Strategies to Improve Drug Delivery Across the Blood-Brain Barrier for Glioblastoma.

PURPOSE OF REVIEW: Glioblastoma remains resistant to most conventional treatments. Despite scientific advances in the past three decades, there has been a dearth of effective new treatments. New approaches to drug delivery and clinical trial design are needed. RECENT FINDINGS: We discuss how the blood-brain barrier and tumor microenvironment pose challenges for development of effective therapies for glioblastoma. Next, we discuss treatments in development that aim to overcome these barriers, including novel drug designs such as nanoparticles and antibody-drug conjugates, novel methods of drug delivery, including convection-enhanced and intra-arterial delivery, and novel methods to enhance drug penetration, such as blood-brain barrier disruption by focused ultrasound and laser interstitial thermal therapy. Lastly, we address future opportunities, positing combination therapy as the best strategy for effective treatment, neoadjuvant and window-of-opportunity approaches to simultaneously enhance therapeutic effectiveness with interrogation of on-treatment biologic endpoints, and adaptive platform and basket trials as imperative for future trial design. New approaches to GBM treatment should account for the blood-brain barrier and immunosuppression by improving drug delivery, combining treatments, and integrating novel clinical trial designs.

Systematic review and meta-analysis of intraventricular antibiotics for neonatal meningitis and ventriculitis.

PURPOSE: We aimed to determine the safety and effectiveness of intraventricular antibiotics in neonates with meningitis and/or ventriculitis and analyze the quality of available evidence. METHODS: DESIGN: Systematic review and meta-analysis. DATA SOURCES: PubMed, EMBASE, LILACS, and SCOPUS up to 17 February 2023. ELIGIBILITY CRITERIA FOR SELECTING STUDIES: Randomized experimental and observational studies were included. The Cochrane methodology was used for systematic reviews. RESULTS: Twenty-six observational studies and one randomized clinical trial involving 272 patients were included. The risk of bias in both pediatric and neurosurgical studies was high, and the quality of evidence was low (evidence level C). In the pediatric studies, no significant differences in mortality were found between intraventricular antibiotics and only systemic antibiotic [25.4% vs 16.1%, OR = 0.96 (0.42-2.24), P = 0.93]. However, when analyzing the minimum administered doses, we found a lower mortality when a minimum duration of 3 days for intraventricular antibiotics was used compared to only systemic antibiotic [4.3% vs 17%, OR = 0.22 (0.07-0.72), P = 0.01]. In the neurosurgical studies, the use of intraventricular antibiotics in ventriculitis generally results in a mortality of 5% and a morbidity of 25%, which is lower than that in cases where intraventricular antibiotics were not used, with an average mortality of 37.3% and a morbidity of 50%. CONCLUSION: Considering the low quality of evidence in pediatric and neurosurgical studies, we can conclude with a low level of certainty that intraventricular antibiotics may not significantly impact mortality in neonatal meningitis and ventriculitis. However, reduced mortality was observed in cases treated with a minimum duration of 3 days of intraventricular antibiotic, particularly the multidrug-resistant or treatment-refractory infections. Higher-quality studies are needed to improve the quality of evidence and certainty regarding the use of intraventricular antibiotics for treating neonatal meningitis and ventriculitis.

Pupillary Responses to Dynamic Negative Versus Positive Facial Expressions of Emotion in Children and Parents: Links to Depression and Anxiety.

Witnessing emotional expressions in others triggers physiological arousal in humans. The current study focused on pupil responses to emotional expressions in a community sample as a physiological index of arousal and attention. We explored the associations between parents' and offspring's responses to dynamic facial expressions of emotion, as well as the links between pupil responses and anxiety/depression. Children (N = 90, MAge = 10.13, range = 7.21-12.94, 47 girls) participated in this lab study with one of their parents (47 mothers). Pupil responses were assessed in a computer task with dynamic happy, angry, fearful, and sad expressions, while participants verbally labeled the emotion displayed on the screen as quickly as possible. Parents and children reported anxiety and depression symptoms in questionnaires. Both parents and children showed stronger pupillary responses to negative versus positive expressions, and children's responses were overall stronger than those of parents. We also found links between the pupil responses of parents and children to negative, especially to angry faces. Child pupil responses were related to their own and their parents' anxiety levels and to their parents' (but not their own) depression. We conclude that child pupils are sensitive to individual differences in parents' pupils and emotional dispositions in community samples.

Interactions among stress, behavioral inhibition, and delta-beta coupling predict adolescent anxiety during the COVID-19 pandemic.

The COVID-19 pandemic brought about unprecedented changes and uncertainty to the daily lives of youth. The range of adjustment in light of a near-universal experience of COVID restrictions highlights the importance of identifying factors that may render some individuals more susceptible to heightened levels of anxiety during stressful life events than others. Two risk factors to consider are temperamental behavioral inhibition (BI) and difficulties in emotion regulation (ER). As such, the current paper focused on BI examined prior to COVID, because of its developmental link to anxiety and ER, as difficulties may be associated with differences in anxiety. We examined a neurocognitive marker of ER processes, delta-beta coupling (DBC). The current paper had two goals: (1) to examine BI in relation to COVID-related worry and social anxiety experienced during the pandemic, and (2) to explore the role of individual differences in early DBC in the relationship between BI and anxiety outcomes 6 months apart during COVID-19 (n = 86; T1 Mage  = 15.95, SD = 1.73; T6 Mage  = 16.43, SD = 1.73). We found support for the moderating role of DBC in the relationship between BI levels and social anxiety disorder (SAD) symptom severity during the pandemic. Here, high BI was predictive of increased SAD symptom levels in adolescents with stronger DBC.

"Nobody Here Likes Her"-The Impact of Parental Verbal Threat Information on Children's Fear of Strangers.

Parental verbal threat (vs. safety) information about strangers may induce fears of these strangers in adolescents. In this multi-method experimental study, utilizing a within-subject design, parents provided standardized verbal threat or safety information to their offspring (N = 77, Mage = 11.62 years, 42 girls) regarding two strangers in the lab. We also explored whether the impact of parental verbal threat information differs depending on the social anxiety levels of parents or fearful temperaments of adolescents. Adolescent's fear of strangers during social interaction tasks was assessed using cognitive (fear beliefs, attention bias), behavioral (observed avoidance and anxiety), and physiological (heart rate) indices. We also explored whether the impact of parental verbal threat information differs depending on the social anxiety levels of parents or fearful temperaments of adolescents. The findings suggest that a single exposure to parental verbal threat (vs. safety) information increased adolescent's self-reported fears about the strangers but did not increase their fearful behaviors, heart rate, or attentional bias. Furthermore, adolescents of parents with higher social anxiety levels or adolescents with fearful temperaments were not more strongly impacted by parental verbal threat information. Longitudinal research and studies investigating parents' naturalistic verbal expressions of threat are needed to expand our understanding of this potential verbal fear-learning pathway.

Adolescent coping and social media use moderated anxiety change during the COVID-19 pandemic.

INTRODUCTION: Adolescence is a sensitive period during which stressors and social disruptions uniquely contribute to anxiety symptoms. Adolescent's coping strategies (i.e., avoidance and approach) during the coronavirus disease 2019 (COVID-19) pandemic may be differentially related to anxiety symptom changes. Further, social media use (SMU) is ubiquitous and may serve as an avenue to deploy avoidant and/or approach coping. METHOD: Participants included 265 adolescents (ages 12-20 years; 55.8% female, 43.8% male) and one parent per adolescent. At two time points separated by ~6 months, adolescents reported on SMU and coping strategies, and parents and adolescents reported demographic information and adolescents' anxiety symptoms. Data were collected online in the United States, from summer 2020 through spring 2021. RESULTS: Increases in avoidant coping predicted increasing anxiety, particularly when approach coping decreased. Decreases in both avoidant coping and SMU coincided with decreasing anxiety. Older adolescents showed decreasing anxiety when avoidant coping declined and SMU increased. CONCLUSION: Coping strategies and SMU predicted patterns of adolescent anxiety symptom change across 6 months during the COVID-19 pandemic. Results highlight that coping and SMU should be contextualized within the time course of stressors.

Effect of the Lys62Ala Mutation on the Thermal Stability of BstHPr Protein by Molecular Dynamics.

We analyzed the thermal stability of the BstHPr protein through the site-directed point mutation Lys62 replaced by Ala residue using molecular dynamics simulations at five different temperatures: 298, 333, 362, 400, and 450 K, for periods of 1 µs and in triplicate. The results from the mutant thermophilic BstHPrm protein were compared with those of the wild-type thermophilic BstHPr protein and the mesophilic BsHPr protein. Structural and molecular interaction analyses show that proteins lose stability as temperature increases. Mutant and wild-type proteins behave similarly up to 362 K. However, at 400 K the mutant protein shows greater structural instability, losing more buried hydrogen bonds and exposing more of its non-polar residues to the solvent. Therefore, in this study, we confirmed that the salt bridge network of the Glu3-Lys62-Glu36 triad, made up of the Glu3-Lys62 and Glu36-Lys62 ion pairs, provides thermal stability to the thermophilic BstHPr protein.

Disorder and amino acid composition in proteins: their potential role in the adaptation of extracellular pilins to the acidic media, where Acidithiobacillus thiooxidans grows.

There are few biophysical studies or structural characterizations of the type IV pilin system of extremophile bacteria, such as the acidophilic Acidithiobacillus thiooxidans. We set out to analyze their pili-comprising proteins, pilins, because these extracellular proteins are in constant interaction with protons of the acidic medium in which At. thiooxidans grows. We used the web server Operon Mapper to analyze and identify the cluster codified by the minor pilin of At. thiooxidans. In addition, we carried an in-silico characterization of such pilins using the VL-XT algorithm of PONDR® server. Our results showed that structural disorder prevails more in pilins of At. thiooxidans than in non-acidophilic bacteria. Further computational characterization showed that the pilins of At. thiooxidans are significantly enriched in hydroxy (serine and threonine) and amide (glutamine and asparagine) residues, and significantly reduced in charged residues (aspartic acid, glutamic acid, arginine and lysine). Similar results were obtained when comparing pilins from other Acidithiobacillus and other acidophilic bacteria from another genus versus neutrophilic bacteria, suggesting that these properties are intrinsic to pilins from acidic environments, most likely by maintaining solubility and stability in harsh conditions. These results give guidelines for the application of extracellular proteins of acidophiles in protein engineering.

Developmental trajectories of behavioral inhibition from infancy to age seven: The role of genetic and environmental risk for psychopathology.

The present study leveraged data from a longitudinal adoption study of 361 families recruited between 2003 and 2010 in the United States. We investigated how psychopathology symptoms in birth parents (BP; Mage  = 24.1 years; 50.5-62.9% completed high school) and adoptive parents (AP; Mage  = 37.8 years; 80.9% completed college; 94% mother-father couples) influenced children's behavioral inhibition (BI) trajectories. We used latent growth models of observed BI at 18 and 27 months, and 4.5 and 7 years in a sample of adopted children (Female = 42%, White = 57%, Black = 11%, Multi-racial = 21%, Latinx = 9%). BI generally decreased over time, yet there was substantial variability in these trajectories. Neither BP nor AP psychopathology symptoms independently predicted systematic differences in BI trajectories. Instead, we found that AP internalizing symptoms moderated the effects of BP psychopathology on trajectories of BI, indicating a gene by environment interaction.

Variability in caregiver attention bias to threat: A Goldilocks effect in infant emotional development?

Attention biases to threat are considered part of the etiology of anxiety disorders. Attention bias variability (ABV) quantifies intraindividual fluctuations in attention biases and may better capture the relation between attention biases and psychopathology risk versus mean levels of attention bias. ABV to threat has been associated with attentional control and emotion regulation, which may impact how caregivers interact with their child. In a relatively diverse sample of infants (50% White, 50.7% female), we asked how caregiver ABV to threat related to trajectories of infant negative affect across the first 2 years of life. Families were part of a multi-site longitudinal study, and data were collected from 4 to 24 months of age. Multilevel modeling examined the effect of average caregiver attention biases on changes in negative affect. We found a significant interaction between infant age and caregiver ABV to threat. Probing this interaction revealed that infants of caregivers with high ABV showed decreases in negative affect over time, while infants of caregivers with low-to-average ABV showed potentiated increases in negative affect. We discuss how both high and extreme patterns of ABV may relate to deviations in developmental trajectories.

Molecular Dynamics Simulations of HPr Proteins from a Thermophilic and a Mesophilic Organism: A Comparative Thermal Study.

The histidine-containing phosphocarrier (HPr) is a monomeric protein conserved in Gram-positive bacteria, which may be of mesophilic or thermophilic nature. In particular, the HPr protein from the thermophilic organism B. stearothermophilus is a good model system for thermostability studies, since experimental data, such as crystal structure and thermal stability curves, are available. However, its unfolding mechanism at higher temperatures is yet unclear at a molecular level. Therefore, in this work, we researched the thermal stability of this protein using molecular dynamics simulations, subjecting it to five different temperatures during a time span of 1 µs. The analyses of the structural parameters and molecular interactions were compared with those of the mesophilic homologue HPr protein from B. subtilis. Each simulation was run in triplicate using identical conditions for both proteins. The results showed that the two proteins lose stability as the temperature increases, but the mesophilic structure is more affected. We found that the salt bridge network formed by the triad of Glu3-Lys62-Glu36 residues and the salt bridge made up of Asp79-Lys83 ion pair are key factors to keep stable the thermophilic protein, maintaining the hydrophobic core protected and the structure packed. In addition, these molecular interactions neutralize the negative surface charge, acting as "natural molecular staples".

The Inhibition of the FGFR/PI3K/Akt Axis by AZD4547 Disrupts the Proangiogenic Microenvironment and Vasculogenic Mimicry Arising from the Interplay between Endothelial and Triple-Negative Breast Cancer Cells.

Vasculogenic mimicry (VM), a process in which aggressive cancer cells form tube-like structures, plays a crucial role in providing nutrients and escape routes. Highly plastic tumor cells, such as those with the triple-negative breast cancer (TNBC) phenotype, can develop VM. However, little is known about the interplay between the cellular components of the tumor microenvironment and TNBC cells' VM capacity. In this study, we analyzed the ability of endothelial and stromal cells to induce VM when interacting with TNBC cells and analyzed the involvement of the FGFR/PI3K/Akt pathway in this process. VM was corroborated using fluorescently labeled TNBC cells. Only endothelial cells triggered VM formation, suggesting a predominant role of paracrine/juxtacrine factors from an endothelial origin in VM development. Via immunocytochemistry, qPCR, and secretome analyses, we determined an increased expression of proangiogenic factors as well as stemness markers in VM-forming cancer cells. Similarly, endothelial cells primed by TNBC cells showed an upregulation of proangiogenic molecules, including FGF, VEGFA, and several inflammatory cytokines. Endothelium-dependent TNBC-VM formation was prevented by AZD4547 or LY294002, strongly suggesting the involvement of the FGFR/PI3K/Akt axis in this process. Given that VM is associated with poor clinical prognosis, targeting FGFR/PI3K/Akt pharmacologically may hold promise for treating and preventing VM in TNBC tumors.

Oral glutamine supplementation increases seizure severity in a rodent model of mesial temporal lobe epilepsy.

Background: Glutamine synthetase (GS) is the only enzyme known to synthesize significant amounts of glutamine in mammals, and loss of GS in the hippocampus has been implicated in the pathophysiology of medication refractory mesial temporal lobe epilepsy (MTLE). Moreover, loss-of-function mutations of the GS gene causes severe epileptic encephalopathy, and supplementation with glutamine has been shown to normalize EEG and possibly improve the outcome in these patients. Here we examined whether oral glutamine supplementation is an effective treatment for MTLE by assessing the frequency and severity of seizures after supplementation in a translationally relevant model of the disease.Methods: Male Sprague Dawley rats (380-400 g) were allowed to drink unlimited amounts of glutamine in water (3.6% w/v; n = 8) or pure water (n = 8) for several weeks. Ten days after the start of glutamine supplementation, GS was chronically inhibited in the hippocampus to induce MTLE. Continuous video-intracranial EEG was collected for 21 days to determine the frequency and severity of seizures.Results: While there was no change in seizure frequency between the groups, the proportion of convulsive seizures was significantly higher in glutamine treated animals during the first three days of GS inhibition.Conclusion: The results suggest that oral glutamine supplementation transiently increases seizure severity in the initial stages of an epilepsy model, indicating a potential role of the amino acid in seizure propagation and epileptogenesis.

Developmental patterns of affective attention across the first 2 years of life.

This study examined patterns of attention toward affective stimuli in a longitudinal sample of typically developing infants (N = 357, 147 females, 50% White, 22% Latinx, 16% African American/Black, 3% Asian, 8% mixed race, 1% not reported) using two eye-tracking tasks that measure vigilance to (rapid detection), engagement with (total looking toward), and disengagement from (latency to looking away) emotional facial configurations. Infants completed each task at 4, 8, 12, 18, and 24 months of age from 2016 to 2020. Multilevel growth models demonstrate that, over the first 2 years of life, infants became faster at detecting and spent more time engaging with angry over neutral faces. These results have implications for our understanding of the development of affect-biased attention.

Do you see what I mean?: Using mobile eye tracking to capture parent-child dynamics in the context of anxiety risk.

Temperamental behavioral inhibition (BI) is a robust endophenotype for anxiety characterized by increased sensitivity to novelty. Controlling parenting can reinforce children's wariness by rewarding signs of distress. Fine-grained, dynamic measures are needed to better understand both how children perceive their parent's behaviors and the mechanisms supporting evident relations between parenting and socioemotional functioning. The current study examined dyadic attractor patterns (average mean durations) with state space grids, using children's attention patterns (captured via mobile eye tracking) and parental behavior (positive reinforcement, teaching, directives, intrusion), as functions of child BI and parent anxiety. Forty 5- to 7-year-old children and their primary caregivers completed a set of challenging puzzles, during which the child wore a head-mounted eye tracker. Child BI was positively correlated with proportion of parent's time spent teaching. Child age was negatively related, and parent anxiety level was positively related, to parent-focused/controlling parenting attractor strength. There was a significant interaction between parent anxiety level and child age predicting parent-focused/controlling parenting attractor strength. This study is a first step to examining the co-occurrence of parenting behavior and child attention in the context of child BI and parental anxiety levels.

Implementation of the diffusion model on dot-probe task performance in children with behavioral inhibition.

Attentional bias to threat, the process of preferentially attending to potentially threatening environmental stimuli over neutral stimuli, is positively associated with behavioral inhibition (BI) and trait anxiety. However, the most used measure of attentional bias to threat, the dot-probe task, has been criticized for demonstrating poor reliability. The present study aimed to assess whether utilizing a sequential sampling model to describe performance could detect adequate test-retest reliability for the dot-probe task, demonstrate stronger cueing effects, and improve the association with neural signals of early attention. One hundred and twenty children aged 9-12 years completed the dot-probe task twice. During the second administration, event-related potentials (ERPs) were obtained as time-sensitive neural markers of attention. BI was not associated with traditional or diffusion model measures of performance. Traditional and diffusion model measures of performance were also not associated with N1, P2, or N2 ERP amplitude. There were main effects of Visit, in which RTs were faster and standard deviation of RT smaller during the second administration due to an increase in drift rate and a decrease in non-decision time. The traditional RT bias score (r = 0.06) and bias scores formed via diffusion model parameters (all r's < 0.40) all demonstrated poor reliability. Results confirm recommendations to move away from using the dot-probe task as the primary or sole index of attentional bias.

The social learning of threat and safety in the family: Parent-to-child transmission of social fears via verbal information.

Parental verbal threat (vs. safety) information regarding the social world may impact a child's fear responses, evident in subjective, behavioral, cognitive, and physiological indices of fear. In this study, primary caregivers provided standardized verbal threat or safety information to their child (N = 68, M = 5.27 years; 34 girls) regarding two strangers in the lab. Following this manipulation, children reported fear beliefs for each stranger. Physiological and behavioral reactions were recorded as children engaged with the two strangers (who were blind to their characterization) in a social interaction task. Child attention to the strangers was measured in a visual search task. Parents also reported their own, and their child's, social anxiety symptoms. Children reported more fear for the stranger paired with threat information, but no significant differences were found in observed child fear, attention, or heart rate. Higher social anxiety symptoms on the side of the parents and the children exacerbated the effect of parental verbal threat on observed fear. Our findings reveal a causal influence of parental verbal threat information only for child-reported fear and highlight the need to further refine the conditions under which acquired fear beliefs persist and generalize to behavior/physiology or get overruled by nonaversive real-life encounters.

Moderating effects of environmental stressors on the development of attention to threat in infancy.

An attention bias to threat has been linked to psychosocial outcomes across development, including anxiety (Pérez-Edgar, K., Bar-Haim, Y., McDermott, J. M., Chronis-Tuscano, A., Pine, D. S., & Fox, N. A. (2010). Attention biases to threat and behavioral inhibition in early childhood shape adolescent social withdrawal. Emotion (Washington, D.C.), 10(3), 349). Although some attention biases to threat are normative, it remains unclear how these biases diverge into maladaptive patterns of emotion processing for some infants. Here, we examined the relation between household stress, maternal anxiety, and attention bias to threat in a longitudinal sample of infants tested at 4, 8, and 12 months. Infants were presented with a passive viewing eye-tracking task in which angry, happy, or neutral facial configurations appeared in one of the four corners of a screen. We measured infants' latency to fixate each target image and collected measures of parental anxiety and daily hassles at each timepoint. Intensity of daily parenting hassles moderated patterns of attention bias to threat in infants over time. Infants exposed to heightened levels of parental hassles became slower to detect angry (but not happy) facial configurations compared with neutral faces between 4 and 12 months of age, regardless of parental anxiety. Our findings highlight the potential impact of the environment on the development of infants' early threat processing and the need to further investigate how early environmental factors shape the development of infant emotion processing.

Endothelium-Dependent Induction of Vasculogenic Mimicry in Human Triple-Negative Breast Cancer Cells Is Inhibited by Calcitriol and Curcumin.

In highly aggressive tumors, cancer cells may form channel-like structures through a process known as vasculogenic mimicry (VM). VM is generally associated with metastasis, mesenchymal phenotype, and treatment resistance. VM can be driven by antiangiogenic treatments and/or tumor microenvironment-derived factors, including those from the endothelium. Curcumin, a turmeric product, inhibits VM in some tumors, while calcitriol, the most active vitamin D metabolite, exerts potent antineoplastic effects. However, the effect of these natural products on VM in breast cancer remains unknown. Herein, we studied the effect of both compounds on triple-negative breast cancer (TNBC) VM-capacity in a co-culture model. The process of endothelial cell-induced VM in two human TNBC cell lines was robustly inhibited by calcitriol and partially by curcumin. Calcitriol promoted TNBC cells' morphological change from spindle-like to cobblestone-shape, while curcumin diminished VM 3D-structure. Notably, the treatments dephosphorylated several active kinases, especially those involved in the PI3K/Akt pathway. In summary, calcitriol and curcumin disrupted endothelium-induced VM in TNBC cells partially by PI3K/Akt inactivation and mesenchymal phenotype inhibition. Our results support the possible use of these natural compounds as adjuvants for VM inactivation in patients with malignant tumors inherently capable of forming VM, or those with antiangiogenic therapy, warranting further in vivo studies.

A computational network perspective on pediatric anxiety symptoms.

BACKGROUND: While taxonomy segregates anxiety symptoms into diagnoses, patients typically present with multiple diagnoses; this poses major challenges, particularly for youth, where mixed presentation is particularly common. Anxiety comorbidity could reflect multivariate, cross-domain interactions insufficiently emphasized in current taxonomy. We utilize network analytic approaches that model these interactions by characterizing pediatric anxiety as involving distinct, inter-connected, symptom domains. Quantifying this network structure could inform views of pediatric anxiety that shape clinical practice and research. METHODS: Participants were 4964 youths (ages 5-17 years) from seven international sites. Participants completed standard symptom inventory assessing severity along distinct domains that follow pediatric anxiety diagnostic categories. We first applied network analytic tools to quantify the anxiety domain network structure. We then examined whether variation in the network structure related to age (3-year longitudinal assessments) and sex, key moderators of pediatric anxiety expression. RESULTS: The anxiety network featured a highly inter-connected structure; all domains correlated positively but to varying degrees. Anxiety patients and healthy youth differed in severity but demonstrated a comparable network structure. We noted specific sex differences in the network structure; longitudinal data indicated additional structural changes during childhood. Generalized-anxiety and panic symptoms consistently emerged as central domains. CONCLUSIONS: Pediatric anxiety manifests along multiple, inter-connected symptom domains. By quantifying cross-domain associations and related moderation effects, the current study might shape views on the diagnosis, treatment, and study of pediatric anxiety.