The deleterious effects of adversity are likely intergenerational, such that one generation’s adverse experiences can affect the next. Epidemiological studies link maternal adversity to offspring depression and anxiety, possibly via transmission mechanisms that influence offspring fronto-limbic connectivity. However, studies have not thoroughly disassociated postnatal exposure effects nor considered the role of offspring sex. We utilized infant neuroimaging to test the hypothesis that maternal childhood maltreatment (CM) would be associated with increased fronto-limbic connectivity in infancy and tested brain-behavior associations in childhood. Ninety-two dyads participated (32 mothers with CM, 60 without; 52 infant females, 40 infant males). Women reported on their experiences of CM and non-sedated sleeping infants underwent MRIs at 2.44 ± 2.74 weeks. Brain volumes were estimated via structural MRI and white matter structural connectivity (fiber counts) via diffusion MRI with probabilistic tractography. A subset of parents (n = 36) reported on children’s behaviors at age 5.17 ± 1.73 years. Males in the maltreatment group demonstrated greater intra-hemispheric fronto-limbic connectivity (b = 0.96, p= 0.008, [95%CI 0.25, 1.66]), no differences emerged for females. Fronto-limbic connectivity was related to somatic complaints in childhood only for males (r = 0.673, p = 0.006). Our findings suggest that CM could have intergenerational associations to offspring brain development, yet mechanistic studies are needed.

OBJECTIVES/GOALS: Current COVID-19 rapid molecular tests require cartridge-reader detection, expensive circuitry, and complex microfluidics making the most accurate tests unavailable to the masses. Here we present a rapid molecular diagnostic leveraging isothermal amplification and paper-based microfluidics for a low-cost ultra-sensitive COVID-19 assay. METHODS/STUDY POPULATION: We designed a reverse transcription recombinase polymerase amplification (RT-RPA) assay for the detection of SARS-CoV-2 and bacteriophage MS2 RNA. RT-RPA is a sequence specific, ultrasensitive, rapid isothermal DNA amplification technique that is well suited to home based testing due to its rapid assay time, robustness, ease of use, and readout options. RT-RPA reagents are added to a tube and incubated at 39°C in a fluorometer. Realtime fluorometer data gives results in under 15 minutes. This assay also provides visual detection via lateral flow readout with results in 23 minutes. RESULTS/ANTICIPATED RESULTS: We have developed a rapid multiplexed nucleic acid amplification assay with an internal process control for SARS-CoV-2 using single-pot RT-RPA. We screened 21 primer combinations to select primers that demonstrated excellent performance and target specificity against common respiratory viruses. We demonstrate the ability to multiplex SARS-CoV-2 and MS2 detection, utilizing MS2 as an internal process control for lysis, reverse transcription, amplification, and readout. We show duplexed detection using both fluorescence readout and visual readout using lateral flow strips. Duplexed fluorescence detection shows a limit of detection of 25 copies per reaction. Duplexed lateral flow readout shows a limit of detection of 50 copies per reaction DISCUSSION/SIGNIFICANCE: We developed a duplexed RT-RPA assay for SARS-CoV-2 with fluorescence or lateral flow readout. Our assay does not re-quire expensive reader, circuity, or fluid handling. The low material cost, temperature, and robustness make it ideal for a more accurate home-based COVID-19 diagnostic.

In this three-generation longitudinal study of familial depression, we investigated the continuity of parenting styles, and major depressive disorder (MDD), temperament, and social support during childrearing as potential mechanisms. Each generation independently completed the Parental Bonding Instrument (PBI), measuring individuals’ experiences of care and overprotection received from parents during childhood. MDD was assessed prospectively, up to 38 years, using the semi-structured Schedule for Affective Disorders and Schizophrenia (SADS). Social support and temperament were assessed using the Social Adjustment Scale – Self-Report (SAS-SR) and Dimensions of Temperament Scales – Revised, respectively. We first assessed transmission of parenting styles in the generation 1 to generation 2 cycle (G1→G2), including 133 G1 and their 229 G2 children (367 pairs), and found continuity of both care and overprotection. G1 MDD accounted for the association between G1→G2 experiences of care, and G1 social support and temperament moderated the transmission of overprotection. The findings were largely similar when examining these psychosocial mechanisms in 111 G2 and their spouses (G2+S) and their 136 children (G3) (a total of 223 pairs). Finally, in a subsample of families with three successive generations (G1→G2→G3), G2 experiences of overprotection accounted for the association between G1→G3 experiences of overprotection. The results of this study highlight the roles of MDD, temperament, and social support in the intergenerational continuity of parenting, which should be considered in interventions to “break the cycle” of poor parenting practices across generations.

ABSTRACT IMPACT: This project successfully implemented a promising team science model by introducing and facilitating best practices to develop high functioning teams working to accelerate health innovations from bench to bedside. OBJECTIVES/GOALS: The goal of this project was to improve the team science knowledge, skills, and attitudes of interdisciplinary engineering students (undergraduate and graduate) who were partnered with health professionals to develop technical solutions to translational health challenges during a year-long Engineering Innovation in Health (EIH) program. METHODS/STUDY POPULATION: We adapted, implemented, and evaluated team science training content and approaches in the EIH program at the University of Washington (UW). EIH faculty and the UW Institute of Translational Health Sciences’ (ITHS) Team Science Core co-developed and delivered highly interactive team science training modules and evaluated their impact with biannual surveys. A student cohort was surveyed prior to the implementation of the team science trainings, which served as a baseline. Descriptive statistics were used to summarize student demographics and survey responses within and between years. Median and interquartile range of responses to Likert-type questions were calculated, and Mann-Whitney U Tests (independent samples Wilcoxin Rank Sum Tests) were used to test for differences within and between years. RESULTS/ANTICIPATED RESULTS: During both the baseline and the team training year, student demographics were similar in terms of gender and past experience working in teams. Team training during the first year of implementation was well-received. Post-implementation surveys of students demonstrated measurable improvement in team dynamics, communication, and effectiveness; including, students reporting higher levels of psychological safety and self-efficacy within their teams. Comparisons within the team training year and between the baseline and team training years identified numerous instances in which differences were statistically significant. DISCUSSION/SIGNIFICANCE OF FINDINGS: Tailored team science training in an interdisciplinary EIH program was successful at improving psychological safety and self-efficacy among undergraduate and graduate students and offers a promising model for similar settings and audiences.

ABSTRACT IMPACT: Recognizing Interdisciplinary Collaborative Research in Promotion and Tenure Processes OBJECTIVES/GOALS: Academic institutions have traditionally focused on individual achievements for promotion. We present our effort on identifying and measuring attitudes on promotion and tenure (PT) criteria that values and rewards interdisciplinary research (IR). We have developed a toolkit to facilitate the recognition of IR in PT processes. METHODS/STUDY POPULATION: Our group reviewed appointment, promotion and tenure (APT) policies from each of the six Health Science Schools and the College of Engineering at the University of Washington (UW) to assess language of objective criteria and attributes of IR to guide APT committees in the evaluation of interdisciplinary researchers. We surveyed faculty about their attitudes relating to IR within the context of promotion and tenure. Interviews of department chairs and administrators about institutional policies and infrastructure that supports or inhibits IR, and current best practices, were conducted. We have developed toolkits for junior faculty, department chairs, external reviewers, and APT committees to facilitate rewarding IR at promotion. RESULTS/ANTICIPATED RESULTS: Review of APT policies found criteria that recognizes IR for APT in three schools. 118 faculty responded to the survey (44% Professor, 26% Associate, and 37% eligible for APT committees). The majority of faculty reported they were currently conducting IR (95%), considered IR important (98%), and believed the UW faculty code should encourage IR (85%). Although a vast majority considered their units supportive of IR (>80%), less than half (43%) reported that their APT criteria provided examples that included participation in IR. Our survey also found that APT committees were challenged about best practices to reward IR, APT external reviewers struggle to evaluate individual vs team contributions, and individual faculty are challenged to describe contributions for APT within context of an interdisciplinary team. DISCUSSION/SIGNIFICANCE OF FINDINGS: IR is conducted and valued by UW faculty; however, current structures, policies, and APT code do not facilitate IR for promotion and tenure. We have developed a toolkit for promotion-eligible faculty, chairs, external reviewers, and APT committees to facilitate IR. Our goal is to modify UW faculty code and unit APT criteria to recognize and reward IR.

Severe temper outbursts (STO) in children are associated with impaired school and family functioning and may contribute to negative outcomes. These outbursts can be conceptualized as excessive frustration responses reflecting reduced emotion regulation capacity. The anterior cingulate cortex (ACC) has been implicated in negative affect as well as emotional control, and exhibits disrupted function in children with elevated irritability and outbursts. This study examined the intrinsic functional connectivity (iFC) of a region of the ACC, the anterior midcingulate cortex (aMCC), in 5- to 9-year-old children with STO (n = 20), comparing them to children with attention-deficit/hyperactivity disorder (ADHD) without outbursts (ADHD; n = 18). Additional analyses compared results to a sample of healthy children (HC; n = 18) and examined specific associations with behavioral and emotional dysregulation. Compared to the ADHD group, STO children exhibited reduced iFC between the aMCC and surrounding regions of the ACC, and increased iFC between the aMCC and precuneus. These differences were also seen between the STO and HC groups; ADHD and HC groups did not differ. Specificity analyses found associations between aMCC–ACC connectivity and hyperactivity, and between aMCC–precuneus iFC and emotion dysregulation. Disruption in aMCC networks may underlie the behavioral and emotional dysregulation characteristic of children with STO.

Mitchell originally proposed that an asymmetric ion flux across an organism's membrane could generate electric fields that drive locomotion. Although this locomotion mechanism was later rejected for some species of bacteria, engineered Janus particles have been realized that can swim due to ion fluxes generated by asymmetric electrochemical reactions. Here we present governing equations, scaling analyses and numerical simulations that describe the motion of bimetallic rod-shaped motors in hydrogen peroxide solutions due to reaction-induced charge auto-electrophoresis. The coupled Poisson–Nernst–Planck–Stokes equations are numerically solved using Frumkin-corrected Butler–Volmer equations to represent electrochemical reactions at the rod surface. Our simulations show strong agreement with the scaling analysis and experiments. The analysis shows that electrokinetic locomotion results from electro-osmotic fluid slip around the nanomotor surface. The electroviscous flow is driven by electrical body forces which are generated from a coupling of a reaction-induced dipolar charge density distribution and the electric field it creates. The magnitude of the electroviscous velocity increases quadratically with the surface reaction rate for an uncharged motor, and linearly when the motor supports a finite surface charge.

An experimental investigation of electrokinetic instabilities (EKIs) of non-dilute colloidal suspensions in microchannels is presented. The addition of charged colloidal particles to a solution can alter the solution's electrical conductivity and permittivity as well as the average particle electrophoretic mobility. In this work, a colloidal (500 nm polystyrene) volume fraction gradient is achieved at the intersection of a Y-shaped polydimethylsiloxane (PDMS) microchannel. The flow becomes unstable when the electroviscous stretching and folding of the conductivity and permittivity interfaces exceed the dissipative effects of viscous forces and particle diffusion. The suspension conductivity as a function of the particle volume fraction is presented. The critical conditions required for flow instability are measured along with a scaling analysis which shows that the flow becomes unstable due to a coupling of applied electric fields and the electrical conductivity and permittivity gradients in the flow. The flow becomes unstable at a critical electric Rayleigh number of Rae = 1.8 × 105 for a wide range of applied electric fields spanning three orders of magnitude and colloid volume fractions varying two orders of magnitude. EKIs of non-dilute colloidal suspensions may be important for applications such as the electrophoretic deposition of micropatterned colloidal assemblies, electrorheological devices and on-chip electrokinetic (EK) manipulation of colloids.

Nanoscale apertures that provide a fluidic path between two reservoirs can be used for numerous applications. These applications include patch-clamp type measurements, Coulter counting and molecular studies. For Coulter counting of nanometer-sized analytes, we have developed a process capable of reproducibly fabricating cylindrical apertures in a silicon-on-insulator substrate with diameters less than 30 nm. The fabrication process utilizes electron beam lithography for the lithographic definition of the apertures enabling accurate control of final device dimensions. Measurements of the conductance of the pores as a function of KCl concentration reveal the presence of a surface conduction mechanism that dominates the conductance of the nanopore and leads to a deviation of the concentration dependence of the conductance from the case of bulk solution. From current traces recorded, the passage of individual particles through the pore can be concluded.

We present a parametric experimental study of convective electrokinetic instability (EKI) in an isotropically etched, cross-shaped microchannel using quantitative epifluorescence imaging. The base state is a three-inlet, one-outlet electrokinetic focusing flow configuration where the centre sample stream and sheath flows have mismatched ionic conductivities. Electrokinetic flows with conductivity gradients become unstable when the electroviscous stretching and folding of conductivity interfaces grows faster than the dissipative effect of molecular diffusion. Scalar images, critical applied fields required for instability, and temporal and spatial scalar energy are presented for flows with a wide range of applied d.c. electric field and centre-to-sheath conductivity ratios. These parameters impose variations of the electric Rayleigh number across four orders of magnitude. We introduce a scaling for charge density in the bulk fluid as a function of local maximum conductivity gradients in the flow. This scaling shows that the flow becomes unstable at a critical electric Rayleigh number ($Ra_{e, \ell} \,{=}\, 205$) and applies to a wide range of applied field and centre-to-sheath conductivity ratios. This work is relevant to on-chip electrokinetic flows with conductivity gradients such as field amplified sample stacking, flow at the intersections of multi-dimensional assays, electrokinetic control and separation of sample streams with poorly specified chemistry, and low-Reynolds number micromixing.

The circumplex model of affect proposes that all affective states arise from cognitive interpretations of core neural sensations that are the product of two independent neurophysiological systems. This model stands in contrast to theories of basic emotions, which posit that a discrete and independent neural system subserves every emotion. We propose that basic emotion theories no longer explain adequately the vast number of empirical observations from studies in affective neuroscience, and we suggest that a conceptual shift is needed in the empirical approaches taken to the study of emotion and affective psychopathologies. The circumplex model of affect is more consistent with many recent findings from behavioral, cognitive neuroscience, neuroimaging, and developmental studies of affect. Moreover, the model offers new theoretical and empirical approaches to studying the development of affective disorders as well as the genetic and cognitive underpinnings of affective processing within the central nervous system.This work was supported in part by NIMH Grants MH01232, MH59139, MH36197, MHK02-74677, and MH068318; a grant from the National Alliance for Research in Schizophrenia and Affective Disorders (NARSAD); NSF Grant BSC-0421702; and funding from the Thomas D. Klingenstein and Nancy D. Perlman Family Fund and the Suzanne Crosby Murphy Endowment at Columbia University.

A novel deposition process has been developed for manufacturing of high quality rare-earth doped glass films with complex composition. This process, Laser Reactive Deposition (LRDTM), comprises photothermal laser pyrolysis using an aerosol feedstock coupled with deposition of the resulting nanoparticle product onto Si substrates with our without an underlying glass film. This paper describes this novel process as well as the structure, properties and performance of the Erdoped, multicomponent silicate glass films produced to date with this process.