bluuebunny/crossref_metadata_2025 huggingface dataset:
Optimal tip position of peripherally inserted central catheter (PICC) using electrocardiogram–a retrospective analysis
Authors: Lee Kichang , Kelly Linda , Sabouri A. S. , Jung Ki T.
Published: May 2022

Background

Correct tip location of a peripherally inserted central catheter (PICC) is crucial to ensure optimal drug delivery and avoid creating potential complications. P‐wave amplitude changes in the intravascular electrogram (ivECG) can guide the PICC tip to the optimal location during the catheter placement procedure, but its accuracy is limited. Therefore, the main objective of this study was to evaluate the ratios of different waves in ivECG and body external ECG (exECG) when the correct tip location was confirmed by chest X‐ray (CXR).

Methods

In this retrospective study, we analyzed the ivECG, exECG, and CXR of 146 patients who underwent a PICC placement procedure. The optimal location of the PICC tip was defined in the lower 1/3 of the superior vena cava (SVC) to the junction of the SVC and right atrium, known as the cavo‐atrial junction (CAJ). The catheter tip to carina distance (TCD) was also measured using vertebral body units (VBU). The PICC tip location was categorized as follows: Zone 1, malposition with TCD < 1.2 VBU; Zone 2, suboptimal with 1.2 VBU ≤ TCD < 1.8 VBU; Zone 3, optimal with 1.8 VBU ≤ TCD < 2.8 VBU; Zone 4 deep with TCD > 2.8 VBU. The amplitude ratios between/within the ivECG and the exECG of the corresponding waves were calculated and compared.

Results

The external/internal ratio of the amplitude of uprising P‐wave (Psp) and declining P‐wave (Ppe) in Zone 3 was significantly higher than in Zone 1 (p < 0.001) and Zone 2 (p = 0.004). The amplitude ratio in the ivECG of uprising P and QR (Pspiv/QRiv), uprising P and RS (Pspiv/RSiv), declining P and QR (Ppeiv/QRiv), and declining P and RS (Ppeiv/RSiv) were significantly lower in Zone 3 compared to Zone 1 (p < 0.001, respectively). There was a moderate correlation between TCD using VBU and ECG height ratios such as Pspiv, Ppeiv, Pspiv/QRiv, and Pspiv/RSiv (p < 0.001, respectively).

Conclusions

The height ratios of ECG waves such as Pspiv, Ppeiv, Pspiv/QRiv, and Pspiv/RSiv can help determine the catheter tip location during the catheter insertion procedure; however, caution is required for accurate positioning of the PICC tip.
Angiotensin‐(1‐7) Restores Gut Barrier Integrity and Ameliorates Systemic Inflammation in Aging
Authors: Chittimalli Kishore , Jahan Jesmin , Sakamuri Anil , Jarajapu Yagna P.
Published: May 2022

Background

Advancing of age decreases the barrier properties of intestinal epithelium which leads to sterile systemic inflammation. Chronic low‐grade chronic inflammation is an underlying cause for the age‐related comorbidities such as cardiovascular disorders. Renin Angiotensin System (RAS) plays an important role in the homeostasis of cardiovascular functions however, the role of RAS in the aging gut is poorly studied. The protective axis of RAS, which constitutes Angiotensin converting enzyme‐2 (ACE2)/ Angiotensin‐(1‐7) (Ang‐(1‐7))/Mas receptor (MasR) is known to be counter‐regulatory to the classical axis, ACE/Ang II/AT1 receptor (AT1R). This study tested the hypothesis that leaky gut with aging is associated with ACE2/ACE imbalance and that activation of MasR with Ang‐(1‐7) would restore gut barrier integrity and ameliorates systemic inflammation.

Methods

Study was carried out in Young (3‐4 months) and Old (20‐24 months) male mice. ACE and ACE2 enzyme activities were evaluated in the colon. Gene and protein expressions of RAS components were determined by western blotting. Ang‐(1‐7) was administered by subcutaneous osmotic pump (1 µg/Kg/min) for four weeks and the gut permeability was evaluated by using FITC‐dextran. Colon wall integrity was evaluated by immunohistochemistry of claudin 1 and occludin. Zonulin‐1, IL‐6, and TNFα were analyzed in the plasma. A779, MasR antagonist, was administered by an osmotic pump in selected experiments (1 µg/Kg/min, subcutaneous).

Results

ACE2 protein and activity were decreased in Old group while that of ACE were increased (n=6) (n=8) in the colon of the Old mice compared to the Young. Protein expression of both AT1R and MasR was higher while that of AT2R was lower in the Old (P<0.05, n=8). Protein expression of ACE2, ACE AT1R and MasR in the Old colons were restored to Young levels by treatment with Ang‐(1‐7). Gut permeability was higher in Old mice (P<0.01 vs Young, n=6) that was reversed by Ang‐(1‐7). The beneficial effect of Ang‐(1‐7) was abolished by concurrent administration of A779. Hematoxylin & Eosin staining revealed that aging was associated with atrophied villi in the gut wall that was normalized by Ang‐(1‐7). Gene expression of claudin 1 and occludin were lower in the Old colon wall that were restored by Ang‐(1‐7) and similar observations were made by immunohistochemistry. Importantly, plasma levels of Zonulin‐1, (P<0.01) IL‐6 and TNFα (P<0.05) (n=8) were higher in the Old group compared to the Young, which were lowered by Ang‐(1‐7) (n=5).

Conclusion

These results suggest that activation of MasR ameliorates aging‐associated systemic inflammation by restoring gut barrier integrity, which at least in part contributes to the cardiovascular‐protective functions.
Exploring the Effect of RNA Binding on TDP‐43 Liquid‐Liquid Phase Separation
Authors: Padilla George , Shorter James , Rubien Jack
Published: May 2022

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that is characterized by progressive deterioration of nerve cells in the brain and spinal cord. Cytoplasmic aggregates of TDP‐43 have been observed in the majority of ALS cases (~97%). In addition to ALS, TDP‐43 aggregates have been associated with other neurodegenerative disease pathologies such as frontotemporal dementia (FTD) and limbic‐predominant age‐related TDP‐43 encephalopathy (LATE). Besides forming aggregates, TDP‐43 can form liquid‐like droplets termed ‘condensates’. TDP‐43 condensates are a consequence of liquid‐liquid phase separation (LLPS), a physical process in which a solution de‐mixes into two phases, a dense phase, and a dilute phase. Aberrant LLPS of TDP‐43 is shown to be neurotoxic in the disease pathology of ALS/FTD. In recent literature, yeast models that are predisposed to form cytoplasmic aggregates of TDP‐43 have increased viability in comparison to their droplet forming counterparts. This suggests that TDP‐43 aggregates may act as a cellular defense mechanism against toxic liquid‐like condensates. GU‐rich RNAs have displayed binding of TDP‐43 that mitigate neurotoxicity by inhibition of LLPS. Specifically, RNA binding of TDP‐43's RNA recognition motif (RRM) domains. Based on these findings, I focus on the dynamics between RNA binding and TDP‐43 phase separation, and the importance of TDP‐43 RRMs in LLPS. Depending on characteristics of RNAs, RNA binding on TDP‐43 RRMs may enhance the impedance of droplet formation. We can better understand how TDP‐43's RRMs facilitate and stabilize RNA binding interface by the introduction of inhibitory residue mutations on TDP‐43's RRM 1 & 2. Through the screening of TDP‐43 RRM mutants and wild‐type with various RNA sequences, we can reveal how RNA interactions are crucial for preventing neurotoxic droplets.
Cardiorespiratory Coupling Response to Chest Wall Strapping in Chronic Obstructive Pulmonary Disease
Authors: Peng Maggie , Amaza Iliya , Eberlein Michael , Bates Melissa
Published: May 2022

Chronic obstructive pulmonary disease (COPD) is a common lung disease characterized by limited expiratory airflow due to lung destruction or airway obstruction. Chest wall strapping (CWS) is a potential therapy technique that restricts the upper abdomen, inducing breathing at lower lung volumes and opening small airways. Although the respiratory impacts of CWS have been examined, the relationship between CWS and the cardiovascular system, which is closely tied to ventilation, has not been well‐established. Additionally, COPD patients have reduced autonomic nervous system activity, which may be reflected in cardiorespiratory coupling. Cardiorespiratory coupling causes increased heart rate during inhalation to accommodate for increased blood return. The strength of this relationship was evaluated during CWS in five COPD and six control patients by analyzing heart rate changes during ventilation cycles. Heart rate and ventilatory data were collected during baseline, CWS, and post‐CWS removal periods. Average heart rate during each inhalation and exhalation was plotted against tidal volume, with the slope of the linear regression representing the strength of the cardiorespiratory coupling relationship (BPM/L). Multi‐factor ANOVA statistics showed a significantly diminished strength in COPD patients compared to control (p < 0.001). There was no significant change in either group post‐CWS, suggesting that the same‐sized breaths will cause the heart to respond similarly regardless of the total lung capacity. A potential consequence of diminished cardiorespiratory coupling in COPD patients is strain on the right side of the heart, which then has to pump larger stroke volumes. Blunted cardiorespiratory coupling is not further impaired by CWS.
Inhibition of CHRM1 reverts docetaxel resistance in castration‐resistant prostate cancer
Authors: Wang Jing , Bland Tyler , Wei Jing , Pu Tianjie , Lin Tzu‐Ping , Wu Boyang
Published: May 2022

Background

Chemotherapy remains the major treatment option for castration‐resistant prostate cancer (CRPC). Docetaxel (DTX)‐based chemotherapy provides significant survival benefit in patients with CRPC. However, the clinical utility of DTX is compromised when resistance is encountered. Therefore, a more thorough understanding of mechanisms underlying DTX resistance may potentially improve survival in patients with CRPC. The objective of this study is to investigate the role and mechanism of cholinergic receptor muscarinic 1 (CHRM1) activation upon chemotherapy resistance in PC and the efficacy of the CHRM1 antagonist dicyclomine (Dic) in treating DTX‐resistant PC. The hypothesis is that CHRM1 is essential for developing chemotherapy resistance in PC cells and driving survival and growth of DTX‐resistant PC cells.

Methods

The CHRM1 expression levels and their association with DTX resistance were determined in PC clinical specimens and cultured cells. Human PC cells subjected to stable knockdown or overexpression of CHRM1 were evaluated for their response to DTX by cell proliferation, colony and tumorsphere formation, and TUNEL assays. Synergistic growth inhibition by combination treatment with DTX and Dic was assessed by cell viability and tumorgenicity assays.

Results

CHRM1 expression was elevated under DTX treatment in CRPC 22Rv1 and DU145 cells, which was paralleled by higher CHRM1 expression in these cells after acquiring resistance to DTX (22Rv1DTXR and DU145DTXR). Immunohistochemical analyses revealed higher CHRM1 protein levels in longitudinally collected tumor tissues post‐ versus pre‐chemotherapy from a PC patient cohort. Enforced expression of CHRM1 increased DTX IC50 by up to 5‐fold, along with reduced cell apoptosis marker c‐PARP1 expression, in 22Rv1 and DU145 cells. Conversely, silencing of CHRM1 in 22Rv1DTXR cells decreased cell proliferation, invasion, and colony and tumorsphere formation. CHRM1 inactivation with Dic in combination with DTX resulted in more cell death of both DTX‐sensitive (22Rv1 and DU145) and ‐resistant (22Rv1DTXR, DU145DTXR and PC‐3DTXR) cells compared to either treatment alone. Further, Dic restored DTX sensitivity to suppress the growth of DTX‐resistant cells and xenograft tumors (22Rv1DTXR and PC‐3DTXR). Mechanistically, CHRM1 activation upregulated phosphoprotein levels of ERK, JNK and p38, which all are key players in the MAPK pathway and potential mediators driving chemoresistance in cancer cells, while Dic treatment reduced their phosphorylation levels in 22Rv1DTXR and DU145DTXR cells.

Conclusion

Our results suggest that CHRM1 contributes to the development of DTX resistance in CRPC, which can be reversed by genetic or pharmacologic inactivation of CHRM1, providing new insights into developing a CHRM1‐targeted therapeutic strategy for overcoming DTX‐based chemotherapy resistance in CRPC.
Molecular Signalling Cascade Time in Sympathetic Stimulation of Angiotensin Renin Axis and Its Correlation With Ultra Low Frequency (ULF) of Heart Rate Variability (HRV)
Authors: Acero Mondragon Edward J. , Cortes Cabrera Jorge Armando , Gallego Correa John Jairo , Urdaneta Paredes Danial Antonio , Chaustre Nieto Luis Carlos , Serrano Contreras Samuel Ernesto , Vasquez Bazurto Maria Paula , Zabala Rodíguez Laura Daniela
Published: May 2022

INTRODUCTION

In Heart Rate Variability (HRV), Ultra Low Frequency Band (ULF) ranges between 0.003 to 0.03 Hz. It is believed that metabolism, body temperature as well as the renin‐angiotensin axis is related to ULF. Within juxtaglomerular cells, all the different biochemical events times may be quantified. Such chain of events start from B1 receptor activation, including renin secretion all the way through angiotensinogen conversion to angiotensin II. The present study’s objective is to correlate sympathetic stimulation of juxtaglomerular cells to ULF bands found on heart rate variability.

MATERIAL AND METHODS

Google Scholar and National Library of Medicine databases were used to search articles published between 1990 and 2021. The time involved in juxtaglomerular cell intracellular signalling after sympathetic stimulation was searched, with the combination of the following terms: 1) ((“Titer (TI) = (FRET), TI = (BRET), TI = (EDANS) and ‘Adrenergic B1 activation time’ ‘Gs protein activation’ ‘Adenylate Cyclase activation time’ ‘cAMP Activation time’ ‘Activation time Protein Kinase A “Renin Secretion Time” Angiotensinogen Conversion Time “Angiotensin I Conversion Time” Angiotensin II Conversion Time “In Vitro Cell Model ‘Y)].” Inclusion criteria for each study consisted in Ligand‐Receptor and renin related secretion cellular cascade timelines. Studies that satisfied the previously mentioned conditions were included. Times for events that ranged from B1 adrenergic receptor activation, renin secretion all the way to angiotensinogen conversion to angiotensin II were estimated. Such times were added together and later converted in Hertz using time and frequency inverse relationship.

RESULTS

Juxtaglomerular cell signalling cascade involving B1 adrenergic receptor activation and its related renin secretion summed around 300 seconds. On the other hand, renin activation time to angiotensin 2 was 89 seconds. Once these values were added, total time was 389 seconds that once converted to frequency yielded a value of 0,00257 Hz. Such value is close to the lower limit of ULF bands that range between 0,003 and 0,03 Hz in HRV domain frequency.

CONCLUSIONS

The biochemical sequence of events that start from renin secretion and ends in angiotensin II production takes approximately 389 seconds. Such time once converted, through the inverse relationship of time and frequency, is 0,00257 Hertz. Such value suggests that ULF band with HRV domain may correspond to renin angiotensin axis activity.
Milk protein and fat in Saanen goats
Authors: Mudruk Semen , Bakhta Alesia , Karpenko Larisa , Kozitcyna Anna , Polistovskaia Polina
Published: May 2022

Dairy industry is one of the leading branches of animal husbandry. Dairy goat breeding is becoming more and more relevant each day; it has already acquired special significance for the agro‐industry of Russian Federation. In addition to the outstanding milk productivity of Saanen goats, it is worth noting the versatility of the milk processing, which is important for farms additionally engaged in manufacturing of goat cheese and other dairy products. Therefore, the chemical composition of goat milk depending on various physiological states of goats is of study interest.

Research was conducted in North‐Western region of Russian Federation, in Saanen goats. Five groups included 30 Saanen goats each, from 1stto 5th lactations. The milk samples were taken once during control milking. Levels of milk fat and proteins were assessed. Results presented as mean±standard error of the mean. Student's t‐test was used after proving normal distribution. Level of significance is labeled as p<0.05.

As a result of presented study, it was revealed that, in goats of 1st lactation the milk protein level was 3.1±0.22% and milk fat was 3.8±0.98%, in animals of 2nd lactation the milk protein level was 2.9±0.21% and fat was 3.7±0.52%, in goats of 3rd lactation the protein level was 2.8±0.23% and milk fat was 3.3±0.58%, in animals of 4th lactation the milk protein level was 2.1±0.23% and fat was 3.3±0.53%, in animals of 5th lactation, the protein level was 3.1±0.27% and milk fat was 3.5±0.48%. The average values according to these indicators were 3.0±0.24% for milk protein, 3.6±0.73% for milk fat. When analyzing the milk protein obtained results level he minimum values were noted in goats of 4th lactation. This marker was significantly (p<0.05) lower by 30% relative to the average protein content in milk. The maximum amount of milk protein was observed in goats of 1st and 5th lactations. There were no significant differences in animals of 1st, 2nd, 3rd and 5th lactations when compared to the average value for the entire lactation period. When analyzing the amount of milk fat it was noted that there were no significant differences in the goats of 1st, 2nd, 3rd and 4th lactations. There was a tendency to milk fat decreasing in animals of 3rd, 4thand 5th lactations relative to the average values for the entire lactation period, and a tendency to milk fat increasing in goats of 1st and 2nd lactations by 5.5% and 2.77%, respectively.

Consequently this data is important to be taken into consideration keeping goats and planning further processing of milk depending on the prevalent number of presented goats in the herd.
Monitoring of vitamin‐mineral metabolism’ indicators in cows of different period of lactation
Authors: Plemyashov Kirill , Korochkina Elena
Published: May 2022

The vitamin‐mineral metabolism of cows has an important role in the body’s functional status of productive animals. Lactation cycle is the stress in the body of animals. For this reason the continuous provision by vitamins and minerals of organism is actual and important point. The aim of the study was to research the influence of vitamin‐mineral bolus’ supplmentation with prolong action on vitamin and mineral metabolism of the cows in the different time of lactation cycle. The studies were carried out in one of the breeding farms of the Leningrad region (Russia). Holstein cows were used aged from 3 to 5 years. The milk production of cows was 5625 kg. 5 groups were formed (n=5 by each). The cows of the 1st experimental group have received two boluses All – mineral plus (the compound: Mg, Zn, Cu, Co, Se, I, Vitamin A, D3, E, Holland Animal Care, Holland), 2d experimental group– two boluses Uno Biotin (the compound: Mg, Zn, Cu, Co, Se, I, Vitamin A, D3, E, Holland Animal Care, Holland), 3d experimental group ‐ one Cattle bolus with Iodine (the compound: Cu, Co, Se, I, Telsol Limited, United Kingdom), 4th experimental group – one Cattle Bullet (the compound: Mg, Zn, Cu, Co, Se, I, Vitamin A, D3, E, Holland Animal Care, Holland). The 5th group was control with ordinary ration vitamin‐mineral diet. All cows of experimental groups have got boluses one time which was 60 days before calving. The prolongation of all boluses was 120 days after receiving except bolus All‐mineral plus. The prolongation of their was 180 days. Blood were collected 4 times: 60 days (before bolus’supplementation) before calving, in the dry period (36 days before calving), in the fresh period (7 days after calving) and in the lactation period (81 days after calving). Serum was analyzed for zinc, copper, iodine and carotin. We were used Student's t‐tests to study differences between groups. Thus, we have observed the statistically increasing of copper’s level in the blood of cows of 1st group in the dry period in 1.7 times (p=0,000479) compared with the data beforebolus’supplementation. The level of zinc was increased in 1.6 times (p=0,000143) in the cows of 1st group in the dry time and in 1.4 times (p=0.039139) in the cows of 2d group in the same time compared with data before bolus’supplementation. The level of iodine was increased by 1.5 in the cows of 1st group in the dry time compared with data beforebolus’supplementation. There was 5,00±0,20 mcg %. The carotin’s level increased in 3.4 times (p=0,003647) in the cows of 1st group in the dry period and in 3.9 times (p=0,012621) in the fresh period compared with data before bolus’supplementation. This point was statistically increased in 1.4 times (p=0,049364) in the cows of 2d group in the dry period and it was 2,71±0,18 mg %. So, we can assume that the supplementation of vitamin‐mineral boluses with prolongation by 180 days has a positive influence on the vitamin and mineral metabolism of cows in the dry and fresh periods.
Histological features of the mitral valve in Anglo‐Nubian goats
Authors: Melnikov Sergey , Shchipakin Mikhail , Khvatov Victor , Bylinskaya Darya , Vasiljev Dmitriy , Stratonov Andrey
Published: May 2022

The mitral valve, also known as the left atrioventricular valve, is located in the left side of the heart between the left atrium and the left ventricle. The valve structures are involved in the completion of the pulmonary circulation and contribute to the continuous circulation of blood in the animal's body. Knowledge of the valve leaflets' histological features in the breed aspect allows the most accurate and individual approach to diagnosing, preventing, and treating cardiac pathologies. Our study aims to research the histological features of the left atrioventricular valve structure in the Anglo‐Nubian goats. The material for the study was ten corpses of Anglo‐Nubian goats aged from one to two years. The material was fixed with a 10% formalin solution for 24 hours, and then it was paraffin‐embedded. The prepared sections were stained with hematoxylin and eosin. The mitral valve of the Anglo‐Nubian goats has anterior and posterior leaflets in its composition, also labeled septal and mural. Each leaflet consists of fibrous plates formed from dense connective tissue with a high amount of cells. The leaflets have three layers in their structure: the atrialis, which faces the left atrial cavity; the fibrosa, which faces the left ventricular cavity; and the spongiosa, which is located between the previous two. The thickness of the atrialis layer of mitral valve leaflets in examined Anglo‐Nubian goats is 175.42±19.0 μm. This layer is characterized by a smooth surface, which is covered with one layer of endothelial cells. Some areas of the histological section are superimposed like scales on top of each other. In contrast to the atrialis, the fibrosa layer has a rough surface due to high outgrowths on it, from which the chordae tendineae begin. The thickness of the layer in the Anglo‐Nubian goats averages 126, 35±16.3 μm. As well as atrialis, fibrosa is covered with one layer of endothelial cells and has a similar histological picture. The spongiosa layer of the mitral valve leaflets is consisting of loose connective tissue. Large amount of glycosaminoglycan is observed on a histological section of this layer. The thickness the spongiosa is 286±46.8 μm. Based on the results of our research, the following conclusions can be drawn: The leaflets of Anglo‐Nubian goats' mitral valve consist of three layers, of which the spongiosa layer is most developed. A large amount of glycosaminoglycan was found on the histological section of the spongiosa layer, which confirms the development of valve leaflets from the cardiac skeleton.
Nuclear Location Bias of HCAR1 Drives Cancer Malignancy by Multiple Routes
Authors: Mohammad Nezhady Mohammad Ali , Cagnone Gael , Chemtob Sylvain
Published: May 2022

Introduction

G‐Protein Coupled Receptors (GPCR) are virtually involved in all physiological processes. HCAR1 (GPR81), as a GPCR, is endogenously activated by lactate and has been shown to promote cancer malignancy via higher level of glycolysis due to Warburg effect. Its expression level is highly elevated in many cancers and negatively correlates with patient’s prognosis. However, its mechanism of action is not well understood.

On the other hand, nuclear localization of several GPCRs have been described albeit it is unusual feature for them. Additionally, it has been shown that nuclear GPCRs can perform functions distinct from their cell surface counterparts in vivo. Here we demonstrate HCAR1 has a nuclear localization and this localization pattern promotes cancer malignancy by multiple routes.

Methods and Results

We determined HCAR1 nuclear localization pattern by cell fractionation, immunofluorescence confocal imaging and TEM. Site‐directed mutagenesis showed ICL3 and phosphorylation of C‐terminal domains are required for nuclear localization. We also demonstrated that this localization is ligand independent and there is a pool of nuclear HCAR1 (N‐HCAR1) in the cells. We show N‐HCAR1 induces intra‐nuclear signaling through Gi and Gßγ by WB and ELISA leading to increased phosphorylation of nuclear AKT and ERK resulting in increased cancer cell survival and proliferation. Our ChIP‐sequencing data shows N‐HCAR1 binds to the genes regulating transcription and promoting expression of genes involved in cell migration and we validated this in cellulo proving N‐HCAR1 promotes migration. We identified N‐HCAR1 interactome by Bio‐ID coupled with mass spectrometry and found, it interacts with proteins involved in ribosome biogenesis, translation and DNA damage repair and our experimental data demonstrates that specifically the N‐HCAR1 promotes these three process in cellulo. Additionally, we identified the transcriptomic signature of N‐HCAR1 and showed it regulated a larger gene network than its plasma membrane counterpart. Concordantly, our in vivo tumor xenografts and tail vein injections proves that tumors without N‐HCAR1 have smaller size and tumor mass and lower metastatic rate as well.

Conclusion

Here we show an unusual localization of a GPCR in the nucleus and provide evidence that N‐HCAR1 specifically promotes various hallmarks of cancer malignancy. The effect of N‐HCAR1 is validate in vivo in tumor xenografts as well. Understanding these mechanisms can provide targets and cues for therapeutic developments.
Effects of Increasing Endocannabinoid 2‐AG Tone on Sensorimotor Gating and Anxiety‐like behavior
Authors: Lampkin Alexius , Bakshi Vaishali
Published: May 2022

The cannabinoid system has been widely studied for its ability to modulate multiple behavioral functions, including addiction‐like behaviors. Our study aimed to determine if increasing endocannabinoid availability would alter sensorimotor gating and anxiety‐like behavior in rats. Both types of behavior are aberrant in psychiatric illnesses such as post‐traumatic stress disorder (PTSD). Sensorimotor gating is a mechanism which protects the brain from being overloaded by sensory stimuli to allow for normal information processing. The functionality of sensorimotor gating can be assessed using the prepulse inhibition (PPI) of startle paradigm, in which a weak prestimulus prior to a startling stimulus inhibits the resultant startle response. We were particularly interested in pursuing an understudied approach, augmentation of endogenous endocannabinoid ‘tone’ via inhibition of monoacylglycerol lipase (MAGL), the breakdown enzyme for the endocannabinoid 2‐AG. To inhibit MAGL, we administered the JZL184 to increase brain levels of 2‐AG. JZL184 did not affect PPI on its own but did reverse the PPI deficit that was produced by the psychotomimetic, amphetamine. Because JZL184 reversed sensorimotor gating deficits, we wanted to further determine if JZL184 possesses anxiety‐reducing effects. The elevated plus maze (EPM) assesses rats’ innate anxiety‐like state by measuring relative time spent exploring novel (fear‐inducing) spaces (the open arms of the EPM) versus remaining in spaces that feel ‘safe’ (the closed arms of the EPM, which are darker and enclosed by walls). While JZL184 (16 mg/kg) did not increase open arm entries or time, it did reduce time spent in the closed arms. Moreover, it increased center time (an intermediate space between the open and closed arms that is moderately fear‐inducing) and stretch‐attends (rats stretch their bodies so that their head can peer into the open arm while their hindpaws remain in a closed arm). Both of those measures are ethological indices of novel‐environment exploration. Taken together, these findings indicate that JZL184 produces a subtle anxiety‐reducing effect in the EPM. To our knowledge, we are the first to demonstrate this effect via ethological measures of EPM exploration. Similarly, our data are the first to show effects of JZL184 on PPI. These results raise the possibility that modulation of the endocannabinoid system, specifically 2‐AG tone, could be a novel pharmacological mechanism to regulate cognitive and emotional processes related to psychiatric disorders like PTSD.
Integration of Biomedical and Clinical Sciences into Collaborative, Team‐taught Sessions
Authors: Ward Peter J. , Asano Shinichi , Benmerzouga Imaan , Eleazer Courtney , Landvoigt Maple , Martin Kathleen , Scopa Kelso Rebecca , Stover Kristin
Published: May 2022

In 2012, the West Virginia School of Osteopathic Medicine shifted to a new curriculum, and the faculty was tasked with integrating course content. Initially, we re‐shuffled existing lectures; however, this was ineffective as the lectures had been designed to fit in a different context and sequence. The objective of the faulty in each department shifted to deep integration of their content to create and an effective and coordinated flow of information. We hypothesized that by creating new sessions we could minimize unplanned repetition of content. We generated two main integrated modalities: 1. integrating biomedical sciences (e.g., Physiology and Histology), 2. integrating biomedical and clinical sciences (e.g., Embryology and Pediatrics). The teaching faculty met and went through several rounds of reorganizing their content. In the case of integrated biomedical science sessions, we found it helpful to proceed in a macroscopic to microscopic manner. For example, we discussed the anatomy of the upper airway and how the muscles of respiration change the pleural pressure physiologically, then proceeding to the microscopic organization of the alveoli and how diffusion is accomplished according to Fick’s law before drawing students' attention to the histologic makeup of the parenchyma of the lungs and how spirometry findings can illustrate the effects of COPD or fibrosis. In the pediatric/embryology sessions, we found the most effective method was to examine embryology chronologically while interspersing the clinical presentation and management of problems associated with each developmental stage. For example, development of the gastrointestinal tract was presented chronologically but was deliberately interspersed with input from clinical faculty about how malformations arise and how they present clinically. In conclusion, by creating team‐taught, collaborative, integrative sessions, we were able to eliminate redundancy of content while presenting the biomedical sciences in a context that made their clinical importance obvious to the learners. Planning and rehearsal were key in making these sessions effective, and we found the transitions between speakers to be the most important aspect. An additional benefit was the student feedback regarding these sessions has been overwhelmingly positive, and they appreciate the relevancy of these sessions to their education. We have started looking at a “next step” in integration, writing integrated exam questions that blend the topic areas.
Interferon‐γ Induces Bcl3 Expression in Ovarian Cancer Cells by JAK1/STAT1 Signaling
Authors: Gaire Bijaya , Padmanabhan Sveta , Vancura Ales , Vancurova Ivana
Published: May 2022

The proto‐oncogene Bcl3 is an atypical member of IkB family, which regulates expression of a number of NFkB target genes, including anti‐apoptotic genes, cytokines, and immune checkpoints. Bcl3 expression is increased in many types of human cancers and is associated with increased tumor growth and malignant potential. Interferon‐γ (IFNγ) is a pleiotropic cytokine that has been used in cancer treatment; in addition, its expression is induced in response to radiation therapy and immune checkpoint blockade during cancer treatment. We have recently shown that IFNγ induces Bcl3 expression in ovarian cancer cells, resulting in their increased proliferation, migration, and invasion. In this study, we have investigated the mechanisms of how IFNγ induces the Bcl3 expression in ovarian cancer cells. Our results demonstrate that the IFNγ‐induced Bcl3 expression is dependent on JAK1 and STAT1 signaling, as well as on p65 NFkB. In addition, we have identified a potential interferon regulatory factor 1 (IRF1) binding site in human Bcl3 promoter and found that the IFNγ‐induced Bcl3 expression in ovarian cancer cells is associated with an increased occupancy of IRF1, Ser‐727 phosphorylated STAT1 and acetylated histone H3 at the IRF1 binding site in Bcl3 promoter. These findings identify Bcl3 as a novel target of IFNγ/JAK1/STAT1 signaling and suggest that targeting the JAK1/STAT1 pathway may suppress the IFNγ‐induced Bcl3 expression in ovarian cancer.
Characterizing the transcriptionally‐activated ensembles recruited by cocaine in the nucleus accumbens
Authors: Thibeault Kimberly , Lopez Alberto , Dryer Joseph , Kondev Veronika , Patel Sachin , Siciliano Cody , Calipari Erin
Published: May 2022

While significant effort has been made to understand the neural basis of addiction, it remains unclear exactly how the brain controls motivated behaviors and how drugs of abuse alter these neural systems to control drug taking and seeking. The goal of this study was to understand how cocaine and associated stimuli are encoded in the brain and how neuronal activation in response to cocaine acts to drive drug seeking. In a given brain region, only a small percentage of cells are activated to any stimulus ‐ termed an “ensemble”. Here we aimed to determine if ensembles transcriptionally activated by cocaine in the nucleus accumbens (NAc) – a brain region central to reward encoding ‐ were capable of driving motivated behaviors in isolation. Using transgenic animals that allowed for the temporally specific tagging of ensembles with optical tools, we were able to record from, identify, and manipulate the neural ensembles that are selectively activated by cocaine experience. Using conditional viruses in combination with these transgenic mice, we expressed opsins in cocaine‐ or saline‐ (for control) activated neuronal ensembles within the NAc. A fiberoptic was placed above the region and animals performed operant conditioning tasks where a nosepoke triggered a laser pulse that would reactivate or inhibit cocaine‐activated ensembles to determine their role in motivated behaviors. Mice were highly motivated to optically reactivate the ensembles that were activated by the first cocaine experience, highlighted by their high rates of responding for reactivation. Repeated cocaine injections in these animals did not change the reinforcing efficacy of ensemble activation, suggesting that the increases in motivated behavior that occur following repeated cocaine exposure may be due to the recruitment of a novel ensemble. Indeed, via immunohistochemistry, we find minimal overlap between the neurons activated by the first cocaine experience and the tenth. Using a similar approach as described above, we conditionally deleted the cocaine‐ or saline‐activated ensembles using a diphtheria toxin and had the mice perform cocaine self‐administration to assess changes in acquisition, performance, or intake of cocaine. We observed no changes in cocaine self‐administration behavior, suggesting that, while the cocaine ensemble is sufficient for reinforcing motivated behavior, it is not necessary for reinforcement learning about cocaine or cocaine‐associated cues. We also show that repeated cocaine exposure recruits a new population of neurons to guide behavior. Overall, we hypothesize that the neural ensemble initially recruited by cocaine may be important for reinforcement learning, but that other ensembles of neurons in the nucleus accumbens may be recruited in order to mediate the transition to addiction.
Effects of stimulants and HIV Proteins on Pyroptosis and Apoptosis Pathways in Human Brain Microvascular Endothelial (hBMVEC)
Authors: Ikedife Joyce C.
Published: May 2022

Human brain microvascular endothelial cells (hBMVEC), together with astrocytes and pericytes, construct the blood‐brain barrier (BBB) to protect the brain from toxins and pathogens via paracellular, transcellular, transporter, and extracellular matrices, and tight junction proteins. The BBB also inhibits many therapeutic substances from entering the brain, making it challenging to design drugs that can migrate through the BBB effectively. The objective of the project was to understand the detrimental effects of psychological stimulants (cocaine, THC, Amphetamine) and HIV Tat proteins on the integrity of the BBB, and to inform the potential therapeutic strategies. The hBMVEC cell culture was treated with the stimulants and two different constructs of HIV Tat proteins. Western blot and RT‐qPCR techniques were used to detect pyroptosis and apoptosis proteins that could lead to endothelial cell death. We found that proteins involved in apoptosis (PEA‐15, caspase‐3, and caspase‐8) were more expressed and upregulated comparing to proteins involved in pyroptosis (caspase‐1, gasdermin D) and cytokine proteins (IL‐1β, IL‐6, IL‐18).
Structure Activity Relationship and Biological Evaluation of Small Molecule Antagonists for G‐Protein Coupled Receptor Associated with Neuropathic Pain
Authors: Olayide Israel I. , Braden Kathryn , Latzo Nicholas , Salvemini Daniela , Arnatt Christopher
Published: May 2022

Opioids are commonly prescribed for the treatment of neuropathic pain; however, their high rates of physical dependency and addiction raise concern. Given their adverse side effects, there is a need for new therapeutics that target non‐opioid receptors. GPR183 (Epstein‐Barr induced gene 2, EBI2) is a G‐protein coupled receptor (GPCR) that plays a role in the transduction of neuropathic pain. SAE‐14 is a novel molecule that is able to antagonize, or inhibit GPR183; thus, preventing the transmission of pain. To gain a better understanding of which substituents improve the binding between SAE‐14 and GPR‐183, it is necessary to synthesize several analogs of the molecule. We have developed a structure‐activity‐relationship for GPR183 antagonism and the results from this gives a better understanding of the functional groups that are necessary for GPR183 antagonism and improve to the compound’s potential therapeutic for neuropathic pain treatment.
Periprostatic Adipose Tissue Thromboinflammation Drives Early Prostatic Neoplastic Alterations in a Rat Model of Mild Metabolic Dysfunction
Authors: Alzaim Ibrahim , Darwiche Nadine , Abou‐kheir Wassim , El‐Yazbi Ahmed F.
Published: May 2022

Emerging evidence links periprostatic adipose tissue (PPAT) inflammation, a consequence of metabolic impairment, and certain features of prostate cancer (PCa) aggressiveness. AT thromboinflammation, mediated by the proteolytic activation of protease‐activated receptors (PARs) particularly by thrombin and Factor Xa (FXa), drives AT dysfunction. Nevertheless, the contribution of PPAT thromboinflammation to early prostatic neoplastic alterations has not been investigated. Here, we utilized a non‐obese prediabetic rat model to assess prostatic neoplasia in association with PPAT thromboinflammation. Mild hypercaloric diet (MHC)‐fed rats exhibited hyperinsulinemia, hypertriglyceridemia, a lower metabolic efficiency, and a higher fat/lean ratio in comparison to regular chow‐fed rats. When compared to regular chow‐fed rats, MHC‐fed rats exhibited PPAT with larger adipocytes, enhanced oxidative stress and increased immune cell influx. Concurrently, prostates from MHC‐fed rats displayed higher number of prostatic intraepithelial neoplasia (PIN), enhanced expression of the proliferation markers Ki67 and PCNA, enhanced expression of cytokeratin 8 (CK8), pronounced fibrosis and increased immune cell influx. Further profiling of macrophage polarization in PPAT and the prostate is underway. Importantly, these prostatic and adipocytic manifestations were attenuated in MHC‐fed rats treated with non‐hemorrhagic doses of rivaroxaban, a direct FXa inhibitor. Ongoing in vitro investigation demonstrates that rivaroxaban reduces the migratory potential of murine prostate cancer cells (Plum‐AD); and the effect of rivaroxaban on the colony forming and sphere forming capacities of Plum‐AD cells are being studied. Further investigation is ongoing to delineate the molecular underpinnings of rivaroxaban‐associated amelioration of PPAT inflammation and the associated prostatic neoplastic alterations.
Stimulation of Osteoblastic Bone Metastasis by Canine Prostate Cancer
Authors: Yuan Shiyu , Kantake Noriko , Elshafae Said M. , Rosol Thomas J.
Published: May 2022

Prostate cancer metastases primarily localize in the bone where they induce a unique osteoblastic response. The mechanisms are not well understood; however, they depend on a complex interplay between prostate cancer cells and the cellular microenvironment in the bone marrow. We hypothesize that prostate cancer cells secreted biological factors that induce new bone formation and either induce or stimulate osteoclastic bone resorption to permit growth of bone metastasis. Dogs are the only other species that develop spontaneous prostate cancer with bone metastases in addition to men. Previously, our laboratory has developed three canine prostate cancer cell lines that mimic human osteoblastic bone metastases when injected into the left cardiac ventricle of nude mice. This study then investigated the effect of three canine prostate cancer cell lines (Ace‐1, Probasco, and LuMa) on bone formation and resorption. Mouse calvaria were treated with conditioned medium (CM) collected from cell lines. A novel calcein uptake assay was developed to measure bone formation and mineralization in vitro. Ace‐1 CM induced predominantly bone resorption in calvaria, while Probasco CM induced marked bone formation, mineralization, and healing of calvaria defects. The expression of osteoblast‐related genes in calvaria showed that Probasco CM stimulated the maturation and differentiation of osteoblasts and inhibited osteoclastogenesis. Both bone modeling and remodeling were involved in Probasco‐induced bone formation and mineralization by inhibiting remodeling with zoledronic acid. RNAseq results from CM treated osteoblasts indicated that the WNT signaling pathway was activated by Probasco compared to either control or Ace‐1. In addition, Calvaria treated with Probasco+hDKK‐1 CM, Probasco transduced with human DKK‐1 cDNA (a WNT antagonist), showed decreased bone formation. In vivo, Our preliminary data showed that DKK‐1 significantly increased the growth of Probasco tumors after intratibial injection of Probasco+hDKK‐1 cells in nude mice, which suggested the autocrine effect of DKK‐1 on tumor cells. In conclusion, Probasco cells induced bone formation and mineralization in vitro that depended on the WNT signaling pathway. Probasco cells serve as a valuable model for studying the mechanisms of osteoblastic bone metastasis in prostate cancer.
Gut Microbial and Metabolic Signatures are Altered in Adolescents with Type 1 Diabetes
Authors: Pon Velayutham Anandh Babu , Mokhtari Pari , Metos Julie M. , Jambal Puujee , Shankar Kartik
Published: May 2022

Background

Evidence suggests that the pathogenesis and development of Type 1 Diabetes (T1D) are driven by both genetic predisposition and immune dysfunction which may be influenced by gut microbiota. The present study aimed to investigate the composition and functional pathways of the gut microbiome in adolescents with T1D, determine the association between gut microbes and clinical and dietary factors in T1D, and identify features predictive of T1D.

Methods

Healthy adolescents (n=35) and adolescents with T1D (n=30) were recruited and microbial profiling in participants’ stool samples was performed using deep shotgun metagenomic sequencing. The bioBakery3 pipeline (Kneaddata, Metaphlan3 and HUMAnN) was used to analyze the reads to assign taxonomy and functional annotations. The differences in taxonomic abundances were identified using Linear Discriminant Analysis (LDA) Effect Size (LEfSe) and ecological measures (diversity) were assessed using standard methods. The association between the abundance of specific bacterial species and the metadata (HbA1C, skin carotenoids, diet and physical activity) was assessed by performing Spearman’s correlation. Random forest modeling (using the MUVR package) was used to identify features predictive of T1D.

Results

Adolescents with T1D exhibited a significant increase in Simpson α‐diversity index as compared to healthy adolescents. β‐diversity was similar between the two groups. LEfSe indicated a significant difference in the microbial community at different taxonomic levels between the two groups. Seven taxa were enriched in adolescents with T1D at the species level that includes Alistipes Shahii, Asaccharobacter celatus, Blautia obeum, Coprococcus eutactus, Coprobacillus cateniformis, Clostridium symbiosum, and Eggerthella lenta. Three species Alistripes Putredins, Dialister invisus, and Eubacterium ventriosum were enriched in healthy adolescents. Further, 33 out of 400 microbial metabolic pathways were significantly different between the two groups. Importantly, biosynthesis of amino acids, vitamins, enzyme cofactors, and electron carrier were downregulated whereas fermentation pathways were upregulated in adolescents with T1D as compared to healthy adolescents. Association analysis indicated that specific bacterial species are correlated with HbA1C, skin carotenoids, dietary factors and physical activity. Most of the bacterial species associated with these measures were different between the two groups. MUVR identified bacterial taxa predictive of T1D status (0.82 sensitivity and 0.63 specificity). The top features in using genus level abundance included Coprococcus, Faecalibaterium and Streptococcus.

Conclusion

The present study indicates an extensive alteration in the composition and functional capacity of gut microbiota in adolescents with T1D providing new insight into microbial and metabolic signatures in adolescents with T1D.