CHRI sis at the NICU: The Medley with Midazolam

Approximately 1.5 million neonates undergo anesthesia for surgical procedures in the United States every year1. Midazolam is a commonly used anesthetic agent used in the Neonatal Intensive Care Unit (NICU). It is used to sedate neonates and facilitates complex procedures such as mechanical ventilation.2 The extensive use of midazolam has raised questions about whether it affects the cognitive development of infants. In 2014, the International Anesthesia Research Society released a statement saying, “Surgeries and procedures requiring anesthetic and sedative drugs that could reasonably be delayed should possibly be postponed because of the potential risk to the developing brain of infants, toddlers, and preschool children”. 3 Although some evidence shows that midazolam exposure could harm an infant’s cognitive development, little is known about what parts of the developing brain are directly affected by midazolam. Additionally, research has yet to uncover whether the effects of midazolam persist into adulthood. In order to examine the consequences of midazolam exposure, a holistic system biology approach should be implemented. Experimental data from four different levels ─ the molecular level, the physical trait level, the behavioral level, and “omics” level would help address these issues. Our objective is to investigate how prolonged exposure to midazolam affects cellular as well as behavioral functions. A rodent model was implemented to study the effects at infanthood, adolescence, and adulthood. Our molecular results revealed that midazolam could potentially cause disturbances in key brain protein levels. Additionally, midazolam could potentially contribute to social deficits as evidenced by behavioral results. Overall, the results all point to midazolam\u27s potential to delay proper neurodevelopment.https://digitalcommons.unmc.edu/surp2021/1028/thumbnail.jp

Peritoneal Mouse as Detected on 18F-FDG PET-CT

We present the case of a 77-year-old male with a history of prostate cancer. Follow up PET-CT and contrast-enhanced CT demonstrated a small peritoneal loose body or “mouse” in the pelvis. This is an uncommon, benign, asymptomatic finding which is usually incidentally discovered. The significance of being aware of this entity is to distinguish it from metastasis, especially in patients with known abdominal and pelvic malignancies

Effect of Combined Methamphetamine and Oxycodone Use on the Synaptic Proteome in an In Vitro Model of Polysubstance Use

Polysubstance use (PSU) generally involves the simultaneous use of an opioid along with a stimulant. In recent years, this problem has escalated into a nationwide epidemic. Understanding the mechanisms and effects underlying the interaction between these drugs is essential for the development of treatments for those suffering from addiction. Currently, the effect of PSU on synapses-critical points of contact between neurons-remains poorly understood. Using an in vitro model of primary neurons, we examined the combined effects of the psychostimulant methamphetamine (METH) and the prescription opioid oxycodone (oxy) on the synaptic proteome using quantitative mass-spectrometry-based proteomics. A further ClueGO analysis and Ingenuity Pathway Analysis (IPA) indicated the dysregulation of several molecular functions, biological processes, and pathways associated with neural plasticity and structural development. We identified one key synaptic protein, Striatin-1, which plays a vital role in many of these processes and functions, to be downregulated following METH+oxy treatment. This downregulation of Striatin-1 was further validated by Western blot. Overall, the present study indicates several damaging effects of the combined use of METH and oxy on neural function and warrants further detailed investigation into mechanisms contributing to synaptic dysfunction

Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early Development

Recently, the long-term use of sedative agents in the neonatal intensive care unit (NICU) has raised concerns about neurodevelopmental outcomes in exposed neonates. Midazolam (MDZ), a common neonatal sedative in the NICU, has been suggested to increase learning disturbances and cognitive impairment in children. However, molecular mechanisms contributing to such outcomes with long-term MDZ use during the early stages of life remain unclear. In this study, we for the first time elucidate the role of brain-derived extracellular vesicles (BDEVs), including mining the BDEV proteome post long-term MDZ exposure during early development. Employing our previously established rodent model system that mimics the exposure of MDZ in the NICU using an increasing dosage regimen, we isolated BDEVs from postnatal 21-days-old control and MDZ groups using a differential sucrose density gradient. BDEVs from the control and MDZ groups were then characterized using a ZetaView nanoparticle tracking analyzer and transmission electron microscopy analysis. Next, using RT-qPCR, we examined the expression of key ESCRT-related genes involved in EV biogenesis. Lastly, using quantitative mass spectrometry-based proteomics, we mined the BDEV protein cargo that revealed key differentially expressed proteins and associated molecular pathways to be altered post long-term MDZ exposure. Our study characterized the proteome in BDEV cargo from long-term MDZ exposure at early development. Importantly, we identified and validated the expression of YWHAH as a potential target for further characterization of its downstream mechanism and a potential biomarker for the early onset of neurodevelopment and neurodegenerative diseases. Overall, the present study demonstrated long-term exposure to MDZ at early development stages could influence BDEV protein cargo, which potentially impact neural functions and behavior at later stages of development

Crisis in the NICU and the Medley with Midazolam

Epidemiologic studies of human patients have revealed a correlation between childhood exposure to general anesthetic and sedative agents and subsequent cognitive deficits. This association is supported by data from animal models, which shows that developmental exposure to both anesthetics and sedatives causes lasting impairments in learning. This study focused on midazolam (MDZ), a common benzodiazepine regularly used as a sedative agent on neonates in the Neonatal Intensive Care Unit (NICU). However, a knowledge gap that remains is how long-term exposure to MDZ during very early stages of life impacts synaptic alterations and neurobiological mechanisms. Elucidation of these mechanisms is of high clinical importance and may develop neuroprotective therapeutic strategies for optimizing outcomes for uniquely vulnerable NICU populations. Using a preclinical rodent model system, we mimicked a dose-escalation regimen from postnatal day 3 (P3) pups until P21 to comprehensively characterize how early-life exposure to MDZ impacts neurodevelopment outcomes at different tiers ─ phenotypic, molecular, behavioral, and high throughput- “omics” levels. Our data demonstrated that repetitive exposure to MDZ at an early age stunts neurodevelopment during the early stages of life disrupts the blood-brain barrier, and alters the synaptic components and neurochemistry, which may be indicative of behavioral deficits at later development. Additionally, our bioinformatics analysis from purified synaptosome identified enrichment of proteins associated with actin-binding and protein depolymerization process. One potential hit identified was alpha adducin (ADD1), belonging to the family of cytoskeleton proteins, upregulated in the MDZ group and whose expression was further validated by western blot. Our study has provided a comprehensive characterization of MDZ effects on development at multiple tiers yielding novel insights on how long-term exposure to MDZ impacts development. Notably, the identification of ADD1 as a potential target and further characterization of its downstream mechanisms can give additional insights into its role as a potential therapeutic for treating neurodevelopmental alterations associated with long-term MDZ use in neonates.https://digitalcommons.unmc.edu/chri_forum/1056/thumbnail.jp

Transgenic Mice for Intersectional Targeting of Neural Sensors and Effectors with High Specificity and Performance

SummaryAn increasingly powerful approach for studying brain circuits relies on targeting genetically encoded sensors and effectors to specific cell types. However, current approaches for this are still limited in functionality and specificity. Here we utilize several intersectional strategies to generate multiple transgenic mouse lines expressing high levels of novel genetic tools with high specificity. We developed driver and double reporter mouse lines and viral vectors using the Cre/Flp and Cre/Dre double recombinase systems and established a new, retargetable genomic locus, TIGRE, which allowed the generation of a large set of Cre/tTA-dependent reporter lines expressing fluorescent proteins, genetically encoded calcium, voltage, or glutamate indicators, and optogenetic effectors, all at substantially higher levels than before. High functionality was shown in example mouse lines for GCaMP6, YCX2.60, VSFP Butterfly 1.2, and Jaws. These novel transgenic lines greatly expand the ability to monitor and manipulate neuronal activities with increased specificity.Video Abstrac

F-18 fluorodeoxyglucose positron emission tomography and/or computed tomography findings of an unusual breast lymphoma case and concurrent cervical cancer: a case report

<p>Abstract</p> <p>Introduction</p> <p>Breast lymphoma accounts for less than 1% of all non-Hodgkin's lymphomas and approximately 0.1% of all breast neoplasms. Most breast lymphomas are classified as diffuse large B-cell lymphomas or as mucosa associated lymphoid tissue lymphomas. Concurrent cases of breast lymphoma and cervical cancer are extremely rare.</p> <p>Case presentation</p> <p>We report a case of a 46-year-old woman of unknown ethnic origin diagnosed with concurrent diffuse large B-cell lymphoma of the breast and squamous cell cancer of the cervix that was detected and followed with F-18 fluorodeoxyglucose (FDG) positron emission tomography and/or computed tomography (PET/CT). The metastatic pattern of this case of breast lymphoma is similar to that of a typical metastatic breast carcinoma. These findings have never been described in the literature. PET/CT also demonstrated an incidentally intense FDG focus in the uterine cervix ultimately leading to the pathologic diagnosis of squamous cell carcinoma of the uterine cervix. An appropriate staging of breast lymphoma and cervical cancer with FDG PET/CT is important because of therapeutic consequence. This case report and review of the literature highlights the role of FDG PET/CT in staging and restaging of both breast lymphoma and cervical cancer.</p> <p>Conclusions</p> <p>We report a case of a breast lymphoma with a metastatic pattern similar to that of typical metastatic breast carcinoma. The FDG PET/CT scan also diagnosed a rare case of concurrent breast lymphoma and cervical cancer. This concurrence has not been reported previously in the medical literature.</p

Using a population-based approach to prevent hepatocellular cancer in New South Wales, Australia: effects on health services utilisation

<p>Abstract</p> <p>Background</p> <p>Australians born in countries where hepatitis B infection is endemic are 6-12 times more likely to develop hepatocellular cancer (HCC) than Australian-born individuals. However, a program of screening, surveillance and treatment of chronic hepatitis B (CHB) in high risk populations could significantly reduce disease progression and death related to end-stage liver disease and HCC. Consequently we are implementing the <it>B Positive </it>pilot project, aiming to optimise the management of CHB in at-risk populations in south-west Sydney. Program participants receive routine care, enhanced disease surveillance or specialist referral, according to their stage of CHB infection, level of viral load and extent of liver injury. In this paper we examine the program's potential impact on health services utilisation in the study area.</p> <p>Methods</p> <p>Estimated numbers of CHB infections were derived from Australian Bureau of Statistics data and applying estimates of HBV prevalence rates from migrants' countries of birth. These figures were entered into a Markov model of disease progression, constructing a hypothetical cohort of Asian-born adults with CHB infection. We calculated the number of participants in different CHB disease states and estimated the numbers of GP and specialist consultations and liver ultrasound examinations the cohort would require annually over the life of the program.</p> <p>Results</p> <p>Assuming a 25% participation rate among the 5,800 local residents estimated to have chronic hepatitis B infection, approximately 750 people would require routine follow up, 260 enhanced disease surveillance and 210 specialist care during the first year after recruitment is completed. This translates into 5 additional appointments per year for each local GP, 25 for each specialist and 420 additional liver ultrasound examinations.</p> <p>Conclusions</p> <p>While the program will not greatly affect the volume of local GP consultations, it will lead to a significant increase in demand for specialist services. New models of CHB care may be required to aid program implementation and up scaling the program will need to factor in additional demands on health care utilisation in areas of high hepatitis B sero-prevalence.</p