bluuebunny/crossref_metadata_2025 huggingface dataset:
Decade Long Study in HIV‐1+ Puerto Ricans Finds Reduction in Average Drug Resistance Per Individual, Changes in Antiretroviral Medication Resistances and Shifts in Mutations Expression
Authors: Cubano Luis , Roger María Luisa , Rishishwar Lavanya , Sepúlveda‐Torres Lycely del C. , Ríos‐Olivares Eddy , Jordan I. King , Boukli Nawal
Published: November 2023

The objective of this study was to determine if changes in the patterns of expression of antiretroviral resistance and associated mutations could be detected in a population of HIV‐1+ Puerto Ricans.

The data collected from 2002 to 2011 was obtained from 4,475 (1,397 female, 2,581 male and 497 anonymous) patients. The samples were analyzed using the TRUGENE HIV‐1 Genotyping Kit and the OpenGene DNA Sequencing System.

The average number of resistances per individual decreased from 6.0 in 2002 to 2.1 in 2011. The average number of reverse transcriptase mutations decreased from 2.8 to 0.9. In contrast the average number of protease inhibitor mutations increased from 2.5 to 3.3.

The most prevalent antiretroviral medication resistances changed from Zalcitabine, which was the dominant one from 2002 to 2006, to Emtricitabine from 2007 – 2010 with declining percentages during that time from 44.7% to 27.9%. In 2010 the predominant resistance was a tied between Efavirenz and Nevirapine (18%). In 2011 the predominant resistance was Saquinavir + Rinonavir at 27.4%

The most prevalent reverse transcriptase mutation was M184V but its percentage of expression decreased from 47.3% in 2004 to 10.1% in 2010. In the year 2011 K103N became the dominant reserve transcriptase mutation in the population examined with 16.1%. In 2011 the percent of M184V was 8.1%.

The most prevalent protease inhibitor mutation during the decade was L63P. Similarly to M184V, L63P percentage of expression decreased but not as dramatically with a decrease from 73.9% in 2002 to 62.9% in 2011.

Data show a descending trend for most of the mutations, accompanied by an increase in the incidence of minority mutations. This was reflected by a shift in the most prevalent antiretroviral medication resistances.

Support or Funding Information

This research was supported by grant G12MD007583 from the National Institutes of Health and grants P031M105050 and P031S130068 from the US Department of Education.
Gαo Proteins Mediate the Antihyperalgesic and Antidepressant‐like Effects, but not the Convulsive Effects, Produced by Delta Opioid Receptor Activation
Authors: Dripps Isaac , Neubig Richard , Rice Kenner , Jutkiewicz Emily
Published: November 2023

The delta opioid receptor (DOPr) is a member of the opioid receptor family of G protein‐coupled receptors (GPCRs). Activation of DOPr induces antihyperalgesia and antidepressant‐like effects in animal models. However, some DOPr agonists cause convulsions, hindering their development as therapeutics in humans. Regulator of G protein signaling (RGS) proteins act as negative modulators of GPCR signaling. Our lab has previously demonstrated that RGS4 acts as a negatively modulator of DOPr‐mediated antihyperalgesia and antidepressant‐like effects without impacting convulsions. These findings indicate that the antihyperalgesic and antidepressant‐like effects of DOPr are G protein‐mediated. However, DOPr is known to couple to multiple Gi/o proteins. Therefore, we sought to determine which G protein mediates these behaviors and whether that G protein also plays a role in generating DOPr‐mediated convulsions.

To this end, we compared behaviors induced by the DOPr agonist SNC80 in Gαo RGS‐insensitive (RGSi) knock‐in, and Gαo heterozygote knockout mice. DOPr‐mediated antihyperalgesia was evaluated using a CNS‐mediated nitroglycerin‐induced thermal hyperalgesia assay. Antidepressant‐like effects were measured using the forced swim test. The potency of SNC80 to produce antihyperalgesia was significantly increased in the Gαo RGSi mice. SNC80 also produced antidepressant‐like effects in the Gαo RGSi mice with enhanced potency and efficacy. In the Gαo heterozygote knockout mice, SNC80‐induced antihyperalgesia was completely abolished while antidepressant‐like effects in the forced swim test were unaltered. There were no changes in the frequency, duration, or time of onset of SNC80‐induced convulsions in either mouse strain.

Taken together, our data demonstrate that DOPr‐mediated antihyperalgesia and antidepressant‐like effects require Gαo. A 50% loss in Gαo was sufficient to abolish the antihyperalgesic, but not the antidepressant‐like effects of DOPr, indicating that DOPr‐mediated antidepressant‐like effects had a lower efficacy requirement than the antihyperalgesic effects. Ostensibly, DOPr‐mediated convulsions are not regulated by Gαo. However, it is possible that the efficacy requirement for convulsions is low enough that they are unaffected by loss of RGS regulation or a 50% reduction in Gαo. DOPr‐mediated convulsions may also be generated by a G protein‐independent signaling mechanism such as the β‐arrestin pathway. Future studies will focus on investigating the efficacy requirements of DOPr‐mediated behaviors and characterizing these behaviors in β‐arrestin2 knockout mice.

Support or Funding Information

Research support was provided by start‐up funds to EMJ.
Peptidomimetics Exhibiting Mu Opioid Receptor (MOPr) Agonist/Delta Opioid Receptor (DOPr) Antagonist Activity as Novel Pain Therapeutics
Authors: Griggs Nicholas , Bender Aaron , Gao Chao , Anand Jessica , Traynor John , Mosberg Henry
Published: November 2023

Drug discovery and development of opioid ligands has largely favored highly selective agonists/antagonists for a single opioid receptor (OPr); opioid analgesics, such as morphine, are primarily selective for and activate the mu opioid receptor (MOPr). However, evidence suggests an interaction with other OPrs may be beneficial in MOPr‐mediated analgesia. In particular, there is evidence that simultaneous activation of MOPr in the absence, or with inhibition, of the delta opioid receptor (DOPr) can reduce the development of morphine tolerance and dependence. Thus one strategy is to design single compounds, which bind to and concurrently activate MOPr while inhibiting DOPr, as potential therapeutics for the improved clinical management of pain. Here we report on the pharmacological evaluation of a series of compounds with a tetrahydroquinoline (THQ) core and with chemically diverse substituents that alter steric and electronic characteristics of the molecules. The compounds are peptidomimetics based on key elements of opioid peptides vital for activity but that have small molecule‐like features to provide for bioavailability, blood brain barrier permeability, and duration of action in vivo. The peptidomimetics were characterized in vitro by ligand binding to provide affinity (Ki) values and for potency and relative efficacy (EC50, % stimulation compared to standard agonist) using the degree of incorporation of GTPγ35S into G proteins in membranes from cells expressing MOPr or DOPr. Peptidomimetics showing an appropriate profile of similar binding affinity for MOPr and DOPR but acting as MOPr agonists and DOPr antagonists were evaluated for their antinociceptive activity and tolerance liability using the mouse warm water tail withdrawal assay. Certain compounds showed maximal analgesic efficacy that lasted for more than 3 hours as well as a reduction in the development of tolerance. Analysis of in vitro and in vivo data and docking peptidomimetics into the MOPr active and DOPr inactive crystal structures revealed structure‐activity relationships to guide further chemical synthesis and drug development of improved analogues with MOPr agonist/DOPr antagonist activity.

Support or Funding Information

Funded by DA‐03910 and the Pharmacological Sciences Training Program (NIGMS‐GM007767)
The 5‐HT4 Receptor Agonists Prucalopride and BIMU8 Suppress Abdominal Nociception in Animal Models
Authors: Lyubashina Olga , Busygina Irina , Sivachenko Ivan , Panteleev Sergey
Published: November 2023

5‐HT4 receptor agonists are primarily used as prokinetic agents to treat patients with irritable bowel syndrome and other functional gastrointestinal disorders associated with hypomotility. Meanwhile, considering the relatively high expression and a neuromodulatory role of 5‐HT4 receptors within brain regions involved in the endogenous control of nociception, it is conceivable that ligands for these receptors, along with prokinetic action on the gut, can affect central processing and perception of abdominal pain. The aim of the presented study was to evaluate the effects of two 5‐HT4 agonists—prucalopride and BIMU8—on abdominal nociception in wakeful mongrel dogs (N = 5) and urethane‐anaesthetized Wistar rats (N = 34). In both models, for inducing abdominal nociception the mechanical distension of the large intestine by inflating a rubber balloon to a pressure of 80 mmHg was applied. In conscious dogs, the rectal distension‐evoked visceromotor (abdominal muscle contractions) and cardioautonomic (changes in heart rate) responses were monitored by electromyographic and electrocardiographic recordings using needle and cutaneous electrodes respectively. These reactions were evaluated before and at fixed intervals for 90 min after intramuscular administration of prucalopride. In anaesthetized rats, the single neuron activity within the caudal ventrolateral medulla was extracellularly recorded by tungsten microelectrodes. The ongoing and colorectal distension‐induced neuronal discharges were quantified prior to and at frequent intervals for 90 min after intravenous injection of BIMU8. In dogs, the nociceptive rectal distension caused pronounced abdominal muscle contractions and tachycardia. Intramuscular administration of prucalopride at doses of 0.05 and 0.1 mg/kg resulted in suppression of both reactions. The effect occurred in 30 min after the injection and was maximal at the end of the recording (90 min). The visceral analgesic action was more pronounced and stable when the drug dose of 0.1 mg/kg was used. In neurophysiological experiments in rats, intravenous BIMU8 at doses of 1 and 2 mg/kg caused substantial decreases in both the ongoing activity of the tested medullary neurons and their reactions to nociceptive colorectal distension. The inhibitory effect occurred in 5 min after the drug administration and maintained for at least 90 min. The dose of 2 mg/kg was significantly more effective at suppressing neuronal activity than the dose of 1 mg/kg. The drug's antinociceptive action was blocked by preliminary intracerebroventricular administration of a selective 5‐HT4 receptor antagonist GR113808. Thus, the 5‐HT4 receptor agonists studied—prucalopride and BIMU8—are able to suppress abdominal nociception in conscious and neurophysiological animal preparations respectively. The visceral analgesic effect can be a component of the therapeutic action of 5‐HT4 receptor agonists in pain‐related functional bowel disorders.
A biased mu‐opioid receptor positive allosteric modulator that does not alter receptor regulation
Authors: Livingston Kathryn E , Traynor John R
Published: November 2023

Currently used opioid analgesics compete with endogenous opioid peptide ligands by binding to the orthosteric site of the mu opioid receptor (MOPr). Activation of this G protein‐coupled receptor (GPCR) affords analgesia but also causes respiratory depression, sedation, constipation, and euphoria leading to a high addictive liability. Our lab is currently pursuing the idea that positive allosteric modulators of MOPr (mu‐PAMs) may be a way to treat pain with a better therapeutic profile. Such compounds, including the lead ligand BMS‐986122, enhance the affinity, potency, and efficacy of opioid agonists to bind MOPr and activate G protein. On the other hand, BMS‐986122 does not stimulate G protein activation, as measured by [35S]GTPγS binding in the absence of an orthosteric ligand. Here, we sought to determine if BMS‐986122 activates non‐G protein mediated pathways downstream of MOPr and to investigate the long‐term consequences of BMS‐986122 exposure on the regulation of MOPr in vitro using C6 rat glioma cells expressing rat MOPr. The results demonstrate that BMS‐986122 initiates phosphorylation of ERK1/2 in a pertussis‐toxin insensitive manner supporting a G protein‐independent mechanism. BMS‐986122 alone did not cause any long‐term adaptations in MOPr signaling. Moreover, in contrast to its ability to enhance agonist‐mediated activation of G protein, BMS‐986122 failed to enhance agonist‐induced desensitization of MOPr as measured by receptor downregulation, receptor phosphorylation, and loss of receptor‐mediated signaling following long‐term agonist exposure. Development of biased allosteric ligands at GPCRs represents a novel avenue for analgesic drug development. Future work will determine the physiological consequences of this biased PAM in mouse models of pain.

Support or Funding Information

Funded by DA039997
Evaluation of Analytical Parameters by Mineralization by Dry Method for Lead Detection in Urine
Authors: Giolo Janaina Aparecida , da Silva Cunha Tiritan Gabrieli , Neves Fernando Tozze Alves
Published: November 2023

Lead is a metal with no physiological function in the human body, but with harmful effects known since ancient times. Into human body, it is metabolized and complexed to macromolecules, being directly absorbed, distributed in several compartments including blood, soft tissues (kidney, bone marrow, liver and brain), and mainly mineralized tissues (bones and teeth), and it is subsequently excreted, remaining in the body for large periods due to its prolonged half‐life. Both blood and urine may represent suitable samples for analysis, depending of convenience and in accordance with the purpose of the toxicological study performed. For acute poisoning cases, the urine represents the sample choice for screening tests. Among the various methodologies for the toxicological screening tests of metals, the mineralization by the dry process technique is considered the most suitable, especially for lead. In literature, there are few analytical parameters described regarding the sensitivity and specificity of this method. Thus, this study aimed to evaluate the analytical parameters of the mineralization by dry method in urine samples, containing five different concentrations of lead (10; 5,0; 2,5; 1,0 and 0,1 ppm). For the tests, we used blank urine samples contaminated with different forms of lead (nitrate, acetate and chloride) in the concentrations established. The trials were carried out in duplicate by the dry mineralization method with 100 mg (0.1g) of potassium iodide. Our results show that it is possible to verify that this methodology is not specific as to the types of lead compounds tested because all of them had a positive reaction to the test (yellowish color); however, there was lower staining intensity in the lead nitrate. Regarding the sensitivity, it was possible to detect the presence of lead in samples from the concentration of 0.1 ppm in all kinds of compounds analyzed. Thus, we conclude the mineralization method can be considered as a sensitive method, but not so specific regarding the differentiation of lead compound types present in a sample. The formation of the yellowish color in reaction to the potassium iodide makes it specific compared to the other heavy metals.

Support or Funding Information

University of Sagrado Coracao (USC), Bauru, Sao Paulo, Brazil.
Activity of hBSEP transporter expressed in human and insect cells
Authors: wright stephen , Wilkerson Matt , Li Jibin , Hidalgo Ismael
Published: November 2023

Membrane transporters involved in the recycling of bile acids are also very important for the disposition of some drugs in the liver. The opportunity to measure the interaction between transporters such as BSEP (ABCB11) and potential drug candidates is very valuable to the drug development process. Overexpression systems, such as the Sf9 insect platform, have been widely used to monitor most of the ABC efflux transporters of clinical interest. While heterologous expression of the human proteins in insect cell membranes often gives high levels of activity some proteins need the lipid environment of their native cells to achieve optimal activity. Attempts by some researchers to overcome this limitation by supplementing the insect membranes with missing components such as cholesterol have met modest success. Although expression in human‐derived cell lines can be challenging, a relevant platform to predict how a human protein might function in vivo should express the protein in a system that mimics, as close as possible, its native environment. We have compared the activity of hBSEP in inverted vesicles derived from either Sf9 or human embryonic kidney (HEK) cells and found that transporter activity is higher in HEK membrane‐derived vesicles but the overall sensitivity to inhibitors (as indicated by IC50 values) is approximately the same.
Organic Cation Transporters in Drug Interactions with Methamphetamine and Metabolites
Authors: Wagner David James , Wang Joanne
Published: November 2023

Methamphetamine (Meth) is the second most commonly abused illicit drug with marijuana being the first. High levels of Meth abuse are reported in individuals receiving treatment for HIV and hepatitis who may be receiving multiple drugs for treatment. Organic cation transporters (OCTs) and multidrug and toxin extrusion transporters (MATEs) are known to be involved in tissue distribution and elimination of a number of these antiretrovirals including lamivudine and zalcitabine. Meth and its main metabolites (amphetamine and p‐hydroxyMeth) are organic cations but their potential effects and drug interaction potential with the OCTs and MATEs are poorly understood. The goal of this study is to determine if Meth and metabolites inhibit renal and hepatic OCTs and MATEs, which may lead to clinically significant drug‐drug interactions with antiretroviral drugs in Meth users. The inhibitory effects of Meth and metabolites are determined using uptake assays in cell lines stably expressing human OCT and MATE transporters and the drug interaction potential is assessed by comparing IC50 values with reported Meth/metabolite concentrations in drug abusers. Meth inhibited hOCT1, hOCT2, hOCT3, hMATE1, and hMATE2k at clinically relevant drug concentrations with the greatest potencies against hOCT1 (IC50 = 21 ± 8.8 μM) and hOCT2 (IC50 = 17 ± 6.8 μM). Amphetamine showed a similar inhibition potency and pattern towards these transporters. p‐hydroxyMeth is a more potent inhibitor of hOCT3, exhibiting approximately 8‐fold greater potency than Meth but was approximately 5‐fold less potent toward hOCT2. Based on the reported blood concentrations of Meth and metabolites in illicit users and the FDA guidance on transporter‐mediated DDIs, our studies suggest potential in vivo drug interactions of Meth with drug substrates of the OCTs and MATEs. This information can be considered when prescribing medications to suspected or known abusers of methamphetamine to mitigate the risk of increased toxicity or reduced therapeutic efficacy.

Support or Funding Information

This study was supported by NIH grants P01DA032507 and T32GM07750.
Tamoxifen affects PLCg1 expression in ER+ breast cancer cells
Authors: Vergne Ralphdy , Ortiz Carmen , Godoy Lenin , Matta Jaime
Published: November 2023

Phospholipase C gamma 1 (PLCg1) can be found in the cytoplasm and in the nucleus of the cells and is involved in signal transduction pathways and proliferation. Previous data from our lab shows that PLCg1 expression in breast cancer cells is higher in comparison to non‐tumorigenic cells. Also, oxidative stress‐induced PLCg1 activation enhances cell survival. Many anticancer drugs can cause oxidative stress as a side effect, such as tamoxifen. Tamoxifen is currently used for the treatment of both early and advanced estrogen receptor positive (ER+) breast cancer in pre‐ and post‐menopausal women. In addition, patients using tamoxifen have a higher probability to develop oxidative stress by consequence of the drug. Our aim is to compare and analyze PLCg1 expression in an ER+ breast cancer cell line (MCF‐7) under the effect of tamoxifen. Our hypothesis is that there will be a high expression of PLCg1 in cells treated with tamoxifen in comparison to untreated cells. To achieve this, we cultured MCF‐7 cells using complete media without phenol red and 4μM tamoxifen. After 24hrs, cells were lysed and protein and RNA were extracted. To determine PLCg1 expression we used a Taqman gene expression assay using GAPDH as housekeeping gene. Our results shows that PLCg1 expression was two times higher in MCF‐7 cells treated with tamoxifen compared to untreated cells at 24 hrs. However, this effect was not statistically significant. This preliminary data indicates that the PLCg1 expression might be involved in ER modulation. Currently, we are developing an MCF‐7 cell line resistant to tamoxifen to further study the changes in PLCg1 protein and gene expression upon acquired resistance.

Support or Funding Information

This work was supported by the UPR‐PRISE Program Grant #R25GM096955 and NIGMS‐NIH grants R25GM082406 (CO), and 9SC1CA182846‐04. We also acknowledge the support of the Molecular and Genomics Core at Ponce Health Sciences University (NIMHD Grant MD007579).
In vitro Basis for Cell Line‐Dependent Interaction between Sorafenib and Dasatinib in Human Non‐Small Cell Lung Carcinoma Cell Lines
Authors: Zhou Qingyu , Choksi Riya , Guo Xiaofang
Published: November 2023

Background

Recent clinical trials have demonstrated a better‐than‐average therapeutic outcome of the multi‐kinase inhibitor sorafenib in patients with chemorefractory non‐small cell lung carcinomas (NSCLCs) as well as the inevitable treatment failure due to the reduced tumor sensitivity to sorafenib over time. In search of strategies to overcome the acquired sorafenib resistance, we evaluated the feasibility of combining sorafenib with dasatinib for the treatment of NSCLCs with different sensitivity to sorafenib.

Methods

The sorafenib and dasatinib combination was evaluated in A549, H358 and H1915 human NSCLC cell lines. The effect of sorafenib and dasatinib alone and in combination on tumor cell growth was determined by the MTT assay using fixed molar concentration ratio design, inhibitory sigmoid Emax model and combination index (CI) theorem. Western blot analysis of changes in targeted signaling pathways were performed to assess sorafenib and dasatinib interactions.

Results

Combination of sorafenib with dasatinib inhibited the in vitro growth of A549, H358 and H1915 cells in an additive or synergistic fashion based on the CI values as defined by Chou and Talalay (mean CI = 0.99, 1.04 and 0.28 for A549, H358 and H1915, respectively). The improved in‐vitro sorafenib cytotoxicity by the combination in those NSCLC cells was associated with the increased inhibition of the AKT/GSK3 and MEK/ERK/p90RSK pathways.

Conclusions

Sorafenib and dasatinib exhibited increased cytotoxicity in NSCLC cell lines when used in combination. Given that the H1915 cell line was originally developed from a brain metastasis from a primary lung carcinoma, future studies are warranted for further assessment of the beneficial therapeutic effect of sorafenib and dasatinib combination in an in vivo orthotopic model of lung cancer metastasis to brain.

Support or Funding Information

This work is supported by the University of South Florida College of Pharmacy start‐up fund and Internal Seed Grant (2012–2013) to Q.Z.
Overexpression of ATP‐binding cassette transporter ABCG2 mediates acquired resistance LDC000067, a selective inhibitor of cyclin‐dependent kinase CDK9
Authors: Lin Ming‐Lu , Wu Chung‐Pu
Published: November 2023

The serine/threonine cyclin‐dependent kinase 9 (CDK9) is a critical elongation factor for RNA polymerase II‐directed transcription, which is important for cancer cell survival. Recently, CDK9 has emerged as a valid therapeutic target for cancer, and many CDK9 inhibitors have been investigated. LDC000067 is a novel small molecule compound that selectively and potently inhibits the activity of CDK9. However, alike most therapeutic drugs, the risk of developing drug resistance to LDC000067 presents a therapeutic challenge. Since the overexpression of ATP‐binding cassette (ABC) drug transporter ABCG2 is one of the most common mechanisms of drug resistance, we thus investigated the effect of ABCG2 on the cellular efficacy of LDC000067. In this study, we discovered that ABCG2‐overexpressing cancer cells and cells transfected with wild‐type ABCG2 were highly resistant to LDC000067. The inhibitory activity of LDC000067 was significantly reduced by the function of ABCG2. More importantly, we found that by inhibiting the function of ABCG2, the reduced apoptosis and sensitivity to LDC000067 treatment in ABCG2‐overexpressing cells can be restored. In conclusion, the development of ABCG2‐mediated drug resistance to LDC000067 in cancer poses a significant therapeutic challenge, and a combination therapy of LDC000067 with a modulator of ABCG2 should be further investigated as a potential treatment approach.
Radiofrequency Renal Denervation Improves Left Ventricular Function and Hemodynamics Through Modulation of Natriuretic Plasma Peptides in the Setting of Heart Failure
Authors: Polhemus David J , Gao Juan , Scarborough Amy , Trivedi Rishi , Bradley Jessica , Goodchild Traci , Smart Frank , Kapusta Daniel R , Lefer David
Published: November 2023

Background

Overactivity of the sympathetic nervous system plays a critical role in the pathogenesis of heart failure. We investigated the effects of renal sympathetic nerve denervation (RDN) on left ventricular (LV) function and vasoactive natriuretic peptide levels in a rodent model of ischemic heart failure.

Methods

Spontaneously hypertensive rats (SHR) were subjected to 45 min of coronary artery ligation and reperfusion for 12 weeks. At 4 weeks post‐reperfusion, SHR underwent either bilateral radiofrequency RDN (n=9) or sham RDN (n=8) procedure. Every 2 weeks, transthoracic echocardiogram was performed using Vevo 2100 (Visualsonics). LV function was analyzed from EKV™ (ECG‐Gated Kilohertz Visualization) generated M‐mode long‐axis images. 12 weeks following reperfusion, plasma brain natriuretic peptide‐32 (BNP) and atrial natriuretic peptide (ANP) were quantified using ELISA technique.

Results

RDN preserved LV ejection fraction (49.4 ± 2.6 vs. 32.1 ± 2.6%, p < 0.05) and LV stroke volume (318 ± 24.5 vs. 215 ± 17.8 μl, p < 0.05) at 12 weeks compared to sham. Heart weight/tibia length was also markedly reduced in the RDN group (41 ± 2.4 vs. 49 ± 1.2 mg/mm, p < 0.05). Despite reduced cardiac enlargement, Plasma BNP levels (377.7 ± 12.0 vs. 166.6 ± 37.3 pg/ml, p < 0.05) and ANP levels (297.0 ± 25.7 vs. 57.0 ± 12.4, p < 0.05) were significantly elevated in the RDN treated group compared to sham.

Conclusion

Radiofrequency RDN improved LV function, reduced remodeling and significantly increased circulating cardioprotective natriuretic peptides. We conclude that RDN exerts beneficial effects in preventing and treating heart failure following coronary ischemia and reperfusion; in part via increasing circulating cardiprotective natriuretic peptides.

Support or Funding Information

P30GM106392 to DK
Ontogeny Influences Pharmacological‐Induced L‐Type Ca2+ Dependent Contractility of Pulmonary Arteries in Sheep
Authors: Wolfe Craig , Brunelle Alexander , Longo Lawrence , Wilson Sean M
Published: November 2023

We and others have shown that K+‐depolarization induced pulmonary arterial (PA) contraction is due to the combined activation of L‐type Ca2+ channels (CaL) and Rho kinase (ROK). In fetal sheep we demonstrated that CaL inhibition with nifedipine (NIF) reduced 125 mM K+‐mediated contraction up to about one‐half, with ROK contributing the remainder, and that the relationship between these two pathways was preserved following antenatal chronic hypoxia (CH). To better define the relationship between these pathways and the impact of antenatal CH, we tested the hypothesis that direct activation of CaL would elicit a NIF sensitive contraction and this contraction would be similar in arteries from fetus or adult. This was examined using wire‐myography approaches on small diameter PA isolated from late gestation fetal (~140 days) or adult sheep. Our results show that direct activation of CaL with 1 mM FPL 64176 (FPL) elicited a contraction that was similar to the maximum K+‐induced depolarization (TKmax) in fetus but was reduced in adults. The contraction due to 1 mM Bay K 8644 (BayK) was reduced relative to High K in both fetus and adult. In fetal and adult PA 10 mM NIF reduced FPL and BayK, but not High K‐induced contraction back to baseline. These data illustrate that there are subtle differences between FPL and BayK induced, NIF‐sensitive CaL‐dependent contraction with ontogeny and that direct CaL activation causes PA constriction in a distinct way compared to K+‐depolarization.

Support or Funding Information

Support from NIH
Western Medicine vs. Oriental Medicine in Hypertension Therapy: Case Study
Authors: Tulp Orien L , Chow Tin , Konyk Carla M , Einstein George P
Published: November 2023

Background

Hypertension [HTN] is a key risk factor for cardiovascular morbidity and mortality and is a leading cause of end‐stage renal disease; approximately 7.1 million deaths per year may be directly attributed to poor control of blood pressure. Advances in the diagnosis and treatment of HTN have had a major impact on management of this chronic disease. To determine the comparative effectiveness of conventional Western treatment vs drug free Oriental Therapy [OT] for HTN, we reviewed the data from two independent studies.

Methods and Research

The prevention and therapy include multi variables that cause or contribute to the development of HTN. The research review is based on a review of the diagnosis and therapy of Western and Oriental medicine. The patients were administered the ARB Sartan [SR] in the EU and Amlodipine [AD] in USA. For patients in China the Chinese oriental medicine of Wu Cao Si Wu Tang formula was administered. For the treatment of high blood pressure [HTN] with this protocol, 21 were men and 9 women, age range 30–67 years old, with mean systolic pressure [SBP] of 160 mmHg or above, and the mean diastolic pressure [DBP] of 95 mmHg upon admission to the study.

Results

The patients used SR in EU and AD in USA stabilized the systolic and diastolic blood pressure effectively, however with frequent side effects. The Chinese formula resulted in an equally effective outcome, defined as resulting SBP of 141 mmHg or less and DBP of 90 or less with normalization of BP and disappearance of symptoms of HTN, similar to that which was observed on the altiplano where abundant daily exercise and traditional Bolivian diet contributed to a healthy lifestyle [Tulp, 1994].

Conclusion

The therapy by SR in EU and AD in USA to stabilize the blood pressure was equally effective to OT. Common side effects of SR included blurred vision, that was reported to be disruptive for driving and reading by patients. The AD side effects, which can be potentially life‐threatening, included hives, swelling of the throat, and difficulty breathing. In contrast, the Wu Cao Si Wu Tang formula is effective and no side effects were observed and appears to be a worthy alternative to present day conventional Western approaches for the treatment of HTN.

Support or Funding Information

Supported by Institutional Resources of USAT and the Einstein Medical Institute
CARDIOVASCULAR EFFECTS INDUCED BY LINALOOL AFTER SUBCHRONIC TREATMENT OF SPONTANEOUSLY HYPERTENSIVE RATS
Authors: Camargo Samuel Barbosa , Simões Letícia Oliveira , de Azevedo Medeiros Carla Fiama , de Melo Jesus Anderson , de Jesus Silva Jailton , Fregoneze Josmara Bartolomei , Silva Darizy Flavia
Published: November 2023

Introduction

Linalool (LIN) is an alcoholic monoterpene with biological activities previously described. This study in order to evaluate the cardiovascular effects in vivo subchronic treatment of spontaneously hypertensive rats (SHR) is focusing on antihypertensive properties of linalool.

Methods

All experiments were performed with SHR aged 9–12 weeks and (−)‐linalool (≥ 97 % purity) was solubilized in cremophor in a ratio of 2:1 and diluted with physiological saline solution (100 mg/kg) and controls solutions captopril (30 mg/kg) and vehicle (saline + cremophor) were administered daily for 21 days by orogastric injections. Each 5 days the arterial pressure and the heart rate were measured by tail cuff method. At the end of treatment, mesenteric artery was isolated to reactivity tests and the heart was removed and placed in an oven to dry at 60° C, for 24 to 48 hours, to obtain the dry weight.

Results

Animals treated with linalool, vehicle and captopril showed no statistically significant differences at the levels of water consumption and feeding and no difference in progressive increasing body weight between the three groups. None of the treated groups had significant changes in the values of heart rate. Animals treated with vehicle showed a progressive increase in the mean arterial pressure (MAP). Captopril induced a significant decrease at MAP, from the fifth day of treatment and (−)‐linalool was able to maintain the initial pressure values, avoiding the progression of hypertension and demonstrating a significant reduction in the blood pressure at fifteenth day. The subchronic treatment with linalool decreases the cardiac mass index similar to that found in animals treated with captopril when compared to the negative control group. Finally, linalool induced beneficial changes in vascular responsiveness.

Conclusion

These results suggest that linalool, after 21 days of subchronic treatment of spontaneously hypertensive rats, demonstrated antihypertensive potential inducing the maintenance and decrease of the mean arterial pressure with no changes in heart rate, being able to reduce cardiac hypertrophy, to decrease vascular reactivity to a vasoconstrictor agent phenylephrine (PHE) and to increase pharmacological potency to a vasodilator agent sodium nitropusside (SNP).

3012 ‐ ASPET Cardiovascular Pharmacology – General/Other

Support or Funding Information

Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Lipocalin‐2 Deficiency Protects against Aldosterone‐Induced Hypertension and Organ Damages by Inhibiting Mineralocorticoid Receptor Signaling
Authors: Yang Kangmin , Luo Cuiting , Cheung Bernard MY. , Félétou Michel , Vilaine Jean‐Paul , Villeneuve Nicole , Xu Aimin , Vanhoutte Paul M. , Wang Yu
Published: November 2023

Lipocalin‐2, an adipokine dysregulated in cardiometabolic syndrome, is causatively involved in the development of obesity‐related endothelial dysfunction and hypertension. The present study investigated the contribution of lipocalin‐2 to aldosterone‐induced pathologies involving mineralocorticoid receptors (MR). Mice were treated during four weeks with vehicle, angiotensin II or challenged with the combination of aldosterone plus uninephrectomy and administration of 1% saline in the drinking water (ANS). In mice without lipocalin‐2 (LKO), the gene expression levels of MR in heart, kidney, adipose tissue and aorta are significantly lower than those of wild‐type (WT) controls. ANS significantly increased arterial blood pressures in WT but not LKO mice. Moreover, lipocalin‐2 deficiency prevented ANS‐induced fibrotic damages in heart and kidneys. Treatment with the MR antagonist, spironolactone, protected mice against ANS‐induced hypertension and organ damages to a similar extent as lipocalin‐2‐deficiency. On the other hand, angiotensin II‐induced elevation of blood pressure was not affected significantly by either lipocalin‐2‐deficiency or MR inhibition. The mechanistic involvement of lipocalin‐2 in aldosterone‐stimulated MR expression and/or activation was further investigated. Collectively, the results demonstrate that lipocalin‐2 plays a critical role in the cardiovascular damage induced by activation of the aldosterone‐MR signaling axis.

Support or Funding Information

This work was supported by the grants from Hong Kong Research Grant Council [General Research (17121714, HKU 763312 and HKU 780613M), Collaborative Research Fund (C7055‐14G)] and an educational grant from Les Laboratories Servier (RS100503).
PRR activates canonical Wnt signaling pathway during pregnancy‐induced hypertension in rats
Authors: Lopez Pedro , Ramirez‐Montero Claudia , Anguiano‐Robledo Liliana , Avila‐Ramirez Massiel A. , Hernandez‐Campos Maria Elena
Published: November 2023

Pregnancy is a physiological condition with profound hemodynamic changes. Pregnancy‐induced hypertension (PIH) is a common and severe complication. There is no a clear cause, but several systems have been involved. Renin Angiotensin Aldosterone System (RAAS) is one of them. Prorrenin renin receptor (PRR) a component of this system could play a role in PIH physiopathology. This receptor has been associated to Wnt signaling pathway, both canonical and non‐canonical. In this work we studied the posible activation of Wnt pathway by PRR in a PIH model in rats. We submitted female Wistar rats to sub‐renal aortic coarctation (SRAC) to induce PIH, set in control, pregnant‐non SRAC and pregnant‐SRAC groups. We measured blood pressure, proteinuria and offspring weight and size. We determined b‐catenin, DVL‐1, HDA4, JNK‐1and PKCa in aorta, heart, and kidney by immunoblot. We found and increased expression of PRR, b‐catenin, HDA4 and DVL‐1 in cardiac, renal and aortic tissue from PIH animals. Similar findings were shown using immunohistochemistry on same tissues. No significant changes were found in JNK‐1 and PKCa in any tissue. These results suggest that canonical Wnt pathway seems to be activated in the studied tissues during PIH, whereas non‐canonical pathway seems not to have a role in it.

Support or Funding Information

Supported by SIP20150457 grant
Motivating Young Investigators to Pursue Cancer Research Careers and Training
Authors: Kidd LaCreis Renee , Hein David W.
Published: November 2023

University of Louisville Cancer Education Program (CEP), funded by a National Cancer Institute (NCI) grant, seeks to educate, and motivate undergraduate and professional students to pursue further training and future careers in cancer research. Over the past 4 years, 121 participants completed the program, including 42% females, 16% African‐Americans, 12% Asians, 2.4% Hispanic/Latino, 2.4% Indian, and 2.4% mixed. All trainees formulated a cancer research project in collaboration with their faculty mentor engaged in cancer research at the Brown Cancer Center. Laboratory research training/mentoring was supplemented with cancer‐related seminars and professional development activities. A blinded survey was used to assess participants’ perspective on program components. Participants ranked the programmatic activities as “poor”, “could be better”, “neutral”, “good”, and “excellent”. Over the past four years, there was a significant improvement in the participants’ interest in cancer research after completing the program (P < 0.0001). The interest level in cancer research improved, remained the same, or decreased among 46.5%, 45.4%, and 8.2%, respectively. A majority of the students perceived the overall program experience (86.4%), quality of the faculty mentor (88.5%), helpfulness of lab mentor (86.3%), and quality of the professional development activities (84.4%) as “good to excellent”. R25 alumni have presented their research at national/international scientific meetings (n = 22), published their research in scholarly journals, and continue their cancer research/oncology training at various post‐graduate degree programs/institutions (n = 32). UofL's Cancer Education Program is well poised to produce the next generation of cancer researchers.

Support or Funding Information

The program is supported by NCI grant R25‐CA134283.
Outcomes of a Dedicated Pharmacy Student Summer Research Program
Authors: Brandl Katharina , Adler David , Kelly Carolyn , Taylor Palmer , Best Brookie
Published: November 2023

Objectives

This study investigated the impact of an optional 12‐week Summer Research Program on the publication outcomes and satisfaction with the required research projects of Pharm.D. students at the Skaggs School of Pharmacy and Pharmaceutical Sciences at UC San Diego (SSPPS).

Methods

PubMed and Google searches provided student publications, and satisfaction surveys submitted by students and preceptors provided their perceptions of the research project value.

Results

Of the studied cohort, the 130 students who fulfilled the requirement through the optional Summer Research Program provided 61 full‐text manuscripts and 113 abstracts. The 305 students who chose the standard pathway provided 35 full‐text manuscripts and 34 abstracts. Students and preceptors in both pathways agreed or strongly agreed that the research project was a valuable experience.

Conclusions

The 12‐week intensive Summer Research Program improved the publication rate of pharmacy students and provided a high overall satisfaction with this independent learning experience.
Distinct EC coupling mechanism drives spatial control of vascular tone in cerebral arteries
Authors: Zechariah Anil , Hald Bjorn Olav , Mazumdar Neil G. , Welsh Donald G.
Published: November 2023

Cerebral arterial networks consist of thousands of segments that work together and in isolation, tuning the magnitude and distribution of brain blood flow. The existence of spatially distinct responses suggests that distinct excitation‐contraction (EC) coupling mechanisms are encoded into each arterial segment. This study sought to define which EC coupling mechanisms enable cerebral arterial segments to work together and in isolation, from one another. Using both theoretical and experimental approaches, we show that multi‐segmental responses require the sharing of charge via gap junctions. This process of “electrical communication” enables arteries to equilibrate membrane potential, synchronize Ca2+ concentration and coordinate myosin light chain phosphorylation among large populations of smooth muscle cells. In contrast, isolated vessel behavior was independent of charge sharing and electromechanical coupling. It depended upon: 1) the voltage‐independent generation of Ca2+ waves; and 2) direct catalytic subunit inhibition of myosin light chain phosphatase. The latter was found to be driven by protein kinase C and the phosphorylation of CPI‐17. Subsequent work revealed that the presence of distinct EC coupling mechanisms enabled multi‐segmental and isolated responses to simultaneously but independently co‐exist in a single artery. As such, the magnitude and distribution of brain blood flow can be concurrently tuned depending on the nature of the originating stimulus. Translationally, these findings indicate that the ‘one‐size‐fits‐all” strategy to treating blood flow abnormalities should be reconsidered and tailored to the underlying dysfunction in EC coupling.

Support or Funding Information

This work is supported by an operating grant to Dr Welsh from the Natural Science and Engineering Council of Canada (NSERC). Dr Welsh is the Roraback Chair in Neuroscience and Vascular Biology at the University of Western Ontario. A. Zechariah is supported by Canadian Institutes of Health Research (CIHR) Postdoctoral scholarship, Alberta Innovates Postdoctoral scholarship and Eyes High Postdoctoral fellowship.
Disruption In PKC‐mediated K Channel Regulation Contributes To Enhanced Serotonin Mediated Carotid Vessel Constriction from the Fawn Hooded Hypertensive Rat
Authors: Kaur Gurjit , Pabbidi Mallikarjuna
Published: November 2023

Previous studies indicate Fawn Hooded Hypertensive (FHH) rats exhibit cerebral damage due to diminished cerebrovascular myogenic function associated with elevated BK channel function. Introgression of the 2.4 Mbp region of BN chromosome 1 into the FHH rat restored myogenic response and BK channel function in a FHH.1BN congenic rat. However, results obtained from resistance vessels cannot be extrapolated to conduit vessels due to structural and functional differences. Thus, the present study addresses the hypothesis that carotid vessels of FHH rats exhibit altered serotonin (5‐HT)‐mediated contraction compared to FHH.1BN rats. Using tension myography to measure conduit vascular function, we found that carotid vessels of FHH rats exhibited greater 5‐HT‐mediated contraction than FHH.1BN rats. Activation of BK channels (NS1619: 5 to 30μM) relaxed carotid vessels to a lesser extent (1.17 times less) in FHH rats. On the other hand, inhibition of BK channels (Paxilline or Iberiotoxin: 100nM) had no effect in either strain. Inhibition of voltage gated calcium channels (VGCC, Diltiazem: 10μM) or store operated calcium (SOC) channels (2‐APB: 75μM) relaxed carotid vessels similarly in both strains. Furthermore, PKC activation (PMA: 100nM) or inhibition (GF109203X: 3μM) increased and decreased 5‐HT‐mediated constriction respectively in FHH.1BN rats but had minimal effect in FHH rats. These results suggest that a mutation in the genes located in 2.4Mbp region in FHH rats contribute to a disruption in PKC and its regulation of K channels, causing enhanced 5‐HT‐mediated contraction.

Support or Funding Information

This research has received funding support from the American Heart Association, Scientist Development Grant (13SDG14000006) and the IOSP Undergraduate Summer Research Fellowship supported by the American Physiological Society and a grant from the National Science Foundation Integrative Organismal Systems Award No. IOS‐ 1238831.
Measurements of Blood Flow, Tissue Oxygen Tension, and Oxygen Consumption in Rat Skeletal Muscle during Recovery from Bouts of Brief, Transient Ischemia
Authors: Dodhy Sami C , Golub Aleksander S , Pittman Roland N
Published: November 2023

In resting muscle, blood flow is regulated to meet the demand for O2 by the respiring tissue. A locally induced ischemia/reperfusion protocol was utilized to investigate the effects of transient ischemia (5, 15, 30, and 60 s duration) in rat skeletal muscle. The results will be utilized in a model that relates blood flow, interstitial oxygenation, and oxygen consumption. We anticipate that this study will aid in furthering our understanding of the local regulation of blood flow and tissue oxygenation. 29 spinotrapezius muscles from male Sprague‐Dawley rats were surgically exteriorized for intravital microscopy. Interstitial PO2 (PISFO2) and arteriolar PO2 (PaO2) measurements were made using phosphorescence quenching microscopy (PQM). Red cell velocity was measured using Optical Doppler velocimetry and used to calculate blood flow (Q). Oxygen consumption (VO2) was measured with a quasi‐continuous, flash‐synchronized, rapidly pressurizing airbag system to both initiate ischemia and sample the rate of change in oxygen tension (dPO2/dt). The average baseline value of arteriolar blood flow was measured as 5.8±0.8 μl/min; as expected, reactive hyperemia was observed during the recovery from ischemia. Flow recovery (dQ/dt) was fastest [3.88±0.64 (μl·min−1)/s] after 60 seconds of ischemia. Average baseline PISFO2 was 62±4 mmHg; PISFO2 decreased to 75%, 34%, 10% and 2% of baseline following 5, 15, 30 and 60 s of ischemia, respectively. Up to 30 s of ischemia, the rate of recovery of PISFO2 steadily increased as the ischemic duration increased; however, it dropped off following 60 seconds of ischemia. The rate of recovery of PISFO2 was fastest (4.2±0.7 mmHg/s) following 30 seconds of ischemia. Average baseline VO2 was 120±19 nl O2/cm3·s and it increased above baseline during the early part of the recovery period. As is consistent with our current understanding, the time course of recovery of PISFO2 represents a balance between oxygen delivery, proportional to the local arteriolar tissue perfusion, and local oxygen consumption. When the ischemic duration increased, the interstitial oxygen tension decreased. An assumption was made in the modeling process that the arteriolar oxygen tension did not change during the course of each trial. Further investigation of both intravascular and perivascular PaO2 using the PQM system and the same ischemia/reperfusion protocol will highlight any differences in the supply of oxygen to the interstitium versus what was already present at baseline.

Support or Funding Information

Supported by AHA Award #14GRNT20380551