 74.6%, 79.7%, 81.2% and 78.4% (p=0.18). There were no significant differences observed in the percentage of blastocyst formation as a function of the length of ovarian stimulation (61.8%, 61.3%, and 61.8%, p=0.95), number of ova retrieved (61.5%, 61.2%, and, 63.3%, p=0.88) or treating physicians (57.6%, 66.5%, 62.3% and 67.0%, p=0.25). In donor ova cycles, the rate of euploid blastocysts was independently associated with the number of ova retrieved. The blasocysts formation frequency was not affected by the length of the ovarian stimulation, number of ova retrieved, or the treating physician.

Recent Advances in the Genetic Manipulation of Methylosinus trichosporium OB3b.

Methanotrophic bacteria utilize methane as their sole carbon and energy source. Studies of the model Type II methanotroph Methylosinus trichosporium OB3b have provided insight into multiple aspects of methanotrophy, including methane assimilation, copper accumulation, and metal-dependent gene expression. Development of genetic tools for chromosomal editing was crucial for advancing these studies. Recent interest in methanotroph metabolic engineering has led to new protocols for genetic manipulation of methanotrophs that are effective and simple to use. We have incorporated these newer molecular tools into existing protocols for Ms. trichosporium OB3b. The modifications include additional shuttle and replicative plasmids as well as improved gene delivery and genotyping. The methods described here render gene editing in Ms. trichosporium OB3b efficient and accessible.

The importance of post-thaw subculture for standardizing cellular activity of fresh or cryopreserved mouse embryonic stem cells.

ObjectiveRemarkable difference in cellular activity was found between early and late subpassaged embryonic stem cell (ESCs) lines, which can be created by subtle changes in cell manipulation protocol. This study subsequently examined whether post-thaw subculture of early subpassaged ESC lines could further affect the activity of the ESCs.MethodsFresh (as a control treatment) or cryopreserved F1 hybrid (B6CBAF1) early ESC lines (C57BL/6xCBA) of the 4 (P4) or the 19 passage (P19) were subcultured once, twice or six times under the same condition. The post-thaw survival of the ESCs was monitored after the post-treatment subculture and the ability of cell proliferation, reactive oxygen species (ROS) generation, apoptosis and mitochondrial ATP synthesis was subsequently examined.ResultsRegardless of the subculture number, P19 ESCs showed better (p<0.05) doubling time and less ATP production than P4 ESCs and such difference was not influenced by fresh or cryopreservation. The difference between P4 and P19 ESC lines became decreased as the post-treatment subculture was increased and the six times subculture eliminated such difference. Similarly, transient but prominent difference in ROS production and apoptotic cell number was detected between P4 and P19 ESCs only at the 1st subculture after treatment, but no statistical differences between two ESC lines was detected in other observations.ConclusionThe results of this study suggest that post-thaw subculture of ESCs under the same environment is recommended for standardizing their cellular activity. The activity of cell proliferation ability and ATP synthesis can be used as parameters for quality control of ESCs.

Stimulated Raman adiabatic control of a nuclear spin in diamond

Coherent manipulation of nuclear spins is a highly desirable tool for both quantum metrology and quantum computation. However, most of the current techniques to control nuclear spins lack of being fast impairing their robustness against decoherence. Here, based on Stimulated Raman Adiabatic Passage, and its modification including shortcuts to adiabaticity, we present a fast protocol for the coherent manipulation of nuclear spins. Moreover, we show how to initialise a nuclear spin starting from a thermal state, and how to implement Raman control for performing Ramsey spectroscopy to measure the dynamical and geometric phases acquired by nuclear spins.

Feasibility and Emotional Impact of Experimentally Extending Sleep in Short-Sleeping Adolescents.

Published experimental sleep manipulation protocols for adolescents have been limited to the summer, limiting causal conclusions about how short sleep affects them on school nights, when they are most likely to restrict their sleep. This study assesses the feasibility and emotional impact of a school-night sleep manipulation protocol to test the effects of lengthening sleep in habitually short-sleeping adolescents. High school students aged 14-18 years who habitually slept 5-7 hours on school nights participated in a 5-week experimental sleep manipulation protocol. Participants completed a baseline week followed in randomized counterbalanced order by two experimental conditions lasting 2 weeks each: prescribed habitual sleep (HAB; sleep time set to match baseline) and sleep extension (EXT; 1.5-hour increase in time in bed from HAB). All sleep was obtained at home, monitored with actigraphy. Data on adherence, protocol acceptability, mood and behavior were collected at the end of each condition. Seventy-six adolescents enrolled in the study, with 54 retained through all 5 weeks. Compared to HAB, during EXT, participants averaged an additional 72.6 minutes/night of sleep (p < .001) and had reduced symptoms of sleepiness, anger, vigor, fatigue, and confusion (p < .05). The large majority of parents (98%) and adolescents (100%) said they would "maybe" or "definitely" recommend the study to another family. An experimental, school-night sleep manipulation protocol can be feasibly implemented which directly tests the potential protective effects of lengthening sleep. Many short-sleeping adolescents would benefit emotionally from sleeping longer, supporting public health efforts to promote adolescent sleep on school nights.

Robust Quaternion-based Cooperative Manipulation without Force/Torque Information

This paper proposes a task-space control protocol for the collaborative manipulation of a single object by N robotic agents. The proposed methodology is decentralized in the sense that each agent utilizes information associated with its own and the object’s dynamic/kinematic parameters and no on-line communication takes place. Moreover, no feedback of the contact forces/torques is required, therefore employment of corresponding sensors is avoided. An adaptive version of the control scheme is also introduced, where the agents’ and object’s dynamic parameters are considered unknown. We also use unit quaternions to represent the object’s orientation. In addition, load sharing coefficients between the agents are employed and internal force regulation is guaranteed. Finally, experimental studies with two robotic arms verify the validity and effectiveness of the proposed control protocol.

0961 SHORT SLEEP INDUCES EMOTIONAL EATING AND WORSE MOOD IN TEENS

Prior research indicates that short sleep causes negative changes in mood, emotion regulation, and increased caloric intake in adolescents. Eating as a way to cope with changes in mood may contribute to the relationship between short sleep and greater caloric intake, yet the impact of sleep on emotional eating has not yet been examined. The present study aims to determine if short sleep increases emotional eating and explores whether any increase is related to sleep-specific changes in mood. Participants were 47 adolescents aged 14–17 who completed an experimental sleep manipulation protocol with a randomized, cross-over design in which teens experienced 5 consecutive nights of short sleep (SS; 6.5 hours in bed) and 5 nights of healthy sleep (HS; 9.5 hours in bed). At the end of each condition, teens completed the Emotional Eating Scale (measuring how often they ate in response to Anxiety/Anger/Frustration, Depressive Symptoms, and Unsettled Feelings), the Profile of Mood States, and a measure of emotion regulation. Compared to HS, during SS teens reported increased emotional eating in response to depressive symptoms (p=.019) but not anxiety/anger/frustration or unsettled feelings (p>.05). They also reported an overall decline in mood (all subscales p<.05) and worse emotion regulation (p=.009). Finally, changes in sad/depressed mood during sleep restriction were significantly related to increased emotional eating in response to sadness (p=.01). Short sleep causes teens to experience a decline in mood, putting them at risk for using maladaptive strategies to cope with this change. Teens report eating more in response to feelings of sadness when sleep restricted, and this increase in emotional eating is related to changes in their mood induced by insufficient sleep. Increases in emotional eating may help explain the relationship between poor sleep and obesity risk in teens. Financial support from the US National Institutes of Health (R01 HL120879).