Addressing the Intracellular Vestibule of the Plasmodial Lactate Transporter PfFNT by p-Substituted Inhibitors Amplifies In Vitro Activity.

Inhibition of the lactate transporter PfFNT is a valid novel mode of action against malaria parasites. Current pyridine-substituted pentafluoro-3-hydroxy-pent-2-en-1-ones act as substrate analogs with submicromolar EC50 in vitro, and >99.7% activity in mice. The recently solved structure of a PfFNT-inhibitor complex visualized the binding mode. Here, we extended the inhibitor layout by series of amine- and anilide-linked pyridine p-substituents to generate additional interactions in the cytoplasmic vestibule. Virtual docking indicated hydrogen bonding to Tyr31 and Ser102. Fluorescence cross-correlation spectroscopy yielded respectively enhanced target affinity. Strikingly, the in vitro activity increased by 1 order of magnitude to 14.8 nM at negligible cytotoxicity. While p-amine substitutions were rapidly metabolized, the more stable p-acetanilide cleared 99.7% of parasites at 4 × 50 mg kg-1 in a mouse malaria model. Future stabilization of the p-substitution against metabolism may translate the gain in in vitro potency to the in vivo situation.

Dual role of BdMUTE during stomatal development in the model grass Brachypodium distachyon.

Grasses form morphologically derived, four-celled stomata, where two dumbbell-shaped guard cells (GCs) are flanked by two lateral subsidiary cells (SCs). This innovative form enables rapid opening and closing kinetics and efficient plant-atmosphere gas exchange. The mobile bHLH transcription factor MUTE is required for SC formation in grasses. Yet whether and how MUTE also regulates GC development and whether MUTE mobility is required for SC recruitment is unclear. Here, we transgenically impaired BdMUTE mobility from GC to SC precursors in the emerging model grass Brachypodium distachyon. Our data indicate that reduced BdMUTE mobility severely affected the spatiotemporal coordination of GC and SC development. Furthermore, although BdMUTE has a cell-autonomous role in GC division orientation, complete dumbbell morphogenesis of GCs required SC recruitment. Finally, leaf-level gas exchange measurements showed that dosage-dependent complementation of the four-celled grass morphology was mirrored in a gradual physiological complementation of stomatal kinetics. Together, our work revealed a dual role of grass MUTE in regulating GC division orientation and SC recruitment, which in turn is required for GC morphogenesis and the rapid kinetics of grass stomata.

Predictors of Positive Airway Pressure Therapy Adherence Among U.S. Active Duty Military Personnel With Obstructive Sleep Apnea.

INTRODUCTION: Obstructive sleep apnea (OSA) is prevalent among U.S. military personnel, but adherence to positive airway pressure (PAP) treatment is suboptimal. This study sought to identify factors that predict the adherence to PAP therapy of active duty military patients newly diagnosed with OSA. MATERIALS AND METHODS: The study was a retrospective cross-sectional analysis of 239 active duty military patients (94% men, mean age 40.0 ± 7.8 years), who were admitted into PAP therapy and participated in a single 90-minute behavioral intervention session. The session included a self-assessment of sleep trouble, sleep distress, impact of sleep on functioning, emotional difficulty from sleep, stress appraisal, and beliefs about OSA and PAP. PAP usage data of the patients were analyzed at 30-, 60-, and 180 days post-intervention using descriptive statistics and multivariable linear regression analysis. RESULTS: Positive airway pressure adherence of at least 4 hours nightly usage decreased slightly over the course of the study; mean number of days used over a 30-day timeframe at the 30-, 60-, and 180-day mark were 15.6, 14.9, and 14.1 days, respectively. On nights used, PAP usage remained consistent at 4.8 to 5.0 hours over the 30-, 60-, and 180-day mark. Age, apnea-hypopnea index, readiness, self-efficacy or confidence in treatment implementation, and worries about sleep were statistically significant predictors of adherence. Furthermore, worries about sleep negatively affected PAP usage hours on nights used, regardless of meeting adherence criterion, in the early phase of treatment and in the long run at the 180-day mark. CONCLUSIONS: Readiness for therapy, self-efficacy, confidence in putting into action the treatment plan, and worries about sleep are modifiable variables that may be targeted in programs to boost PAP adherence and usage among the military population. Future studies should explore the predictive aspects of each of these variables and identify interventions to improve them.

Human cytomegalovirus induces neuronal gene expression for viral maturation.

Viral invasion of the host cell causes some of the most dramatic changes in biology. Human cytomegalovirus (HCMV) extensively remodels host cells, altering nuclear shape and generating a cytoplasmic viral-induced assembly compartment (vIAC). How these striking morphology changes take place in the context of host gene regulation is still emerging. Here, we discovered that histone variant macroH2A1 is essential for producing infectious progeny. Because virion maturation and cellular remodeling are closely linked processes, we investigated structural changes in the host cell upon HCMV infection. We discovered that macroH2A1 is necessary for HCMV-induced reorganization of the host nucleus, cytoskeleton, and endoplasmic reticulum. Furthermore, using RNA-seq we found that while all viral genes were highly expressed in the absence of macroH2A1, many HCMV-induced host genes were not. Remarkably, hundreds of these HCMV-induced macroH2A1-dependent host genes are associated with neuronal synapse formation and vesicle trafficking. Knock-down of these HCMV-induced neuronal genes during infection resulted in malformed vIACs and smaller plaques, establishing their importance to HCMV infection. Together, our findings demonstrate that HCMV manipulates host gene expression by hijacking a dormant neuronal secretory pathway for efficient virion maturation.

Atypical semantic cognition in schizotypal personality disorder and borderline personality disorder.

Increased schizotypal traits have previously been associated with atypical semantic cognition in community samples. However, no study has yet examined whether adults diagnosed with schizotypal personality disorder (SPD) display atypical semantic fluency and memory. We hypothesized that 24 adults diagnosed with SPD would name more idiosyncratic words on the semantic fluency task and show decreased semantic recall for animal and fruit category words compared with 29 participants with borderline personality disorder (BPD) and a community sample of 96 age-matched controls. We examined whether atypical semantic cognition was specifically associated with disorganized and eccentric speech and thinking, or more broadly with pathological personality traits and personality functioning. Our main hypothesis was confirmed, as the SPD participants named more idiosyncratic words and recalled fewer semantically related words compared with controls. Surprisingly, participants with BPD likewise named more atypical words compared with controls. More idiosyncratic semantic fluency was associated with more eccentric speech and thinking. Increased idiosyncratic semantic fluency and reduced semantic recall were both coupled to increased detachment and lowered personality functioning, while reduced semantic recall further was related to increased interpersonal problems. Our findings suggest that persons with SPD, and to a lesser degree BPD, show atypical semantic cognition, which is associated with eccentric speech and thinking, and more broadly with impaired personality function, social withdrawal, and emotional flatness. The idiosyncratic semantic cognition may worsen difficulties with social reciprocity seen in SPD and BPD.

Evaluation of Student Academic Performance After Curricular Content Reduction.

OBJECTIVE: To evaluate the effect of curricular content reduction in an integrated course sequence spanning 3 years of a Doctor of Pharmacy curriculum on student examination scores and course grades. METHODS: This 2-year, prepost study compared student overall average and final examination scores and overall course grades after the transition from a 5-day to a 4-day week of an integrated learning experience (ILE) course sequence. In addition, an anonymous, optional 23-item survey was distributed to first to third year pharmacy students asking about the 4-day week change, how they utilized the non-ILE day, and additional demographic and social characteristics to identify factors influencing success on examination and course performance during the 4-day week. RESULTS: There were 533 students included in the overall analysis, with no significant differences in overall course grades in the 5-day vs 4-day week. Examination scores were not significantly different after the transition, except in 2 of 12 courses where scores were higher and final examination scores were not significantly different, except for higher final examination scores in 1 course during the 5-day week. Significant positive influencers of top quartile of examination performance included prepharmacy grade point average ≥ 3.5, age 25 to 29 years, and prepharmacy coursework at the parent institution, whereas using the non-ILE day primarily to sleep negatively influenced outcomes. CONCLUSION: Curricular density is a prevalent problem and addressing it at a program level is essential. Reducing curricular content and hours at our institution did not adversely impact student examination and course performance and slight improvement was noted in some areas.

Nirsevimab: Expansion of Respiratory Syncytial Virus Prevention Options in Neonates, Infants, and At-Risk Young Children.

OBJECTIVE: Review available data from clinical trials of nirsevimab for efficacy and safety in the setting of respiratory syncytial virus (RSV) prophylaxis in infants and children, while exploring nirsevimab's role in clinical practice and highlighting continuing questions. DATA SOURCES: A literature search of PubMed was conducted utilizing the phrases "nirsevimab" and "medi8897." Additional references were identified through found references. Organizational guidelines, medication labeling, and regulatory organization presentations were utilized. STUDY SELECTION AND DATA EXTRACTION: Relevant clinical trials investigating nirsevimab in infants and children were included as well as other references on pharmacology, pharmacokinetics, and pharmacoeconomics. DATA SYNTHESIS: Nirsevimab, a once-a-season monoclonal antibody, demonstrated a 79.5% (95% CI, 65.9-87.7; P < 0.00001) lower incidence of RSV-associated medically attended lower respiratory tract infections (MA RSV-associated LRTI) and 77.3% (95% CI, 50.3-89.7; P = 0.0002) reduction in hospitalizations for RSV-associated MA-LRTI across 2 placebo-controlled studies. Nirsevimab demonstrated comparable safety to placebo with minor dermatologic reactions being the most common adverse event (0.9% vs 0.6%). RELEVANCE TO PATIENT CARE AND CLINICAL PRACTICE IN COMPARISON WITH EXISTING AGENTS: Nirsevimab was approved by the US Food and Drug Administration, and recommended by the Advisory Committee on Immunization Practices and American Academy of Pediatrics for broad administration to infants entering their first RSV season and at risk patients during their second RSV season. Questions remain over efficacy in infants born < 29-week gestation and other economical considerations. CONCLUSIONS: Nirsevimab demonstrated clinical efficacy in reducing RSV-associated MA-LRTI and RSV-associated hospitalizations in infants and was well tolerated.

Diagnosis and Treatment of ADHD in the Pediatric Population.

Attention-deficit/hyperactivity disorder (ADHD) is one of the most common neurodevelopmental disorders in childhood with approximately 6 million children (age 3 to 17 years) ever diagnosed based on data from 2016-2019. ADHD is characterized by a constant pattern of inattention and/or hyperactivity-impulsivity symptoms that interferes with development or functioning. Specific criteria from the Diagnostic and Statistical Manual of Mental Disorders, 5th edition Text Revision assist with the diagnosis with multiple guidelines available providing non-pharmacologic and pharmacologic recommendations for the treatment of ADHD in the pediatric population. While all guidelines similarly recommend behavioral and/or stimulant therapy as first-line therapy based on age, not all stimulant products are equal. Their differing pharmacokinetic profiles and formulations are essential to understand in order to optimize efficacy and safety for patients. Additionally, new stimulant products and non-stimulant medications continue to be approved for use of ADHD in the pediatric population and it is important to know their differences in formulation, efficacy, and safety to other products currently available. Lastly, due to drug shortages, it is important to understand product similarities and differences to select alternative therapy for patients.

Cholesterol-binding motifs in STING that control endoplasmic reticulum retention mediate anti-tumoral activity of cholesterol-lowering compounds.

The cGAS-STING pathway plays a crucial role in anti-tumoral responses by activating inflammation and reprogramming the tumour microenvironment. Upon activation, STING traffics from the endoplasmic reticulum (ER) to Golgi, allowing signalling complex assembly and induction of interferon and inflammatory cytokines. Here we report that cGAMP stimulation leads to a transient decline in ER cholesterol levels, mediated by Sterol O-Acyltransferase 1-dependent cholesterol esterification. This facilitates ER membrane curvature and STING trafficking to Golgi. Notably, we identify two cholesterol-binding motifs in STING and confirm their contribution to ER-retention of STING. Consequently, depletion of intracellular cholesterol levels enhances STING pathway activation upon cGAMP stimulation. In a preclinical tumour model, intratumorally administered cholesterol depletion therapy potentiated STING-dependent anti-tumoral responses, which, in combination with anti-PD-1 antibodies, promoted tumour remission. Collectively, we demonstrate that ER cholesterol sets a threshold for STING signalling through cholesterol-binding motifs in STING and we propose that this could be exploited for cancer immunotherapy.

Transcutaneous auricular vagus nerve stimulation for the treatment of myoarthropatic symptoms in patients with craniomandibular dysfunction - a protocol for a randomized and controlled pilot trial.

BACKGROUND: Temporomandibular disorders (TMD) are a collective term for pain and dysfunction of the masticatory muscles and the temporomandibular joints. The most common types of TMD are pain-related, which may impact the psychological behavior and quality of life. Currently, the most popular methods for the treatment of TMD patients are occlusal splint therapy, often in combination with physical- and/or pharmacotherapy. However, due to the complexity of etiology, the treatment of chronic TMD remains a challenge. Recently, CE-certified systems for non-invasive VNS (transcutaneous auricular vagus nerve stimulation, taVNS) have become available and show positive effects in the treatment of chronic pain conditions, like migraine or fibromyalgia, with which TMD shares similarities. Therefore, it is the main purpose of the study to evaluate the feasibility of daily taVNS against chronic TMD and to assess whether there is an improvement in pain severity, quality of life, and kinetic parameters. METHODS: This study is designed as a single-blinded, double-arm randomized controlled trial (RCT) in a 1:1 allocation ratio. Twenty adult patients with chronical TMD symptoms will be enrolled and randomized to stimulation or sham group. In the stimulation group, taVNS is performed on the left tragus (25 Hz, pulse width 250 µs, 28 s on/32 s off, 4 h/day). The sham group will receive no stimulation via a non-functional identical-looking electrode. Validated questionnaire data and clinical parameters will be collected at the beginning of the study and after 4 and 8 weeks. The compliance of a daily taVNS of patients with chronical TMD will be evaluated via a smartphone app recording daily stimulation time and average intensity. Additionally, the treatment impact on pain severity and quality of life will be assessed with different questionnaires, and the effect on the mandibular mobility and muscle activity will be analyzed. DISCUSSION: This is the first clinical trial to assess the feasibility of taVNS in patients with chronic TMD symptoms. If taVNS improves the symptoms of TMD, it will be a significant gain in quality of life for these chronic pain patients. The results of this pilot study will help to determine the feasibility of a large-scale RCT. TRIAL REGISTRATION: This study has been registered in the DRKS database (DRKS00029724).

Dietary acrylamide-linked burden of cancers in four sub-sahara African countries: A review and data synthesis.

Acrylamide (AA) is a food processing byproduct that forms at high temperatures and is classified as a probable human carcinogen. Previous studies have linked AA to kidney, uterus, and ovary cancer burdens, but its study in African countries remains underexplored. This study systematically used six recent articles on dietary AA concentration data from scholarly databases using specific search terms. We also collected health metrics secondary data from the Institute of Health Metrics and Evaluation and other sources for the period 2015-2019. We used a Monte-Carlo simulation to integrate the dietary AA exposure, risks, and health metrics to estimate the cancer burdens. The results showed that the modal healthy life years lost ranged from 0.00488 (Ghana) to 0.218 (Ethiopia) per 100,000 population. The median statistic indicated 1.2 and 26.10 healthy life years lost for Ghana and Ethiopia, respectively, due to the three cancer types. The four-country study areas' total disability-adjusted life years (DALYs) were 63.7 healthy life-year losses. Despite the limitations of the non-standardized age-related food consumption data and the few inclusive articles, the probabilistic approach may account for the uncertainties and provide valid conclusions.

Best practices for the execution, analysis, and data storage of plant single-cell/nucleus transcriptomics.

Single-cell and single-nucleus RNA-sequencing technologies capture the expression of plant genes at an unprecedented resolution. Therefore, these technologies are gaining traction in plant molecular and developmental biology for elucidating the transcriptional changes across cell types in a specific tissue or organ, upon treatments, in response to biotic and abiotic stresses, or between genotypes. Despite the rapidly accelerating use of these technologies, collective and standardized experimental and analytical procedures to support the acquisition of high-quality data sets are still missing. In this commentary, we discuss common challenges associated with the use of single-cell transcriptomics in plants and propose general guidelines to improve reproducibility, quality, comparability, and interpretation and to make the data readily available to the community in this fast-developing field of research.

Influence of ceramic thickness and dental substrate on the survival rate and failure load of non-retentive occlusal veneers after fatigue.

OBJECTIVE: To investigate the effect of ceramic thickness and dental substrate (enamel vs. dentin/enamel) on the survival rate and failure load of non-retentive occlusal veneers. MATERIALS AND METHODS: Human maxillary molars (n = 60) were divided into five test-groups (n = 12). The groups (named DE-1.5, DE-1.0, DE-0.5, E-1.0, E-0.5) differed in their dental substrate (E = enamel, DE = dentin/enamel) and restoration thickness (standard: 1.5 mm, thin: 1.0 mm, ultrathin: 0.5 mm). All teeth were prepared for non-retentive monolithic lithium-disilicate occlusal veneers (IPS e.max Press, Ivoclar). Restorations were adhesively cemented (Syntac Classic/Variolink II, Ivoclar) and exposed to thermomechanical fatigue (1.2 million cycles, 1.6 Hz, 49 N/ 5-55°C). Single load to failure was performed using a universal testing-machine. A linear-regression model was applied, pairwise comparisons used the Student-Newman-Keuls method (p < 0.05). RESULTS: Three dentin-based occlusal veneers (one DE-1.0, two DE-0.5) revealed cracks after fatigue exposure, which corresponds to an overall-survival rate of 95%. Load to failure resulted in the following ranking: 2142 N(DE-0.5) > 2105 N(E-1.0) > 2075 N(E-0.5) > 1440 N(DE-1.5) > 1430 N(DE-1.0). Thin (E-1.0) and ultrathin enamel-based occlusal veneers (E-0.5) revealed high failure loads and surpassed the standard thickness dentin-based veneers (DE-1.5) significantly (p = 0.044, p = 0.022). CONCLUSION: All tested monolithic lithium disilicate occlusal veneers obtained failure loads above physiological chewing forces. Thin and ultrathin enamel-based occlusal veneers outperformed the standard thick dentin-based occlusal veneers. CLINICAL SIGNIFICANCE: Minimally invasive enamel-based occlusal veneer restorations with non-retentive preparation design may serve as a conservative treatment option.

Co-occurrence of borderline and schizotypal personality disorders: a scoping review.

BACKGROUND: The historical concept of borderline conditions refers to the pathology on the border between neurosis and psychosis. In DSM-III the conditions were divided into specific but also somewhat overlapping diagnostic criteria for Borderline Personality Disorder (BPD) and Schizotypal Personality Disorder (SPD). This phenomenological overlap, which results in co-occurrence of the two diagnoses, remains a clinical challenge to this day. METHODS: To address this issue we examined the co-occurrence of SPD and BPD according to the established DSM-IV/-5 diagnostic criteria. A literature search was conducted including studies that employed a structured interview with defined BPD and SPD criteria. RESULTS: Studies from 20 samples were included (i.e. 15 patients, 3 community and 2 forensic samples). For patients diagnosed primarily with BPD, 1-27% also met the criteria for SPD and for patients diagnosed primarily with SPD, 5 - 33% showed co-occurrence with BPD. In the forensic samples, co-occurrence for primary BPD was 10% and 67 - 82% for primary SPD. In the community samples, co-occurrence for primary BPD was 29% and 50% for primary SPD. The pattern of co-occurrence across community samples was particularly heterogeneous. CONCLUSION: The identified co-occurrences for BPD and SPD were considerably sample-dependent, and samples and measurements were generally too heterogeneous for a precise meta-analysis. Forensic and community samples generally showed higher co-occurrences, but these findings were characterized by potential methodological limitations.

Trusting information systems in everyday work events - effects on cognitive resources, performance, and well-being.

In today's data-intensive work environments, information systems are crucial for supporting workers. However, workers often do not rely on these systems but resort to workarounds. We argue that trust is essential for workers' reliance on information systems, positively affecting workers' cognitive resources, performance, and well-being. Moreover, we argue that the organisational context (accountability, distractions) and user-related factors qualify trust-outcome associations by affecting workers' trust calibration. In a preregistered study, we asked N = 291 employed users of information systems to re-experience prior everyday usage events (event reconstruction method) and assess event-specific trust in the system, work outcomes, and context conditions. Results confirmed the assumed association between trust in the information system and workers' ratings of both performance and well-being. Moreover, workers' technology competence and need for cognition - but not contextual conditions - qualified trust-outcome associations. Our results offer specific suggestions for achieving successful use of information systems at work.

This preregistered study with 291 employees demonstrates the importance of trust in information systems for workers' performance and well-being in everyday work events. User-related factors, namely, users' technology competence and need for cognition, further qualified these effects. Practical implications for active trust management are drawn.

T-cell derived extracellular vesicles prime macrophages for improved STING based cancer immunotherapy.

A key phenomenon in cancer is the establishment of a highly immunosuppressive tumour microenvironment (TME). Despite advances in immunotherapy, where the purpose is to induce tumour recognition and hence hereof tumour eradication, the majority of patients applicable for such treatment still fail to respond. It has been suggested that high immunological activity in the tumour is essential for achieving effective response to immunotherapy, which therefore have led to exploration of strategies that triggers inflammatory pathways. Here activation of the stimulator of interferon genes (STING) signalling pathway has been considered an attractive target, as it is a potent trigger of pro-inflammatory cytokines and types I and III interferons. However, immunotherapy combined with targeted STING agonists has not yielded sustained clinical remission in humans. This suggests a need for exploring novel adjuvants to improve the innate immunological efficacy. Here, we demonstrate that extracellular vesicles (EVs), derived from activated CD4+ T cells (T-EVs), sensitizes macrophages to elevate STING activation, mediated by IFNγ carried on the T-EVs. Our work support that T-EVs can disrupt the immune suppressive environment in the tumour by reprogramming macrophages to a pro-inflammatory phenotype, and priming them for a robust immune response towards STING activation.

Reply to Freckmann et al. Comment on "Hochfellner et al. Accuracy Assessment of the GlucoMen® Day CGM System in Individuals with Type 1 Diabetes: A Pilot Study. Biosensors 2022, 12, 106".

We thank Dr. Freckmann et al. [...].

A few small steps or giant leap? De-densify the curriculum in one move.

INTRODUCTION: We describe a two-year, stepwise curricular change plan with a goal to address student and faculty wellbeing by reducing curricular density and rescheduling courses. METHODS: At year five of a new practice-ready curriculum, we strategically assessed and redesigned our integrated learning experience (ILE) course sequence to minimize curricular density with consideration for student and faculty wellbeing. This stepwise approach to curricular revision involved converting an in-class day (three hours) to asynchronous delivery in 12 core didactic courses (first- through third-professional years) for one year followed by a transition to a four-day vs. five-day course week the following year. Additional scheduling changes were implemented to improve the use of physical classroom space and overlapping student activities. RESULTS: Whereas thinking a gradual transition would be more accepted, moving straight to a four-day ILE week would have been more beneficial for faculty workload and wellbeing and decreased student confusion. However, the rescheduling of courses freed up classroom space allowing all classes to be held in person in a physical classroom and alleviated overlapping curricular activities during the week. CONCLUSIONS: The stepwise change of including an asynchronous day resulted in confusion and stress during the transition to a four-day class week and additional faculty work. However, adjusting the course schedule increased uncommitted student and faculty time, alleviated classroom constraints, and eliminated overlapping curriculum activities. Learning about our process and approach to reduce curricular density, what worked well, and lessons learned may allow other programs to be more efficient when making curricular changes.

Mentorship Landscape and Common Practices in an Academic Pharmacy Association.

OBJECTIVES: To explore the landscape of mentorship within professional associations in pharmacy academia, including reviewing available literature and describing currently available programs within the American Association of Colleges of Pharmacy, and recommend key considerations for the development of mentorship programs within professional associations. FINDINGS: A literature review of mentorship programs within professional associations for pharmacy academics was conducted, with a total of 5 articles identified and summarized. Additionally, a survey was conducted to determine the landscape of available mentorship programs within American Association of Colleges of Pharmacy affinity groups to capture unpublished experiences. Information regarding common characteristics and assessment methods was collected for groups that have mentorship programs, while needs and barriers were collected for those who did not. SUMMARY: Literature, while limited, supports positive perceptions of mentorship programs within professional associations. Based on the responses and working group experience, several recommendations are proposed for mentorship program development, including the need for clearly defined goals, relevant program outcomes, association support to reduce redundancies and promote participation, and, in some cases, implementation of an association-wide program to ensure access to mentorship.