The C-terminal transactivation domain of MITF interacts promiscuously with co-activator CBP/p300.

The microphthalmia-associated transcription factor (MITF) is one of four closely related members of the MiT/TFE family (TFEB, TFE3, TFEC) that regulate a wide range of cellular processes. MITF is a key regulator of melanocyte-associated genes, and essential to proper development of the melanocyte cell lineage. Abnormal MITF activity can contribute to the onset of several diseases including melanoma, where MITF is an amplified oncogene. To enhance transcription, MITF recruits the co-activator CREB-binding protein (CBP) and its homolog p300 to gene promoters, however the molecular determinants of their interaction are not yet fully understood. Here, we characterize the interactions between the C-terminal MITF transactivation domain and CBP/p300. Using NMR spectroscopy, protein pulldown assays, and isothermal titration calorimetry we determine the C-terminal region of MITF is intrinsically disordered and binds with high-affinity to both TAZ1 and TAZ2 of CBP/p300. Mutagenesis studies revealed two conserved motifs within MITF that are necessary for TAZ2 binding and critical for MITF-dependent transcription of a reporter gene. Finally, we observe the transactivation potential of the MITF C-terminal region is reliant on the N-terminal transactivation domain for function. Taken together, our study helps elucidate the molecular details of how MITF interacts with CBP/p300 through multiple redundant interactions that lend insight into MITF function in melanocytes and melanoma.

Quantum-cascade-laser-absorption-spectroscopy diagnostic for temperature, pressure, and NO X 2 Π 1/2 at 500 kHz in shock-heated air at elevated pressures.

The design, validation, and application of a quantum-cascade-laser-absorption-spectroscopy diagnostic for measuring gas temperature, pressure, and nitric oxide (NO) in high-temperature air are presented. A distributed-feedback quantum-cascade laser (QCL) centered near 1976c m -1 was used to scan across two transitions of NO in its ground electronic state (X 2 Π 1/2). A measurement rate of 500 kHz was achieved using a single QCL by: (1) performing current modulation through a bias-tee, and (2) targeting closely spaced transitions with a large difference in lower-state energy. The diagnostic was validated in a mixture of 95% argon and 5% NO, which was shock-heated to ≈2000 to 3700 K. The average mean percent differences between laser-absorption-spectroscopy (LAS) measurements and predictions from shock-jump relations for temperature, pressure, and NO mole fraction were 3.1%, 4.1%, and 6.5%, respectively. The diagnostic was then applied to characterize shock-heated air at high temperatures (up to ≈5500K) and high pressures (up to 12 atm) behind either incident or reflected shocks. The LAS measurements were compared to theoretical predictions from shock-jump relations, pressure sensors mounted in the wall of the shock tube, and equilibrium values of the NO mole fraction. The average mean percent differences between LAS measurements and their aforementioned reference values were 3.2%, 10.8%, and 10.4% for temperature, pressure, and NO mole fraction, respectively. Last, a comparison between a measured NO mole fraction time history and a time-stepped homogeneous reactor simulation performed using two different chemical kinetics mechanisms is presented.

Pulse-burst spontaneous Raman thermometry of unsteady wave phenomena in a shock tube.

A high-speed temperature diagnostic based on spontaneous Raman scattering (SRS) was demonstrated using a pulse-burst laser. The technique was first benchmarked in near-adiabatic ${{\rm H}_2} \text{-} {\rm air}$ flames at a data-acquisition rate of 5 kHz using an integrated pulse energy of 1.0 J per realization. Both the measurement precision and accuracy in the flame were within 3% of adiabatic predictions. This technique was then evaluated in a challenging free-piston shock tube environment operated at a shock Mach number of 3.5. SRS thermometry resolved the temperature in post-incident and post-reflected shock flows at a repetition rate of 3 kHz and clearly showed cooling associated with driver expansion waves. Collectively, this Letter represents a major advancement for SRS in impulsive facilities, which had previously been limited to steady state regions or single-shot acquisition.

Resource Use After Unwanted Sexual Experiences in Undergraduates: A Comprehensive Evaluation of Factors Related to the Decision to Seek Help.

The risk for sexual victimization in college is high, and although there can be significant mental and physical consequences for the survivor, resource utilization for this issue is extremely low. The current study used the frameworks of both the ecological model and the theory of planned behavior to examine factors related to resource use in undergraduate survivors of unwanted sexual experiences. The sample consisted of 217 undergraduates who reported at least one unwanted sexual experience while in college (89% female, 76.5% Caucasian, M age 19.6). Data were collected in an online survey from a private liberal arts college, and were analyzed using decision tree modeling. Results suggested that resource use was less related to demographic or event characteristics, but rather related to victim response to the event as well as perceptions about control over potential resource use. In addition to key assault prevention efforts, it is thus important to also support survivor mental health and self-efficacy postassault.

Coronal Remodeling Potential of Pediatric Distal Radius Fractures.

BACKGROUND: Distal radius fractures in the pediatric population are common injuries with a remarkable capability to remodel. The degree of angulation that can reasonably be expected to remodel is controversial though, particularly when it comes to angulation in the coronal plane. The purpose of this study was to quantify the rate of coronal remodeling, via the distal radius physis, present in a retrospective cohort of skeletally immature patients with coronally angulated distal radius fractures. METHODS: A retrospective chart review was performed to identify skeletally immature patients treated for an angulated distal radius fracture with over 10 degrees of angulation in the coronal plane during the healing process at a single institution by either a pediatric orthopaedic surgeon or an orthopaedic trauma surgeon from 2009 to 2018. Coronal angulation was measured at every visit where radiographs were available from time of injury to the final follow-up visit to determine the rate of remodeling. RESULTS: In total, 36 patients with distal radius fractures with a mean age of 7.93 years (range, 4 to 12 y) at the time of injury were identified. The median peak angulation during the healing process in the coronal plane was 17 degrees (range, 12.4 to 30.4 degrees). The mean follow-up period was 6.4 months from the time of maximum angulation to the final visit. The median time from cast removal to final follow-up was 6.59 months (range, 2.5 to 8.72 mo). At final follow-up, the median coronal angulation was 3.35 degrees (range, 0.24 to 14.0 degrees). At the 95% confidence level, remodeling rates ranged from 2.00 to 2.59 degrees per month. The mean rate of coronal angulation remodeling from maximum angulation to final follow-up was 2.30 degrees per month. CONCLUSIONS: Distal radius fractures have a large capacity to remodel in the pediatric population. This remodeling occurs in a predictable and reliable manner, even in the coronal plane. On the basis of this study, coronal angulation was shown to remodel at a rate of at least 2 degrees per month for the first 6 months following the injury, which should likely continue at a similar rate for the first year after the injury. Repeat manipulation is not indicated in skeletally immature patients where the maximum coronal angulation is <24 degrees, which provides a conservative estimate of the amount of remodeling that can be expected to occur in the first year after fracture. LEVEL OF EVIDENCE: Level III-retrospective comparative study.

Bone accrual over 18 months of participation in different loading sports during adolescence.

This study investigated the impact and non-impact sports on bone mineral density accrual in adolescents over 18 months. The impact sports were beneficial for bone health (accrual of bone density). In contrast, swimmers had similar or lower bone mineral density compared with the control group depending on the skeletal site. PURPOSE: To investigate the impact and non-impact sports on bone mineral density (BMD) accrual in adolescents over a period of 18 months METHODS: The sample was composed of 71 adolescents, avarage age of 12.7 (± 1.7) years old at baseline. Bone outcomes were compared according to the loading of the sports practiced (impact sports, n = 33 [basketball, karate, and judo], non-impact sport, n = 18 [swimming], and control group, n = 20). Areal bone mineral density (aBMD) was measured by dual-energy X-ray absorptiometry (DXA) and bone mineral apparent density (BMAD) estimated through equation. The results were compared between the groups using analysis of variance and analysis of covariance. RESULTS: Adjusted aBMD at lower limbs, whole body less head (WBLH), and adjusted WBLH BMAD were significantly greater in the impact sport group than the non-impact sport group at all time points. Adjusted upper limbs aBMD was significantly higher at the impact sports group compared to the non-impact sport group at 9 months and 18 months, besides compared to the control group at baseline and 18 months. Non-impact sport group presented a significant lower adjusted aBMD compared with control group at lower limbs and WBLH at 9 months, and at 9 months and 18 months in WBLH BMAD. There was a significant interaction (time × sport group) at upper limbs (p = 0.042) and WBLH aBMD (p = 0.006), and WBLH BMAD (p < 0.001). CONCLUSION: Impact sports were more beneficial on accumulating aBMD and BMAD over a period of 18 months, while non-impact group (swimmers) had similar and lower aBMD and BMAD compared with the control group.

Minimally Invasive Medial Plate Osteosynthesis of High-Energy Pediatric Tibia Fractures.

OBJECTIVES: We report on the largest series to date of minimally invasive medial plate osteosynthesis of high-energy tibia fractures in skeletally immature patients. DESIGN: Retrospective. SETTING: Level 1 trauma center. PATIENTS: Skeletally immature patients with high-energy tibia fractures treated with minimally invasive medial plate osteosynthesis between 2006 and 2017. We identified 28 fractures in 26 patients. MAIN OUTCOME MEASURE: We reviewed the record for patient, injury, and treatment factors as well as for complications and reoperation. We assessed the relationships between age, implant selection, and implant removal using χ and t test analyses. RESULTS: Mean age was 12 years (4-15) with OTA/AO classifications including 42A (18), 42B (6), and 42C (4). There were 10 open fractures (1 type I, 2 type II, and 7 type III). Mean follow-up was 37.6 months (minimum 7). Mean time to full weight-bearing and union was 2 and 3 months, respectively. Plate stock (2.7, 3.5, and 4.5 mm) and length (6-20 holes) varied independent of age, P = 0.19. There were 2 superficial infections, one 15 mm leg length discrepancy, and one case of fasciotomy at the time of plate fixation for trauma-related compartment syndrome. Implant removal was more likely in younger patients (10.9 vs. 13.2 years, P = 0.04) and not associated with plate stock, P = 0.97. There were no neurovascular injuries, losses of reduction, or rotational deformities. Angulation was universally <5 degrees in the coronal plane and <10 degrees in the sagittal plane. CONCLUSIONS: Minimally invasive medial plate osteosynthesis of high-energy pediatric tibia fractures may be performed with a low rate of complications and should be considered in this select patient population with multiple injuries, high-energy mechanisms, and significant soft-tissue trauma. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.

Bone Mineral Density and Sports Participation.

The objective of our study was to identify differences in bone mineral density (BMD) according to different sports among adolescents, as well as to identify assessments of relevant mediators. The sample consisted of 429 adolescents with a mean age of 13.3 ± 1.7 yr. The sports modalities that we evaluated were judo, karate, kung fu, swimming, basketball, and soccer. Nonsports group was composed of 111 adolescents. Bone density was assessed using dual-energy X-ray absorptiometry. Regarding BMD values, female engaged in karate had higher BMD values than girls from the no sports group (p = 0.01). Boys engaged in soccer and karate presented the highest BMD values than all the other groups (p = 0.01). The most relevant mediators were the body mass index and lean soft tissue for boys and girls. Different sports presented different results regarding BMD gains, which were influenced by body mass index and whole-body lean soft tissue.

Impact of sports participation on incidence of bone traumatic fractures and health-care costs among adolescents: ABCD - Growth Study.

Objective: To analyze the risk of bone traumatic fractures according to the engagement in sports, as well as to identify the potential impact of sports participation and traumatic fractures on health-care costs among adolescents. Methods: This is a longitudinal 12-month study of 285 adolescents of both sexes in Brazil. We assessed the occurrence of traumatic fractures and health-care services (hospitalizations, medicine use, medical consultations, and exams) by phone contact every month for 12 months. Adolescents were divided into four groups according to sport characteristics: non-sport (n = 104), non-impact sport (swimming [n = 34]), martial arts (n = 49 [judo, karate, kung-Fu]) and impact sports (n = 98 [track-and-field, basketball, gymnastics, tennis, and baseball]). Results: The incidence of new fractures was 2.1%. The overall costs accounted during the 12-month follow-up were U$ 3,259.66. Swimmers (US$ 13.86) had higher health-care costs than non-sport (US$ 1.82), martial arts (US$ 2.23), and impact sports (US$ 2.32). Conclusion: swimming seems to be related to higher health-care costs among adolescents.

Parents' Lifestyle, Sedentary Behavior, and Physical Activity in Their Children: A Cross-Sectional Study in Brazil.

BACKGROUND: This study investigated associations between different types of sedentary behavior (SB) and physical activity (PA) in parent and their child, including the moderating effects of parent and child sex. METHODS: In total, 1231 adolescents, 1202 mothers, and 871 fathers were evaluated. The SB (TV viewing + computer + video game); different types of PA (leisure-time PA, occupational PA, and total PA); and the socioeconomic level were evaluated by questionnaire. The relationship between adolescents' SB and PA with parental characteristics was estimated by linear regression. RESULTS: The SB of male adolescents was correlated to the father's SB (ß = 0.26; 95% confidence interval [CI], 0.13-0.39) and mother's SB (ß = 0.18; 95% CI, 0.06-0.31). A similar relationship was observed between SB of female adolescents and the father's SB (ß = 0.31; 95% CI, 0.19-0.42) and mother's SB (ß = 0.29; 95% CI, 0.20-0.38]). The SB of girls was inversely related to mother's occupational PA (ß = -2.62; 95% CI, -3.66 to -0.53]). The PA of the boys and girls was correlated with their fathers and mothers PA. All the results were adjusted for age and parent's socioeconomic level. CONCLUSIONS: SB and PA of parents were associated with SB and PA of their children, regardless of gender. Strategies for health promotion should consider the family environment to increase PA and reduce SB.

Sports Participation Decreases the Incidence of Traumatic, Nonsports-Related Fractures Among Adolescents.

OBJECTIVES: To investigate the effects of different sports on the incidence of traumatic fractures (TF; sport-related fractures and those occurring in daily activities) among adolescents during the 9-month follow-up period. METHODS: The adolescents were contacted in 11 different locations (3 public/private schools and 8 sports clubs), and the final sample was divided into 3 groups: control (n = 121), swimming (n = 51), and impact sports (n = 142). The incidence of TF was calculated by considering the exposure to sports (TF/1000 h). RESULTS: In the overall sample, the incidence of TF was 1.29 TF/1000 hours of sports exposure, while the incidence of sport-related TF was 0.39 TF/1000 hours of sports exposure. Adolescents engaged in sports (P = .004), independently of type (P = .001), for 3 or more days per week (P = .004) and more than 60 minutes per day (P = .001) had lower incidence of TF. Adolescents engaged in more than 300 minutes per week of sport (0.17 TF/1000 h) had lower incidence than those who did not (2.06 TF/1000 h [P = .001]). A similar finding was observed for sport-related TF (≥300 min/wk: 0.08 TF/1000 h vs 300 min/wk: 0.615 TF/1000 h [P = .02]). CONCLUSION: Adolescents engaged in sports showed a lower incidence of TF than nonengaged adolescents.

Monolith columns for liquid chromatographic separations of intact proteins: A review of recent advances and applications.

In this article we survey 256 references (with an emphasis on the papers published in the past decade) on monolithic columns for intact protein separation. Protein enrichment and purification are included in the broadly defined separation. After a brief introduction, we describe the types of monolithic columns and modes of chromatographic separations employed for protein separations. While the majority of the work is still in the research and development phase, papers have been published toward utilizing monolithic columns for practical applications. We survey these papers as well in this review. Characteristics of selected methods along with their pros and cons will also be discussed.

Implementation and Evaluation of Shared Medical Appointments for Type 2 Diabetes at a Free, Student-Run Clinic in Alamance County, North Carolina.

INTRODUCTION: Type 2 diabetes is a significant problem among uninsured patients. Shared medical appointments (SMA) have been shown to improve outcomes in type 2 diabetes. We hypothesized that the SMA model could be adapted for a non-profit clinic in North Carolina that serves uninsured patients with diabetes that have incomes at/below 150% of the federal poverty line. RESEARCH DESIGN AND METHODS: We implemented and sustained a patient-driven, student-led SMA model that incorporated the monthly rotations of students, physician assistant, and undergraduate students as well as pharmacy residents and an endocrinologist who collectively provide diabetes care at the free clinic. SMA groups are 'open' cohorts and include 4-12 patients scheduled for the monthly clinic. Teams of transdisciplinary trainees work together to perform triage, medication reconciliation, brief history, and physical exam, after which patients participate in the SMA. The endocrinologist evaluates SMA patients individually during and after the visit. RESULTS: Between November 2015 and January 2017, we enrolled 29 patients in SMA. There was high variability in HbA1c at baseline. Among eight type 2 diabetes patients seen in endocrine clinic and with complete data one year before and after SMA implementation, the mean (SD) HbA1c before SMA was 9.7% ± 1.7% (83±7 mmol/ mol); mean HbA1c after SMA was 9.2% ± 1.8% (77 ± 8mmol/mol). The median HbA1c before SMA was 9.5% (80 mmol/mol); median HbA1c after SMA was 8.9% (74 mmol/mol). Overall, 6/8 patients showed decreased HbA1c after SMA although there was variability between individuals in response of glycemic control to SMA. SMA increased clinic efficiency and offered an opportunity to integrate transdisciplinary trainees. Trainees gain experience with novel models of care and the complexities of the patient experience of diabetes. CONCLUSIONS: We hope this observation encourages others to implement such programs to enhance the evidence-base for SMA to address health disparities and increase the quality of free diabetes care.

Growing Neural PC-12 Cell on Crosslinked Silica Aerogels Increases Neurite Extension in the Presence of an Electric Field.

Externally applied electrical stimulation (ES) has been shown to enhance the nerve regeneration process and to influence the directionality of neurite outgrowth. In addition, the physical and chemical properties of the substrate used for nerve-cell regeneration is critical in fostering regeneration. Previously, we have shown that polyurea-crosslinked silica aerogels (PCSA) exert a positive influence on the extension of neurites by PC-12 cells, a cell-line model widely used to study neurite extension and electrical excitability. In this work, we have examined how an externally applied electric field (EF) influences the extension of neurites in PC-12 cells grown on two substrates: collagen-coated dishes versus collagen-coated crosslinked silica aerogels. The externally applied direct current (DC) bias was applied in vitro using a custom-designed chamber containing polydimethysiloxane (PDMS) embedded copper electrodes to create an electric field across the substrate for the cultured PC-12 cells. Results suggest orientation preference towards the anode, and, on average, longer neurites in the presence of the applied DC bias than with 0 V DC bias. In addition, neurite length was increased in cells grown on silica-crosslinked aerogel when compared to cells grown on regular petri-dishes. These results further support the notion that PCSA is a promising material for nerve regeneration.

Track and Field Practice and Bone Outcomes among Adolescents: A Pilot Study (ABCD-Growth Study).

BACKGROUND: Osteoporosis is considered a public health problem with high worldwide prevalence. One approach to prevention is through the promotion of physical activity, especially exercise, during adolescence. METHODS: This study compared bone variables in different body segments in adolescents according to participation in track and field. The study included 34 adolescents (22 boys), of whom 17 were track and field athletes and 17 were control subjects. Bone mineral density (BMD, g/cm2) and bone mineral content (BMC, g) were analyzed using dual energy X-ray absorptiometry (total body stratified by body segments). Peak height velocity was used to estimate somatic maturation. RESULTS: Athletes had higher BMD (P=0.003) and BMC (P=0.011) values in the lower limbs and higher whole body BMD (P=0.025) than the control group. However, when adjusted for confounding factors, the difference was not maintained. The groups had similar lean soft tissue values (P=0.094). Training overload was positively correlated with BMD in the upper limbs (r=0.504; 95% confidence interval, 0.031-0.793). Although track and field athletes had higher BMD and BMC values in the lower limbs, these differences were not significant when adjusted for confounding factors. CONCLUSIONS: Track and field participation in adolescence appears to influence BMD and BMC in lower limbs, and fat-free mass seems to mediate this effect. Also, higher training loads were found to be positive for bone health in upper limbs.

Two-dimensional liquid chromatography consisting of twelve second-dimension columns for comprehensive analysis of intact proteins.

A comprehensive two-dimensional liquid chromatography (LCxLC) system consisting of twelve columns in the second dimension was developed for comprehensive analysis of intact proteins in complex biological samples. The system consisted of an ion-exchange column in the first dimension and the twelve reverse-phase columns in the second dimension; all thirteen columns were monolithic and prepared inside 250 µm i.d. capillaries. These columns were assembled together through the use of three valves and an innovative configuration. The effluent from the first dimension was continuously fractionated and sequentially transferred into the twelve second-dimension columns, while the second-dimension separations were carried out in a series of batches (six columns per batch). This LCxLC system was tested first using standard proteins followed by real-world samples from E. coli. Baseline separation was observed for eleven standard proteins and hundreds of peaks were observed for the real-world sample analysis. Two-dimensional liquid chromatography, often considered as an effective tool for mapping proteins, is seen as laborious and time-consuming when configured offline. Our online LCxLC system with increased second-dimension columns promises to provide a solution to overcome these hindrances.

Multiple-channel ultra-violet absorbance detector for two-dimensional chromatographic separations.

In recent years, much research has gone into developing online comprehensive two-dimensional liquid chromatographic systems allowing for high peak capacities in comparable separation times to that of one-dimensional liquid chromatographic systems. However, the speed requirements in the second dimension (2nd-D) still remain one challenge for complex biological samples due to the current configuration of two column/two detector systems. Utilization of multiple 2nd-D columns can mitigate this challenge. To adapt this approach, we need a multiple channel detector. Here we develop a versatile multichannel ultraviolet (UV) light absorbance detector that is capable of simultaneously monitoring separations in 12 columns. The detector consists of a deuterium lighthouse, a flow cell assembly (a 13-channel flow cell fitted with a 13-photodiode-detection system), and a data acquisition and monitoring terminal. Through the use of a custom high optical quality furcated fiber to improve light transmission, precise machining of a flow cell to reduce background stray light through precision alignment, and sensitive electronic circuitry to reduce electronic noise through an active low pass filter, the background noise level is measured in the tens of µAU. We obtain a linear dynamic range of close to three orders of magnitude. Compared to a commercialized multichannel UV light absorbance detector like the Waters 2488 UV/Vis, our device provides an increase in channel detection while residing within the same noise region and linear range.

Two-dimensional chromatographic analysis using three second-dimension columns for continuous comprehensive analysis of intact proteins.

We develop a new two-dimensional (2D) high performance liquid chromatography (HPLC) approach for intact protein analysis. Development of 2D HPLC has a bottleneck problem - limited second-dimension (second-D) separation speed. We solve this problem by incorporating multiple second-D columns to allow several second-D separations to be proceeded in parallel. To demonstrate the feasibility of using this approach for comprehensive protein analysis, we select ion-exchange chromatography as the first-dimension and reverse-phase chromatography as the second-D. We incorporate three second-D columns in an innovative way so that three reverse-phase separations can be performed simultaneously. We test this system for separating both standard proteins and E. coli lysates and achieve baseline resolutions for eleven standard proteins and obtain more than 500 peaks for E. coli lysates. This is an indication that the sample complexities are greatly reduced. We see less than 10 bands when each fraction of the second-D effluents are analyzed by sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE), compared to hundreds of SDS-PAGE bands as the original sample is analyzed. This approach could potentially be an excellent and general tool for protein analysis.

Backscatter particle image velocimetry via optical time-of-flight sectioning.

Conventional particle image velocimetry (PIV) configurations require a minimum of two optical access ports, inherently restricting the technique to a limited class of flows. Here, the development and application of a novel method of backscattered time-gated PIV requiring a single-optical-access port is described along with preliminary results. The light backscattered from a seeded flow is imaged over a narrow optical depth selected by an optical Kerr effect (OKE) time gate. The picosecond duration of the OKE time gate essentially replicates the width of the laser sheet of conventional PIV by limiting detected photons to a narrow time-of-flight within the flow. Thus, scattering noise from outside the measurement volume is eliminated. This PIV via the optical time-of-flight sectioning technique can be useful in systems with limited optical access and in flows near walls or other scattering surfaces.

Miniaturized high-performance liquid chromatography instrumentation.

Miniaturized high performance liquid chromatography (HPLC) has attracted increasing attention for its potential in high-throughput analyses and point-of-care applications. In this review we highlight the recent advancements in HPLC system miniaturization. We focus on the major components that constitute these instruments along with their respective advantages and drawbacks as well as present a few representative miniaturized HPLC systems. We discuss briefly some of the applications and also anticipate the future development trends of these instrumental platforms.