bluuebunny/crossref_metadata_2025 huggingface dataset:
Is Naloxone use in Southeast Alabama Rising? Evaluation of Naloxone Administration in an Emergency Department Setting
Authors: Parker Robert Wayne , Thorn Morgan , Patel Divyesha
Published: April 2020

Background

Fatalities caused by drug‐overdose have increased greatly in recent years across the US and is now the leading cause of injury‐related death in the United States. Drug related deaths with cointoxicants are being seen far too often and the increase in prevalence of opioid overdoses with possible concomitant medication use is a public health concern that needs to be addressed. Naloxone (Narcan ®) is an opioid antagonist approved in November 2015 by the FDA to help combat the effects of opioid overdose. Awareness of naloxone’s availability over the counter and education on opioid related overdoses for communities in the United States has been shown to improve opioid overdose deaths. In this study, we document naloxone administration in the Southeast Health Emergency Department to help identify high risk sub‐populations in this region that present with signs of potential opioid overdose and the connection between overdoses for opioids or a cointoxicant.

Study Design

A retrospective study was conducted evaluating the administration of naloxone in individuals arriving in the emergency department at Southeast Health from August 01, 2011 to August 31, 2018. Patient cases were collected based on time, route, and dose of naloxone given, and data was then selected based on ICD 10 codes and documented positive reaction to naloxone. Patients who were not between ages 18–89 or were incarcerated or pregnant prior to their hospital were excluded from this study. Further data was collected on patients’ socioeconomic status, medical history, and the course of their encounter.

Results

Of the 721 charts reviewed, 261 were deemed true overdoses and 51 were possible overdoses. While there were more overdoses in individuals with a history of chronic pain or mental illness, this was not statistically significant. The data showed most patients who overdosed were taking both opioids and benzodiazepines.

Conclusions

It is apparent that disabled/retired females over 65 with Medicare or Medicaid were the most likely to be a true opioid overdose.

Due to the patient’s mental status upon arrival, not all charts had complete histories or medication records. There were no clear documentation standards, so this varied from by provider and year, making it difficult to determine if there was a true overdose. Data was also dependent on patient honesty, relying on them to admit a history of abuse or what substances they had taken. The number of cases of overdose is likely under reported. Southeast Health is not the only hospital in the area. Emergency responders or home naloxone may have been administered decreasing the number of emergency consults. When considering patients insurance status, only the primary insurance listed was recorded. A patient could have also had a Medicaid plan indicating low income, but this was not noted.

Age Distribution

Figure 1

Naloxone and Past Medical History

Figure 2
From the Bench to the Bedside, the Determination of the Etiology of Dry Eye Using Spectroscopy
Authors: Borchman Douglas
Published: April 2020

Purpose

Meibum lipids are believed to be important to tear film stability, but tear film lipid compositional, structural and functional relationships are not well established.

Methods

NMR, Fourier transform infrared and Bruster angle spectroscopies were used to quantify meibum lipid composition, structure, and film lipid rheology, respectively.

Results

The molar ratio of Cholesterylesters/Wax ester increased with age, in cohorts aged 1 to 19 years old, respectively. The same molar ratio decreased for cohorts aged 20 to 88 years old with and without meibomian gland dysfunction (MGD) that leads to dry eye, respectively. The molar ratio of =C‐CH2 /ester increased with age for cohorts aged 1 to 19 years old, respectively. The same molar ratio decreased for cohorts aged 20 to 88 years old, with and without MGD, respectively. Compared with meibum from donors without dry eye, meibum from donors with MGD contained less straight‐chains, more iso‐branched chains, and the same amount of anteisobranched hydrocarbon chains. Human meibum order increases with age between 0 and 25 years old, meibomian gland dysfunction and with donors who have had hematopoietic stem cell transplantation. Interestingly, tear film stability also increases in the same order. When symptoms of dry eye were ameliorated with treatment, lipid order was restored. Principal component analysis scores correlated with age and dry eye by 93 %.

Conclusions

The changes in meibum composition with age and MGD contribute to the increase in lipid order of the same samples from donors without compared with those with MGD. Higher lipid order is associated with a higher reciprocal compressibility modulus that results in a tear film lipid layer that is not as compressible and not as viscoelastic. Strong lipid‐lipid interactions could contribute to the aggregation of tear film lipids and contribute to a decrease in tear film spreading and dry eye.

Support or Funding Information

Major support was obtained from the National Institute of Health [grant number EYO RO126180] and an unrestricted grant from Research to Prevent Blindness Inc. New York, NY, USA [grant number GN151619B].

Lipid order or stiffness correlates with tear film stability with age and dry eye symptoms.

Figure 1
Renal Interstitial Injection of a‐Trinositol Increases Renal Interstitial Hydrostatic Pressure (RIHP) but Does Not Modify its Low Frequency Oscillatory Pattern
Authors: Rodríguez-Martínez Manuel , López-Rodriguez Juan Francisco , Flores-Sandoval Omar , Silva-Ramírez Ana Sonia , Guerrero-Ojeda Vianney
Published: April 2020

RIHP increases after renal interstitial medullary injection (RIMI) of a 5μ bolus of saline solution (SS) + 2 μM α‐Trinositol (α –T, drug that increases collagen fiber network contraction, FASEB J 31:701.4: 2017). To assess whether this increase is induced by changes in the frequency and/or amplitude of oscillatory pattern of RIHP we evaluate in hydropenic rats the time course of RIHP before and after either an acute sham (CTR group, n = 9) or an acute (1s) real RIMI of a 5μ bolus of either vehicle (0.9% SS, SS group, n = 9) or SS + 2 μM α ‐ T (α – T group, n = 9). Left subcapsular RIHP (RIHP) was continuously measured (1 Hz) in anesthetized rats with right nephrectomy, acute left renal denervation, hormonal clamp and renal perfusion pressure control (100 mmHg), before (baseline phase = BL = 30 min) and 30 min after (experimental phase = EXP) sham or real RIMI of 5 μl bolus of either SS or SS + α‐T. The table
shows the 30 min average BL RIHP (1800 s, s = samples or seconds), ΔRIHP (EXP RIHP – BL RIHP) values (mean ± SEM) obtained 30 min after sham or real RIMI during 300 s. It also shows the RIHP frequency (F) of major oscillations (cycles/min = cpm) and the standard deviation (SD, an oscillatory amplitude index) of global RIHP oscillation before and 30 min after RIMI. Three were no differences among groups in BL RIHP, F or SD. For ΔRIHP data there were time * group interaction effect (P < 0.009); * P = 0.05, ** P < 0.001 vs. BL; • P < 0.04 vs. SS group, ¨P < 0.001 vs. CTR group. For SD data ° P <0.001 between phases.

CONCLUSIONS

The results show that RIHP oscillates at an equally low frequency (≈ 0.03 Hz) before and after sham or real RIMI but with lower amplitude after sham or real RIMI in any of the groups. This suggests that α‐T increases RIHP but not through modify the frequency or amplitude of its low frequency oscillatory pattern.

Support or Funding Information

Own financial resources of Integrative Physiology Lab.

RIHP

RIHP

RIHP

(mmHg)

Frequency

SD

Delta

cpm

(mmHg)

BL

EXP‐BL

BL

EXP

BL

EXP

30 min

at min 30

last 5 min

at min 30

30 min

at min 30

Group

1800 s

300 s

300 s

300 s

1800 s

300 s

CTR

3.0 ± 0.10

−1.1 ± 0.16

2.2 ± 0.26**

2.1 ± 0.26

0.54 ± 0.07

0.20 ± 0.04°

SS

3.1 ± 0.03

−0.15 ± 0.36

2.1 ± 0.44

2.2 ± 0.46

0.50 ± 0.06

0.18 ± 0.03°

a‐T

3.1 ± 0.06

1.26 ± 0.51

2.2 ± 0.32*•♦

2.2 ± 0.25

0.35 ± 0.04

0.19 ± 0.04°
Ablation of RAGE Attenuated Cardiac Fibrosis by Inhibition of Autophagy medicated Endothelial‐to‐mesenchymal transition
Authors: ZHANG LU , HE JIAQI , LIANG BIRONG , XIAN SHAOXIANG , WANG LINGJUN
Published: April 2020

Endothelial‐to‐mesenchymal transition (EndMT) has been found involved in the progression of heart failure (HF), which maybe one of the source of myofibroblasts. Emerging evidence support the notion that EndMT was regulated by autophagy. Our previous studies demonstrated deletion of receptor for advanced glycation end products (RAGE) inhibited autophagy and attenuated cardiac fibrosis in HF. Thus, here we investigated whether reduction of cardiac fibrosis by ablation of RAGE was through inhibition autophagy medicated EndMT. Using transverse aortic constriction (TAC) in mice as a model of pressure overload induced HF, we demonstrated that EndMT and autophagy were induced simultaneously at 8 weeks after TAC. Co‐localization of CD31 and α‐smooth muscle actin (α‐SMA) was observed in myocardium. Knockout of RAGE displayed reduced cardiac fibrosis and significantly improved cardiac function in TAC mice. Importantly, knockout of RAGE deceased the expression of autophagy related proteins (LC3BII/I and Beclin1), which was accompanied with the decrease of co‐expression of CD31 and α‐SMA in TAC mouse hearts. Moreover, 3‐methyladenine (3‐MA) and chloroquine (CQ), inhibitors of autophagy, attenuated EndMT and cardiac fibrosis in TAC mice. Taken together autophagy inducing EndMT may contribute to the development of cardiac fibrosis. Inhibition of this process by RAGE deletion lead to cardiac function improvement and reduction of cardiac fibrosis. Thus, we concluded that inhibition of the RAGE‐autophagy‐EndMT axis could be a novel strategy for treatment of heart failure.

Support or Funding Information

The National Natural Science Foundation of China (81373570, 81673920, 81473621, 81673796 and 81973776). The Natural Science Foundation of Guangdong Province(2016A030311030).

Reduction of cardiac fibrosis especially cardiovascular fibrosis after TAC by inhibition of RAGE and autophagy (Scale bar= 1,000 μm. n = 4. Values are shown as means ± SEM, *P < 0.05 vs Sham group, #P < 0.05 vs TAC group.)

Figure 1

Deceased expression of autophagy and EndMT after TAC by inhibition of RAGE and autophagy. (n = 4. Values are shown as means ± SEM, *P < 0.05 vs Sham group, #P < 0.05 vs TAC group.)

Figure 2
The Behavioral Effects of Omitting the α1T gene in Drosophila
Authors: Tran Kristen , Cheng Ryan , Luu Richard , Iniguez Jorge
Published: April 2020

Voltage gated calcium channels regulate calcium influx and vesicular release of neurotransmitter at chemical synapses. In Drosophila, there are three genes, Dmca1D, 1A, and α1T encoding Cav1, Cav2 and Cav3‐type channels respectively. Studies have shown that the α1T channels are important in regulation of action potential firing in Drosophila at the neuromuscular junction and the central nervous system. The α1T has been shown to be expressed across the adult fly brain. These include sensory neuropils, motor‐associated neuropils, and neuropils associated with learning and memory. We explored the role the α1T gene in adult Drosophila plays in motor‐associated activity using a locomotor assay. Our data suggests that the α1T gene knockout results in a defect in regulating excitability in adult Drosophila that manifests in altered motor activity. Using a T‐maze assay, the Drosophila are exposed to two different odors separately: 3‐Octanol with no electric current, and 4‐Methylcyclohexanol (MCH) administered with electric currents. Exposure to MCH with electric currents will create a negative association towards MCH and condition the flies to favor 3‐Octanol. The flies without the α1T gene are expected to be greatly impacted in their capability to build a negative connotation towards MCH, when compared to flies with the α1T gene.

Support or Funding Information

Build PoderNational Institute of HealthPasadena City College

T‐Maze

This figure illustrates how the Drosophila will favor the 3‐Octanol odor over the 4‐Methylcyclohexanol (MCH) odor.

Figure 1
The phases that separate RNA and transcription
Authors: Schwartz Jacob C.
Published: April 2020

RNA‐binding proteins recruited to nascent transcripts have the ability to regulate transcription and to spontaneously assemble into granule bodies through a process referred to as phase separation. These proteins, such as FUS (Fused in sarcoma) and EWS (Ewing sarcoma related 1), are endowed with these two abilities through domains that have low‐complexity amino acid sequences and are intrinsically disordered. Despite recent progress dissecting the biophysical principles underlying protein phase separation, its contribution to RNA‐processing and transcription has remained obscure. We have found that these proteins do assemble into granules that are associated with RNA polymerase II in cells. The formation of these granules and their association with the polymerase is dependent on transcription activity itself. Surprisingly, when transcription and phase separation is recombined in vitro, we have uncovered that at least one such protein, FUS, not only affecting transcription through its direct interactions with the transcription machinery but also by sequestering RNA structures that can hinder progress of the polymerase. Taken together, the role these proteins play and their peculiar RNA‐binding and phase separating properties provide a sophisticated mechanism to concentrate transcription and RNA‐processing machinery at genes and possibly to coordinate crosstalk between transcription and the maintenance of chromosome integrity.

Support or Funding Information

National Institutes of Health (NIH), American Cancer Society (ACS)

Figure 1
Functional antagonism between p53 and WWOX in vivo leads to protein aggregation in the brain
Authors: Chang Nan-Shan , Chou Pei-Yi , Lin Sing-Ru , Lee MIng-Hui , Schultz Lori , Sze Chun-I
Published: April 2020

Tumor suppressor WW domain‐containing oxidoreductase WWOX is a newly defined risk factor for Alzheimer’s disease (AD) in 2019. We discovered WWOX in 2000 and have been working on how WWOX blocks AD progression since then. We reported that WWOX starts to downregulate in the brain cortex and hippocampus in the middle‐aged normal individuals. Consequently, a likely scenario of slow activation of a protein aggregation cascade occurs with age as shown in the normal and AD brains as follows: 1) TRAPPC6AΔ, TIAF1 and SH3GLB2 aggregation, 2) caspase activation and APP degradation, and 3) final aggregation of amyloid β and Tau. The protein aggregation event may last 20 to 30 years and even longer. Here, we identified a tumor suppressor WWOX/TIAF1/p53 triad, which is potent in cancer suppression by blocking cancer cell migration, anchorage‐independent growth and SMAD promoter activation, and causing apoptosis. Yet, when p53 functionally antagonizes with WWOX in the mouse brain, p53 blocks WWOX‐mediated inhibition of inflammatory immune response induced by cancer, and this leads to protein aggregation in the brain as seen in AD and other neurodegeneration. The aggregated proteins include APP, amyloid β, neurofibrillary tangles (or tangled tau), p‐ERK and α‐tubulin. Together, we have discovered for the first time that WWOX limitation of neurodegeneration is blocked by p53 under inflammatory conditions in vivo.

Support or Funding Information

Ministry of Science and Technology, Taiwan, Department of Defense, USA, and National Health Research Institutes, Taiwan.

Summary illustration. Three scenarios account for cancer growth or suppression: 1) loss of WWOX, p53 and TIAF1 increases cancer growth and metastasis; 2) stabilized p53/WWOX/TIAF1 triad suppresses cancer growth, inhibits metastasis, and induces apoptosis; 3) functional antagonism between p53 and WWOX allows cancer cell growth and yet induces inflammation for causing neural protein aggregation as seen in the Alzheimer’s disease.

Figure 1
OvoceutinTM Attenuates Diarrhea Caused by Porcine Epidemic Diarrhea Virus in Piglets
Authors: Talukder Jamil , Srivastava Ajay , Rajamanickam Darwin , Lall Rajiv
Published: April 2020

Background

Porcine epidemic diarrhea virus (PEDV), a coronavirus, infects the small intestinal epithelium of a pig. Older hogs mostly get sick and lose body weight after being infected, whereas newborn piglets usually die within five days of contracting the virus due to enteritis, severe diarrhea, and dehydration. PEDV is highly infectious, resulting in significant morbidity and mortality in piglets ‐ approaching 100% in suckling piglets, which leads to substantial economic burden on pig farms. Controlling enteritis, diarrhea, and dehydration in piglets remains a great challenge to reduce mortality. Lactoferrin and ovotransferrin are iron‐binding multifunctional glycoproteins belonging to the transferrin family, which not only prevent diarrhea but also function as antibacterial, anti‐inflammatory, anti‐viral, etc. In addition, ovotransferrin (OvoceutinTM) has been shown to bind with Marek’s disease virus to block binding with host cell in poultry. Thus, Ovoceutin may play important roles in enteritis and diarrhea in PEDV‐piglets.

Aim

Determine the effects of Ovoceutin on diarrhea caused by PEDV in piglets.

Methods

Pigs on a farm were diagnosed with PEDV in Wisconsin, USA. Newborn piglets showing severe diarrhea were divided into five groups with five piglets in each group. Ovoceutin was made at an in‐house facility of Vets Plus, Inc. The control group received only electrolytes in 50ml of lukewarm water (EW); treated groups received 1 or 2.5g of Ovoceutin and water, or along with EW, twice a day for five consecutive days. Consistency of feces was scored as watery to solid (1–5), and physical conditions were scored as laying down (1), no response to caretaker (2), or walking (3).

Results

The control group (EW) showed poor survival rate (40%) while 2.5g of Ovoceutin‐water treated group had a higher survival rate (80%), better fecal score, and physical conditions. EW‐2.5g of Ovoceutin group showed the highest survival rate (100%), best fecal, and physical condition scores.

Conclusions

We conclude from these results that OvoceutinTM attenuates diarrhea in PEDV‐piglets and can be used to improve piglets’ digestive health.

Support or Funding Information

Research and Development Fund, Vets Plus, Inc.

Effect of OvoceutinTM in diarrhea caused by Porcine Epidemic Virus Diarrhea in newborn piglets.

Figure 1
3D pdb Structures as Native 3D Models into MS PowerPoint
Authors: Ouellet Jonathan
Published: April 2020

An effective way to transmit knowledge is to share an image. In fields such as astronomy, engineering, morphology or biochemistry, the plain 2D paper format has limitations on the visualization of objects in 3D.

Innovations such as 3D PDF from Adobe allowed publishers to integrate 3D objects into pdf while many researchers create 360‐degrees rotation movies of their molecule of interest. Still, the lack of easy insertion of 3D objects in common files makes the 3D visualization hard to share.

In January 2018, Microsoft Office (MSO) introduced a new option within its Insert Menu by adding 3D Models. Stock 3D models can be added, rotated and animated. More importantly, multiple 3D file formats can be incorporated.

This focus of this presentation is to demonstrate a “How to” guide to easily convert a pdb structure to a 3D format which is compatible with MSO. Such 3D model are lightweight, easy to create and very easy to share.

Procedure to import pdb structures into MS PowerPoint

Figure 1
Maternal Exposure to Non‐nutritive Sweeteners Impacts Progeny’s Metabolism and Microbiome
Authors: Van Stichelen Stephanie Olivier , Rother Kristina I. , John Hanover A.
Published: April 2020

Non‐nutritive sweeteners (NNS e.g. Splenda, Sweet’N’Low, Equal, Stevia, …) are highly prevalent in the diet of United States adults and children and are found in beverages, yogurt, cocoa powder, coffee sweeteners, medicine and kid electrolyte solution (Pedialyte). They are marketed as sugar alternatives providing sweet taste with few or no calories. First thought to be harmless, a growing number of research studies now questioning the safety of NNS consumption. In the past decades, NNS have been shown to alter gut hormonal secretion, glucose absorption, appetite, kidney function, insulin secretion, adipogenesis and gut microbiota. The wide range of action associated by NNS exposure is likely linked to their binding to the sweet taste receptor, found not only on the tongue but also in the intestines, lungs, bones, adipose tissue and testis.

However, to date, the effects of NNS on early development are incompletely understood. In critical stage such as embryonic/ fetal development, small exposures can be serious as demonstrated for alcohol consumption or smoking. While there are many specific recommendations during pregnancy or lactation for various products, there are no guidelines for NNS consumption. Furthermore, it is interesting to know that only one of four parents are able to correctly identify NNS on food product suggesting that many are consuming NNS involuntarily Interestingly, at least 10 different studies demonstrated an association between increased BMI and NNS consumption suggesting a significant association in children. Furthermore, despite known transmission through amniotic fluid and breast milk, the impact of NNS on early metabolic development remains largely unknown.

To answer this question, we exposed pregnant and lactating mice to NNS (sucralose, acesulfame‐K) at doses relevant for human consumption (Acceptable Daily Intake). While the pups’ exposure seemed minimal, metabolic changes were drastic, indicating extensive down‐regulation of hepatic detoxification mechanisms and changes in bacterial metabolites. Microbiome profiling confirmed a significant increase in firmicutes and a striking decrease of Akkermansia muciniphila. Similar microbiome alterations in humans have been linked to metabolic disease and obesity. Our finding corroborates the existing literature but required considerably shorter NNS exposure (only ~40 days). Therefore, we concluded that pregnancy is a vulnerable time for exposing offspring to NNS. While our findings need to be reproduced in humans, they suggest that NNS consumption during pregnancy and lactation have adverse effects on infant metabolism and potential predisposition to metabolic deregulation later in life.

Summary figure of the effect of sucralose and acesulfame‐K consumption during pregnancy and lactation.

Figure 1
Cardiac Autonomic Neuropathy and Hemodynamic Dysfunction as a Consequence of Mild Hypercaloric Intake: Modification by Phosphate Supplementation
Authors: Ajouz Ghina MohammadNabil , Dwaib Haneen Salem , Mougharbil Nahed , Mroueh Ali , Refaat Marwan , Obeid Omar , El-Yazbi Ahmed
Published: April 2020

Cardiac autonomic neuropathy (CAN) has been linked to negative outcomes in diabetic patients, where it is associated with a high risk of cardiac arrhythmia and sudden death. Our previous research showed that prediabetic rats develop CAN in absence of overt signs of metabolic derangement or change in body weight. This was associated with vascular dysfunction and perivascular adipose tissue (PVAT) inflammation, which preceded inflammatory changes in other adipose pools, possibly linked to increased Uncoupling protein 1 (UCP1) and hypoxia‐inducible factor (HIF1‐a) expression. In parallel, phosphate supplementation is a simple intervention to improve body weight, energy metabolism, and glucose tolerance. Moreover, inorganic phosphate acts as a UCP1 inhibitor. Here, we examined the role of phosphate supplementation in ameliorating the early signs of cardiovascular dysfunction in metabolically challenged rats. Male SD rats (5–6 weeks) were randomly allocated into four groups fed either control diet (group 1) or high‐calorie (HC) diet for 12 weeks with three different levels of dietary phosphate; group 2: low phosphate (LP)‐ 0.37 mg/KCal, group 3: intermediate phosphate (IP)‐0.7 mg/KCal, group 4: high phosphate (HP)‐ 1.5 mg/KCal. Daily food intake, body weight, and body composition were recorded. Serum insulin was measured by ELISA. Invasive hemodynamic measurements were done and baroreceptor sensitivity (BRS) was assessed by the vasoactive method. After sacrifice, organs were collected for immunohistochemistry and western blotting. No changes in caloric intake, body weight, blood pressure, and blood glucose levels were observed. However, rats on HC diet showed an increase in serum insulin level and fat/lean ratio. Consistent with our previous work, HC‐feeding was associated with significant reduction in parasympathetic BRS in both the LP and IP groups. This was also accompanied by a reduced left ventricular systolic function. On the cellular level, Oil Red O stain showed a significant increase in the diameter of PVAT adipocytes in LP condition, together with increased expression of HIF‐1α, IL‐1β and UCP‐1. Of note, inflammatory changes observed in the PVAT were not detected in neither white nor brown adipose tissue. Furthermore, higher macrophage infiltration and total ROS score in the PVAT, heart mid‐section and brainstem were observed in LP when compared to control. Our present results highlight a role of phosphate supplementation in the amelioration of the HC induced parasympathetic dysfunction. This could be due to the reported ability of dietary phosphate to improve the assimilation of fat and reduce adipose tissue hypoxia possibly via UCP1 inhibition.

Support or Funding Information

Funded by an AUB CRS grant

IL1‐UCP1 for perivascular adipose tissue (PVAT), white adipose tissue (WAT), and brown adipose tissue (BAT)

Figure 1
Mast Cell Concentrations Correlate with Hepatic Fibrosis in Cancer Patients
Authors: Lombardo Jamie Lynne , Taggart Melissa , Rashid Asif
Published: April 2020

Background

The pathogenesis of liver fibrosis in non‐alcoholic steatohepatitis (NASH) has been studied in animal models and humans, with some researchers finding a direct correlation between the concentration of mast cells and the stage of fibrosis in NASH. This suggests that mast cells play a key role in the progression of fibrosis in NASH patients.

This study investigates patients with liver fibrosis in a more varied clinical setting with exposure to hepatotoxic chemotherapy regimens and checkpoint inhibitor therapy with various underlying causes of hepatic fibrosis including alcohol use, chronic hepatitis C or Hepatitis B infection, and NASH. The goal is to determine if correlations between mast cell concentration (MCC) and fibrosis remain in patients with multifactorial liver injury.

Design

Hepatic tissue from seventy‐seven patients (13 resections and 64 biopsies), was retrospectively selected from the database of a large cancer center, and reviewed. Of the cases selected, 38 were male and 39 were female. The gender breakdown by stage of fibrosis is detailed in figure
1. Patients ranged in age from 28–87. The stage of fibrosis was determined using the Batts‐Ludwig criteria. Potential injury‐inducing factors including chemotherapy regimen, checkpoint inhibitor therapy use, and any background liver disease were noted. Six of the patients had multiple primary cancers, 47 had cancer plus underlying liver disease, 18 had liver disease alone, 11 were taking checkpoint inhibitors, 30 were on chemotherapy, 7 were taking hormone therapy, and 6 had one primary cancer with no liver involvement or underlying liver disease.

A tryptase immunohistochemical stain (clone AA1, Dako) was performed on a representative section from each case to evaluate MCC. Mast cells in five 40× fields (1 mm2) in both portal tracts and lobular areas were counted and separately recorded. MCC was then compared to the stage of fibrosis using an independent student’s t‐test calculation (Graphpad Prism statistics software).

Results

MCC in portal areas significantly correlated with each histologic stage of fibrosis (p‐values < 0.05, Fig
1). Lobular MCC showed a statistically significant correlation between stage 0 and stage 1–2 (p‐value 0.0043) but did not correlate with fibrosis in later stages (Fig.
1). Though a positive trend is seen in MCC compared to the stage of fibrosis in both portal tracts and lobular areas, a stronger association is apparent in the portal tracts (Fig
2).

Conclusion

In medically complex patients from a large cancer center with exposure to various chemotherapy types and checkpoint inhibitor therapy, mast cell concentrations within portal areas directly correlate with the stage of hepatic fibrosis. This finding further supports the role of mast cells in the development of hepatic fibrosis.

Support or Funding Information

MD Anderson Department Chair Research Fund for Fellows

Figure 1Figure 2
Quantitative Sudies of Spontaneous Electrical Activities in Detrusor Smooth Muscle Cells towards Understanding Urinary Bladder Overactivities
Authors: Mahapatra Chitaranjan , Manchanda Rohit
Published: April 2020

Context

The urinary incontinence (UI) is defined as the involuntary loss of urine and associated with the enhanced spontaneous contractions of the detrusor smooth muscle (DSM). The spontaneously evoked action potentials (sAPs) in DSM cells initiate and modulate these contractions. The DSM is strongly innervated, connecting approximately 16000 afferent and efferent axons from ganglion neurons. It generates sAPs due to the stochastic nature of purinergic neurotransmitter release from the parasympathetic nerve.

Objectives

The aim of this current study is to understand the putative relationship between the fluctuating ion channel conductances and stochastically release of ATP in generating sAPs.

Methods

The neurotransmitter current was considered as an independent excitatory conductance in the model where gex(t) and Eex are the one‐variable stochastic process conductance and the reversal potential respectively. In addition, Dex and λ1(t) are known as the diffusion coefficients and Gaussian white noise. The point‐conductance is incorporated into a single DSM cell model based on single cylindrical compartment.

Results

The elicited AP consists an after depolarization and after hyperpolarization phase. The AP peak amplitude and duration are about 15 mV and 40 ms respectively. The stochastic nature of purinergic neurotransmitter is designed by a point process model. Then, the random injection of point process model is conducted to elicit a series of sAPs and membrane depolarization for 5 seconds. The membrane resting potential is held at − 50 mV with a 3 mV of fluctuation. The stochastically depolarization up to 20 mV activates the T‐type Ca2+ channel first and then the L‐type Ca2+ channel to generate action potential. The Figure
1 A illustrates series of AP generation in the DSM cell due to stochastically depolarizations. The model does not evoke any AP and depolarization, when the conductance of T‐type Ca2+ channel is set to zero (Figure
1 B).

Conclusions

From the results, it is found that the T‐type Ca2+ channel blocker can be used as a new pharmacological target for the UI. In addition, an extended multidimensional models will a provide windows of insight into the factors that govern excitability and contraction in both normal and unstable bladder.

Support or Funding Information

This work is supported in part by Department of Biotechnology (DBT), India (grant number BT/PR12973/MED/122/47/2016).

Effects of T‐type Ca2+ channel blockers on AP generation.

Figure 1
Understanding the Structure and Function of Type I Acyl Carrier Protein and its Interaction with Growing Acyl Chain
Authors: Garima  , Sundd Monica
Published: April 2020

Fatty acids are central to the formation of biological membranes and play important roles in metabolism. There are two major classes of fatty acid synthases (FAS); Type I present in mammals and yeast that utilizes large multifunctional enzyme complexes, while type II, harbored by prokaryotes and plants, utilizing discrete monofunctional enzymes. Acyl Carrier Protein (ACP) is a universal and highly conserved player in both the pathways. ACP is modified from apo to holo form by enzyme 4′‐Phosphopantetheinyl Transferase (PPT). PPT enzyme is crucial for growth of many pathogens and is therefore regarded as a potential drug target. Structural and biochemical studies on type II ACP highlight various aspects of ACP‐PPT and ACP‐acyl chain interaction. However, acyl chain binding and dynamic interactions of type I ACP are not well understood. We therefore intend to understand the function of type I ACP, their interaction with bound acyl chain and cognate and non cognate transferases. Literature highlights the importance of type I FAS in the survival and pathogenicity of fungal pathogens like C. albicans and C. parasilopsis. For our studies, we employed type I ACP from Saccharomyces cereviceae and its putative PPT enzyme. Using NMR spectroscopy we show that the growing acyl chain was sequestered up to a certain carbon chain length in the hydrophobic core of the protein employing a unique motif. In conclusion, our studies provide some interesting insights into the structure and function of type I ACP and the molecular mechanism it employs to protect the growing acyl chain. Our findings can provide new avenues for structure based drug designing.

Support or Funding Information

Department of Biotechnology, India
Simultaneous Non‐invasive Recording of Skin Sympathetic Nerve Activity
Authors: Kusayama Takashi , Wong Johnson , Liu Xiao , He Wenbo , Doytchinova Anisiia , Adams David E. , Chen Lan S. , Everett Thomas H. , Chen Peng-Sheng
Published: April 2020

Our research laboratory recently invented a new method (neuECG) to simultaneously and non‐invasively record electrocardiogram (ECG) and skin sympathetic nerve activity (SKNA) from humans using standard ECG patch electrodes. These methods are non‐invasive and stable over long periods of time. However, because the neuECG analyses required custom written software, these methods have not been widely used or tested by the scientific community. The purpose of the present study was to develop a method for neuECG recording and analyses using commercially available hardware and software. We used ADInstruments PowerLab to record bipolar electrical signals from the skin of thorax and arms. Wide bandwidth and high sampling rate (10,000/s) were used for recording. The data were then analyzed offline using LabChart Pro 8 software. The digitized neuECG signals are amplified and band‐pass filtered between 0.05 Hz and 150 Hz to display ECG and between 500 Hz and 1,000 Hz to show the SKNA (μV). We full‐wave rectify then integrate all digitized SKNA signals using leaky integrator with 100 ms time constant and display the results over time to simulate the display methods of microneurography (iSKNA, μV.s). For quantitative analyses, we divide the absolute sum of the voltage samples by the number of digitized samples in the chosen window to obtain the aSKNA (μV). We can also examine the distribution of the aSKNA and determine a threshold to separate baseline activity from the burst activity. As compared with the microneurography studies, the neuECG does not require invasive procedures to impale electrodes in nerves and is voltage calibrated. The latter characteristics allow investigators to compare aSKNA, iSKNA and burst duration between subjects or groups. Figure shows the actual neuECG recording. SKNA band‐pass filtered from the digitized signal was converted to iSKNA and aSKNA. Down arrows show premature ventricular contractions. The dotted line indicates the commencement of cold water pressor test that elevated the aSKNA (red double headed arrows) and heart rate. We conclude that neuECG can be recorded and analyzed using commercially‐available hardware and software. These new methods may facilitate the testing and use of this novel technology in experimental biology research.

Support or Funding Information

This study was supported in part by NIH Grants 1OT2OD028190, R01 HL139829, R56 HL71140, TR002208‐01 (Dr. Chen), R42DA043391 (Dr Everett), a Charles Fisch Cardiovascular Research Award endowed by Dr Suzanne B. Knoebel of the Krannert Institute of Cardiology (Drs. Kusayama and Everett), a Medtronic‐Zipes Endowment, and the Indiana University Health‐Indiana University School of Medicine Strategic Research Initiative (Dr. Chen).

Example of actual neuECG recording

Figure 1
Small Molecule TREX1 Inhibitors as Novel Immunotherapies
Authors: Hemphill Wayne , Harvey Scott , Gumpena Rajesh , Simpson Sean , Grayson Jason , Smalley Terrence , Salsbury Fred , Hollis Tom , Perrino Fred
Published: April 2020

Mutations in the 3′ ‐> 5′ exonuclease TREX1 are associated with a spectrum of autoimmune phenotypes in humans and mice. In those affected, pathology is driven by persisting DNA which would normally be degraded by functional TREX1 enzyme. This undegraded DNA stimulates the nucleic acid sensing pathway, cGAS‐ STING, whose subsequent signaling generates a type‐I interferon response. This type‐I interferon response ultimately results in the chronic immune system activation typical of those affected. TREX1 and the cGAS‐ STING pathway have also been implicated in the tumor microenvironment, where TREX1 is believed to degrade tumor‐derived DNA destined to trigger cGAS‐STING signaling. If this tumor‐derived DNA is not degraded, the stimulated cGAS‐STING pathway promotes IFN‐dependent antitumor immunity. Consequently, small molecule stimulators of the cGAS‐STING pathway have shown promise as immunotherapies in animal models. Similarly, mice devoid of TREX1 exonuclease activity exhibit T‐cell dependent resistance to tumor growth. Collectively, these findings indicate that stimulation of the cGAS‐STING pathway activates the immune system, and that TREX1 inactivation represents a novel strategy for this purpose. Furthermore, they demonstrate the immunotherapy potential of such approaches. Thus, we hypothesize that small molecule inhibitors of TREX1 will activate the cGAS‐STING pathway. Studies described here identify small molecule inhibitors of TREX1, determine their mechanism of inhibition, and directly test the hypothesis that they can activate the cGAS‐STING pathway in murine cells. Findings from these studies will generate new structural insights into how TREX1 exonuclease activity may be affected, and have already identified several candidate chemotypes for development into novel immunotherapies.

Support or Funding Information

This work was supported by the NIH, NIAID, NIGMS, Lupus Research Alliance, Wake Forest Innovations, and a Sandy Lee Cowgill Memorial Scholarship.

D18N mice display T‐cell dependent tumor resistance.

5×106 H31ml tumor cells were injected subcutaneously into WT and D18N mice, and tumor volume and survival tracked daily. Data in ‘B’ are separate study from ‘A’. For each study, eight to 16 mice were examined in two to four independent experiments. (A) D18N mice have decreased tumor growth in a mouse tumor model. (B) D18N model tumor‐resistance is T‐cell dependent. WT and D18N were treated with αCD8, αCD4, or isotype‐control antibodies. Arrows indicate D18N mice without treatment, or with CD8+ / CD4+ T‐cell depletion.

Figure 1
Preliminary study of in‐situ 3D bioprinted nano‐silicate biopolymer scaffolds for muscle repair in VML defects
Authors: Ahuja Neelam , Awad Kamal , Fiedler Matthew , Aswath Pranesh , Brotto Marco , Varanasi Venu
Published: April 2020

Introduction

The traumatic or surgical loss of skeletal muscle with resultant functional impairment (volumetric muscle loss (VML)) is a challenging clinical problem. The current standard treatment care for VML is autologous tissue grafts and physical therapy, which still faces the challenge of donor site morbidity in large defect cases. 3D printing techniques for tissue engineering allows the precise control of micropatterning by determining the dimensions of filaments, the size and shape of pores and the percentage of porosity of the scaffold. Our hypothesis is using 3D printing of biopolymer‐nanosilicate scaffolds in‐situ will enhance muscle regeneration in VML defects.

Materials and Methods

Methacrylated gelatin (MAG) biopolymer Laponite (Lp) nano‐silicate scaffolds were prepared as a bio‐ink using a solution‐precipitation process. Our study includes designing and optimization of 3D printing parameters for printing MAG‐Lp scaffolds and in‐vitro evaluation of cell viability (live‐dead), proliferation, and differentiation of mouse myoblast precursor cells on these scaffolds. We further analyzed the in‐situ (directly in the defect) 3D printing (using a robocaster) in a rat VML model in tibialis anterior muscle to evaluate the feasibility, precision, flexibility and attachment of the scaffold to the defect.

Results

Silicon ion enhanced cell proliferation and differentiation (fusion index by 1.5 times and overexpression of myogenic factors) in mouse myoblast precursor cells as compared to control. Qualitative live/dead analysis showed more abundance of live myoblast precursor cells (green) and decreased number of dead cells (red) on MAG‐Lp surface as compared to MAG (Fig.
1). We also show successful printing abilities of MAG‐Lp scaffolds in the volumetric muscle loss defect (Fig.
2) along with increased attachment to the adjacent tissues. The animal also showed ability to stand on/flex the surgical limb, with sufficient grip strength to perform normal activity, immediately after recovering from the surgery.

Conclusion

Our preliminary findings conclude that 3D printing of MAG‐Lp can enhance myogenesis in myoblasts. We further conclude that successful proof of concept of an in‐situ bioprinting method of reconstructive scaffolds can be used for immediate repair and loading of muscles for VML injury treatment.

Support or Funding Information

NIH/NIDCR (R03DE023872‐01, 1R56DE027964‐01A1‐01), Texas STARs award, Departmental Start‐up Funds, University of Texas at Arlington, College of Nursing and Health Innovation, Arlington, TX

showing live/dead assay on MAG and MAG‐Lp scaffolds. Green represents live cells, whereas red represents dead cells.

Figure 1

showing In‐situ 3D printing biopolymer scaffold in a rat volumetric muscle loss model in tibialis anterior muscle (20–25% muscle loss) along with UV/photo curing

Figure 2
Histopathological Impact Of Nematodes On The Gastric Mucosa Of The American Alligator (Alligator Mississippiensis)
Authors: Yesudas Rekha , Young Bruce
Published: April 2020

Nematodes are adapted to inhabit a vast array of niches. They have evolved tremendous strategies to survive and succeed in a variety of hosts and host tissues. Intestines represent an ideal habitat for a large number of parasites, compared to the stomach. In the present study we examined the stomach of 14 sub‐adult, wild‐caught American alligators (Alligator mississippiensis) from the coastal region of Louisiana. Nematodes were found in four of the alligators (~29%). Two different parasites were found; Dujardiascaris waltoni (Nematoda: Heterocheilidae) and Ortleppascaris sp.? (Nematoda: Ascaridoidea). D. waltoni was found loose among the stomach contents, whereas Ortleppascaris sp.? was found embedded in the stomach wall. Ortleppascaris was associated with multifocal lesions in the gastric mucosa. These lesions were roughly 4 mm in diameter and housed multiple parasites. The lesions caused a clearly visible penumbra of mucosal damage. As the parasites invaded the gastric mucosa, they induced a granulomatous inflammation of the mucosa and submucosa. There was a marked eosinophilic necrosis, a cellular response that produces the penumbra around the invasive nodules. The impacted worms burrowed into the mucosa, and were surrounded by an eosinophilic exudation, resulting in an elevated nodule of worms and mucosa. This presumably represent proliferating host mucosal tissue responding to the parasitic challenge. Alligators have “typical” vertebrate gastric pits and glands in the stomach mucosa. Mucosal burrows of the nematodes led to a loss of the gastric pits in the infected zone and digestive efficiency of the host. Previous studies have described keratitis and enteritis associated with nematode infections in crocodilians, but similar reports are lacking from Alligator mississippiensis. Alligators and crocodiles have remarkably effective immune systems, which enable them to fight against microbial infections and allow quick wound healing. We hope to expand our understanding of host‐parasite relations in this important species.

Host response/eosinophil exudation in infected gastric mucosa

Figure 1

Nematodes burrowed in to the stomach mucosa

Figure 2
Extract of Foeniculum vulgare seed effect on UV‐induced photoaging‐associated ion channel and collagenase
Authors: Woo Joohan , Kim Yung Kyu , Kim Woo Kyung
Published: April 2020

Skin aging is mediated by genetics, hormonal changes, metabolic processes, and environmental stress. UV radiation is concerned as a major risk factor for skin aging, including wrinkles, and melanoma. Studies suggested that calcium ion channels, such as TRPV1 and ORAI1, are related with melanogenesis, skin wrinkling, and inflammation. Foeniculum vulgare has been used to treat inflammatory diseases. Recent studies showed that it protects against UV radiation‐induced skin photoaging. However, its effect on the ion channels related to UV‐induced photoaging is unclear. Therefore, this study aimed to evaluate the antagonistic effects of F. vulgare extract, fractions, and constituents on TRPV1.

Electrophysiological analysis results showed that, at 100 μM, F. vulgare extract and fractions potently inhibited capsaicin‐induced TRPV1 [methanol extract (91%), n‐hexane (94%), and methylene chloride (100%) at −60 mV]. The methanol extract of F. vulgare inhibited MMP‐1 (collagenase) production (35% at 3.75 μg/mL), compared with control. These results suggested that the compounds in F. vulgare extract and fractions possessed anti‐aging effects that can decrease wrinkles and maintain elasticity by disrupting MMP‐1 production activated by the MAPK pathway. Unfortunately, compounds isolated from the hexane and methylene chloride fractions could not suppress ITRPV1. Surprisingly, even though capsaicin increased ITRPV1, the γ‐terpinene, limonene, scopoletin, and bergapten isolated from these two fractions slightly potentiated ITRPV1. Although 4 compounds of F. vulgare exerted an agonistic effect on TRPV1, the compounds with antagonistic effect have not been discovered. Our study suggested that F. vulgare had potential therapeutic effects on UV‐induced photoaging for therapeutic and cosmetic applications.

Effect of F. vulgare extract and fractions on human TRPV1 (hTRPV1) current under capsaicin (CAP) treatment

Figure 1

Effects of F. vulgare extract and fractions on UV‐induced melanogenesis

Figure 2