bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of High Glucose and Salinity on Embryological Development of Zebrafish, Brachyodanio rerio
Authors: Thompson Ebony , Hensley Justin , Taylor Renfang
Published: May 2022

Diabetes is a disease in which the blood glucose, or blood sugar, levels are too high. Glucose comes from the daily diet. Over time, having too much glucose in the blood can damage the eyes, the kidneys, the nerves, cause heart disease and stroke. Up to date, it is the leading disease around the world. Similarly, high blood pressure, heart disease, and kidney disease are also known to be attributed to high sodium levels. However, the effects of high salinity on embryological development post‐fertilization are yet to be researched. Danio rerio, commonly called the zebrafish, is a small freshwater fish. Its small size, short life cycle and high fecundity favor its lab uses as a test organism. It has been widely used for genetic development, transgenesis, toxicology and other genetic research due to the high conservation of gene function between zebrafish and humans. Zebrafish embryos are also transparent making them a preferable choice for developmental study. In this experiment, the effects on embryological (morphological and genetic) development were examined in the presence of a high glucose environment and high salinity conditions were examined. Zebrafish eggs were treated with normal media and high glucose for 5 days from 3 hours post‐fertilization(hpf) to 96hpf separately as control and experimental groups. RNA is extracted from control and experimental group to determine five genes’ change in high glucose group using PCR. Day 5 Larva is fixed in 10% formalin for H&E staining to detect organogenesis changes under high glucose conditions. Three progenitor genes that regulate organ development will be detected for protein level changes in experimental group by immunohistochemistry. Zebrafish eggs were treated with normal media and high salt (2%) for 3 days from 3 hpf to 72hpf separately as control and experimental groups. Day 1 and Day 2 larvae are fixed in 10% formalin for H&E staining. Glucose delayed the zebrafish embryo development by slowing the hatch rate about 24 hours. The brain, heart, and tail started showing morphology smaller in the glucose group compared to the control group at 24 hpf. Heartbeat is slower in the glucose group compared to the control group on day 2 and 3 with a statistically significant difference. The brain, heart, and tail started showing morphology smaller in the salt group compared to the control group at 48 hpf. These conditions contribute to significant physiological differences that may provide insight on functionality post‐embryological development. Testing to find consistent trends in morphology and genetic differences in high salt conditions, and genetic alterations for high glucose conditions are being further researched in this experiment.
The Effect of Sulindac on Senescence in RPE Cells: Role of cell protective mechanisms Against Oxidative Damage
Authors: Manoharan Samuel P. , Weissbach Herbert P. , Allani Shailaja
Published: May 2022

Age‐related macular degeneration (AMD) is the leading cause of blindness in industrialized countries. One of the potential contributors to AMD pathology is retinal pigment epithelial (RPE) cell senescence. Through experimental trial, it is shown that sulindac, an FDA approved non‐steroidal anti‐inflammatory drug, protects normal cells such as retinal, cardiac, and neuronal cells against oxidative damage through a preconditioning mechanism. The hypothesis is that sulindac can mitigate RPE cell senescence because of its cellular protective properties. To test this, in vitro, an RPE senescence model was established using tert‐butyl hydrogen peroxide (TBHP) as an oxidizing agent. The cells were treated with TBHP for 12 hours followed by further incubation for four days. Senescence cells were identified using a Beta‐galactosidase assay. The effect of sulindac on senescence was determined by pre‐treating cells with varying concentrations of sulindac for 24 h prior to TBHP treatment.

Treatment with TBHP induces senescence in the RPE cells under the conditions used. The Preliminary data shows that there was a reduced number of senescent cells in the samples treated with sulindac compared to the control groups. These findings indicate that oxidative damage induces cell senescence, and sulindac can lower the amount of senescence by protecting the RPE cells from oxidative damage. To summarize, the results suggest that sulindac can be used as a therapeutic agent targeting senescence induced by oxidative damage.
Effect of Hyperinsulinemia on Cerebral Autoregulation and Myogenic Control of Cerebral Blood Flow in Healthy Young Adults
Authors: Shariffi Brian , Lloyd Iman N. , Wagner Jadee , Harper Jennifer L. , McMillan Neil J. , Smith James A. , Manrique‐Acevedo Camila , Padilla Jaume , Limberg Jacqueline K.
Published: May 2022

Objective

Hyperinsulinemia has marked vasodilatory effects within the skeletal muscle vasculature in healthy young adults; however, the magnitude of insulin‐stimulated cerebrovascular vasodilation is less clear. Differences between circulations may be due to a greater importance of the cerebral circulation to maintain a constant level of blood flow (i.e., cerebral autoregulation) during hyperinsulinemia. We hypothesized cerebral autoregulation would increase during hyperinsulinemia. Autoregulatory mechanisms, such as myogenic control, play important roles in mediating cerebral vasomotor responses. Thus, we further sought to examine the contribution of the myogenic mechanism to the control of cerebral blood flow during hyperinsulinemia.

Methods

Middle cerebral artery blood velocity (MCAv) and femoral artery blood velocity (FAV) were measured with Doppler ultrasound and femoral artery blood flow (FBF) was calculated in 17 healthy adults (11M/6F, 28±2 yrs, 24±1 kg/m2) at baseline and following a hyperinsulinemic (40 mU/m2body surface area/min), euglycemic infusion. Finger plethysmography was used to assess mean blood pressure (MBP). Normalized transfer function gain between changes in MBP and MCAv were estimated in the 0.02‐0.07 Hz (very low frequency, VLF) and 0.07–0.20 Hz (low frequency, LF) ranges using the Welch method. Interactions between MBP and MCAv or FAV were determined using cross‐correlation analyses.

Results

Intravenous insulin infusion increased plasma insulin concentrations (6±1 to 50±3 μIU/mL, p<0.01) with blood glucose maintained at baseline levels (p=0.33). There was a significant increase in FBF (121±15 to 151±19 mL/min, p=0.03) during hyperinsulinemia, with no change in MCAv (61±4 to 59±4 cm/s, p=0.28) or MBP (86±2 to 89±2 mmHg, p=0.10). There was no effect of hyperinsulinemia on normalized transfer function gain in the VLF (p=0.25) or LF ranges (p=0.46). Hyperinsulinemia had no effect on the association between MBP and MCAv (p=0.15) or FAV (p=0.82).

Conclusions

Hyperinsulinemia in healthy young adults elicits peripheral vasodilation, but no change in MCAv. Contrary to our hypothesis, the sensitivity of the cerebrovascular circulation to changes in MBP (i.e., cerebral autoregulation) is not altered during systemic hyperinsulinemia in healthy adults. Furthermore, the contribution of the myogenic mechanism to the control of blood flow in the peripheral and cerebral circulations is unaffected by hyperinsulinemia. Whether these results translate to disease states (e.g., diabetes) and the implications for cerebrovascular health warrant further exploration.
RNA Modifications Destabilize a Pyrimidine‐Motif RNA●DNA‐DNA Triple Helix
Authors: Schiefelbein Grace E. , Kunkler Charlotte N. , Brown Jessica A.
Published: May 2022

The formation of pyrimidine‐motif RNA●DNA‐DNA (R●D‐D) triple helices (or triplexes), in which ‘●’ and ‘‐’ represent Hoogsteen and Watson‐Crick interactions, has been proposed for over 60 years, but the stability of these structures with RNA modifications has yet to be studied. Eleven RNA modifications were chosen for this study based on their presence in human transcripts and their effects on human health: 5‐methylcytidine (m5C), 5‐methyluridine (m5U), pseudouridine (Ψ), 2ʹ‐O‐methyladenosine (Am), 2ʹ‐O‐methylcytidine (Cm), 2ʹ‐O‐methylguanosine (Gm), 2ʹ‐O‐methyluridine (Um), 3‐methyluridine (m3U), 4‐thiouridine (s4U), inosine (I), and N6‐methyladenosine (m6A). Several of these had been previously found to stabilize or destabilize other nucleic acid duplex and triplex structures. Using both native gel‐shift assays and microscale thermophoresis, the relative stability of a single modified position in a pyrimidine‐motif R●D‐D triple helix was measured at neutral pH. All eleven modifications were found to either have no effect or to destabilize the R●D‐D triple helix, ranging from 2‐fold to complete disruption of binding. For each of the canonical R●D‐D base triples (U●A‐T and C●G‐C), the 2ʹ‐O‐methyl modifications were found to be the least destabilizing, whereas those that directly interfered with the Hoogsteen interactions, such as m3U, were the most destabilizing modifications. As the formation of R●D‐D triple helices in promoter regions of DNA leads to transcriptional enhancement and repression, this study reveals that some RNA modifications could potentially inhibit R●D‐D triplex formation as another level of transcriptional regulation.
Hyperoxia‐Induced Preretinal Membrane Formation and Retinal Detachment in the Mouse
Authors: Diaz Gabriela M. , Heck Madison , Cuff Evan , Martin Mark , Guthridge Bryanna , Lee Tyler , Penev Boris , Gerhart Jacquelyn , George‐Weinstein Mindy , Bravo‐Nuevo Arturo
Published: May 2022

Background

Retinal detachment (RD) is a vision‐threatening consequence of several conditions including retinopathy of prematurity (ROP). Numerous studies have established the connection of high oxygen concentration altering retinal development in preterm infants, leading to RD and blindness in later stages. The goal of this study was to develop a reproducible, non‐invasive model mouse of human ROP that could be used to study the cellular events leading to the formation of tractional preretinal membranes and RD.

Hypothesis

Placing C57BL/6J mice in hyperoxic conditions for one week starting at birth will induce a non‐traumatic sequence of retinopathy leading to tractional RD that mimics the pathophysiology of detachment seen in ROP.

Methods

C57BL/6J newborn mice were placed into a hyperoxic environment (65% oxygen saturation) for 7 days then reared in ambient air. Retinas were observed at weeks 4, 5, and 7 via spectral‐domain optical coherence tomography (OCT). Eyes were processed for histology and immunostaining for the myofibroblast marker alpha‐smooth muscle actin (α‐SMA).

Results

Four weeks after birth, 100% of mice placed in hyperoxia exhibited retinal blebs, 91% had developed preretinal membranes, and 9% had RD. At 5 weeks, 100% and 27% had preretinal membranes and RD, respectively. At week 7, the number of mice with RD increased to 33%. Preretinal membranes arose at the surface of the retina above the optic nerve head. RD occurred between the neurosensory retina and the underlying retinal pigment epithelium. Myofibroblasts were detected with the antibody to α‐SMA in the preretinal membranes and above areas of RD.

Conclusion

The hyperoxia‐induced retinopathy model was confirmed to induce retinal pathology in newborn mouse pups. Retinal pathology progressed during a 7‐week time period (i.e. retinal blebs to preretinal membranes and folds to retinal detachment). This noninvasive model replicates the oxygen‐induced retinopathy seen in human ROP. The model is useful for investigating the origin of cells that contribute to the formation of tractional retinopathy, especially PRMs and RD. The presence of α‐SMA‐positive cells overlying areas of retinopathy suggests the contractile property of myofibroblasts induces tractional RD.
An Interactive Pharmacogenetics Lesson Using PharmGKB To Individualize Pharmacotherapy Recommendations
Authors: Mosquera Andrea , Aleksunes Lauren M.
Published: May 2022

PharmGKB is a user‐friendly online pharmacogenomics database that annotates clinical guidelines for optimizing drug therapy based on patient genotypes. The majority of guidelines in PharmGKB are based on the interindividual variability in cytochrome P450 (CYP) enzymes responsible for metabolizing drugs. By individualizing patient therapy according to CYP genotypes, there is potential to improve pharmacotherapy outcomes and minimize the risk of adverse events. Although there is growing interest amongst educators and students to integrate pharmacogenetic content into health science curricula, there is a lack of educational resources that teach students how to utilize resources such as PharmGKB to inform clinical decision making. The purpose of this study was to assess and disseminate a case‐based interactive lesson on pharmacogenetic‐based drug dosing principles that uses PharmGKB. Seventy‐one high school students participated in a two‐hour Zoom session as part of a four‐day Toxicology, Health, and Environmental Disease Program. The lesson was divided into a didactic lecture on pharmacogenetics followed by a group‐based analysis of hypothetical case scenarios. Case questions focused on genetic variation in CYP enzymes in the context of different clinical scenarios. Examples included the use of codeine for analgesia (CYP2D6) and clopidogrel for stroke prevention (CYP2C19). Assessment of student understanding was measured by percent gain in correct answers between pre‐ and post‐lesson polling questions. The first question focused on pharmacology concepts while the last two focused on genetics concepts. Students had high pre‐test scores on the pharmacology question having learned these concepts earlier in the program. For the additional two questions, there was a positive gain (42%) demonstrating an increase in knowledge during the lesson. Seventy‐eight percent of students would be ‘extremely likely’ or ‘very likely’ to recommend this activity. We propose that an interactive, group‐based activity can be used to teach basic principles of pharmacogenetics and empower students and educators to effectively use online drug resources.
Uncovering Anti‐Arrhythmic Potential of Stellate Ganglion Purinergic Receptors
Authors: Bussey Carol T. , Davis Harvey , Bogert Emma , Paterson David J. , Paton Julian F.
Published: May 2022

Cardiovascular disease affects over 30% of people worldwide and is one of the leading causes of death each year. Elevated sympathetic nerve activity is a common feature of cardiovascular disease, contributing to end‐organ damage, morbidity and mortality. Recent findings indicate that short‐circuiting sympathetic nerve overactivity by removal of the stellate ganglion can eradicate arrhythmias, emphasising the need for novel therapeutic targets to correct signalling non‐invasively.

We have revealed upregulation of the P2X3 purinergic receptor in stellate ganglia of Spontaneously Hypertensive (SHR) compared to Wistar rats (16 week‐old), via RNASeq transcriptional profiling. We hypothesise that these purinergic receptors within cardiac stellate ganglia play a role in the excessive sympathetic drive to the heart in cardiovascular disease and can initiate cardiac arrhythmias. Thus, we have investigated the functional role of purinergic receptors in the stellate ganglion.

Administration of ATP to acutely isolated post‐ganglionic sympathetic neurones from the stellate ganglia of Wistar rats (4‐6 week‐old) evokes a significant increase (Median; 0.21) in [Ca2+]i as measured by Fura‐2AM imaging (n=15, Wilcoxon matched‐pairs test; p<0.0001). This ATP‐induced calcium transient was inhibited by specific P2X3 receptor antagonism with either NF‐10 or AF‐130.

Cardiac effects of stellate ganglion purinergic stimulation were investigated in the working heart‐brainstem preparation of Wistar rats (4‐6 week‐old). Increasing doses of ATP (50‐250 µg) delivered via microinjection (50‐250 µL) directly to the right stellate ganglion produced either a tachycardia (18 ± 6 bpm, n=6) or bradycardia (‐24 ± 12 bpm, n=5); these responses were typically found at distinct sites within the ganglion. Preliminary data from SHR suggest that higher doses of ATP (≥250µg) may be arrhythmogenic (n=3).

P2X3 purinergic receptors are present in the sympathetic stellate ganglion and contribute to calcium ion flux and cardiac chronotropic regulation. Overexpression of P2X3 receptors is likely to contribute to excessive cardiac sympathetic activity and the development of hypertension and cardiac arrhythmias, making them a promising novel therapeutic target.
Teaching Reproductive Anatomy through Abdominal Hysterectomy on a Cadaver
Authors: Fogarthy Tannice , Richter Emily , Ravikumaran Nair Rakesh
Published: May 2022

Introduction

Medical journals have stated there is a decrease in anatomical knowledge amongst medical students which then hinders their knowledge of pathologic processes. Performing a hysterectomy requires knowledge of the uterine structures and the ability to properly identify them. Performing a hysterectomy would allow for 4th year medical students interested in the field of obstetrics and gynecology to learn these structures as well as learn the steps of a hysterectomy prior to residency.

Objective

To perform an abdominal hysterectomy as a means to enhance pelvic anatomical knowledge and introduce 4th year medical students to the operative technique.

Material and Methods

Two fourth year medical students at HWCOM (CO2022) participated. The abdomen was opened, and its contents retracted to obtain access to the genitourinary structures. The round ligament was ligated and divided opening the retroperitoneal space. The utero‐ovarian ligament was then clamped, divided and ligated. The ovary was left attached to the infundibulopelvic ligament. The same steps were repeated on the opposite side. The anterior vaginal wall was then exposed by dissecting the bladder off the anterior wall of the lower uterine segment and cervix. The uterine vessels were skeletonized bilaterally at the lower uterine segment then clamped and ligated. The remaining parts of the broad ligament was then clamped, divided and ligated to the cervix. The broad ligament was then detached from the lateral cervix further exposing the upper vagina. The vagina was then divided below the cervix with a scalpel, and the uterus and cervix were then removed. The vaginal apex was then closed with a figure of 8 stitch.

Results/Images

There were 8 main steps as illustrated by the pictures in the appendix for visual aid and future instruction of medical students.

Conclusion and Significance/Implication

Performing a hysterectomy on a cadaver required knowledge of uterine anatomy, its surrounding structures and the steps of a hysterectomy. This can be a method to teach medical students uterine anatomy. Future implications of this practice on students first live hysterectomy are yet to be explored but worthwhile as a future study.
Nearest Neighbor Density Analysis of the Stellate Ganglia
Authors: Yang Ronald , Fefferman Ann , Tse Shiaomeng , Park Danny , Wisco Jonathan J.
Published: May 2022

Introduction

The so‐called heart‐brain axis is the neurological and physiological connection between the autonomic systems of both organs. The stellate ganglia and superior cervical ganglia are the location of the sympathetic neuronal cells that innervate the vasculature of the heart and brain, respectively. The paravertebral chain connects the ganglia on either side of the neck. Myocardial infarction is thought to cause remodeling of the sympathetic neurons in the stellate ganglia. In a recent qualitative study, the stellate ganglia were shown to remodel with regard to cell body number, size, density, and clustering in the presence of various cardiac diseases. We developed a tool to automatically quantify the nearest neighbor density of neurons weighted by size in pathology specimens of the stellate ganglia. This is the first step to quantifying sympathetic nervous system changes in the heart‐brain axis of cardiovascular pathology.

Methods

Biopsies of the stellate ganglia were collected bilaterally from an opportunity sample of 17 willed body donors (34 total specimens) with and without histories of self‐reported myocardial infarction (MI) and/or cerebrovascular accident (CVA). The ganglia were stained with luxol fast blue and cresyl violet which allowed for histological analysis of local neurons. Images of the ganglia at its middle coronal slice after staining were processed using multiple libraries in python, primarily PIL and OpenCV. The images first underwent multiple filters: a high contrast filter and a smoothing filter. After this filter, the image passed through a color threshold that obtained the location of each neuron; the locations of each neuron were used to make a binary image. The binary image was passed through various parameters such as neuron area, radius of the minimum enclosing circle, and a ratio of the area of the neuron and area of the minimum enclosing circle. A heatmap was generated from this last image through a convolution calculation that takes into account the location of the neuron, size, and proximity of surrounding neurons‐ all variables that are needed to calculate nearest neighbor density. Quantitative and qualitative measurements are generated from this: a graphical map of high and low density areas and number of neurons per given threshold of density.

Results

A pipeline was created that generates heatmaps of the nearest neighbor density based on the location and size of neurons on the stellate ganglia. Quantitative data is also measured based on how many neurons are located in varying levels of density structures.

Conclusions

Heatmaps of the nearest neighbor density and size of the neurons potentially show signs of cardiac diseases‐ regions of high density or patterns of where highly dense regions are located are indicators. Developments within computation pathology is an easily accessible and pioneering way for new technological innovation.
The Molecular Chaperone, GRP170, Protects Against Acute Kidney Injury and ER Stress in Mice
Authors: Buck Teresa M. , Porter Aidan , Ruiz Wily G. , Clayton Dennis R. , Mutchler Stephanie M. , Ray Evan C. , Marciszyn Allison L. , NKashama Lubika J. , Subramanya Arohan R. , Gingras Sebastien , Kleyman Thomas , Apodaca Gerard , Hendershot Linda M. , Brodsky Jeffrey L.
Published: May 2022

GRP170 is an Hsp70‐like, molecular chaperone localized to the endoplasmic reticulum (ER). Two separate functions have been described for GRP170. First, GRP170 acts as a nucleotide exchange factor (co‐chaperone) for the ER lumenal, Hsp70, BiP. Second, GRP170 possess “holdase” activity, and independently binds to aggregation prone regions of proteins to maintain solubility. We previously demonstrated that GRP170 regulates the quality control of the epithelial sodium channel, ENaC. ENaC is responsible for sodium reabsorption in the distal nephron and regulates salt/water homeostasis, and therefore, blood pressure. To better understand how GRP170 impacts kidney function we generated an inducible, nephron specific, GRP170 KO mouse. Loss of GRP170 results in rapid weight/volume loss, electrolyte imbalance and significantly elevated aldosterone levels. The GRP170 KO animals also demonstrate many of the hallmarks of acute kidney injury (AKI) including elevated plasma BUN and creatinine levels. Loss of GRP170 also results in induction of the unfolded protein response (UPR) as shown by upregulation of UPR targets by qPCR (sXbp1, BiP, CHOP and ATF4) and western blotting. We hypothesize that sustained induction of the UPR leads to kidney injury associated with our model, alternatively, misregulation of ion channel trafficking and volume loss may contribute to the pathogenic phenotype. To begin to understand how loss of an ER localized chaperone results in profound kidney injury we treated our GRP170 KO mice with the UPR inhibitor, TUDCA, or a high‐salt diet. Preliminary data suggest both UPR induction and electrolyte imbalance may contribute to kidney injury associated with loss of GRP170.
Wasp Venom Relieves Alzheimer's Phenotypes in 5xFAD Transgenic Mice through Inhibition of Aβ Aggregation
Authors: Jeong Yoon Ah , Chae Ji Hyeong , Jang Chan Ho , Kim Yoonsu , Lee Jinkyung , Kwon Min Jeong , Kwon Eun Jeong , Kim Hyo Jung , Oh Jisun , Kim Jong‐Sang
Published: May 2022

Alzheimer’s disease (AD), a neurodegenerative disorder, has been known to be mostly commonly diagnosed in people over the age of 65. However, there is no drug that can cure AD. This study aimed to investigate if wasp venom (WV) can mitigate phenotypes of AD using a 5xFAD transgenic mouse model. The mice were administered intraperitoneally with WV obtained from Vespa velutina at a concentration of 250 or 400 μg/kg body weight (BW) once a week for a total of 14 weeks. WV treatment was shown to alleviate memory impairments in 5xFAD mice in behavioral tasks, including passive avoidance, Y‐maze and Morris water maze tests. In addition, we found that WV treatment decreased the level of 8‐hydroxy‐2′‐deoxyguanosine, a biomarker of oxidative DNA damage, in mouse plasma and the level of malondialdehyde, an indicative of lipid peroxidation, in liver and cerebral cortex, indicating that WV prevents oxidative damage. In particular, WV treatment increased expression of antioxidant‐associated proteins, a nuclear factor erythroid‐derived 2‐related factor 2 (Nrf2) and heme oxygenase‐1 (HO‐1), in the cerebral cortex and decreased inflammation‐associated protein, cyclooxygenase‐2 (COX‐2), amyloid beta (Aβ) plaque‐associated proteins, C‐terminal fragment (C99), and beta‐site amyloid precursor protein cleaving enzyme 1 (BACE1, β‐secretase) in the hippocampal tissue homogenates. Furthermore, we confirmed that WV treatment reduced the accumulation of Aβ plaque in the hippocampal area via thioflavin S staining. Taken together, these findings suggest that WV could ameliorate AD symptoms and delay AD onset and progression.
Intracranial Cerebrovascular Reactivity by Traditional and Novel Methods in Young, Middle, and Old Aged Healthy Males and Females
Authors: Koep Jodie L. , Coombes Jeff S. , Barker Alan R. , Taylor Chloe E. , Pizzey Faith K. , Ruediger Stefanie L. , Bond Bert , Bailey Tom G.
Published: May 2022

Objective

Lower cerebrovascular reactivity (CVR) to hypercapnia is associated with increased risk of age‐related cognitive decline and all‐cause mortality. Despite the important clinical implications of CVR, evidence of the physiological changes across the healthy adult lifespan is conflicting, with limited findings in females. The aim of this study was to assess the effect of age and sex on CVR. We hypothesized: 1) CVR to be lower in middle‐aged and older compared to young adults, and 2) a higher CVR in young females compared with young males, and a higher CVR in older males than older females.

Methods

Young (n=25, 12 female, age 27±1.5y), middle‐age (n=31, 18 female, age 54±7.3y) and older healthy participants (n=20, 10 female, age 70±2.5y) completed CVR via inhalation of 5% carbon dioxide (CO2) for 5 minutes. Traditional CVR was characterized by the peak middle cerebral artery velocity (MCAv) response per mmHg increase in end‐tidal CO2 (traditional cm/s‐1mmHg), and using a novel mono‐exponential model to characterize the dynamic onset response of the MCAv. Independent samples ANOVA was used to compare main and interaction effects of age and sex.

Results

Traditional CVR was not different between young, middle and older groups (2.0±0.7 vs 1.8±0.9 vs 1.9±0.9 cm/s‐1mmHg; P=0.102). Dynamic onset (novel) responses revealed the time constant of the MCAv response was significantly slower in older and middle‐aged compared to young adults (29±16 vs 27±20 vs 14±9 seconds, P=0.03). No effects of sex were observed for traditional or novel CVR (P≥0.14), but young females had a higher baseline MCAv than young males (70.8 vs 78.6 cm/s‐1P=0.04), which was not observed in middle and older age adults.

Conclusions

CVR is preserved in ageing adults, but the speed of the MCAv response is blunted in older and middle‐aged adults compared to young adults. Utilizing dynamic onset responses reveal underlying differences in cerebral regulation with healthy ageing that were not detected by traditional methods.
The Association Between Quality of Diet, Frequency of Caffeine Consumption, Mental Distress, and Illicit ADHD Drug Use
Authors: Khan Humza , Umeozor Devon , Zubin Jared , Horowitz Julia , Yuvanavattana Natalie , Scott Nicole , Hinkley Courtney
Published: May 2022

The illicit use of Attention Deficit Hyperactivity Disorder (or ADHD) medications as study drugs is becoming common among undergraduate students. These “study drugs” are predominantly used to improve memory, focus, and attention with the end goal of boosting academic performance; however, studies on illicit drug use have hinted towards negative side effects that have the potential to compromise both mental and physical health. The relationship between lifestyle habits (i.e. diet and exercise) and illicit ADHD medication use is still unexplored; thus, the purpose of this study was to identify relationships between exercise patterns, caffeine consumption, dietary habits, and illicit ADHD medication use. An anonymous survey was sent to several universities via online platforms using Google Forms. A total of 1187 students completed the survey. The survey was comprised of the validated Food‐Mood Questionnaire, and included questions that addressed ADHD medication use frequency, academic performance, education on the effects of ADHD medication, and physical health. Our results reveal a statistically significant positive correlation between frequency of caffeine use and frequency of other drug use (including depressants and psychostimulants such as Adderall) (p < 0.01). Additionally, a positive correlation was detected between frequency of caffeine consumption and various hallmarks of mental distress, including feelings of worthlessness, depression, and restlessness (p < 0.01, 0.05, and 0.01 respectively). Additionally, there was a negative correlation between consumption of nutritious, healthy foods and illicit ADHD medication usage (p < 0.05). Considering all, a potential vicious cycle may exist. Those who illicitly use psychostimulants develop poorer dietary habits negatively impacting brain function, which may trigger higher consumption of caffeine as a stimulant.
Prognostic Potential of a PSAT1‐Associated Gene Signature in Identifying High‐Risk Patients in Early‐Stage EGFR‐Mutant Lung Cancer
Authors: Biyik‐Sit Rumeysa , Clem Brian F.
Published: May 2022

De novocellular production of serine may contribute to tumor growth by providing precursors for macromolecular production and one‐carbon metabolism. Accordingly, multiple cancers exhibit increased expression of serine biosynthetic enzymes. For example, elevated PSAT1 (phosphoserine aminotransferase 1), which catalyzes the second step in converting 3‐phosphohydroxypyruvate to phosphoserine, is associated with poorer clinical outcomes. Several reports have implicated PSAT1 in the proliferation, migration, invasion, and chemo‐resistance of malignant cells. Our previous study found that PSAT1 translocated into the nucleus within EGFR‐activated NSCLC cells and was associated with poorer clinical outcomes of patients with EGFR‐mutant lung cancer, suggesting a relevant function for PSAT1 in EGFR‐driven tumorigenesis. In this study, we designed a bioinformatic approach to identify a PSAT1‐associated gene signature for EGFR‐mutant lung cancer (Fig. 1). Since PC9 cells are frequently utilized as an EGFR‐mutant lung cancer model, we speculated that a subset of PSAT1‐regulated genes independently contributes to EGFR‐driven lung tumorigenesis. For this, we transcriptionally profiled PSAT1 silenced PC9 cells. Analysis of the RNASeq identified 490 genes (279‐down, 211‐up) that were differentially expressed upon PSAT1 silencing (PSAT1‐mediated DEG). We next performed a comparative analysis between PSAT1‐mediated DEG and publicly available microarray datasets obtained from EGFR‐mutant patient tumors. Twenty‐five genes (13‐down, 12‐up) were differentially expressed in all EGFR‐mutant lung cancer datasets. These were classified as a PSAT1‐associated gene signature for EGFR‐mutant lung tumors and used to assess the relationship between this gene signature and both relapse‐free (RFS) and overall survival (OS) by the survival prediction tool from BRB‐ArrayTools. Thirteen and seventeen genes from the 25‐gene signature were found to predict the OS and RFS rates of EGFR‐mutant lung cancer, respectively. However, this signature failed to predict patient outcomes for either wild‐type or KRAS‐mutant lung tumors. Furthermore, the expression profile of RFS high‐risk patients with stage I EGFR‐mutant tumor phenocopies the high‐risk stage II tumor, supporting the prognostic power of PSAT1‐mediated gene signature for these EGFR‐mutant tumors. As early‐stage cancers dominate the gene signature obtained from human EGFR‐mutant lung tumors, analysis of genes involved in late‐stage tumor progression and metastasis is limited. Therefore, differentially expressed genes between parental PC9‐P and brain‐metastatic PC9‐BrM3 were extracted from the Nguyen et al.study (2009). Comparative analysis with RNA‐seq data found a potential PSAT1‐associated metastatic gene signature. In summary, our genomic approach yielded a PSAT1‐associated gene signature with prognostic value in EGFR‐mutant NSCLC patient outcomes.
Profile of insulin resistance in adipose tissue of diet induced obese mice in response to anti‐obesity treatment
Authors: Rizk Nasser M. , Babiker Abeer , ElSaddig Ahmed Ali Maram , MOhamed Omnia , Elsayegh Dina , El‐Gamal Abdulrahman
Published: May 2022

Background

Adipose tissue is an insulin‐responsive organ that contributes to both glucose and lipid metabolism. Insulin resistance (IR) is when insulin‐dependent tissues lose their sensitivity to insulin, resulting in high blood glucose and insulin due to impairment of the negative feedback mechanism. Insulin resistance progress to metabolic disorders like obesity and type 2 diabetes. Sulforaphane (SFN) is a potential anti‐obesity drug under investigation.

Aim

This study aims to understand the effect of SFN on insulin resistance pathways in adipose tissues of diet induced obese (DIO) mice.

Methods

Wild‐type CD1 male mice were fed a high‐fat diet (HFD) for 16 weeks to induce obesity. Mice were divided into two groups DIO‐SFN and DIO‐V. DIO mice have been injected via IP route by SFN (5 mg/kg BW) or Vehicle (25μl) for 4 weeks. Daily body weight measurements have been recorded, and glucose tolerance test (GTT) at day 21 after injection and insulin tolerance test (ITT) at day 25 after injection. Subsequently, mice were sacrificed, blood and tissue samples were collected and snap‐frozen instantly. Total RNA was extracted from epididymal white adipose tissue (eWAT). Plasma leptin, insulin, and glucose level were measured, and 84 IR genes in adipose tissues were examined using RT‐PCR array technology.

Results

SFN treatment to DIO mice causes a significant reduction in body weight by 18.4%. Furthermore, SFN causes a significant reduction in circulating leptin concentrations in DIO‐SFN compared to the vehicle‐treated group. SFN is associated with the improvement of GTT and ITT. SFN causes dysregulation of eight genes related to inflammation, adipocytokine signaling, Il‐6, and HMGB1 pathways. SFN treatment causes significant changes in functions related to adipose tissue insulin resistance such as inhibition of chemotaxsis, inflammatory response and lipid accumulation.

Conclusion

This study demonstrated that SFN treatment effectively reduces body weight and can ameliorate IR in eWAT via inhibition of pathways mainly related to inflammation, such as adipocytokine signaling, Il‐6, and HMGB1. SFN can be considered a future anti‐obesity drug.
Study on exosome proteomics related to Parkinson's disease
Authors: Duan Hubin , Hao Chunyan , Li Mingli , Xue wen , Zhu Xiaolin , Qiao Binjun , Zhang Jinbao , Qiao Xiaohong
Published: May 2022

Background

Parkinson disease (PD) is a serious neurodegenerative disease with high increasing rate. Exosomes, carrying numerous biological materials including proteins and lipids, function as crucial players in multiple diseases including PD.

Methods

Exosomes were isolated from serum samples of PD patients and healthy people, which was validated through transmission electron microscope (TEM) image and protein expression detection of cluster of differentiation protein 9 (CD9) and tumor susceptibility gene 101 protein (TSG101) (exosome markers) via western blot assay. Differentially expressed proteins (DEPs) in serum exosomes were identified through iTRAQ‐LC‐MS/MS. DEPs were annotated and analyzed by Gene Ontology (GO) and KEGG Pathway databases. Protein‐protein interaction networks of these DEPs were established through STRING database.

Results

A total of 42 DEPs (24 up‐regulated, 18 down‐regulated) were identified in serum exosomes of PD patients versus healthy control group. GO‐biological_process analysis showed that most of DEPs were involved in the regulation of immune system process, vesicle‐mediated transport, response to stress and signal transduction. Moreover, some crucial proteins (e.g. antichymotrypsin (SERPINA3) and hemopexin (HPX)) associated with the pathology of PD were identified through interaction analysis. Western blot and ELISA assays further validated that SERPINA3 and HPX were highly expressed and phospholipid transfer protein (PLTP) was low expressed in serum exosomes of PD patients versus control group.

Conclusion

Proteomics analysis for serum exosomes of PD patients and healthy people could provide some potential biomarkers and vital information of intercellular communication and pathogenic mechanisms in PD.
The Analysis of Phagocytic Myeloid Cells in Low and High Fiber Fed Mice after Three Weeks of Hypoxia
Authors: Pakhomov Nikolai , Kostyunina Daria , McLoughlin Paul , Baugh John
Published: May 2022

Pulmonary hypertension (PH) is a devastating disease that is characterized by increased blood pressure (BP) in the pulmonary vasculature. Immune cells participate in adverse cardiovascular remodelling of pulmonary vessels and could represent targets for PH treatment. It has been shown that patients with PH have increased numbers of interstitial macrophages and dendritic cells in the lungs that originate from circulating immune cells. These myeloid immune cells migrate to the vessel wall and promote vascular hypertrophy. Attenuation of immune cell invasion into the vessel wall could potentially slow the progression of PH. One of the possible ways to reduce inflammation in the cardiovascular system is dietary modification with increased content of soluble fiber. Soluble fiber increases the level of short chain fatty acids (SCFAs) in the plasma which demonstrate anti‐inflammatory properties in systemic hypertension and cardiac overload. In different models of cardiovascular diseases high fiber (HFi) diet and SCFAs improve cardiovascular functions reducing the level of immune cells at the site of injury. The effects of HFi diet on the pulmonary circulation are unknown. This study establishes the effects of low (LFi) and HFi diet on macrophage and dendritic cell composition in the lungs in the model of hypoxia‐induced PH.

C57bl/6j mice were fed with LFi and HFi diet five weeks before and three weeks during hypoxic exposure (10% O2). After that hematocrit, cecum, spleen and body weight were measured. Flow cytometry was used to identify the proportion of alveolar and interstitial macrophages and dendritic cells in the lungs.

Hypoxic animals had increased hematocrit level compared to normoxic controls. Hematocrit did not differ between LFi fed animals and HFi fed animals in hypoxic conditions but in normoxia hematocrit was higher in LFi diet group. Cecum weight was significantly higher in HFi fed mice than in LFi fed mice whereas total body weight and spleen weight was not affected by diet composition. The percentage of alveolar macrophages (CD11b+CD11c+CD64+) did not differ between groups. Hypoxia increased the proportion of dendritic cells (CD11b+MHCll+CD24+) and reduced the proportion of interstitial macrophages (CD11b+MHCll+CD64+) among CD11b+MHC ll+ cells. Dietary fiber did not affect the proportion of dendritic cells and interstitial macrophages in normoxic and hypoxic animals.

This data demonstrates that three weeks of hypoxia changes the proportion of CD11b+ dendritic cells and interstitial macrophages in the mouse lungs however soluble dietary fiber did not affect it.
The role of mineralocorticoid receptor in the potassium switch: effect on NCC regulation+
Authors: Li Na , Wang Dongmei , Reina Patricia , van Bemmelen Miguel , Belaid Houwayda , Maillard Marc , McCormick James , Ellison David , Staub Olivier
Published: May 2022

Thiazide sensitive sodium‐chloride cotransporter (NCC) is a major salt transport pathway in the apical membrane of the nephron distal convoluted tubule. While we know the importance of its function in maintaining sodium‐potassium homeostasis, the regulation of NCC is complicated and the mechanisms are not well understood. The aim of this study is to evaluate the role of mineralocorticoid receptors (MRs) in the regulation of NCC. Here, we generated DCT‐specific MRs Knockout (KO) mice, in which MRs were deleted in cells expressing NCC, taking advantage of a cre recombinase under the control of the NCC gene. Under standard and Low‐Na+ diets, these DCT‐specific MRs‐KO mice display normal Na+/K+ balance but exhibit problems with Cl‐ regulation. These KO mice presented hypochloremia under normal diet but unexpectedly, hyperchloremia after 6d on low Na+ diet. The expression of NCC and phosphorylated NCC were both decreased in KO mice compared to that in control mice, under both standard and low‐Na+ diet. This decrease in NCC protein levels is related to a decrease in transcription levels. Under standard diet, decreased NCC expression is compensated by an increase in αENaC and pendrin expression, which is not the case under low‐salt diet. Under a low‐K+ diet, KO mice exhibit hypokalemia and volume depletion with increased blood CO2 levels and Renin mRNA levels. NCC expression and its phosphorylation are both increased in control and KO groups under low‐K+ diet compared to standard diet but, the levels of NCC protein and mRNA are still decreased in the KO group compared to the control group mice. Taken together, these results demonstrate that MRs is not necessary for the complete regulation of NCC under low‐Na+ and low‐K+ diet. On the other hand, MRs appears to be indispensable to assure proper NCC expression in adult DCT renal tubules.
Incidence and Prevalence of Alprazolam and Clonazepam Consumption at the Hospital Level in Ecuador
Authors: Muñoz Daniel , Zambrano Gema , Orellana‐Manzano* Andrea
Published: May 2022

Background

The consumption of benzodiazepines has increased worldwide due to their therapeutic properties such as anticonvulsants, sedatives, and muscle relaxants. Both men and women were consuming them at different ages. This study aims to analyze the incidence and prevalence of Alprazolam and clonazepam consumption at the IESS La Libertad Hospital to identify which patients treated in La Libertad are more likely to consume BZD

Method

The sample size was determined by patients who consumed Alprazolam or clonazepam from 2016 to 2020. A total of 1204 patients was selected who had a prescription from the psychiatrist. It was an exploratory‐descriptive methodology with an AS400 computer system, which is the official cybernetic system of the primary care Hospital at the national level

Results

Clonazepam Lq. Oral 2.5 mg/ml presents a greater incidence with 28.6%, followed by Alprazolam sd. Oral 0.5 mg with 10.65% and finally with Clonazepam tablets with 8.68%. The prevalence of consumption was Clonazepam LQ. Oral 2.5ml / mg 2.34%; Clonazepam sd, oral 0.67% and Alprazolam sd. Oral 0.99% had a higher prevalence of the consumption of Clonazepam vs. Alprazolam. Additionally, the increase in medical prescription was evaluated in the five years of study. The study was shown that in 2020, there was an increase of consumption with 21%, in 2019 with 16%, 2018 with 16%.

Conclusion

, it was determined that of the 1204 people who had prescriptions for psychotropic drugs, 58% are women, with a lower rate than men with 42% of prescriptions—validated by the VigiAccess portal. These results indicate the increase in the consumption of psychotropic drugs at the national level that will help carry out a better study for the annual reprogramming of drugs related to the acquisition of psychotropic drugs—achieving an optimal supply, delivering the complete treatment on time.
Binge drinking‐like behavior in juvenile rats causes long‐lasting cardiorespiratory dysfunction that persists until adulthood
Authors: Díaz Hugo S. , Andrade David C. , Toledo Camilo , Pereyra Katherine V. , Schwarz Karla G. , Del Rio Rodrigo
Published: May 2022

Harmful alcohol abuse is associated with serious psychosocial and health damage, including cardiac and neurological morbidities. Americans drink 40% more than the global average and adolescents are the heaviest drinkers, displaying a pathological consumption pattern termed “binge drinking”, consisting in short episodes of heavy alcohol intake followed by long withdrawal periods. Despite adolescent binge drinking (ABD) drives severe impairments in neurological and cardiorespiratory function, there are no current health policies to prevent ABD or its physiological consequences. Indeed, little is known about the long‐term consequences of ABD in adulthood physiology. Therefore, we aimed to investigate the potential harmful effects of ABD on cardiac and respiratory (patho)physiology in the adult life. Sixteen male juvenile Sprague‐Dawley rats underwent ABD‐like alcohol administration protocol or control treatment, consisting of 2‐days “ON” and 2‐days “OFF” doses of ethanol (3.0 g/kg, 25% w/v in saline administered i. p.) beginning on post‐natal day (PND) 25 until PND 38. Once rats reached adulthood (PND80), respiratory and cardiac function were respectively assessed by whole‐body plethysmography and intraventricular pressure‐volume loops. Compared to controls, adult rats exposed to ABD displayed no differences in resting blood pressure, heart rate and pulmonary ventilation, but they showed a significant (p<0.05) increase in the breath‐to‐breath interval variability (40.8 ± 2.8 vs. 25.6 ± 2.3 ms; ABD vs. Control, respectively) and in the incidence of apneas/hypoapneas (AHI: 5.83 ± 0.40 vs. 2.50 ± 0.37 events/h; ABD vs. Control, respectively). Intraventricular cardiac function analysis using PV‐loops revealed a reduction in left ventricular compliance in ABD animals, evidenced by the augmented slope of the end diastolic pressure‐volume relationship (EDPVR: 0.0303 ± 0.006 vs. 0.009 ± 0.001 mmHg/µL; ABD vs. Control). Also, we found that ABD induced cardiac autonomic imbalance estimated by low/high frequency ratio of heart rate variability (LFHRV/HFHRV: 1.59 ± 0.23 vs. 1.01 ± 0.08) and increased arrhythmia incidence (53.6 ± 12.7 vs. 21.7 ± 5.7 events/h; ABD vs. Control). Our results shows that binge drinking in adolescence causes long‐lasting cardiorespiratory dysfunction that includes at least cardiac diastolic dysfunction, autonomic imbalance, and generation of irregular breathing all of them persisting until the adulthood.

1Laboratory of Cardiorespiratory Control, Department of Physiology, Pontificia Universidad Católica de Chile, Santiago, Chile. 2Biomedical Department, Facultad of Health Science, Universidad de Antofagasta. 3Centro de Excelencia de Biomedicina en Magallanes (CEBIMA), Universidad de Magallanes, Punta Arenas, Chile. 4Centro de Envejecimiento y Regeneración CARE‐UC, Pontificia Universidad Católica de Chile, Santiago, Chile.