bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Rho‐kinase inhibitor, fasudil, has antidepressant‐like efficacy in adolescent mice
Authors: Shapiro Lauren , Gourley Shannon 
Published: November 2023

Adolescence represents a critical period of neurodevelopment, defined by structural reorganization and synaptic maturation within the prefrontal cortex. Although these processes are critical for the transition to adulthood, structural instability may open a window of vulnerability to neuropsychiatric disorders including depression. Interventions that facilitate activity‐dependent neural remodeling, as occurs during adolescence, may be advantageous. Here we evaluated the therapeutic‐like potential of Rho‐kinase (ROCK) inhibition, which can expedite activity‐dependent dendritic spine plasticity. The brain‐penetrant ROCK inhibitor fasudil had antidepressant‐like effects in the forced swim test in adolescent mice and was comparable to ketamine and fluoxetine. Fasudil also decreased the latency to approach a palatable food in the novelty suppressed feeding task, a rapid antidepressant‐like effect. Within the adolescent ventromedial prefrontal cortex (vmPFC), fasudil increased levels of the post‐synaptic marker PSD‐95, while pruning dendritic spines, resulting in adult‐like spine densities. Fasudil stimulated several neurotrophin‐related signaling factors in the vmPFC, including increasing the ratio of full‐length:truncated tyrosine kinase receptor B (TrkB). Subsequent experiments utilizing viral vector‐based and pharmacological manipulations indicated, however, that the antidepressant‐like actions of fasudil are nonetheless attributable to the inhibition of the neuronal ROCK isoform, ROCK2.Support or Funding InformationThis work was supported by F31MH109208, Children's Healthcare of Atlanta, the Molecular and Systems Pharmacology Training Grant 5T32GM008602‐17, the Katherine Deschner Family NARSAD Award and a BRAINS Award: MH101477. The Yerkes Center is supported by the Office of Research Infrastructure Programs/OD P51OD011132. The Emory Viral Vector Core is supported by P30NS055077.
β2‐adrenergic receptor control of endosomal PTH receptor signaling via Gβγ
Authors: Jean‐Alphonse Frederic , Wehbi Vanessa , Chen Jingming , Taboas Juan , Xiao Kunhong , Vilardaga Jean‐Pierre 
Published: November 2023

Cells express several G protein‐coupled receptors (GPCRs) at their surfaces transmitting simultaneous extracellular hormonal and chemical signals into cells. A comprehensive understanding of mechanisms underlying the integrated signaling response induced by distinct GPCRs is thus required. Here we found that the β2‐adrenergic receptor, which induces short cAMP response, prolongs nuclear cAMP and PKA activation by promoting endosomal cAMP production in PTH receptor signaling through the stimulatory action of Gβγ subunits on adenylate cyclase type 2.Support or Funding InformationThis work was supported by the National Institute of Diabetes and Digestive and Kidney Diseases (NIDKK) of the US National Institutes of Health (NIH) under Award numbers R01 DK087688 and R01 DK102495, and K01AR062598, and the Costwold Foundation Fellowship Award.
Structural Basis for Regulation of GPR56 by its Alternatively Spliced Extracellular Domains
Authors: Arac Demet 
Published: November 2023

Emerging evidence shows adhesion G‐protein‐coupled receptors (aGPCRs) play critical roles in diverse neurobiological processes including myelination. aGPCRs have large alternatively spliced extracellular regions (ECRs) that likely mediate intercellular signaling; however, the precise roles of ECRs remain unclear. The aGPCR GPR56/ADGRG1 regulates oligodendrocyte development. Accordingly, human GPR56 mutations cause myelination defects and reduced volume of white matter. Using biochemistry, protein engineering, structural biology and in vivo assays, we set out to identify the mechanisms underlying the physiological and pathophysiological functions of GPR56 in myelination. First, we engineered a small protein, termed monobody (Mb), which binds tightly and specifically to the ECR of GPR56. We solved the crystal structure of the ECR of GPR56 bound to this Mb at 2.5Å resolution, revealing for the first time the domain architecture and atomic structure of a complete aGPCR ECR. The structure reveals two domains in the ECR of GPR56: a previously unidentified domain at the N‐terminus and an unusually short, but autoproteolytically active GPCR‐Autoproteolysis INducing (GAIN) domain. Using in vitro and in vivo functional approaches, we observed the effects of several structure‐guided mutations and thereby identified a surface‐exposed conserved patch on the N‐terminal domain necessary to rescue CNS myelination in GPR56‐hypomorph zebrafish. Finally, we measured the effect of Mb binding on GPR56 signaling in vitro and observed a decrease in basal activity, classifying the Mb as an allosteric inverse‐agonist. Our results suggest that the ECR of GPR56 has an essential role in regulating GPR56 function in oligodendrocyte development and myelination in an intricate and multifaceted manner.Support or Funding InformationNIGMSBrain Research FoundationBig Ideas Generator
Delta opioid receptor antagonism/inverse agonism and its effect on mu opioid receptor signaling in SH‐SY5Y cells
Authors: Griggs Nicholas , Mosberg Henry , Traynor John 
Published: November 2023

Opioid analgesics are prescribed with success to combat chronic and severe pain, yet there are many clinical and public health problems associated with the use of these medications that include constipation, respiratory depression and physical dependence. Academic scientists to pharmaceutical companies are developing opioid drugs, G‐protein biased agonists and allosteric modulators to name a few, in an effort to identify safer and more effective analgesics. Previous studies have reported that the co‐administration of naltrindole (a delta opioid receptor antagonist) with morphine (mu opioid receptor agonist) in vivo maintains analgesia yet reduces the development of morphine tolerance and dependence. Following this observation, in collaboration with the Mosberg lab we have discovered and characterized several mu agonist/delta antagonist “bifunctional compounds” that lack tolerance and dependence liabilities in vivo. The goal of my studies is to use these bifunctional compounds, as well as selective mu and delta opioid receptor ligands, in an attempt to explain this phenomenon on a molecular level. The SH‐SY5Y human neuroblastoma cell line is an ideal system in which to assess mu and delta receptor cross‐talk because it endogenously expresses both receptor types. My first objective was to demonstrate mu opioid receptor tolerance at the level of the G protein (a decrease in GTPγ35S binding) using the selective agonist DAMGO. Compared to untreated cells, DAMGO (1μM, 6 hours) treated cells showed a 50% reduction in DAMGO‐mediated GTPγ35S binding and a slight decrease in basal GTPγ35S binding. I then found that the addition of the delta inverse agonist ICI 174,864 (1μM, 6 hours) prevented DAMGO‐mediated tolerance. To further study this response I used another delta antagonist TIPPΨ and three bifunctional compounds. Overall the results suggest that delta opioid receptor inhibition is advantageous for sustained mu opioid receptor function in a cellular model of opioid tolerance. Experiments are ongoing to decipher how delta blockade alters mu‐opioid receptor signaling.Support or Funding InformationDA‐03910 and NIGMS‐GM007767
Aryl Hydrocarbon Receptor Activation Differentially Alters Cholera Toxin‐Specific IgA Levels in Feces and Serum
Authors: King Aspen , Gawronska Elleanour , Metten Madeline , DeKrey Gregory 
Published: November 2023

Secretory IgA plays a critical role in neutralizing enteric toxins and protecting against some intestinal pathogens. In mice, activation of the aryl hydrocarbon receptor (AhR) with 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD) has been shown to suppress antigen‐specific systemic IgM and IgG responses, whereas the impact on mucosal IgA responses is less clear. The goal of this study was to determine if TCDD exposure could alter fecal levels of antigen‐specific IgA after oral immunization. Female C57Bl/6 mice were administered peanut oil (vehicle) or TCDD (2.5, 10 or 40 μg/Kg) orally. On the following day, each animal was immunized with whole Cholera toxin (CT, 10 μg, po) with two subsequent immunizations being given at 10‐day intervals. Antibody levels were measured by ELISA in fecal extracts each week, and in serum at termination (week 4). We found that total serum IgA levels were significantly decreased (P < 0.05) in TCDD‐treated animals, relative to controls. In contrast, no change in total fecal IgA levels was observed in TCDD‐treated mice at any time point measured (weeks 1–4). A significant and dose‐dependent decrease in CT‐specific serum IgA, IgM and IgG1 was observed in TCDD‐treated animals (P ≤ 0.05). In feces, CT‐specific IgA levels were significantly decreased (P ≤ 0.05) in TCDD‐treated mice at week 2, but not at later time points (weeks 3 and 4). These results demonstrate that intestinal IgA responses can recover from AhR‐mediated suppression more rapidly than systemic responses.Support or Funding InformationSupported by the University of Northern Colorado.
Calabash Clay (Nzu) Affects Pregnancy in Whistar Albino Rats
Authors: Aprioku Jonah Sydney , Ogwo‐ude Ezinne Margaret 
Published: November 2023

Calabash clay (Nzu) is a natural product, locally prepared from clay, sand and other non‐nutritional components with or without salt. Nzu has been reported to contain heavy metals and is widely used as a traditional remedy for morning sickness during pregnancy and breastfeeding in most African countries, including Nigeria. The present study was carried out in female Whistar albino rats to explore its effect on pregnancy. Pregnant rats weighing 150–170 g were randomized into 3 groups (n=8 per group) and orally administered 0, 400 or 800 mg/kg body weight of Nzu. The treatments were started on the first day of pregnancy and continued till delivery. Rats were laprotomised after delivery under deep diethylether anaesthesia and the two horns of uteri were examined to determine the implantation sites and presence of resorption sites. Maternal body weight and number of pups delivered were recorded. Data obtained were analysed using students t Test where appropriate and expressed as mean±SEM. Animal experiments were conducted in accordance with our institutional ethical guidelines on animal research. There was maternal body weight gain over time in control rats (p < 0.0001;p = 0.0026), but in treated groups, body weight increased only between gestation day 0 (GD0) and GD10 (p = 0.0022;p = 0.0005) but not afterwards, p = 0.1479;p = 0.7497 (Fig. 1). Additionally, whereas, all control rats gave birth, only 6 (62.5 %) and 1 (12.5 %) of Nzu (400 and 800 mg/kg) treated rats gave birth. Besides, 2 (5 %) of the births that occurred in 400 mg/kg Nzu treated rats were still births (Table 1). Further, number of implantations in Nzu treated rats were lower (p = 0.0262; p = 0.0004) compared with control, as well as number of pups delivered, p = 0.0084; p < 0.0001 (Table 2). The number of pups delivered in Nzu treated rats were fewer than the number of implantations observed, whereas both were equal in control. The results demonstrate that maternal exposure to Nzu dose‐dependently affects pregnancy outcome by potentially inhibiting maternal weight gain, fetal growth, and causing resorption or loss of pregnancy.
Effect of Calabash clay (Nzu) on pregnancy outcome in Whistar albino rats

Dose (mg/kg)
No. of rats that gave birth
Total no. of implantations
Total no. of live pups delivered
No. of still births
No. of resorptions

Control
8
75
75
0
0

400
5
47
38
2
7

800
1
19
8
0
11

Calabash clay (Nzu) exposure during pregnancy reduces number of implantations and pups delivered in Whistar albino rats

Dose (mg/kg)
No. of implantations
No. of live pups

Control
9.38±0.46
9.38±0.46

400
5.88±1.59*
5.00±1.55**

800
2.38±1.56***
1.00±1.00***

P < 0.05,
P < 0.01,
P < 0.001, compared to control.
Values in mean±SEM, n = 8
In vivo and In Vitro Effects of Work Site Fracking Sand Dust (FSD) Inhalation on Rat Lung: Dose‐ and Time‐Response Evaluation
Authors: Fedan Jeffrey S. , Thompson Janet A. , Russ Kristen A. , Newcomer Dean , McKinney Walter , Cumpston Amy M. , Jackson Mark C. , Reynolds Jeffrey S. 
Published: November 2023

A fine dust is generated when sand is manipulated during operations at drilling sites that utilize the hydraulic fracturing, or “fracking,” process, potentially putting workers at risk of inhalation. Previously we investigated the effects of inhalation of 10 and 30 mg/m3 FSD (mass median aerodynamic diameter,1.75 μm; count median aerodynamic diameter, 227nm) 1, 7 and 27 d after exposure (6 h/d for 4 d) and observed modest changes in airway reactivity (7 d; in terms of lung resistance and dynamic compliance) and inflammation (1 d). In the present investigation, additional animals were exposed to the same inhalation conditions to further study its effects on aspects of lung function. Pulmonary function was examined in anesthetized rats using the Flexivent system (single compartment and constant phase models). There were no changes in respiratory system resistance (Rrs), respiratory elastance (Ers), tissue damping (G), tissue elastance (H), Newtonian resistance (Rn) or hysteresivity (η) after any post‐exposure period in rats exposed to either dose of FSD. To investigate whether reactivity of airway smooth muscle or the modulatory effect of epithelium on airway reactivity was affected by FSD inhalation, methacholine (MCh) was applied to the intraluminalor extraluminal baths of the isolated, perfused trachea preparation (IPT). FSD inhalation had no effect on reactivity(EC50 values) or contractile responses to MCh at any FSD dose or post‐exposure time. The possible effects of FSD inhalation on epithelial ion transport was investigated in vitro using Ussing chambers. Neither dose of FSD affected transepithelial potential difference (Vt) or resistance (Rt), or short‐circuit current (Isc) at any time point. Whereas there was no effect of FSD on responses to apical amiloride (Na+ channel blocker), apical NPPB(Cl− channel blocker) or basolateral ouabain (Na+,K+‐pump inhibitor) at the 10 mg/m3 dose, responses to amiloride were significantly inhibited at all post‐exposure times in tracheas from rats exposed to 30 mg/m3 FSD, suggesting that epithelial Na+ transport was impaired by FSD. The results indicate that FSD is generally without appreciable biological effects on pulmonary function broadly speaking, either in vivo or in vitro; however, the dust does interfere with Na+ transport at the 30 mg/m3 dose in a manner that is sustained for at least one month. Such a change could result in hydration of the airway surface liquid.Support or Funding InformationNIOSH
Differential gut microbiota modulates the susceptibility to acetaminophen‐induced hepatotoxicity
Authors: Cho Sung‐joon , Chlipala George , Green Stefan , Lee Hyunwoo , Jeong Hyunyoung 
Published: November 2023

Acetaminophen (APAP) is a commonly used analgesic and antipyretic. APAP‐induced hepatotoxicity is the most frequent cause of acute liver failure in the US. Individual susceptibility to APAP hepatotoxicity is highly variable, and the risk factors for developing severe APAP hepatotoxicity are not well understood. The objective of this study is to investigate a potential role of gut microbiota in modulating susceptibility to APAP toxicity by using a mouse model. C57BL/6 mice from two different vendors [Jackson (JAX) and Taconic (TAC)] were cohoused with mice from the same or the other vendor for 4 weeks, creating the following 4 groups: JAX, TAC, CoJAX (i.e., JAX mice cohoused with TAC) and CoTAC (i.e., TAC mice cohoused with JAX). In parallel, separate 4 groups of mice were co‐housed (n=6–7/group) as untreated controls. Fecal samples were collected at the end of cohousing and subject to next‐generation sequencing analysis of bacterial 16S rRNA gene amplicons. The mice were treated with APAP (600 mg/kg, oral, n=19–21/group) and sacrificed at 24 h post‐dosing, followed by collection of the liver and serum samples. Hepatotoxicity was determined by measuring serum levels of liver enzymes (i.e., alanine aminotransferase, ALT). Results from gut microbiome analysis showed the differences in gut microbiota composition between JAX and TAC at the end of cohousing and indicated the transfer of gut microbiota between cohoused mice. After APAP dosing, JAX mice exhibited 16‐fold higher hepatotoxicity as compared to TAC mice. This difference was abrogated upon cohousing, suggesting that differential gut microbiota between JAX and TAC may modulate susceptibility to APAP hepatotoxicity. Correlation analysis between ALT and different gut microbiota taxa revealed that the relative abundance of bacteria from the genera Mucispirillum, Turicibacter and Ruminococcus in fecal samples prior to APAP dosing is associated with increased APAP hepatotoxicity. The basal mRNA expression levels of Cyp2e1 and Cyp1a2 in the liver did not differ among 4 groups of mice, and the basal hepatic glutathione level was 1.3‐fold higher in JAX compared to TAC, suggesting that neither differential glutathione nor Cyp2e1/Cyp1a2 level is responsible for the difference in APAP hepatotoxicity. Taken together, our data suggest that differential gut microbiota may modulate susceptibility to APAP‐induced hepatotoxicity. This finding may shed a light on identifying potential risk factors for APAP hepatotoxicity.Support or Funding InformationThis study was supported by Chicago Biomedical Consortium.
Cyproterone Acetate Enhances TRAIL‐induced Androgen‐independent Prostate Cancer Cell Apoptosis via Upregulation of Death Receptor 5
Authors: Chen Linjie , Wolff Dennis W. , Xie Yan , Lin Ming‐Fong , Tu Yaping 
Published: November 2023

ObjectiveVirtually all prostate cancer deaths occur due to obtaining the castration‐resistant phenotype after prostate cancer cells escaped from the apoptosis and/or growth suppression initially induced by androgen receptor blockade. TNF‐related apoptosis‐inducing ligand (TRAIL) was an attractive cancer therapeutic agent due to its minimal toxicity to normal cells and remarkable apoptotic activity in tumor cells. However, most localized cancers including prostate cancer are resistant to TRAIL‐induced apoptosis, thereby creating a therapeutic challenge of inducing TRAIL sensitivity in cancer cells. Herein the effects of cyproterone acetate, an antiandrogen steroid, on the TRAIL‐induced apoptosis of androgen receptor‐negative prostate cancer cells are reported.MethodsCell apoptosis was assessed by both annexin V/propidium iodide labeling and poly (ADP‐ribose) polymerase cleavage assays. Changes of gene expression were determined by western blot and RT‐PCR assays. The effect of cyproterone acetate on the gene promoter activity was determined by luciferase reporter assay.ResultsCyproterone acetate dramatically increased the susceptibility of androgen receptor‐negative prostate cancer PC‐3 and DU145 cells to TRAIL‐induced apoptosis. Further investigation of the TRAIL‐induced apoptosis pathway revealed that cyproterone acetate exerted its effect by selectively increasing death receptor 5 (DR5) expression. Cyproterone acetate treatment increased DR5 gene promoter activity, which could be abolished by mutation of a consensus binding domain of transcription factor CCAAT‐enhancer‐binding protein homologous protein (CHOP) in the DR5 gene promoter. Cyproterone acetate increased CHOP expression by inducing endoplasmic reticulum (ER) stress / unfolded protein response (UPR) and siRNA silencing of CHOP significantly reduced cyproterone acetate ‐induced DR5 upregulation and TRAIL sensitivity in prostate cancer cells.ConclusionTogether, our study shows a novel effect of cyproterone acetate on prostate cancer cell apoptosis pathways and raises the possibility that a combination of TRAIL with cyproterone acetate could be a promising strategy for treating castration‐resistant prostate cancer.Support or Funding InformationThis work was supported by grants from the National Institutes of Health (5P20GM103489 and R01HL116849), Nebraska State LB595 research program.
Pipoxolan exhibits antitumor activity toward oral squamous cell carcinoma cell through the reactive oxygen species mediated apoptosis
Authors: Hsiao Jun‐Han , Chou Pei‐Yu , Sheu Ming‐Jyh 
Published: November 2023

Pipoxolan is a well‐known smooth muscle relaxant. However, its apoptotic effects and mechanism in oral squamous cell carcinoma (OSCC) have not been clarified. In the present study, we investigated whether pipoxolan could induce apoptosis in OSCC and explored its potential mechanisms. Cell cytotoxicity were evaluated by MTT assay and apoptosis were measured by Annexin V/PI double staining and flow cytometry analysis. Apoptotic‐related proteins were assessed by Western blotting. Our results demonstrated that pipoxolan treatment caused a time‐dependent decrease the protein expressions of Fas/CD95, cytosolic cytochrome c, active form of caspase‐8, ‐9 and ‐3, hydrolysis of PARP, and Bcl‐2. However, Bax was increased. Clearly, an apoptosis was associated with an increased Bax/Bcl‐2 ratio. Pipoxolan markedly suppressed the expressions of PI3K and phosphorylation of Akt. Reactive oxygen species (ROS) generation and mitochondrial membrane potential (MMP) were measured with fluorescent probes. Furthermore, following exposure of TW206 cells with pipoxolan, a time‐dependently decrease in the MMP and an increase in the ROS. Since high levels of ROS were produced early in the drug treatment, intracellular ROS seemed to play a key role in the pipoxolan‐induced apoptosis. Obviously, apoptosis was significantly abrogated by the free radical scavenger N‐acetyl‐L‐cysteine (NAC). Also, pipoxolan‐induced apoptosis can be blocked by pan‐caspase inhibitor (Z‐VAD‐FMK). The abovementioned data suggested that pipoxolan acted against TW206 and HSC‐3 in vitro via both intrinsic, extrinsic apoptotic signaling pathways, and exhibited in PI3K/Akt signaling pathway. Thus, pipoxolan might be a candidate therapeutic agent for OSCC.Support or Funding InformationMOST104‐2320‐ B‐039‐ 007