Cross-Platform Synaptic Network Analysis of Human Entorhinal Cortex Identifies TWF2 as a Modulator of Dendritic Spine Length.

Proteomic studies using postmortem human brain tissue samples have yielded robust assessments of the aging and neurodegenerative disease(s) proteomes. While these analyses provide lists of molecular alterations in human conditions, like Alzheimer's disease (AD), identifying individual proteins that affect biological processes remains a challenge. To complicate matters, protein targets may be highly understudied and have limited information on their function. To address these hurdles, we sought to establish a blueprint to aid selection and functional validation of targets from proteomic datasets. A cross-platform pipeline was engineered to focus on synaptic processes in the entorhinal cortex (EC) of human patients, including controls, preclinical AD, and AD cases. Label-free quantification mass spectrometry (MS) data (n = 2260 proteins) was generated on synaptosome fractionated tissue from Brodmann area 28 (BA28; n = 58 samples). In parallel, dendritic spine density and morphology was measured in the same individuals. Weighted gene co-expression network analysis was used to construct a network of protein co-expression modules that were correlated with dendritic spine metrics. Module-trait correlations were used to guide unbiased selection of Twinfilin-2 (TWF2), which was the top hub protein of a module that positively correlated with thin spine length. Using CRISPR-dCas9 activation strategies, we demonstrated that boosting endogenous TWF2 protein levels in primary hippocampal neurons increased thin spine length, thus providing experimental validation for the human network analysis. Collectively, this study describes alterations in dendritic spine density and morphology as well as synaptic proteins and phosphorylated tau from the entorhinal cortex of preclinical and advanced stage AD patients.SIGNIFICANCE STATEMENT Proteomic studies can yield vast lists of molecules that are altered under various experimental or disease conditions. Here, we provide a blueprint to facilitate mechanistic validation of protein targets from human brain proteomic datasets. We conducted a proteomic analysis of human entorhinal cortex (EC) samples spanning cognitively normal and Alzheimer's disease (AD) cases with a comparison of dendritic spine morphology in the same samples. Network integration of proteomics with dendritic spine measurements allowed for unbiased discovery of Twinfilin-2 (TWF2) as a regulator of dendritic spine length. A proof-of-concept experiment in cultured neurons demonstrated that altering Twinfilin-2 protein level induced corresponding changes in dendritic spine length, thus providing experimental validation for the computational framework.

Per-pathogen virulence of HIV-1 subtypes A, C and D.

HIV-1 subtypes differ in their clinical manifestations and the speed in which they spread. In particular, the frequency of subtype C is increasing relative to subtypes A and D. We investigate whether HIV-1 subtypes A, C and D differ in their per-pathogen virulence and to what extend this explains the difference in spread between these subtypes. We use data from the hormonal contraception and HIV-1 genital shedding and disease progression among women with primary HIV infection study. For each study participant, we determine the set-point viral load value, CD4+ T cell level after primary infection and CD4+ T cell decline. Based on both the CD4+ T cell count after primary infection and CD4+ T cell decline, we estimate the time until AIDS. We then obtain our newly introduced measure of virulence as the inverse of the estimated time until AIDS. After fitting a model to the measured virulence and set-point viral load values, we tested if this relation varies per subtype. We found that subtype C has a significantly higher per-pathogen virulence than subtype A. Based on an evolutionary model, we then hypothesize that differences in the primary length of infection period cause the observed variation in the speed of spread of the subtypes.

County incidence and geospatial trends of early-onset hypertensive disorders of pregnancy in Kentucky, 2008-2017.

BACKGROUND: Early-onset hypertensive disorders of pregnancy (eHDP) are associated with more severe maternal and infant outcomes than later-onset disease. However, little has been done to evaluate population-level trends. Therefore, in this paper, we seek to address this understudied area by describing the geospatial and temporal patterns of county-level incidence of eHDP and assessing county-level demographics that may be associated with an increased incidence of eHDP. METHODS: Employing Kentucky certificates of live and stillbirth from 2008-2017, this ecological study detected county-level clusters of early-onset hypertensive disorders of pregnancy using SaTScan, calculated average annual percent change (AAPC) with a join point analysis, and identified county-level covariates (% of births to women ≥ 35 years of age, % with BMI ≥ 30 kg/m2, % currently smoking, % married, and % experienced eHDP) with a fixed-effects negative binomial regression model for longitudinal data with an autoregressive (AR) correlation structure offset with the natural log of the number of births in each county and year. RESULTS: County-level incidence of eHDP had a non-statistically significant increase of almost 3% (AAPC: 2.84, 95% CI: -4.26, 10.46), while maternal smoking decreased by almost 6% over the study period (AAPC:-5.8%, 95%CI: -7.5, -4.1), Risk factors for eHDP such as pre-pregnancy BMI ≥ 30 and proportion of births to women ≥ 35 years of age increased by 2.3% and 3.4% respectively (BMI AAPC:2.3, 95% CI: 0.94, 3.7; ≥ 35 years AAPC:3.4, 95% CI: 0.66, 6.3). After adjusting for race, county-level proportions of college attainment, and maternal smoking throughout pregnancy, counties with the highest proportion of births to women with BMI ≥ 30 kg/m2 reported an eHDP incidence 20% higher than counties with a lower proportion of births to mothers with a BMI ≥ 30 kg/m2 and a 20% increase in eHDP incidence (aRR = 1.20, 95% CI: 1.00, 1.44). We also observed that counties with the highest proportion vs. the lowest of mothers ≥ 35 years old (> 6.1%) had a 26% higher incidence of eHDP (RR = 1.26, 95%CI: 1.04, 1.50) compared to counties with the lowest incidence (< 2.5%). We further identified two county-level clusters of elevated eHDP rates. We also observed that counties with the highest vs. lowest proportion of mothers ≥ 34 years old (> 6.1% vs. < 2.5%) had a 26% increase in the incidence of eHDP (RR = 1.26, 95% CI: 1.04, 1.50). We further identified two county-level clusters of elevated incidence of eHDP. CONCLUSIONS: This study identified two county-level clusters of eHDP, county-level covariates associated with eHDP, and that while increasing, the average rate of increase for eHDP was not statistically significant. This study also identified the reduction in maternal smoking over the study period and the concerning increase in rates of elevated pre-pregnancy BMI among mothers. Further work to explore the population-level trends in this understudied pregnancy complication is needed to identify community factors that may contribute to disease and inform prevention strategies.

Investigating the Relation between Electronic Cigarette Use and Sleep Duration in Kentucky Using the BRFSS, 2016-2017.

OBJECTIVES: Like traditional cigarettes, electronic cigarettes (e-cigarettes) contain nicotine, which is known to negatively influence sleep quality. Few studies, however, have examined the relation between e-cigarettes and sleep quality using population-based survey data because of the relatively recent appearance of these products on the market. This study investigated the relation between e-cigarette and cigarette use and sleep duration in Kentucky, a state with high rates of nicotine dependence and related chronic diseases. METHODS: Data from two consecutive years of the Behavioral Risk Factor Surveillance System survey-2016 and 2017-were analyzed using χ2 statistics and multivariable Poisson regression analyses to control for socioeconomic and demographic variables, the presence of other chronic diseases, and traditional cigarette use. RESULTS: This study was conducted using responses from 18,907 Kentucky adults aged 18 years and older. Overall, almost 40% reported short (<7 hours) sleep duration. After adjusting for other covariates, including the presence of chronic diseases, those who had currently or formerly used both traditional and e-cigarettes had the highest risk of short sleep duration. Those who currently or formerly smoked only traditional cigarettes also had significantly higher risk, unlike those who had only used e-cigarettes. CONCLUSIONS: Survey respondents who used e-cigarettes were more likely to report short sleep duration, but only if they currently or formerly smoked traditional cigarettes. Those who used both products, regardless of whether current or former, were more likely to report short sleep duration than those who had used just one of these tobacco products.

Estimating the Population Attributable Fraction of Asthma Due to Electronic Cigarette Use and Other Risk Factors Using Kentucky Behavioral Risk Factor Survey Data, 2016-2017.

Introduction: Electronic nicotine delivery systems ENDS have become popular in the United States among both new users of nicotine and those seeking less harmful alternatives to traditional cigarettes. Users often perceive ENDS as being less harmful than traditional cigarettes. This study investigated the relationship between use of ENDS and asthma in a representative sample of adults. Methods: For this cross-sectional study, we used data from the Kentucky Behavioral Risk Factor Surveillance System telephone survey data from 2016-2017. Using a weighted multivariable logistic regression analysis, we identified important covariates to adjust for to calculate the population attributable fraction (PAF) of asthma due to ENDS and other modifiable risk factors factors (cigarette use, obesity, education, and employment). The confidence intervals for the PAFs were estimated using bootstrap methods of variance estimation. Results: We found that 10.6% of those aged 18-30 reported currently had asthma. After adjusting for noted covariates, ENDS use did not significantly increase the odds of asthma. In the final PAF model, the PAF of asthma due to ENDS was 0.4% (95% CI: -5.41, 6.21). Conclusion: While these findings suggest only modest effects of ENDS use on asthma prevalence, future research including older age groups and more long-term users might produce different results.

Racial differences in college-student drinking.

Racial differences in drinking motives, protective behavioral strategies (PBSs), alcohol consumption, and alcohol-related problems were examined among college student drinkers (N = 443: 296 [66.8%] White, 147 [33.3%] Black). Survey participants were recruited from large undergraduate sociology classes and residence halls at the university. Key differences between Black and White college students in drinking behaviors, reasons for drinking (i.e., motives), and the use of PBSs were observed. These racial differences have implications for the implementation of prevention/intervention programs intending to reduce alcohol consumption and alcohol-related problems among college students.

Spatiotemporal Analysis of Lung Cancer Histological Types in Kentucky, 1995-2014.

Recent metabolic and genetic research has demonstrated that risk for specific histological types of lung cancer varies in relation to cigarette smoking and obesity. This study investigated the spatial and temporal distribution of lung cancer histological types in Kentucky, a largely rural state with high rates of smoking and obesity, to discern population-level trends that might reflect variation in these and other risk factors. The Kentucky Cancer Registry provided residential geographic coordinates for lung cancer cases diagnosed from 1995 through 2014. We used multinomial and discrete Poisson spatiotemporal scan statistics, adjusted for age, gender, and race, to characterize risk for specific histological types-small cell, adenocarcinoma, squamous cell, and other types-throughout Kentucky and compared to maps of risk factors. Toward the end of the study period, adenocarcinoma was more common among all population subgroups in north-central Kentucky, where smoking and obesity are less prevalent. During the same time frame, squamous cell, small cell, and other types were more common in rural Appalachia, where smoking and obesity are more prevalent, and in some high poverty urban areas. Spatial and temporal patterns in the distribution of histological types of lung cancer are likely related to regional variation in multiple risk factors. High smoking and obesity rates in the Appalachian region, and likely in high poverty urban areas, appeared to coincide with high rates of squamous cell and small cell lung cancer. In north-central Kentucky, environmental exposures might have resulted in higher risk for adenocarcinoma specifically.

Effect of Local Smoke-Free Ordinances on Smoking Prevalence in Kentucky, 2002-2009.

OBJECTIVES: Many local communities in Kentucky, a state with one of the highest smoking prevalence rates in the United States, have enacted smoke-free ordinances that prohibit smoking in workplaces and enclosed buildings open to the public. Research has shown that such ordinances are clearly beneficial for public health, but their influence on smoking prevalence in the populations they cover remains unclear. This study explores the effect of local smoke-free ordinances on smoking prevalence in Kentucky. METHODS: We used a database of smoke-free ordinances maintained by the Kentucky Center for Smoke-Free Policy, Kentucky Behavioral Risk Factor Surveillance System survey data, and US Census data. We estimated the proportion of Kentucky adults living in counties with smoke-free ordinances of varying strength; examined bivariate associations between smoke-free ordinances and smoking prevalence; and fit regression models that adjusted for various county-level demographic, socioeconomic, and geographic factors. RESULTS: Smoking prevalence was approximately 5% lower in counties with smoke-free ordinances, even after adjusting for other relevant factors, including a trend in decreasing prevalence throughout the study region. There was a slight dose-response effect related to the strength of smoke-free ordinances after adjustment for these covariates. Smoke-free ordinances appear to have a modest effect on smoking prevalence across the span of several years. CONCLUSIONS: Findings demonstrate that although smoking prevalence fell throughout the state during the study period, counties with smoke-free ordinances experienced a greater decline. Future research should examine the strength of smoke-free ordinances in greater detail to better understand their influence on smoking prevalence.

Safety and efficacy of antibiotics among acutely decompensated cirrhosis patients.

BACKGROUND AND AIM: Infection is a leading precipitant of acute-on-chronic liver failure. This study aims to determine the safety and efficacy of antibiotics within acute-on-chronic liver failure. METHODS: Retrospective study of 457 acute-on-chronic liver failure patients admitted to the University of Arizona Health Network between January 1 and December 31, 2014. Eligibility criteria were as follows: at least 18 years of age and 6 months follow-up, data available to calculate systemic inflammatory response syndrome (SIRS), and acute-on-chronic liver failure. This study collected patient's clinical features and historical data. Key data points were infection, antibiotic use, and SIRS. This study used Cox proportional hazards to model the effects of clinical factors on risk of death. RESULTS: A total of 521 of 1243 met the inclusion criteria, and 64 had missing data, leaving 457 patients. Infection resulted in higher hazard (hazard ratio [HR] = 1.6, confidence interval [CI]: 1.1-1.3, P = 0.01). Patients with infections and antibiotics, compared with non-infected patients without antibiotics, had higher hazard (HR = 1.633, CI: 1.022-2.609, P = .04). Of those infected patients with antibiotics, SIRS patients experienced higher hazard (HR = 1.9, CI: 1.1-3.0, P = .007). Multivariable Cox proportional hazards associated the following with higher hazard: SIRS (HR = 1.866, CI: 1.242-2.804, P = 0.003), vancomycin (HR = 1.640, CI: 1.119-2.405, P = 0.011), Model for End-Stage Liver Disease (HR = 1.051, CI: 1.030-1.073, P < 0.001), gastrointestinal bleeding (HR = 1.727, CI: 1.180-2.527, P = 0.005), and hepatic encephalopathy (HR = 1.807, CI: 1.247-2.618, P = 0.002). CONCLUSION: Overall, treatment of infection with antibiotics did not improve survival; however, patients not meeting SIRS criteria had better outcomes, and vancomycin was associated with poorer survival among acute-on-chronic liver failure patients.

Measuring Alcohol Marketing Engagement: The Development and Psychometric Properties of the Alcohol Marketing Engagement Scale.

Ample evidence exists in support of the influence of media, both traditional and electronic, on perceptions and engagement with alcohol marketing. We describe the development, calibration, and evidence for technical quality and utility for a new measure, the Alcohol Marketing Engagement Scale. Using two samples of college undergraduates (n1 = 199, n2 = 732), we collected field test responses to a total of 13 items. Initial support for scale validity is presented via correlations with attributes previously shown to be related to alcohol engagement. While the joint map of estimated scale locations of items and respondents indicates the need for further scale development, the results of the present analyses are promising. Implications for use in research are discussed.

DUI recidivism by intervention adherence: a multiple risk factor approach.

BACKGROUND: Multi-component impaired driving interventions can reduce driving under the influence (DUI) recidivism rates; however, outcomes are better for those who complete the interventions and are adherent with program expectations. Research is needed to examine the differences between DUI offenders who are adherent vs. non-adherent to intervention efforts. OBJECTIVE: The current study utilized a multi-risk factor model to predict recidivism among first-time DUI offenders enrolled in an intervention program. Differences between offenders who were adherent (including program completion) vs. non-adherent with the intervention were examined. METHOD: Using data from the Mississippi Alcohol Safety Education Program (MASEP) and state administrative records, the current study examined both recidivism rates and rates of intervention completion for all individuals enrolled in the program. The sample was predominantly White (58.8%) and male (80.2%). RESULTS: Individuals who were adherent with the intervention were significantly less likely to recidivate within 3 years. Prior traffic or other criminal violations positively predicted recidivism rates. The likelihood of recidivism varied, with males, African Americans, and younger individuals with less education at greater risk of recidivism. Adherence with the intervention was more common for African American and older participants. CONCLUSION: The MASEP participants who were adherent with the intervention were significantly less likely to recidivate than those who were non-adherent. However, variance in the multi-component intervention completion rates suggests that the program resonates better with specific population subsets. We argue for researchers and policymakers to further explore how specific population subsets react to varying intervention programs to maximize efforts to reduce impaired driving.

Parental and emerging adult psychopathology: Moderated mediation by gender and affect toward parents.

Current research indicates that children tend to view parents with psychopathology more negatively and children who hold negative perceptions of parents are at a greater risk for psychopathology. Yet, less research examines how parental psychopathology influences offspring psychopathology through affect toward parents. The current study tested a model that examined the associations among parental psychopathology, positive affect toward parents, and emerging adult psychopathology. Associations were expected to be partly indirect via positive affect toward parents and emerging adult gender was expected to moderate these associations. Results indicated gender-moderated mediation with significant effects found for males but not females. Results from the current study emphasize the importance of examining affect toward parents as a risk factor for emerging adult psychopathology. Additionally, results of the current study demonstrate the importance of examining the role of emerging adult gender as a potential moderator in these relationships.

Comparison of digital recruitment strategies for Alzheimer's disease patients.

Objectives: Clinical trials studying Alzheimer's Disease (AD) face the challenge of recruiting participants with significant barriers to entering research studies. The objective of this study is to compare digital recruitment strategies' ability to recruit older adults with cognitive impairment (CI). Methods: Older adults with CI were recruited for a clinical trial studying vestibular therapy in reducing falls and improving balance and cognition in older adults with CI. Potential participants were recruited via two different digital recruitment methods, a direct messaging campaign using established patient records and a social media campaign. Potential participants then filled out surveys to determine eligibility for the study. Results: The direct messaging campaign contacted 3060 potential participants and the social media campaign resulted in 8265 instances of unique engagement. Of the number of people reached, the direct messaging campaign had a higher percentage of people who submitted the survey compared to the social media campaign (8.3% vs. 1.2%, p < 0.001). There was no significant difference in age, race, ethnicity, education, household income, and insurance status between the recruitment groups (p > 0.05). Direct messaging recruitment proved more cost-effective at $21.74 per eligible participant compared to the social media campaign at $859.58 per eligible participant. Conclusion: This study found that direct messaging recruitment using established patient records was more cost-effective compared to social media recruitment for this clinical trial. In this sample size, similar demographics were reached by both recruitment methods. Future studies should continue to explore the use of social media and alternative methods to recruit representative participant populations for ongoing AD research.

Parental Discipline During Emerging Adulthood: A Person-Centered Approach.

Although most parental discipline research examines the effects of discipline in children and adolescents, recent research has demonstrated that emerging adults continue to receive parental discipline. Importantly, a newly validated instrument for assessing discipline specifically during emerging adulthood has been created. Scales from this instrument include maternal and paternal approval, disappointment, and abuse, and these scales were associated with other parenting behaviors and psychological outcomes during emerging adulthood. However, a person-centered approach has not been conducted with this instrument. Given that discipline occurs at an idiographic level and that group norms inform such behaviors, a person-centered approach would identify highly informative emerging adult profiles based on patterns of discipline they receive from their parents. Thus, the current study utilized latent profile analysis (LPA) of 1110 participants attending a Southern United States university to identify emerging adult discipline profiles. These groups were then associated with parental and emerging adult psychological problems to gain an understanding of how these factors relate to different patterns of discipline across gender. Results best supported four profiles labeled as approving, distant, disappointed, and abusive. These groups reported increasingly higher parental and personal psychological problems across approving, distant, disappointed, and abusive profiles. Gender moderated some of these effects. Implications and future directions are discussed.

Assessing attitudes towards pediatric research participation across diverse populations: Psychometric properties of a novel tool.

BACKGROUND: Clinical trials play a crucial role in advancing medical knowledge and improving health outcomes. However, there is a recognized need for greater representation of marginalized groups to ensure that research findings can be generalized and effectively applied to all individuals. While the Pediatric Research Participation Questionnaire (PRPQ) was developed to assist pediatric clinical trials research by identifying benefits and barriers to research participation among children with chronic medical conditions, there is still limited insight into the structure of the PRPQ when administered in diverse samples, including the general pediatric population. Therefore, the current study examined the factor structure of the PRPQ in a general pediatric population to investigate whether rural-urban differences exist in the PRPQ factor structure. METHODS: Caregivers (N = 600) of children under age 18 completed the PRPQ in a population-based survey in Mississippi. Sampling was stratified to ensure equal representation in rural (n = 300) and urban areas (n = 300). Exploratory and confirmatory factor analyses were conducted to determine the factor structure of the PRPQ. RESULTS: A five-factor structure was identified, compromising: social pressure, direct benefit, reasons for participation, mistrust in research/researchers, reasons against participation. While results were similar among urban participants, a three-factor structure emerged for rural participants. CONCLUSIONS: This study contributes to the broader understanding of research participation among underrepresented groups. The findings suggest that clinical researchers should consider tailoring recruitment strategies to increase clinical trial participation among children in rural areas. Understanding factors that influence pediatric research participation, particularly among marginalized communities, is crucial for developing effective recruitment and retention strategies.

Quantitative vestibular assessment: The development and validation of a novel, remote video head impulse test against in-clinic measurements.

Objectives: To develop a novel remote head impulse test (rHIT), and to provide preliminary data validating the rHIT vestibular-ocular reflex (VOR) gains against the in-clinic vHIT. Methods: A convenience sample of 10 patients referred for vestibular assessment at our institution was recruited. In-clinic vHIT was used to quantify lateral VOR gains. Patients subsequently underwent an rHIT protocol, whereby patients performed active, lateral head rotations while their eyes and heads were recorded using a laptop camera and video-conferencing software. The vHIT and rHIT VOR gains were compared using paired t-tests, and a Pearson correlation coefficient between the gains was calculated. Absolute accuracy, sensitivity, and specificity of the rHIT were additionally calculated. Results: Of the 10 patients recruited, 4 were male, and the average ± standard deviation (SD) age was 61.4 ± 15.3 years. As determined by the vHIT, 2 patients had normal bilateral VOR gains, 6 with unilateral vestibular hypofunction, and 2 with bilateral vestibular hypofunction. The correlation between the rHIT and vHIT gains was 0.73 (p < .001). The rHIT exhibited an absolute accuracy of 75.0%, sensitivity of 70.0%, and specificity of 80.0%. When ears had a vHIT VOR gain less than 0.40, the rHIT exhibited 100.0% accuracy. Conversely, 60.0% of deficient ears with vHIT VOR gains greater than 0.40 were incorrectly categorized by the rHIT. Conclusion: The rHIT may be better suited for detecting more severe vestibular deficiencies. Future iterations of the rHIT should aim to increase the video frame-rate capabilities to detect subtler VOR impairments. Level of Evidence: 4.

The Intersectionality between Race, Ethnicity, and Residential-Level Socioeconomic Status in Disparities of Head and Neck Cancer Outcomes: A SEER Study.

BACKGROUND: Head and neck cancer (HNC) mortality differs by race, ethnicity, and socioeconomic status (SES). However, it is unclear whether the relationship between race/ethnicity and HNC-specific mortality varies according to the residence-level SES. METHODS: Data from the Surveillance Epidemiology and End Results database included participants with primary HNC between 2006 and 2017 (followed through 2018) to assess the joint association of race/ethnicity and census-tract level SES Yost-index groups (quintiles) with all-cause and HNC-specific mortalities. Relative survival rates at 1, 5, and 10 years were calculated. Multivariable Cox proportional hazard regression models estimated hazard-ratios and 95% confidence intervals for all-cause mortality, and Fine-Gray subdistribution hazard models for HNC-specific mortality. Cumulative incidence curves for HNC-specific deaths were estimated. RESULTS: 76,095 patients were included in the analysis: 63.2% were <65 years, 73.4% male, and 11.3% non-Hispanic (NH) Black. Most patients (58.3%) were diagnosed at regional or distant stages and 20.6% died of HNC. The five-year relative survival rate increased with SES group, with 51.6% in the lowest SES group, and 74.1% in the highest SES group. NH-Black patients had higher risk of all-cause and HNC-specific mortality than NH-White patients, regardless of the SES group. NH-Asian/Pacific Islander and Hispanic patients had higher risk of HNC-specific mortality in some SES groups. CONCLUSIONS: NH-Black patients of all SES strata had significantly worse outcomes. Other factors, such as healthcare quality, may be associated with persistent disparities. IMPACT: The study highlights the persistence of significant racial disparities in HNC survival across socioeconomic categories. There is need to consider additional factors underlying these disparities.

A Molecular Basis of Human Brain Connectivity.

Neuroimaging is commonly used to infer human brain connectivity, but those measurements are far-removed from the molecular underpinnings at synapses. To uncover the molecular basis of human brain connectivity, we analyzed a unique cohort of 98 individuals who provided neuroimaging and genetic data contemporaneous with dendritic spine morphometric, proteomic, and gene expression data from the superior frontal and inferior temporal gyri. Through cellular contextualization of the molecular data with dendritic spine morphology, we identified hundreds of proteins related to synapses, energy metabolism, and RNA processing that explain between-individual differences in functional connectivity and structural covariation. By integrating data at the genetic, molecular, subcellular, and tissue levels, we bridged the divergent fields of molecular biology and neuroimaging to identify a molecular basis of brain connectivity. One-Sentence Summary: Dendritic spine morphometry and synaptic proteins unite the divergent fields of molecular biology and neuroimaging.

Endogenous recapitulation of Alzheimer's disease neuropathology through human 3D direct neuronal reprogramming.

Alzheimer's disease (AD) is a neurodegenerative disorder that primarily affects elderly individuals, and is characterized by hallmark neuronal pathologies including extracellular amyloid-ß (Aß) plaque deposition, intracellular tau tangles, and neuronal death. However, recapitulating these age-associated neuronal pathologies in patient-derived neurons has remained a significant challenge, especially for late-onset AD (LOAD), the most common form of the disorder. Here, we applied the high efficiency microRNA-mediated direct neuronal reprogramming of fibroblasts from AD patients to generate cortical neurons in three-dimensional (3D) Matrigel and self-assembled neuronal spheroids. Our findings indicate that neurons and spheroids reprogrammed from both autosomal dominant AD (ADAD) and LOAD patients exhibited AD-like phenotypes linked to neurons, including extracellular Aß deposition, dystrophic neurites with hyperphosphorylated, K63-ubiquitin-positive, seed-competent tau, and spontaneous neuronal death in culture. Moreover, treatment with ß- or γ-secretase inhibitors in LOAD patient-derived neurons and spheroids before Aß deposit formation significantly lowered Aß deposition, as well as tauopathy and neurodegeneration. However, the same treatment after the cells already formed Aß deposits only had a mild effect. Additionally, inhibiting the synthesis of age-associated retrotransposable elements (RTEs) by treating LOAD neurons and spheroids with the reverse transcriptase inhibitor, lamivudine, alleviated AD neuropathology. Overall, our results demonstrate that direct neuronal reprogramming of AD patient fibroblasts in a 3D environment can capture age-related neuropathology and reflect the interplay between Aß accumulation, tau dysregulation, and neuronal death. Moreover, miRNA-based 3D neuronal conversion provides a human-relevant AD model that can be used to identify compounds that can potentially ameliorate AD-associated pathologies and neurodegeneration.

Conventional PCR Detection and Real-Time PCR Quantification of Reniform Nematodes.

Reniform nematode (Rotylenchulus reniformis) is a relatively recent introduction into the continental United States that can cause major yield losses on a variety of important crops including cotton and soybeans. DNA sequences from the internal transcribed spacer (ITS) region of this nematode were used to design primers for conventional and real-time PCR, as well as a TaqMan probe. These primers amplified DNA of reniform nematode isolates from a wide geographic range but did not detect genetically related species or other pathogenic nematodes found in production fields including Meloidogyne incognita and Heterodera glycines. Both SYBR green and TaqMan assays reliably quantified as little as 100 fg of reniform nematode DNA, and could be used to quantify as few as five reniform nematodes. An inexpensive and rapid DNA extraction protocol for high throughput diagnostic assays is described.