bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification and Evaluation in silico and in vitro of Small Molecules in the KRas4B‐PDEδ Molecular Complex in Pancreatic Cancer
Authors: Aguirre Diana Casique , Gutiérrez Ponciano García , de León Bautista Mercedes Piedad , Mejía Miguel Ángel Vargas 
Published: November 2023

Pancreatic cancer (PDAC) is the most lethal of all cancer types, characterized by an average survival of 6 months and a five‐years survival rate of less than 5%. To increase the survival rate of pancreatic cancer patients is necessary to search for improved tumor markers for an earliest diagnosis and new molecular targets for drug development. In most cases, PDAC is initiated by the mutant of GTPase KRas4B, which has been shown to drive pancreatic neoplasia. Furthermore, it has been demonstrated that nearly 95% of PDAC shown mutational activated KRas4B. Certainly, KRas4B protein has been used as a principal target in anticancer drugs development. Therefore, the identification of new organic molecules that recognize the KRas4B‐PDEδ molecular complex is of great interest for its stabilization for avoiding the activation of KRas4B in the cytoplasmic membrane. Thus, the objective of this work is to identify and evaluate new small molecules that stabilize the KRas4B‐PDEδ complex by using docking techniques. First, we constructed a homology model of KRas4B‐PDEδ complex and this revealed a set of exposed non‐conserved residues at the interface, which are considered an important target for ligand identification. Then, we carried out a docking simulation between the extensive database of drug‐like compounds from ENAMINE and the non‐conserved site at the Kras4B‐PDEδ model using a previous reported protocol. The in vitro evaluation of fourteen compounds with high scores from docking showed a significant specific effect in three of them on the viability of pancreatic cancer cell line MIAPaCa‐2 but not in the pancreatic cell line hTERT‐HPNE used as a control. Interestingly, the compounds decreased Ras‐GTP concentration in the cell line MIAPaCa‐2 in more than 50%, indicating a reduced activity. In conclusion, our results open the possibility to develop drugs against to PDCA; however, we need to perform more experiments in order to establish the specific mechanism of action of the compounds developed in this work.Support or Funding InformationThis work was sponsored by the CONACYT and ISSSTE project number: 002.2015 ISSSTE
A Retrospective Study of Semen Quality in Infertile Men with Varicoceles: Urological Expertise in Designing and Conducting a Testicular Safety Evaluation Study
Authors: Yono Makoto , Inoue Yumi , Ito Kazuya , Kimura Megumi , Furukawa Kohichiro , Hori Masaharu , Tsuji Shigeki , Tanaka Takanori , Sakata Yukikuni , Irie Shin , Latifpour Jamshid 
Published: November 2023

ObjectivesDesigning and conducting a phase 1 study to evaluate the potential for testicular toxicity of a new drug requires urological expertise. Because varicoceles are present in 15–20% of the normal male population and associated with progressive testicular damage and a consequent reduction in fertility, we evaluated semen quality in infertile men with varicoceles, in comparison with control subjects. Information from this study may provide insight into the design and conduct of semen analyses in clinical trials of new drugs.MethodsIn a retrospective study, the records of 12 infertile patients with clinical varicoceles (mean age, 32.6 years) and 8 fertile men without clinical varicoceles (mean age, 32.0 years) between October 2005 and March 2015 were evaluated. Serum follicle‐stimulating hormone (FSH), luteinizing hormone (LH) and testosterone levels and semen parameters (semen volume, sperm concentration, total sperm number and total motility) were assessed in all the subjects. Two semen samples were collected at 4‐week intervals for initial evaluation and semen analyses were performed according to the WHO laboratory manual. Sperm concentration was measured by the same urologist using Makler counting chamber.ResultsAt initial evaluation, inter‐ and intra‐individual variations in semen quality were observed in both the infertile men with grade 1 (palpable during Valsalva maneuver, but not otherwise) varicoceles and the fertile men, and both groups had similar profiles of hormone levels and semen parameters. On the other hand, the infertile men with grade 2 (palpable at rest, but not visible) or grade 3 (visible and palpable at rest) varicoceles had significantly higher FSH and LH levels and lower testosterone levels and semen quality compared with the fertile men. There was a significant increase in FSH levels and a decrease in semen quality in the infertile men with grade 1 varicoceles over a 4‐year period.ConclusionsOur data indicate the presence of large biological variations in semen quality in the normal male population. These findings are considered to have consequences for the interpretation of semen analyses in clinical trials to evaluate testicular safety. Because a significant decline in semen quality over time was observed in the infertile men with grade 1 varicoceles and these small varicoceles could not be detected by hormone levels and semen parameters, men with varicoceles should be excluded from testicular safety evaluation studies by palpation and Valsalva maneuver.
Real‐time, continuous electrochemical monitoring of therapeutic drugs in vivo
Authors: Vieira Philip A , Arroyo‐Curras Netzahualcoyotl , Ploense Kyle L , Somerson Jacob , Plaxco Kevin W , Kippin Tod E 
Published: November 2023

Personalized medicine takes into account individual patient factors that impact the efficacy of pharmaceutical therapies. Measuring the pharmacokinetic/toxicokinetic profile of a specific drug for each patient requires a cost‐effective, selective, robust and minimally invasive sensor. To that end, we have developed an electrochemical aptamer‐based indwelling sensor that achieves continuous, real‐time pharmacokinetic measurements of therapeutic drugs in living animals. Our preliminary experiments demonstrate that these sensors are selective to specific drug targets, durable and stable. We tested sensors selective to aminoglycosides, doxorubicin and cocaine, both in vitro and in vivo, and successfully measured fluctuating concentrations of therapeutics in real‐time. We carried out continuous measurements in Sprague‐Dawley rats (n=6) treated intravenously with separate drugs to create metabolic profiles specific to each animal. We found that our sensors are sensitive to therapeutic doses of antibiotic aminoglycosides (including kanamycin and gentamicin), the antitumor chemotherapy drug doxorubicin, and cocaine. The sensors are stable for up to 6hrs (in vivo) and 12hrs (in vitro), with minimal signal decay over time. This work is a first step in achieving personalized medicine by tracking the pharmacokinetics of selective agents in vivo.Support or Funding InformationThis work was supported by the Keck Foundation (to TEK) and the Otis Williams Fellowship (to NAC).
T‐type Voltage‐gated Ca2+ Channels Do Not Contribute To The Negative Feedback Regulation Of Myogenic Tone In Murine Superior Epigastric Arteries
Authors: Mullan Brendan , Pettis Jessica , Jackson William F 
Published: November 2023

T‐type voltage gated Ca2+ channels (VGCC) have been hypothesized to control spontaneous transient outward currents (STOCs) through large‐conductance Ca2+‐activated K+ channels (BKCa) and contribute to the negative‐feedback regulation of myogenic tone. We tested this hypothesis in superior epigastric arteries (SEAs) isolated from male C57BL‐6 mice. SEAs were isolated and enzymatically dissociated to obtain single smooth muscle cells for whole‐cell recording of paxilline‐sensitive (PAX, 1 μM) STOCs at −30 mV, or cannulated and studied by pressure myography (80 cm H2O, 37°C). The T‐type blocker Ni2+ (30 μM) had no effect on STOC amplitude (20.1±1.7 pA vs. 20.6±1.7 pA; n = 12, p > 0.05), but increased STOC frequency (0.79±0.15 Hz vs. 1.21±0.22 Hz; n = 12, p < 0.05). While Ni2+ produced concentration‐dependent constriction of isolated, pressurized SEAs (logEC50 = −5.6±0.1; Emax = 40±3% constriction), block of BKCa with PAX had no effect on vasoconstriction induced by 30 μM Ni2+ (before PAX = 44±11% constriction vs. in the presence of 1μM PAX = 47±11% constriction; n = 6, p > 0.05). In contrast to Ni2+, the non‐selective T‐type blocker, mibefradil, produced vasodilation (logEC50‐ = −6.9±0.2; Emax = 74±8% dilation), whereas the putative T‐type blocker, ML218, had no significant effect on myogenic tone between 10nM and 10μM (n=6–7, p > 0.05). Our data do not support a role for T‐type VGCC in the negative‐feedback regulation of myogenic tone and suggest that Ni2+ may constrict murine SEAs by means other than block of T‐type VGCC.Support or Funding InformationSupported by PO1‐HL070687 and ASPET‐SURF to B. Mullan
Time to Blood Pressure Control According to Drug Class in Patients with Hypertension
Authors: Johnson Micah William , Sprissler Ryan , Olson Thomas P , Beenken Greg William , Snyder Eric M 
Published: November 2023

IntroductionHypertension is one of the most important preventable contributors to disease and death worldwide and represents the most common condition seen in the primary care setting. Approximately one in three adults living in the US has hypertension. Despite the prevalence and impact of hypertension, little data exists on the average time to blood pressure (BP) control in US patients and time to control according to common BP medication classes.PurposeTherefore, the purpose of this study was to assess the average time to BP control in adults with controlled hypertension, according to drug class.MethodsFor this study we assessed how long (in months) and how many clinic visits were needed in order to control BP according to common medication classes in patients with existing hypertension. The study was performed at two clinical sites where patients came in for one visit for an in‐clinic assessment of BP and for completion of a patient survey regarding their history of BP. The primary care physician performed a chart review for a detailed objective BP history.ResultsNinety‐nine patients completed all aspects of the study (age=58±0.8yrs., %female=46, height=169±1.0cm, weight=86±1.4kg, BMI=29.9±0.4m/kg2). The average initial BP was 151±2mmHg systolic (SBP), 91±1mmHg diastolic, and 111±1mmHg mean arterial (MAP). Patients demonstrated a significant drop in BP with therapy (SBP=16mmHg, DBP=10mmHg, MAP=12mmHg). Patients were prescribed an average of 1.8 BP medications (62% on ACE‐inhibitor, 42% on diuretic, 33% on β‐blocker, 27% on angiotensin receptor blocker, and 16% on Ca+ channel blocker). The average time to BP control was 14.9 months or 3.6 clinic visits. Patients on a diuretic had the longest time to BP control, while patients on a Ca+ channel blocker had the shortest time to control (diuretic=19.5±10.4, angiotensin receptor blocker=22.8±13.1, ACE‐inhibitor=22.2±11.4, β‐blocker=24.5±16.9, Ca+ channel blocker=9.9±4.5 months, pANOVA<0.05). In contrast, patients on a Ca+ channel blocker had significantly more clinic visits to attain BP control (diuretic=4.5±0.6, ACE‐inhibitor=3.1±0.4, angiotensin receptor blocker=3.9±0.6, β‐blocker=4.9±0.7, Ca+ channel blocker=5.1±0.7 visits).ConclusionThese results suggest important differences in the time to BP control, and the number of office visits necessary for BP control according to common classes of hypertension therapies. Funding was provided by Geneticure, LLCSupport or Funding InformationFunded by Geneticure
ROMK Inhibitor Actions in the Nephron Probed with Diuretics
Authors: Kharade Sujay V. , Flores Daniel , Lindsley Craig , Satlin Lisa , Denton Jerod S. 
Published: November 2023

More than a billion people worldwide are afflicted with essential hypertension and diuretics are usually the first line of treatment. Diuretics acting on specific nephron segments to inhibit Na+ reabsorption have been used clinically for decades; however, drug interactions, tolerance, and derangements in serum K+ complicate their use to achieve target blood pressure. ROMK is an attractive new diuretic target, in part, because its inhibition is postulated to indirectly inhibit the bumetanide‐sensitive Na+‐K+‐2Cl− co‐transporter, NKCC2, and the amiloride‐sensitive epithelial Na+ channel, ENaC. ROMK inhibition is also postulated to prevent urinary K+ wasting caused by NKCC2 inhibition. The development of small‐molecule ROMK inhibitors has created opportunities for exploring the physiological responses to ROMK inhibition. The present study evaluated how ROMK inhibition alone or in combination with NKCC2, ENaC, or the hydrochlorothiazide (HCTZ) target NCC, alters fluid and electrolyte transport in the nephron. The ROMK inhibitor VU591 failed to induce diuresis when administered orally to rats. However, another inhibitor, termed Compound A, induced a robust natriuretic diuresis without kaliuresis. Compound A produced additive effects on urine output and Na+ excretion when combined with HCTZ, amiloride or benzamil but not when co‐administered with bumetanide, suggesting that the major diuretic target site is the thick ascending limb (TAL). Interestingly, Compound A inhibited the kaliuretic response to NKCC2 inhibition, an effect we attribute to inhibition of ROMK‐mediated K+ secretion in the TAL and CD. Compound A had no effect on heterologously expressed flow‐sensitive BK channels (Slo1/b1). In conclusion, Compound A represents an important new pharmacological tool for investigating the renal consequences of ROMK inhibition and therapeutic potential of ROMK as a diuretic target.Support or Funding InformationDK082884
PDI via Nrf2 Contributes to the Regulation of Renal AT1 Receptor Function and Blood Pressure
Authors: Wang Xitao , Asghar Mohammad 
Published: November 2023

Renal angiotensin II AT1 receptor (AT1R) plays a vital role in regulation of blood pressure (BP). We recently report that oxidative stress causes exaggerated renal AT1R function and high BP in an aging rat model. This indicates that redox balance is crucial for normal AT1R function and BP. Nrf2 transcription factor is a master regulator of redox‐homeostasis. We hypothesized that protein disulfide isomerase (PDI), an enzyme involved in diverse biological reaction, by regulating Nrf2, regulates redox homeostasis, renal AT1R function and BP. To test the hypothesis, we treated adult Sprague‐Dawley rats with bacitracin, an PDI inhibitor (100mg/kg) for 14 days followed by determination of different parameters. We found that bacitracin treated rats, compared to the saline treated adult rats, showed exaggerated AT1R function, in terms of increased natriuresis and diuresis with AT1R blocker candesartan, and higher systolic BP. Bacitracin treated rats also exhibited increased oxidative stress marker in the renal tissues such as protein carbonyls. Kidney injury marker KIM‐1 level was also elevated in the urine of bacitracin treated rats. Interestingly, bacitracin treatment caused decreased accumulation of nuclear Nrf2 levels suggesting its impaired activity. These data suggest that PDI via Nrf2 pathway plays a critical role in redox‐homeostasis, renal AT1R function and BP.Support or Funding InformationSource of Research Support: NIH/NIA, AG039836
Identification of a biased agonist for P2Y2 receptor to suppress pro‐thrombotic gene expression in human coronary artery endothelial cells
Authors: Zhang Lingxin 
Published: November 2023

Recently we reported that in human coronary artery endothelial cells (HCAEC), the P2Y2 nucleotide receptor (P2Y2R) regulates the pro‐thrombotic gene tissue factor (TF) expression through positive and negative signaling mechanism. Here, we report the identification of a P2Y2R ligand JZS0312 which selectively activates the negative, but not the positive pathways, leading to decreased TF expression. Unlike the endogenous agonist UTP, JZS0312 alone had no effect on TF mRNA transcription, but it unexpectedly inhibited UTP‐induced TF expression. Mechanistic study showed that JZS0312 induced intracellular Ca2+ mobilization in HCAEC which exhibited the same efficacy as UTP and was inhibited by AR‐C118925, a P2Y2R‐selective antagonist. However, when evaluated in their activation of the MAPK pathways, JZS0312 only activated the ERK1/2, but not JNK and p38, whereas UTP activated all the three, suggesting JZS0312 as a biased ligand for the P2Y2R. This notion was supported by the fact that JZS0312‐induced ERK1/2 activation was inhibited by AR‐C1189258 and it had no effect on ERK1/2 in P2Y2R‐null cells. In addition, JZS0312 selectively activated the TF gene repressor Fra‐1, but not the positive AP‐1 subunits c‐Jun and ATF‐2, whereas UTP activated all of them. Furthermore, luciferase activity assay indicates that JZS0312 suppressed TF gene promoter activity via ERK1/2 activation, while UTP increased overall TF promoter activity. These findings reveal that JZS0312 acts as a biased agonist in human endothelial P2Y2R, in which it selectively activates the ERK1/2‐Fra1 pathway, leading to suppression of TF gene transcription. Thus, JZS0312 represents the first identified biased ligand in the P2Y receptor family, opening new avenue for potential drug discovery in relevant inflammatory and thrombotic diseases.Support or Funding InformationScientist Development Grant, AHA
Use of Web‐logs (Blogs) to Promote Student Ownership
Authors: Oliver Kendra H , Picard Danielle , Wikswo John , Brame Cynthia 
Published: November 2023

Blended or hybrid learning is a teaching approach in which some course elements—e.g., content delivery or discussions—occur online while other course elements occur face‐to‐face. Current best practices suggest that the digital environment can be used to guide instruction and to enhance student‐student exchange of ideas. We explore the benefits of online student interactions within an introductory science course as a means of enhancing face‐to‐face interaction and personalization of the learning experience. Weblogs (blogs) have the potential to increase contemplation, community, and collaboration within the classroom through an intimate online framework. In particular, we are interested in building student project ownership that is grounded in student interaction and builds natural learning development through “doing.” However, the effectiveness of blogs to promote ownership has not been fully examined.We investigated the potential of blogs to promote project ownership. Ownership was tested after the addition of two elements to the blog design: instructor curation of the blog and awarding a grade percentage based on student participation. We explored how these modifications alter student educational experience and project ownership. This was measured using de‐identified student data to quantitatively examined the amount of online student‐student interactions. Additionally, we qualitatively analyzed blog content, the quality of students’ final writing samples, and student perception of project ownership via a survey at the end of the semester. Our overall study goal is to develop student fluency in scientific discourse and science project ownership by constructing an online, student‐driven learning blog community. These findings will elucidate the potential for blogs to build student project ownership, driving learner engagement, and academic success.Support or Funding InformationBlended Online Learning Design Fellowship, Center for the Integration of Research, Teaching, and Learning, Center for Teaching
Evaluation of Smooth Muscle Cell Function in Cultured Microvascular Networks
Authors: Motherwell Jessica M. , Azimi Mohammad S. , Katakam Prasad V.G. , Murfee Walter L. 
Published: November 2023

Ex vivo biomimetic models are valuable for investigating the underlying mechanisms of angiogenesis. Common models currently in use include 2D or 3D culture systems and microfluidic devices. However, these systems are limited by their lack of complexity compared to intact‐tissue scenarios. Based on an alternative top‐down approach, our laboratory has recently introduced the rat mesentery culture model as a tool to investigate cell‐cell interactions during angiogenesis in viable, intact microvascular networks. The objective of this study was to determine if vascular smooth muscle cells within the rat mesenteric microvascular networks maintain functionality during culture. Adult Wistar rat mesenteric tissues were harvested and cultured in Modified Eagle's Medium (MEM) + 10% serum. At 0 or 24 hours, tissues were exposed to vasoconstrictors (20nM Endothelin‐1 and 50mM KCl) for 5 minutes. Diameters of arterioles (22μm–27μm) were compared before and after treatment. Vessel constriction measured at the point of maximum diameter change along a vessel was observed at 0‐ and 24‐hours for both vasoconstrictor treatments. The percentage of constriction for arterioles in networks treated with endothelin‐1 at 0‐ and 24‐hour culture were 26 ± 6.2% (n=5 tissues) and 25 ± 3.9% (n=4 tissues) respectively (mean ± SEM). The percentage of constriction for arterioles treated with KCl at 0‐ and 24‐hour culture were 38 ± 4.6% (n=5 tissues) and 25 ± 4.4% (n=4 tissues) respectively (mean ± SEM). We also observed vasoconstriction of venules for both treatment groups at 0‐ and 24‐hour culture. After washing of 50mM KCl vessels were observed to return to baseline measurements supporting the vasoconstriction effect is readily reversible. These results suggest that the vascular smooth muscle cells maintain their functionality in cultured microvascular networks and supports the physiological relevance of the rat mesenteric culture model.Support or Funding InformationNIH 5‐P20GM103629‐04
Attenuated Rapid‐Onset Vasodilation with Greater Force Production in Skeletal Muscle of Caveolin‐2−/− Mice
Authors: Fernando Charmain , Liu Yajun , Sowa Greg , Segal Steven 
Published: November 2023

Caveolin‐2 (Cav2) is a major protein component in caveolae, minute invaginations in plasma membranes of endothelial cells. While absence of Cav2 increases pulmonary endothelial cell proliferation and inhibits tumor growth (PMC133690; PMC4233177), the role of Cav2 in the microcirculation of skeletal muscle is unknown. We tested the hypothesis that genetic deletion of Cav2 would alter the reactivity of microvascular resistance networks controlling muscle blood flow. Internal diameter (ID) responses to physiological agonists and to contractile activity were studied at 35°C in the left gluteus maximus muscle (GM) of anesthetized Cav2−/− and littermate wild‐type (WT) mice (n=5 males/group; 6 months old). For feed arteries (FA), first‐ (1A), second‐ (2A) and third‐order (3A) arterioles, resting IDs (means: 45, 35, 25, 12 μm) and maximal IDs (65, 55, 45, 30 μm) were not different between groups, nor were cumulative endothelium‐dependent dilations to acetylcholine (ACh, 10−9–10−5 M); all right‐shifted ~1 log unit during nitric oxide synthase inhibition (L‐NAME, 10−4M). Constriction‐response curves to norepinephrine (NE, 10−9–10−5 M) overlapped for respective vessel branches. During twitch contractions (2 & 4 Hz; 30 s), steady‐state dilations (range, 5–20 μm) were again similar between groups. However, for single maximal tetanic contractions (100 Hz; 100, 250 & 500 ms), rapid onset vasodilation (ROV) increased with contraction duration but was reduced by 20–50% across vessel branches of Cav2−/− vs. WT (P<0.05). Remarkably, maximal force production (mN/mm2) was ~40% greater in Cav2−/− (152 ± 14) vs. WT (110 ± 3). Thus, while microvascular reactivity is maintained to ACh, NE and moderate contractile activity, diminished ROV with enhanced force production reveals a novel phenotype for Cav2 deletion in mouse skeletal muscleSupport or Funding InformationNIH R37‐HL041026