Impact of Coronavirus Disease-era Clinical Trial Reform on Cancer Trial Access in Rural/Underserved Regions of the Midwest.

OBJECTIVES: The coronavirus disease 2019 pandemic refocused the cancer community on bringing clinical trials closer to patients and increasing access for traditionally underserved communities. Pandemic-era deregulation increased flexibility with telemedicine visits, less frequent testing, and the ability to have tests done locally. This study evaluates the impact of 2020 cancer clinical trial reform on trial accessibility in rural/underserved regions of the Midwest. METHODS: Publicly available clinicaltrials.gov data was accessed from January 1, 2018 to September 30, 2022 for the 3 leading causes of new cancer cases in Kentucky, Tennessee, Illinois, and Indiana. Interventional trials were categorized based on location using corresponding "Rural-Urban Commuting Area" codes (urban/metropolitan, suburban/micropolitan, small town/rural, and isolated/rural) and categorized as pre versus postpandemic (using March 15, 2020, when national regulatory guidelines were modified). Locations of trial offerings from pre and postpandemic dates were analyzed by paired t test. Comparison of trial location category by state and cancer type was analyzed by 1-way analysis of variance with pairwise multiple comparisons made using the Tukey-Kramer method. RESULTS: Pandemic-era deregulation had no impact on increasing trial availability in suburban and small-town/rural locales ( P = 0.1259). Only 18% of trials were offered outside of urban areas, with 15% in suburban and 3% in small town/rural areas. Results varied by state ( P < 0.0001) with Illinois offering the most suburban and small-town trial availability (27%) compared with Kentucky, Indiana, and Tennessee (18%, 6%, and 2%, respectively). Trial availability in rural versus urban areas did not differ by cancer type ( P = 0.07197). CONCLUSIONS: More work must be done to increase access to cancer clinical trials in rural and suburban areas of the United States.

Comparative site-specific N-glycoproteome analysis reveals aberrant N-glycosylation and gives insights into mannose-6-phosphate pathway in cancer.

N-glycosylation is implicated in cancers and aberrant N-glycosylation is recognized as a hallmark of cancer. Here, we mapped and compared the site-specific N-glycoproteomes of colon cancer HCT116 cells and isogenic non-tumorigenic DNMT1/3b double knockout (DKO1) cells using Fbs1-GYR N-glycopeptide enrichment technology and trapped ion mobility spectrometry. Many significant changes in site-specific N-glycosylation were revealed, providing a molecular basis for further elucidation of the role of N-glycosylation in protein function. HCT116 cells display hypersialylation especially in cell surface membrane proteins. Both HCT116 and DKO1 show an abundance of paucimannose and 80% of paucimannose-rich proteins are annotated to reside in exosomes. The most striking N-glycosylation alteration was the degree of mannose-6-phosphate (M6P) modification. N-glycoproteomic analyses revealed that HCT116 displays hyper-M6P modification, which was orthogonally validated by M6P immunodetection. Significant observed differences in N-glycosylation patterns of the major M6P receptor, CI-MPR in HCT116 and DKO1 may contribute to the hyper-M6P phenotype of HCT116 cells. This comparative site-specific N-glycoproteome analysis provides a pool of potential N-glycosylation-related cancer biomarkers, but also gives insights into the M6P pathway in cancer.

Remnant preservation does not affect accuracy of tibial tunnel positioning in single-bundle ACL reconstruction.

PURPOSE: Remnant preservation, in anterior cruciate ligament (ACL) reconstruction, has potential biological advantages. However, graft positioning remains vital to functional outcome and the prevention of failure. The aim of this study was to investigate the accuracy and precision of tibial tunnel positioning in remnant preservation single-bundle hamstring reconstruction. METHODS: Fifty consecutive adult patients, with isolated ACL rupture, were recruited to a prospective study. Remnant preservation was performed in all cases where > 25% of the native ACL was present. Three-dimensional computer tomography was preformed 3-6 months post-operatively to assess tibial tunnel position (using a grid-based measurement). Accuracy and precision of this technique were assessed against published anatomical data in direct comparison with the group where remnant preservation could not be performed. RESULTS: Two patients withdrew following surgery. In the remaining groups (31 remnant preservation; 17 non-remnant preservation), no difference was demonstrated in tunnel position (40.4 ± 6.7% (anterior-to-posterior) and 47.4 ± 1.5% (medial-to-lateral) vs. 38.8 ± 4.9% and 46.7 ± 1.5%, respectively; n.s.), accuracy (6.1% vs. 4.8%; n.s.) or precision (3.9% vs. 2.8%; n.s.). CONCLUSIONS: Remnant preservation can be safely performed without compromising tunnel position. Therefore, the potential benefits of this technique can be utilised, in clinical practice, without sacrificing the ability to optimize tibial tunnel positioning. LEVEL OF EVIDENCE: III.

A comparison of femoral tunnel placement in ACL reconstruction using a 70° arthroscope through the anterolateral portal versus a 30° arthroscope through the anteromedial portal: a pilot 3D-CT study.

BACKGROUND: Graft malposition is a risk factor for failure of anterior cruciate ligament reconstruction. A 70° arthroscope improves visualisation of the medial wall of the lateral femoral condyle without switching portals. We investigated whether the use of this arthroscope affected the accuracy and precision of femoral tunnel placement. METHODS: Fifty consecutive adult patients were recruited. Following one withdrawal and two exclusions, 47 patients (30 in group 1 (70° arthroscope), 17 in group 2 (30° arthroscope)) underwent three-dimensional computed tomography imaging using a grid-based system to measure tunnel position. RESULTS: No difference was found in the accuracy or precision of tunnels (mean position: group 1 = 33.3 ± 6.0% deep-shallow, 27.2 ± 5.2% high-low; group 2 = 31.7 ± 6.9% deep-shallow, 29.0 ± 6.2% high-low; not significant). A post-hoc power analysis suggests a study of 106 patients would be required. CONCLUSIONS: This pilot study suggests that tunnel position is not affected by the arthroscope used. An appropriately powered study could investigate this finding alongside other potential benefits of using a 70° arthroscope for this procedure. TRIAL REGISTRATION: ClinicalTrials.gov, NCT02816606. Registered on 28 June 2016.

Cerebrospinal fluid ß-amyloid42 and neurofilament light relate to white matter hyperintensities.

White matter hyperintensities (WMHs) are associated with poorer brain health, but their pathophysiological substrates remain elusive. To better understand the mechanistic underpinnings of WMHs among older adults, this study examined in vivo cerebrospinal fluid biomarkers of ß-amyloid42 deposition (Aß42), hyperphosphorylated tau pathology, neurodegeneration (total tau), and axonal injury (neurofilament light [NFL]) in relation to log-transformed WMHs volume. Participants free of clinical stroke and dementia were drawn from the Vanderbilt Memory & Aging Project (n = 148, 72 ± 6 years). Linear regression models adjusted for age, sex, race/ethnicity, education, intracranial volume, modified Framingham Stroke Risk Profile (excluding points assigned for age), cognitive diagnosis, and APOE-ε4 carrier status. Aß42 (ß = -0.001, p = 0.007) and NFL (ß = 0.0003, p = 0.01) concentrations related to WMHs but neither hyperphosphorylated tau nor total tau associations with WMHs reached statistical significance (p-values > 0.21). In a combined model, NFL accounted for 3.2% of unique variance in WMHs and Aß42 accounted for an additional 4.3% beyond NFL, providing novel evidence of the co-occurrence of at least 2 distinct pathways for WMHs among older adults, including amyloid deposition and axonal injury.

Lower cardiac index levels relate to lower cerebral blood flow in older adults.

OBJECTIVE: To assess cross-sectionally whether lower cardiac index relates to lower resting cerebral blood flow (CBF) and cerebrovascular reactivity (CVR) among older adults. METHODS: Vanderbilt Memory & Aging Project participants free of stroke, dementia, and heart failure were studied (n = 314, age 73 ± 7 years, 59% male, 39% with mild cognitive impairment). Cardiac index (liters per minute per meter squared) was quantified from echocardiography. Resting CBF (milliliters per 100 grams per minute) and hypercapnia-induced CVR were quantified from pseudo-continuous arterial spin-labeling MRI. Linear regressions with ordinary least-square estimates related cardiac index to regional CBF, with adjustment for age, education, race/ethnicity, Framingham Stroke Risk Profile score (systolic blood pressure, antihypertensive medication use, diabetes mellitus, current cigarette smoking, left ventricular hypertrophy, prevalent cardiovascular disease [CVD], atrial fibrillation), APOE ε4 status, cognitive diagnosis, and regional tissue volume. RESULTS: Lower cardiac index corresponded to lower resting CBF in the left (ß = 2.4, p = 0.001) and right (ß = 2.5, p = 0.001) temporal lobes. Results were similar when participants with prevalent CVD and atrial fibrillation were excluded (left temporal lobe ß = 2.3, p = 0.003; right temporal lobe ß = 2.5, p = 0.003). Cardiac index was unrelated to CBF in other regions assessed (p > 0.25) and CVR in all regions (p > 0.05). In secondary cardiac index × cognitive diagnosis interaction models, cardiac index and CBF associations were present only in cognitively normal participants and affected a majority of regions assessed with effects strongest in the left (p < 0.0001) and right (p < 0.0001) temporal lobes. CONCLUSIONS: Among older adults without stroke, dementia, or heart failure, systemic blood flow correlates with cerebral CBF in the temporal lobe, independently of prevalent CVD, but not CVR.

Exploring linguistic correlates of social anxiety in romantic stories.

The current study used computerized linguistic analysis of stories about either going on a date or taking a walk down a street to examine linguistic correlates of social anxiety in a sample of undergraduate students. In general, linguistic analysis revealed associations of social anxiety with several linguistic variables, including negative emotion, affect, and anxiety words. Participants higher in social anxiety wrote fewer affect words. The relationship between social anxiety and anxiety words depended on gender, whereas the relationship between social anxiety and negative emotion words depended on both gender and the nature of primes (supraliminal vs. subliminal) received. Overall, our findings highlight the potential utility and benefits of using linguistic analysis as another source of information about how individuals higher in social anxiety process romantic stimuli.

Identification of Direct Kinase Substrates Using Analogue-Sensitive Alleles.

Identifying the substrates of protein kinases remains a major obstacle in the elucidation of eukaryotic signaling pathways. Promiscuity among kinases and their substrates coupled with the extraordinary plasticity of phosphorylation networks renders traditional genetic approaches or small-molecule inhibitors problematic when trying to determine the direct substrates of an individual kinase. Here we describe methods to label, enrich, and identify the direct substrates of analogue-sensitive kinases by exploiting their steric complementarity to artificial ATP analogues. Using calcium-dependent protein kinases of Toxoplasma gondii as a model for these approaches, this protocol brings together numerous advances that enable labeling of kinase targets in semi-permeabilized cells, quantification of direct labeling over background, and highly specific enrichment of targeted phosphopeptides.

Investigation of pre-pubertal sex differences in wheel running and social behavior in three mouse strains.

Sex differences in social behaviors exist in mammals during adulthood, and further evidence suggests that sex differences in behavior are present before sexual maturity. In order to model behavioral disorders in animals, it is important to assess baseline sex-related behavioral differences, especially when studying disorders for which sex-related behavioral effects are expected. We investigated the effect of sex on behavior in 3 strains of pre-pubertal mice (C57BL/6, CFW, and CF1) using a wheel-running assay. We found no significant sex differences in latency to run on the wheel or total duration of wheel running within each strain. During the social interaction test, there were no differences between sexes in latency or total duration of contact or following between a subject and novel mouse. We also evaluated behavioral patterns of wheel running and stereotypical behaviors, such as burrowing and grooming. Both sexes showed characteristic wheel running behavior, spending the majority of each trial interacting with the wheel when it was free and more time performing other activities (e.g., stereotypical behaviors, general locomotion) when it was jammed. These results provide evidence that, among various strains of pre-pubertal mice, baseline sex-related behavioral differences are not strong enough to influence the measured behaviors.

CK2 activates kinesin via induction of a conformational change.

Kinesin is the canonical plus-end microtubule motor and has been the focus of intense study since its discovery in 1985. We previously demonstrated a time-dependent inactivation of kinesin in vitro that was fully reversible by the addition of purified casein kinase 2 (CK2) and showed that this inactivation/reactivation pathway was relevant in cells. Here we show that kinesin inactivation results from a conformational change that causes the neck linker to be positioned closer to the motor domain. Furthermore, we show that treatment of kinesin with CK2 prevents and reverses this repositioning. Finally, we demonstrate that CK2 treatment facilitates ADP dissociation from the motor, resulting in a nucleotide-free state that promotes microtubule binding. Thus, we propose that kinesin inactivation results from neck-linker repositioning and that CK2-mediated reactivation results from CK2's dual ability to reverse this repositioning and to promote ADP release.

Combining docking site and phosphosite predictions to find new substrates: identification of smoothelin-like-2 (SMTNL2) as a c-Jun N-terminal kinase (JNK) substrate.

Specific docking interactions between mitogen-activated protein kinases (MAPKs), their regulators, and their downstream substrates, are crucial for efficient and accurate signal transmission. To identify novel substrates of the c-Jun N-terminal kinase (JNK) family of MAPKs, we searched the human genome for proteins that contained (1), a predicted JNK-docking site (D-site); and (2), a cluster of putative JNK target phosphosites located close to the D-site. Here we describe a novel JNK substrate that emerged from this analysis, the functionally uncharacterized protein smoothelin-like 2 (SMTNL2). SMTNL2 protein bound with high-affinity to multiple MAPKs including JNK1-3 and ERK2; furthermore, the identity of conserved amino acids in the predicted docking site (residues 180-193) was necessary for this high-affinity binding. In addition, purified full-length SMTNL2 protein was phosphorylated by JNK1-3 in vitro, and this required the integrity of the D-site. Using mass spectrometry and mutagenesis, we identified four D-site-dependent phosphoacceptor sites in close proximity to the docking site, at S217, S241, T236 and T239. A short peptide comprised of the SMTNL2 D-site inhibited JNK-mediated phosphorylation of the ATF2 transcription factor, showing that SMTNL2 can compete with other substrates for JNK binding. Moreover, when transfected into HEK293 cells, SMTNL2 was phosphorylated by endogenous JNK in a D-site dependent manner, on the same residues identified in vitro. SMTNL2 protein was expressed in many mammalian tissues, with a notably high expression in skeletal muscle. Consistent with the hypothesis that SMTNL2 has a function in skeletal muscle, SMTNL2 protein expression was strongly induced during the transition from myoblasts to myotubes in differentiating C2C12 cells.

On the relationship among social anxiety, intimacy, sexual communication, and sexual satisfaction in young couples.

This study was conducted to better understand why socially anxious individuals experience less sexual satisfaction in their intimate partnerships than nonanxious individuals, a relationship that has been well documented in previous research. Effective communication between partners is an important predictor of relationship satisfaction. Sexual communication, an important aspect of communication between romantic partners, is especially sensitive for couples given the vulnerability inherent in being open about sexual issues. Because socially anxious individuals characteristically report fear of evaluation or scrutiny by others, we hypothesized that the process of building intimacy by sharing personal information about oneself with one's partner, including when this information relates to one's sexuality and/or the sexual domain of the relationship, would be particularly difficult for socially anxious individuals. The present study examined fear of intimacy and sexual communication as potential mediators of the relationship between higher social anxiety and lower sexual satisfaction. Self-report data were collected from 115 undergraduate students and their partners in monogamous, heterosexual, committed relationships of at least 3 months duration. Multilevel path modeling revealed that higher social anxiety predicted higher fear of intimacy, which predicted lower satisfaction with open sexual communication, which, in turn, predicted lower sexual satisfaction. Additionally, there was evidence of mediation as there were significant indirect effects of the antecedent variables on sexual satisfaction. The path model had excellent fit. Implications for social anxiety, intimate relationships, and couples therapy are discussed.

Romantic relationships: do socially anxious individuals benefit?

Psychological health and interpersonal functioning mutually influence each other. Social anxiety has a pervasive effect on interpersonal functioning, resulting in smaller social networks, increased likelihood of being single or divorced, and less intimacy in relationships. However, little is known about how relationships affect socially anxious individuals in return. We utilized a structured interview to assess how romantic relationships were perceived as influencing three aspects of psychological health (well-being, social anxiety and comfort in social situations) and whether these patterns differed as a function of social anxiety in an undergraduate sample. The perceived importance of several reasons for these effects, including those that could be characterized as both protective and harmful, was also assessed. Relationships were perceived as having contributed positively in each domain. However, when positive and negative reasons were examined separately, socially anxious individuals reported benefiting more from the positive reasons and being harmed more by negative reasons. Further, social anxiety was associated with endorsing certain reasons as important.

The longitudinal relationship between fear of positive evaluation and fear of negative evaluation.

Available research suggests that fear of negative evaluation and fear of positive evaluation are related but distinct constructs that each contribute to social anxiety, implying a need to focus on these fears in treatment. Yet, this research is almost entirely based on cross-sectional data. We examined the longitudinal relationship between fears of positive and negative evaluation over three time points in a sample of undergraduate students. We tested competing models consistent with two basic positions regarding these fears: (1) that fear of positive evaluation only appears to affect social anxiety because it arises from the same, single underlying trait as fear of negative evaluation, and (2) fears of positive and negative evaluation are correlated, but clearly distinct, constructs. The best-fitting model was an autoregressive latent-trajectory model in which each type of fear had a separate trait-like component. The correlation between these trait-like components appeared to fully account for the relationships between these constructs over time. This investigation adds to the evidence in support of the second position described above: fear of positive evaluation is best interpreted as a separate construct from fear of negative evaluation.

Interrupted aortic arch in the adult.

Interrupted aortic arch (IAA) is a rare condition in infants that occurs approximately three times per million births. It is considered incompatible with life after the ductus arteriosus closes if it is not surgically corrected. A review found 37 reported cases not identified until the patient was over the age of 18, and analysis of these cases and of our own case is reported.

Computational prediction and experimental verification of new MAP kinase docking sites and substrates including Gli transcription factors.

In order to fully understand protein kinase networks, new methods are needed to identify regulators and substrates of kinases, especially for weakly expressed proteins. Here we have developed a hybrid computational search algorithm that combines machine learning and expert knowledge to identify kinase docking sites, and used this algorithm to search the human genome for novel MAP kinase substrates and regulators focused on the JNK family of MAP kinases. Predictions were tested by peptide array followed by rigorous biochemical verification with in vitro binding and kinase assays on wild-type and mutant proteins. Using this procedure, we found new 'D-site' class docking sites in previously known JNK substrates (hnRNP-K, PPM1J/PP2Czeta), as well as new JNK-interacting proteins (MLL4, NEIL1). Finally, we identified new D-site-dependent MAPK substrates, including the hedgehog-regulated transcription factors Gli1 and Gli3, suggesting that a direct connection between MAP kinase and hedgehog signaling may occur at the level of these key regulators. These results demonstrate that a genome-wide search for MAP kinase docking sites can be used to find new docking sites and substrates.

Multidisciplinary approach decreases length of stay and reduces cost for ventricular assist device therapy.

High implantation costs and long postoperative length of stay (LOS) in debilitated patients complicate ventricular assist device (VAD) therapy. Between July 2000 and February 2005, 30 patients received a VAD at our institution. Of those, 20 patients were successfully discharged from the hospital with VADs. In August 2003, a multidisciplinary team was formed consisting of all services for VAD patients to replace a single-discipline (cardiac surgery) system. This team evaluated potential VAD candidates and identified optimal timing for implantation. These 20 VAD patients were divided into two groups according to the initiation of multidisciplinary team; the traditional group (n=7, July 2000-July 2003) and the multidisciplinary group (n=13, August 2003-February 2005). Patient demographics were not different. The LOS decreased from 61 to 15 days (P<0.01), especially LOS on the floor decreased from 35 to 7 days (P=0.03). The floor cost was significantly reduced ($47,111 vs. $8742, P<0.01), leading to a decrease in total postoperative cost ($202,238 vs. $161,744, P<0.01). The 30-day readmission rate decreased (5/7 patients vs. 1/13 patients, P<0.01). A multidisciplinary approach significantly decreased LOS and cost after VAD therapy, mostly by decreasing the cost of routine non-ICU care, without increasing the readmission rate.

Polycythemia and hyperviscosity of the newborn.

Polycythemia and hyperviscosity of the newborn are well-known conditions that are surrounded by controversy. The patient population most affected by polycythemia is the term or near-term infant. The true incidence of this condition is not known since the majority of infants are likely to be asymptomatic, normal newborns. Diagnosis is largely based on hematocrit values and symptoms, which can range from subtle to severe, and not on measures of viscosity. Hematocrits are not routinely drawn in this population, most likely related to the controversy surrounding the treatment of the asymptomatic infant. Presenting symptoms may be subtle and are not always attributed to polycythemia. Knowledge of the etiology, pathophysiology, and clinical signs and symptoms may contribute to the early identification and treatment of infants with polycythemia and hyperviscosity syndrome.