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Publication The L-DBF vaccine cross protects mice against different serotypes after prior exposure to the pathogen(American Society for Microbiology, 2023-12-12) Picking, Wendy LShigellosis is endemic to low- and middle-income regions of the world where children are especially vulnerable. In many cases, there are pre-existing antibodies in the local population and the effect of prior exposure should be considered in the development and testing of vaccines against infection. Our study shows that L-DBF-induced immune responses are not adversely affected by prior exposure to this pathogen. Moreover, somewhat different cytokine profiles were observed in the lungs of vaccinated mice not having been exposed to , suggesting that the immune responses elicited by infection and L-DBF vaccination follow different pathways.Publication Emerging Trends and Translational Challenges in Drug and Vaccine Delivery(MDPI, 2024-01-11) Agrahari, VibhutiDrug and vaccine delivery have received considerable attention in recent years. Many rationally designed innovative approaches are being explored to address the challenges related to safety, efficacy, patient compliance, and cost-effective means for existing and new therapeutics. The extensive assessment of drug delivery involves pre-formulation and physicochemical characterization, mechanistic biochemical pathways at the molecular level, pharmacological and toxicological evaluations, and detailed preclinical investigations. Recent advancements have evolved to address the limitations that emerged with the evolution of novel therapeutic modalities from simple small molecules to more complex macromolecules, including nucleic acids, peptides, proteins, antibodies, and conjugates [1]. There’s immense interest in exploring the in vitro and in vivo behavior of drugs and vaccines to overcome biological barriers to reach target sites, and in expeditious translation from the lab to a manufacturing scale [2]. This Special Issue on “Emerging Trends and Translational Challenges in Drug and Vaccine Delivery” is the collection of those efforts by several researchers to address the unmet need of advanced drug and vaccine delivery systems. The studies published in this Special Issue are summarized below and are valuable for the readers of Pharmaceutics and the scientific community working in the field of drug and vaccine delivery. The first paper in this collection by Alkholief et al. demonstrated the use of dexamethasone-sodium-phosphate (DEX)-chitosan nanoparticles (CSNPs) coated with hyaluronic acid (HA) as a controlled release ocular delivery vehicle for the treatment of endotoxin-induced-uveitis (EIU) in a rabbit model [3]. The CSNPs were stable at 25 °C for 3 months and in vitro studies showed a similar DEX release in a range of 74–77% for uncoated and HA-coated nanoparticles. Drug-loaded CSNPs were safe for ocular applications and showed a noTable 10-fold increase in transcorneal flux and permeability of DEX in the case of HA-CSNPs vs. DEX-aqueous solution (DEX-AqS). The findings suggest improved delivery properties and promising anti-inflammatory effects of DEX-CSNPs in EIU rabbits with ocular bioavailability, with the half-life and ocular MRT0-inf of DEX being significantly higher than DEX-AqS. Another study focused on extracellular nanovesicles (EVs) that have great potential as drug delivery systems for precision therapy but are limited due to technical challenges to purify and characterize the EVs. To address this issue, Nguyen et al. developed a 3D inner filter-based technique for the simple extraction of apoplastic fluid from blueberries, enabling EV purification [4]. The high drug loading capability and properties to modulate the release of proinflammatory cytokine IL-8 and total glutathione have enabled blueberry-derived EVs (BENVs) to be a promising edible multifunctional nano-bio-platform for future immunomodulatory therapies. Vaccination is the most effective way to prevent infectious diseases but suffers from fading immunity requiring frequent boosters to maintain the immune response. In a novel approach, Kooji et al. demonstrated the effectiveness of a single injection with sustained-release microspheres as an alternative to the conventional multiple injection (prime-boost) immunization schedule of bovine serum albumin in terms of eliciting the same levels of IgG antibody response in mice [5]. The microspheres were designed based on two novel biodegradable multi-block copolymers with an opportunity to tailor the release profile in a range of 4 to 9 weeks by varying the polymer ratios. Adjuvants are ingredients used in many vaccines to elicit a stronger immune response. In a recent study, Liang et al. demonstrated the use of formulated phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), a component of oil-in-water vaccine adjuvant emulsion (known as a stable emulsion or SE), as non-canonical agonists for murine and human TLR4 [6]. The effects of DMPC on human cells were proven but were less pronounced than the composition of emulsion oil and were dependent on the saturation, size, and headgroup of the phospholipid. The next article is focused on drug loaded-microneedles, which are minimally invasive systems capable of painless delivery and offer dose-sparing benefits with a potential to replace hypodermal needles and oral routes of delivery. In this study, Faizi et al. developed a deferasirox-nanosuspension (DFS-NS) loaded with dissolving microneedles (DMN) for intradermal delivery for effective treatment of iron overload [7]. DFS-NSs were formulated by the wet media milling procedure using PVA and showed a 3-fold higher dissolution rate vs. pure DFS. The skin deposition studies showed significantly higher drug deposition from DFS-NSs loaded with polymeric dissolving microneedles (NS-DMN) as compared to DFS-NS transdermal patches without needles (DFS-NS-TP) or pure DFS-DMNs. Hence, the authors showed that loading DFS-NSs into novel DMN devices can be effectively used for transdermal delivery of sparingly soluble drugs, i.e., DFS in aqueous systems. In another study, Peng et al. demonstrated the development of amphotericin B (AMB)- and levofloxacin (LVX)-loaded chitosan films for potential use in antimicrobial wound dressings [8]. An HPLC method developed by the authors measured 100% and 60% release of LVX and AMB, respectively, from the chitosan film after a week. An ex vivo deposition study showed that 20.96 ± 13.54 and 0.35 ± 0.04 of LVX and AMB, respectively, were deposited in porcine skin 24 h after application. Further, the films were able to inhibit the growth of Candida albicans and Staphylococcus aureus, demonstrating their antimicrobial applications. Wang et al. in their recent review discussed the translational challenges and prospective solutions for implementing biomimetic delivery systems (BDSs) for therapeutic delivery [9]. BDSs are based on complex designs of biological structures and have emerged as a powerful tool for drug and vaccine delivery. This review provides recent advances in the development of BDSs, discusses the challenges faced in the translation of BDs from research to clinical applications, and presents emerging solutions, emphasized by real-world case studies. Luo et al. provide insights into the development of organs-on-chips (OCs) and their impact on precision medicine and advanced system simulation [10]. OCs are devices with micro-physiological systems containing small tissues grown inside microfluidic chips with controlled cell microenvironments to study the pathophysiology and effect of drugs on the human body. OCs represent a faster, economical, and precise approach to study drug safety, efficacy, disease modelling and treatments with a potential to complement/replace traditional preclinical cell cultures, animal studies, and even human clinical trials. Ingle and Fang in their recent review present an overview of the stability and delivery challenges of commercial nucleic acid (NA)-based therapeutics, including DNA, RNA, oligonucleotides, siRNA, miRNA, mRNA, small activating RNA, and gene therapies [11]. The review highlights NA-based therapeutics approved by the European Medicines Agency (EMA) and US Food and Drug Administration (US FDA) with a focus on the current progress in improving the stability, delivery, cost, and regulatory acceptance of these therapeutics. There is significant interest in developing approaches to overcome the blood–brain barrier (BBB) for treatment of central nervous system (CNS) diseases. Meyer et al. in their recent review described novel developments to enable the treatment of CNS diseases with targeted drug delivery [12]. The review focuses on unfolding the full potential of novel therapeutic entities, i.e., gene therapy and degradomers, using innovative delivery systems for possible application in the treatment of CNS diseases. In conclusion, this Special Issue converses through the translation of therapeutic delivery from discovery to large-scale production for pharmaceutical and biotechnology applications. The discussed strategies, including the use of polymeric nanoparticles, extracellular nanovesicles, sustained release microspheres, microneedles, polymeric biofilms, biomimetic delivery systems, new adjuvants, and organ-on-chips, possess great potential in addressing the limitations of drug and vaccine delivery. The editors express their gratitude for the interest and cooperation of the contributors and believe this Special Issue of Pharmaceutics would be an interesting addition to the scientific community engaged in drug delivery research.Publication Partner-seeking and limbic dopamine system are enhanced following social loss in male prairie voles (Microtus ochrogaster)(International Behvaioural and Neural Genetics Society, 2023-12-22) Smith, Adam SDeath of a loved one is recognized as one of life's greatest stresses, and 10%-20% of bereaved individuals will experience a complicated or prolonged grieving period that is characterized by intense yearning for the deceased. The monogamous prairie vole (Microtus ochrogaster) is a rodent species that forms pair bonds between breeding partners and has been used to study the neurobiology of social behaviors and isolation. Male prairie voles do not display distress after isolation from a familiar, same-sex conspecific; however, separation from a bonded female partner increases emotional, stress-related, and proximity-seeking behaviors. Here, we tested the investigatory response of male voles to partner odor during a period of social loss. We found that males who lost their partner spent significantly more time investigating partner odor but not non-partner social odor or food odor. Bachelor males and males in intact pairings did not respond uniquely to any odor. Furthermore, we examined dopamine (DA) receptor mRNA expression in the anterior insula cortex (aIC), nucleus accumbens (NAc), and anterior cingulate (ACC), regions with higher activation in grieving humans. While we found some effects of relationship type on DRD1 and DRD2 expression in some of these regions, loss of a high-quality opposite-sex relationship had a significant effect on DA receptor expression, with pair-bonded/loss males having higher expression in the aIC and ACC compared with pair-bonded/intact and nonbonded/loss males. Together, these data suggest that both relationship type and relationship quality affect reunion-seeking behavior and motivational neurocircuits following social loss of a bonded partner.Publication The combination of propylene glycol and vegetable glycerin e-cigarette aerosols induces airway inflammation and mucus hyperconcentration(Nature Research, 2024-01-23) Salathe, MatthiasDespite concerns over their safety, e-cigarettes (e-cigs) remain a popular tobacco product. Although nicotine and flavors found in e-cig liquids (e-liquids) can cause harm in the airways, whether the delivery vehicles propylene glycol (PG) and vegetable glycerin (VG) are innocuous when inhaled remains unclear. Here, we investigated the effects of e-cig aerosols generated from e-liquid containing only PG/VG on airway inflammation and mucociliary function in primary human bronchial epithelial cells (HBEC) and sheep. Primary HBEC were cultured at the air-liquid interface (ALI) and exposed to e-cig aerosols of 50%/50% v/v PG/VG. Ion channel conductance, ciliary beat frequency, and the expression of inflammatory markers, cell type-specific markers, and the major mucins MUC5AC and MUC5B were evaluated after seven days of exposure. Sheep were exposed to e-cig aerosols of PG/VG for five days and mucus concentration and matrix metalloproteinase-9 (MMP-9) activity were measured from airway secretions. Seven-day exposure of HBEC to e-cig aerosols of PG/VG caused a significant reduction in the activities of apical ion channels important for mucus hydration, including the cystic fibrosis transmembrane conductance regulator (CFTR) and large conductance, Ca-activated, and voltage-dependent K (BK) channels. PG/VG aerosols significantly increased the mRNA expression of the inflammatory markers interleukin-6 (IL6), IL8, and MMP9, as well as MUC5AC. The increase in MUC5AC mRNA expression correlated with increased immunostaining of MUC5AC protein in PG/VG-exposed HBEC. On the other hand, PG/VG aerosols reduced MUC5B expression leading overall to higher MUC5AC/MUC5B ratios in exposed HBEC. Other cell type-specific markers, including forkhead box protein J1 (FOXJ1), keratin 5 (KRT5), and secretoglobin family 1A member 1 (SCGB1A1) mRNAs, as well as overall ciliation, were significantly reduced by PG/VG exposure. Finally, PG/VG aerosols increased MMP-9 activity and caused mucus hyperconcentration in sheep in vivo. E-cig aerosols of PG/VG induce airway inflammation, increase MUC5AC expression, and cause dysfunction of ion channels important for mucus hydration in HBEC in vitro. Furthermore, PG/VG aerosols increase MMP-9 activity and mucus concentration in sheep in vivo. Collectively, these data show that e-cig aerosols containing PG/VG are likely to be harmful in the airways.Publication Antipsychotics in child and adolescent patients with major depressive disorder: A retrospective analysis of prescribing patterns(The Mental Health Clinician, 2024-02) Moeller, Karen ECareful consideration should be taken when using off-label antipsychotics in children due to limited studies on efficacy. Future research is warranted to assess the efficacy and safety of these agents in children and adolescents.Publication Consumer perception, knowledge, and uses of cannabidiol(The Mental Health Clinician, 2023-10-02) Nguyen, Cambrey; Moeller, Karen E; McGuire, Michael; Melton, Brittany L.Introduction The legalization of cannabidiol (CBD) across the United States, in varying degrees, has made CBD easily accessible to consumers for complementary and medical purposes. However, there is a paucity of scientific evidence on the benefits and risks of commercially available CBD. In the literature, 2 studies have gathered consumer perceptions and attitudes on cannabis products, specifically CBD, using survey-based questionnaires. This study aimed to build on the aforementioned studies in obtaining consumer perception and knowledge of CBD products using a national survey-based questionnaire. Methods Respondents were recruited through an anonymous, nationwide, online survey administered through Qualtrics in the United States from March 28 to April 30, 2021. The survey consisted of demographics, perceived efficacy and safety of CBD, and resources to obtain CBD information. The survey responses were reported using descriptive statistics along with median and interquartile range for the Likert portion. Results A total of 1158 respondents accessed the survey. The median age was 43 and 50% of respondents were female. The uses for CBD included neurological disorders, pulmonary conditions, gastrointestinal disorders, and chronic pain. The most commonly reported safety concern related to taking CBD was anxiety. Participants agreed that CBD is safe when used responsibly for medical use, and social media was the main source used to obtain information about CBD. Discussion Respondents who used CBD for a condition thought it was helpful; however, most of the adverse effects were rated as moderate to severe, requiring medical attention from a health care professional, hospital, or emergency room visit. Keywords: cannabidiol, consumer, perception, drug resources, surveyPublication Characterization of antibiotic overuse for common infectious disease states at hospital discharge(Cambridge University Press, 2023-12-12) Klatt, MeganAntibiotic overuse was prevalent upon hospital discharge for these three common infectious disease states. Transitions of care should be prioritized as an area for antimicrobial stewardship intervention.Publication Population pharmacokinetic analysis of enrofloxacin and its active metabolite ciprofloxacin after intravenous injection to cats with reduced kidney function(Wiley Open Access, 2023-09-20) Foster, Jonathan D.; Abouraya, Mahmoud; Papich, Mark G.; Muma, Nancy A.Background It is unknown if enrofloxacin accumulates in plasma of cats with reduced kidney function. Hypothesis To determine if enrofloxacin and its active metabolite ciprofloxacin have reduced clearance in azotemic cats. Animals Thirty‐four cats hospitalized for clinical illness with variable degree of kidney function. Methods Prospective study. After enrofloxacin (dose 5 mg/kg) administration to cats, sparse blood sampling was used to obtain 2 compartment population pharmacokinetic results using nonlinear mixed‐effects modeling. Plasma enrofloxacin and ciprofloxacin concentrations were measured and summed to obtain the total fluoroquinolone concentration. A model of ciprofloxacin metabolism from enrofloxacin was created and evaluated for covariate effects on clearance, volume of distribution, and the metabolic rate of ciprofloxacin generation from enrofloxacin. Results Body weight was the only covariate found to affect total fluoroquinolone volume of distribution (effect 1.63, SE 0.19, P < .01) and clearance (effect 1.63, SE 0.27, P < .01). Kidney function did not have a significant effect on total fluoroquinolone clearance (median 440.8 mL/kg/h (range 191.4‐538.0 mL/kg/h) in cats with normal kidney function, 365.8 mL/kg/h (range 89.49‐1092.0 mL/kg/h) in cats with moderate kidney dysfunction, and 308.5 mL/kg/h (range 140.20‐480.0 mL/kg/h) in cats with severe kidney dysfunction (P = .64). Blood urea nitrogen concentration influenced the metabolic generation of ciprofloxacin from enrofloxacin (effect 0.51, SE 0.08, P < .01), but other markers of kidney function did not. Conclusions and clinical importance. Adjustment of enrofloxacin dosage is not indicated for azotemic cats.Publication Mitochondrial Interaction with Serotonin in Neurobiology and Its Implication in Alzheimer’s Disease(IOS Press, 2023-11-01) Tian, Jing; Du, Eric; Guo, LanAlzheimer’s disease (AD) is a lethal neurodegenerative disorder characterized by severe brain pathologies and progressive cognitive decline. While the exact cause of this disease remains unknown, emerging evidence suggests that dysregulation of neurotransmitters contributes to the development of AD pathology and symptoms. Serotonin, a critical neurotransmitter in the brain, plays a pivotal role in regulating various brain processes and is implicated in neurological and psychiatric disorders, including AD. Recent studies have shed light on the interplay between mitochondrial function and serotonin regulation in brain physiology. In AD, there is a deficiency of serotonin, along with impairments in mitochondrial function, particularly in serotoninergic neurons. Additionally, altered activity of mitochondrial enzymes, such as monoamine oxidase, may contribute to serotonin dysregulation in AD. Understanding the intricate relationship between mitochondria and serotonin provides valuable insights into the underlying mechanisms of AD and identifies potential therapeutic targets to restore serotonin homeostasis and alleviate AD symptoms. This review summarizes the recent advancements in unraveling the connection between brain mitochondria and serotonin, emphasizing their significance in AD pathogenesis and underscoring the importance of further research in this area. Elucidating the role of mitochondria in serotonin dysfunction will promote the development of therapeutic strategies for the treatment and prevention of this neurodegenerative disorder.Publication Population pharmacokinetic analysis of enrofloxacin and its active metabolite ciprofloxacin after intravenous injection to cats with reduced kidney function(Wiley Open Access, 2023-09-20) Foster, Jonathan D.; Abouraya, Mahmoud; Papich, Mark G.; Muma, Nancy A.Background It is unknown if enrofloxacin accumulates in plasma of cats with reduced kidney function. Hypothesis To determine if enrofloxacin and its active metabolite ciprofloxacin have reduced clearance in azotemic cats. Animals Thirty‐four cats hospitalized for clinical illness with variable degree of kidney function. Methods Prospective study. After enrofloxacin (dose 5 mg/kg) administration to cats, sparse blood sampling was used to obtain 2 compartment population pharmacokinetic results using nonlinear mixed‐effects modeling. Plasma enrofloxacin and ciprofloxacin concentrations were measured and summed to obtain the total fluoroquinolone concentration. A model of ciprofloxacin metabolism from enrofloxacin was created and evaluated for covariate effects on clearance, volume of distribution, and the metabolic rate of ciprofloxacin generation from enrofloxacin. Results Body weight was the only covariate found to affect total fluoroquinolone volume of distribution (effect 1.63, SE 0.19, P < .01) and clearance (effect 1.63, SE 0.27, P < .01). Kidney function did not have a significant effect on total fluoroquinolone clearance (median 440.8 mL/kg/h (range 191.4‐538.0 mL/kg/h) in cats with normal kidney function, 365.8 mL/kg/h (range 89.49‐1092.0 mL/kg/h) in cats with moderate kidney dysfunction, and 308.5 mL/kg/h (range 140.20‐480.0 mL/kg/h) in cats with severe kidney dysfunction (P = .64). Blood urea nitrogen concentration influenced the metabolic generation of ciprofloxacin from enrofloxacin (effect 0.51, SE 0.08, P < .01), but other markers of kidney function did not. Conclusions and clinical importance. Adjustment of enrofloxacin dosage is not indicated for azotemic cats.Publication The Anti-Cancer Activity of the Naturally Occurring Dipeptide Carnosine: Potential for Breast Cancer(MDPI, 2023-11-08) Maugeri, Salvatore; Sibbitts, Jay; Privitera, Anna; Cardaci, Vincenzo; Pietro, Lucia Di; Leggio, Loredana; Iraci, Nunzio; Lunte, Susan M.; Caruso, Giuseppe; Tikkanen, Ritva; Gillet, GermainCarnosine is an endogenous dipeptide composed of β-alanine and L-histidine, possessing a multimodal pharmacodynamic profile that includes anti-inflammatory and anti-oxidant activities. Carnosine has also shown its ability to modulate cell proliferation, cell cycle arrest, apoptosis, and even glycolytic energy metabolism, all processes playing a key role in the context of cancer. Cancer is one of the most dreaded diseases of the 20th and 21st centuries. Among the different types of cancer, breast cancer represents the most common non-skin cancer among women, accounting for an estimated 15% of all cancer-related deaths in women. The main aim of the present review was to provide an overview of studies on the anti-cancer activity of carnosine, and in particular its activity against breast cancer. We also highlighted the possible advantages and limitations involved in the use of this dipeptide. The first part of the review entailed a brief description of carnosine’s biological activities and the pathophysiology of cancer, with a focus on breast cancer. The second part of the review described the anti-tumoral activity of carnosine, for which numerous studies have been carried out, especially at the preclinical level, showing promising results. However, only a few studies have investigated the therapeutic potential of this dipeptide for breast cancer prevention or treatment. In this context, carnosine has shown to be able to decrease the size of cancer cells and their viability. It also reduces the levels of vascular endothelial growth factor (VEGF), cyclin D1, NAD+, and ATP, as well as cytochrome c oxidase activity in vitro. When tested in mice with induced breast cancer, carnosine proved to be non-toxic to healthy cells and exhibited chemopreventive activity by reducing tumor growth. Some evidence has also been reported at the clinical level. A randomized phase III prospective placebo-controlled trial showed the ability of Zn–carnosine to prevent dysphagia in breast cancer patients undergoing adjuvant radiotherapy. Despite this evidence, more preclinical and clinical studies are needed to better understand carnosine’s anti-tumoral activity, especially in the context of breast cancer.Publication Protection against Aβ-induced neuronal damage by KU-32: PDHK1 inhibition as important target(Frontiers Media, 2023-11-14) Pal, Ranu; Hui, Dongwei; Menchen, Heather; Zhao, Huiping; Mozziconacci, Olivier; Wilkins, Heather; Blagg, Brian S. J.; Schöneich, Christian; Swerdlow, Russell H.; Michaelis, Mary L.; Michaelis, Elias K.A feature of most neurodegenerative diseases is the presence of “mis-folded proteins” that form aggregates, suggesting suboptimal activity of neuronal molecular chaperones. Heat shock protein 90 (Hsp90) is the master regulator of cell responses to “proteotoxic” stresses. Some Hsp90 modulators activate cascades leading to upregulation of additional chaperones. Novobiocin is a modulator at the C-terminal ATP-binding site of Hsp90. Of several novobiocin analogs synthesized and tested for protection against amyloid beta (Aβ)-induced neuronal death, “KU-32” was the most potent in protecting primary neurons, but did not increase expression of other chaperones believed to help clear misfolded proteins. However, KU-32 reversed Aβ-induced superoxide formation, activated Complex I of the electron transfer chain in mitochondria, and blocked the Aβ-induced inhibition of Complex I in neuroblastoma cells. A mechanism for these effects of KU-32 on mitochondrial metabolism appeared to be the inhibition of pyruvate dehydrogenase kinase (PDHK), both in isolated brain mitochondria and in SH-SY5Y cells. PDHK inhibition by the classic enzyme inhibitor, dichloroacetate, led to neuroprotection from Aβ25-35-induced cell injury similarly to KU-32. Inhibition of PDHK in neurons would lead to activation of the PDH complex, increased acetyl-CoA generation, stimulation of the tricarboxylic acid cycle and Complex I in the electron transfer chain, and enhanced oxidative phosphorylation. A focus of future studies may be on the potential value of PDHK as a target in AD therapy.Publication Repeated passive visual experience modulates spontaneous and non-familiar stimuli-evoked neural activity(Nature Research, 2023-11-27) Niraula, Suraj; Hauser, William L.; Rouse, Adam G.; Subramanian, JaichandarFamiliarity creates subjective memory of repeated innocuous experiences, reduces neural and behavioral responsiveness to those experiences, and enhances novelty detection. The neural correlates of the internal model of familiarity and the cellular mechanisms of enhanced novelty detection following multi-day repeated passive experience remain elusive. Using the mouse visual cortex as a model system, we test how the repeated passive experience of a 45° orientation-grating stimulus for multiple days alters spontaneous and non-familiar stimuli evoked neural activity in neurons tuned to familiar or non-familiar stimuli. We found that familiarity elicits stimulus competition such that stimulus selectivity reduces in neurons tuned to the familiar 45° stimulus; it increases in those tuned to the 90° stimulus but does not affect neurons tuned to the orthogonal 135° stimulus. Furthermore, neurons tuned to orientations 45° apart from the familiar stimulus dominate local functional connectivity. Interestingly, responsiveness to natural images, which consists of familiar and non-familiar orientations, increases subtly in neurons that exhibit stimulus competition. We also show the similarity between familiar grating stimulus-evoked and spontaneous activity increases, indicative of an internal model of altered experience.Publication Comparison of inpatient psychiatric medication management in gender diverse youth with cisgender peers(AAPP, 2023-08-07) Carrillo, Nina; McGurran, Maren; Melton, Brittany L.; Moeller, Karen E.Gender diverse (GD) youth, defined as transgender (TG) and nonconforming (NC) adolescents, have a 2- to 3-fold increased risk of having a mental health diagnosis, particularly a depressive or anxiety disorder, compared with cisgender (CG) peers.1,2 Additionally, psychologic distress is commonly reported in GD youths due to harassment, victimization, and bullying by peers.3,4 Approximately 30% to 50% of GD patients report serious psychologic distress leading to attempted suicide, with the highest risk in transmasculine patients.5-7 Mental health evaluations should be routine care for all adolescents.8 However, GD individuals often report stigma and discrimination in health care services secondary to lack of insurance coverage for gender-affirming care, and negative experiences with health care providers.3,9-11 Studies evaluating the treatment of psychiatric disorders in GD youth are limited. Hisle-Gorman et al12 conducted a large retrospective study assessing psychotropic prescribing in GD youth. Researchers identified 3754 military-dependent GD youth (ages <18 years) and matched them to CG siblings (N = 6603) to assess differences in mental health diagnoses, mental health visits, and outpatient psychotropic prescriptions. They found GD youth were approximately 5 times more likely to have a mental health diagnosis and at least 2 times more likely to use mental health services and be prescribed psychotropics. Literature suggests that gender-affirming care, addressing psychologic, surgical, and hormonal therapies, is medically necessary to alleviate gender dysphoria and reduce psychiatric comorbidities.13-16 In the study by Hisle-Gorman et al,12 researchers additionally assessed the effects of gender-affirming hormone therapy (GAHT) on mental health outcomes in the TG subgroup.12 Among the 963 TG youth receiving GAHT, mental health care use did not significantly change after initiation of GAHT; however, psychotropic medication use increased in all categories except stimulants, migraine agents, and lithium.12 Psychotropic medications are not benign agents and can have short- and long-term effects in children and adolescents. Antidepressants have been associated with reduced growth velocity, reduced bone mineral density, and a potential risk for diabetes mellitus secondary to weight gain in pediatric patients.17 All antidepressants carry a black box warning for increased suicidality in adolescent patients.18 Additionally, antipsychotics have increased risks for metabolic syndromes, specifically hyperlipidemia, diabetes mellitus, and cardiovascular disease.19-22 Our study aims to compare psychotropic prescribing in GD youth to their CG peers with the same primary psychiatric diagnosis during an inpatient psychiatric hospitalization.Publication Early social isolation stress increases addiction vulnerability to heroin and alters c-Fos expression in the mesocorticolimbic system(Springer, 2022-01-08) Singh, Archana; Xie, Yang; Davis, Ashton; Wang, Zi-JunRationale Adverse psychosocial factors during early childhood or adolescence compromise neural structure and brain function, inducing susceptibility for many psychiatric disorders such as substance use disorder. Nevertheless, the mechanisms underlying early life stress-induced addiction vulnerability is still unclear, especially for opioids. Objectives To address this, we used a mouse heroin self-administration model to examine how chronic early social isolation (ESI) stress (5 weeks, beginning at weaning) affects the behavioral and neural responses to heroin during adulthood. Results We found that ESI stress did not alter the acquisition for sucrose or heroin self-administration, nor change the motivation for sucrose on a progressive ratio schedule. However, ESI stress induced an upward shift of heroin dose-response curve in female mice and increased motivation and seeking for heroin in both sexes. Furthermore, we examined the neuronal activity (measured by c-Fos expression) within the key brain regions of the mesocorticolimbic system, including the prelimbic cortex (PrL), infralimbic cortex (IL), nucleus accumbens (NAc) core and shell, caudate putamen, and ventral tegmental area (VTA). We found that ESI stress dampened c-Fos expression in the PrL, IL, and VTA after 14-day forced abstinence, while augmented the neuronal responses to heroin-predictive context and cue in the IL and NAc core. Moreover, ESI stress disrupted the association between c-Fos expression and attempted infusions during heroin-seeking test in the PrL. Conclusions These data indicate that ESI stress leads to increased seeking and motivation for heroin, and this may be associated with distinct changes in neuronal activities in different subregions of the mesocorticolimbic system.Publication Opioid induces increased DNA damage in prefrontal cortex and nucleus accumbens(Elsevier, 2023-03-14) Wang, Yunwanbin; Singh, Archana; Li, Guohui; Yue, Shuwen; Hertel, Kegan; Wang, Zi-JunOpioid use disorder (OUD) is a chronic disease characterized by compulsive opioid taking and seeking, affecting millions of people worldwide. The high relapse rate is one of the biggest challenges in treating opioid addiction. However, the cellular and molecular mechanisms underlying relapse to opioid seeking are still unclear. Recent studies have shown that DNA damage and repair processes are implicated in a broad spectrum of neurodegenerative diseases as well as in substance use disorders. In the present study, we hypothesized that DNA damage is related to relapse to heroin seeking. To test our hypothesis, we aim to examine the overall DNA damage level in prefrontal cortex (PFC) and nucleus accumbens (NAc) after heroin exposure, as well as whether manipulating DNA damage levels can alter heroin seeking. First, we observed increased DNA damage in postmortem PFC and NAc tissues from OUD individuals compared to healthy controls. Next, we found significantly increased levels of DNA damage in the dorsomedial PFC (dmPFC) and NAc from mice that underwent heroin self-administration. Moreover, increased accumulation of DNA damage persisted after prolonged abstinence in mouse dmPFC, but not in NAc. This persistent DNA damage was ameliorated by the treatment of reactive oxygen species (ROS) scavenger N-acetylcysteine, along with attenuated heroin-seeking behavior. Furthermore, intra-PFC infusions of topotecan and etoposide during abstinence, which trigger DNA single-strand breaks and double-strand breaks respectively, potentiated heroin-seeking behavior. These findings provide direct evidence that OUD is associated with the accumulation of DNA damage in the brain (especially in the PFC), which may lead to opioid relapse.Publication Insulin-like growth factor 1 regulates excitatory synaptic transmission in pyramidal neurons from adult prefrontal cortex(Elsevier, 2022-10-01) Yue, Shuwen; Wang, Yunwanbin; Wang, Zi-JunInsulin-like growth factor 1 (IGF1) influences synaptic function in addition to its role in brain development and aging. Although the expression levels of IGF1 and IGF1 receptor (IGF1R) peak during development and decline with age, the adult brain has abundant IGF1 or IGF1R expression. Studies reveal that IGF1 regulates the synaptic transmission in neurons from young animals. However, the action of IGF1 on neurons in the adult brain is still unclear. Here, we used prefrontal cortical (PFC) slices from adult mice (∼8 weeks old) to characterize the role of IGF1 on excitatory synaptic transmission in pyramidal neurons and the underlying molecular mechanisms. We first validated IGF1R expression in pyramidal neurons using translating ribosomal affinity purification assay. Then, using whole-cell patch-clamp recording, we found that IGF1 attenuated the amplitude of evoked excitatory postsynaptic current (EPSC) without affecting the frequency and amplitude of miniature EPSC. Furthermore, this decrease in excitatory neurotransmission was blocked by pharmacological inhibition of IGF1R or conditional knockdown of IGF1R in PFC pyramidal neurons. In addition, we determined that IGF1-induced decrease of EPSC amplitude was due to postsynaptic effect (internalization of a-amino-3-hydroxy-5-methyl-4- isoxazolepropionic acid receptors [AMPAR]) rather than presynaptic glutamate release. Finally, we found that inhibition of metabotropic glutamate receptor subtype-1 (mGluR1) abolished IGF1-induced attenuation of evoked EPSC amplitude and decrease of AMPAR expression at synaptic membrane, suggesting mGluR1-mediated endocytosis of AMPAR was involved. Taken together, these data provide the first evidence that IGF1 regulates excitatory synaptic transmission in adult PFC via the interaction between IGF1R-dependent signaling pathway and mGluR1-mediated AMPAR endocytosis.Publication Activity of Delafloxacin and Comparator Fluoroquinolones against Multidrug-Resistant Pseudomonas aeruginosa in an In Vitro Cystic Fibrosis Sputum Model(MDPI, 2023-06-20) Craddock, Vaughn D.; Steere, Evan L.; Harman, Hannah; Britt, Nicholas S.Delafloxacin (DLX) is a recently approved fluoroquinolone with broad activity against common cystic fibrosis (CF) pathogens, including multidrug-resistant Pseudomonas aeruginosa (MDR-Psa). Delafloxacin has been previously shown to have excellent lung and biofilm penetration and enhanced activity at lower pH environments, such as those that would be observed in the CF lung. We analyzed six Psa strains isolated from CF sputum and compared DLX to ciprofloxacin (CPX) and levofloxacin (LVX). Minimum inhibitory concentrations (MICs) were determined for DLX using standard culture media (pH 7.3) and artificial sputum media (ASM), a physiologic media recapitulating the CF lung microenvironment (pH 6.9). Delafloxacin activity was further compared to CPX and LVX in an in vitro CF sputum time-kill model at physiologically relevant drug concentrations (Cmax, Cmed, Cmin). Delafloxacin exhibited 2- to 4-fold MIC reductions in ASM, which corresponded with significant improvements in bacterial killing in the CF sputum time-kill model between DLX and LVX at Cmed (p = 0.033) and Cmin (p = 0.004). Compared to CPX, DLX demonstrated significantly greater killing at Cmin (p = 0.024). Overall, DLX demonstrated favorable in vitro activity compared to alternative fluoroquinolones against MDR-Psa. Delafloxacin may be considered as an option against MDR-Psa pulmonary infections in CF.Publication Opposing Effects of ApoE2 and ApoE4 on Glycolytic Metabolism in Neuronal Aging Supports a Warburg Neuroprotective Cascade against Alzheimer’s Disease(MDPI, 2023-01-25) Zhang, Xin; Wu, Long; Swerdlow, Russell H.; Zhao, LiqinApolipoprotein E4 (ApoE4) is the most recognized genetic risk factor for late-onset Alzheimer’s disease (LOAD), whereas ApoE2 reduces the risk for LOAD. The underlying mechanisms are unclear but may include effects on brain energy metabolism. Here, we used neuro-2a (N2a) cells that stably express human ApoE isoforms (N2a-hApoE), differentiated N2a-hApoE neuronal cells, and humanized ApoE knock-in mouse models to investigate relationships among ApoE isoforms, glycolytic metabolism, and neuronal health and aging. ApoE2-expressing cells retained robust hexokinase (HK) expression and glycolytic activity, whereas these endpoints progressively declined with aging in ApoE4-expressing cells. These divergent ApoE2 and ApoE4 effects on glycolysis directly correlated with markers of cellular wellness. Moreover, ApoE4-expressing cells upregulated phosphofructokinase and pyruvate kinase with the apparent intent of compensating for the HK-dependent glycolysis reduction. The introduction of ApoE2 increased HK levels and glycolysis flux in ApoE4 cells. PI3K/Akt signaling was distinctively regulated by ApoE isoforms but was only partially responsible for the ApoE-mediated effects on HK. Collectively, our findings indicate that human ApoE isoforms differentially modulate neuronal glycolysis through HK regulation, with ApoE2 upregulating and ApoE4 downregulating, which markedly impacts neuronal health during aging. These findings lend compelling support to the emerging inverse-Warburg theory of AD and highlight a therapeutic opportunity for bolstering brain glycolytic resilience to prevent and treat AD.Publication Evaluation of Latex Immunoturbidimetric Assay Thresholds and HIT in Cardiothoracic Surgery(SAGE Publications, 2023-04-17) Hernandez, Jessica; Patel, Hetal; Biddlecome, Phil; Kildea, Megan; Dwivedi, Ruti; Sridhara, Shashank; Silvestry, Scott; Cavarocchi, Nicholas; Francis, John L.; Ventura, DavideBackground Heparin-induced thrombocytopenia (HIT) is a common differential diagnosis in cardiothoracic surgery. The latex immunoturbidimetric assay (LIA) is an enhanced immunoassay that has recently been introduced for the detection of total HIT immunoglobulin and retains a higher specificity of 95% compared to the enzyme-linked immunosorbent assay. Objectives To investigate if a semiquantitative relationship exists between increasing LIA levels beyond the current positivity threshold and its correlation to positive serotonin release assay results in cardiothoracic surgery. Methods This was a multicenter, observational cohort of cardiothoracic surgery patients initiated on anticoagulation with heparin-based products. To conduct sensitivity and specificity analysis of LIA values, HIT positive was defined as a LIA value ≥1 unit/mL and HIT negative was defined as a LIA level <1 unit/mL. A receiver operating characteristic (ROC) analysis was utilized to evaluate the predictive performance of the LIA. Results At manufactures’ cutoffs of ≥1.0 unit/mL, LIA sensitivity and specificity was 93.8% and 22%, respectively, yielding a false positive rate of 78%. At a higher cutoff of 4.5 units/mL, LIA sensitivity and specificity was 75% and 71%, respectively, yielding a false positive rate of 29% and an area under the ROC curve of 0.75 (P = .01; 95% confidence interval: 0.621-0.889). Bivalirudin was initiated in 84.6% of false positive LIA results. Conclusion This study suggests that the diagnostic accuracy of the LIA can be optimized by increasing the LIA positivity threshold. Proposing a higher LIA cutoff, may mitigate unwarranted anticoagulation and bleeding outcomes.