bluuebunny/crossref_metadata_2025 huggingface dataset:
Upregulation of Matrix Degrading Enzymes and Lubricin in the Pathogenesis of Joint Disease Requiring Arthroplasty
Authors: Wanderling Chris , Emanuele Nicholas , Carlsgaard Peter , Lilles Jeff , Schnettler Tim , Syed Daneyal , Guler Nil , Hopkinson William , Hoppensteadt Debra , Fareed Jawed
Published: November 2023

Introduction

The prevalence of total knee and hip arthroplasty (TKA and THA) has been increasing; it is projected that TKA and THA procedures will increase to over 4 million procedures by 2030. Since TKA and THA are performed on individuals who experience joint degeneration, it is of interest to study the matrix degrading enzymes (MDE) such as elastase, collagenase‐1, and heparanase. Lubricin is a synovial/joint glycoprotein‐detectable in circulation‐ that plays a vital role in joint mobility by reducing friction in the joint and may have matrix‐stabilizing properties. Previous studies have investigated MDEs in TKA and THA patients, particularly involving prosthesis loosening, yet the role of lubricin is unclear. Simultaneous MDE and lubricin measurement may provide insight on the role of lubricin and these MDEs in the pathogenesis of joint diseases.

Methods

Blood samples were obtained from patients undergoing THA or TKA procedures as de‐identified samples day one pre‐op and day one post‐op. Patient blood samples were collected in 3.2% sodium citrate to obtain plasma. Patient and control plasma samples were stored at −80°C. Plasma samples were analyzed utilizing commercially available ELISA kits for elastase, collagenase‐1, heparanase, and lubricin.

Results

In comparison to normal samples, collagenase and elastase levels were significantly higher in the pre‐op samples (p<0.0001). However heparanase levels were not significantly different between the normal and patient group (p<0.748). On post‐op day 1, a further increase in the collagenase and elastase levels were noted again (p<0.0001). No further increase was evident in the heparanase level. Lubricin levels were markedly higher in the patient groups in comparison to the normal (p<0.0001). On post‐op day 1 marked enhancement of circulating lubricin was noted.

Discussion

The increased levels of lubricin suggest that, joint matrix deteriorates during joint degeneration damaging the capsule allows the joint fluid into systemic circulation. This may be the primary cause of the increased level of lubricin in circulation. Since DJD is a systemic disease with multiple pathogenic factors, it may result in the upregulation of collagenase and elastase. The role of heparanase remains unclear and requires further exploration. It is likely that lubricin may regulate the functional behavior of matrix degrading proteins. These studies also suggest that beside matrix degrading proteins, lubricin may also have a prognostic role in DJD.
Extracellular matrix (ECM)‐stiffness regulated cancer cell (A549) migration
Authors: Gweon Bomi , Li Zeyuan , Shin Yujin , Yoo Hongki , Shin Jennifer H.
Published: November 2023

Extracellular matrix (ECM)‐stiffness is one of the potent regulator of cell physiology such as proliferation, morphology, and migration. From previous studies, it has been known that tumor develops increased collagen deposition at the surrounding tissues. The presence of this dense collagen matrix is shown to promote tissue‐stiffening and, further, metastatic outcomes. However, little is known about how this mechanical stimulation, ECM‐stiffening, are transduced into cells and derive tumor progression. To investigate the relationship, we grew human lung carcinoma cells (A549) on the collagen coated ECM synthesized by polyacrylamide with physiologically relevant range stiffness (0.3 ~ 150 kPa). We chose A549 cells because A549s are reported to be sensitive to ECM‐stiffness. To correlate the ECM‐stiffness to metastatic behavior of cells, we observed migration of cells under microscope for about 24 hours equipped with incubator. In order to track the path of individual cell movement, we kept the cell population in sparse condition during this 24 hours. When we measured the migration speed, cells on soft substrate (0.3 kPa) migrated relatively slower compare to the ones on stiffer substrate (19 kPa and 153 kPa). Cells, also, tend to spread slightly less on soft substrate (0.3 kPa). To determine if these alterations in cell physiological induced by ECM‐stiffness change is related to cytoskeletal contractility, we additionally measured cellular traction.

Support or Funding Information

This work was supported by the National Research Foundation of Korea Grant funded by the Korean Government (NRF‐2014R1A6A3A04059713 and NRF‐2013S1A2A2035518).
Inhibitory Effect of Luteolin on Hepatic Stellate Cell Activation Is STAT3 Dependent
Authors: Wang Xiaofu , Bohanon Fredrick J. , Lopez Omar Nunez , Kandathiparampil Arline J , Zhou Jia , Radhakrishnan Ravi S
Published: November 2023

Introduction

STAT3 is activated in a majority of cancer and other fast growing cells, such as activated hepatic stellate cells (HSC), which are the primary effector of liver fibrosis. Tyr705 phosphorylation of STAT3 is a key step for its function, so it has become a widely explored target for new drug development. Luteolin is an important member of the flavonoid family and has anti‐tumorigenic, anti‐oxidant, and anti‐inflammatory properties. Previously, we have reported that luteolin has a significant anti‐fibrogenic effect on activated HSC. However, its molecular mechanisms remain unclear. In this present study, we determined the role of STAT3 on luteolin‐induced HSC inactivation.

Methods

The activated human and rat hepatic stellate cell lines LX‐2 and HSC‐T6 were used for this study. Cellular proteins were analyzed by immunoblotting and immunofluorescence. Cell proliferation following STAT3 inhibitors treatment was assessed with Alamar Blue assay.

Results

Luteolin treatment significantly inhibited pSTAT3 (Tyr705) protein expression (p<0.001) in both a time‐ and dose‐dependent manner by immunoblot assay and confirmed by immunofluorescence staining. Additionally, luteolin suppressed pSTAT3 nuclear translocation (p=0.009) and its transcriptional activity (p=0.03). The downstream effectors of STAT3 pathway, c‐Myc and cyclin D1, were inhibited by luteolin treatment (p<0.0001). Importantly, treatment with STAT3 inhibitors, Stattic and SH‐4‐54, resulted in a marked decrease in LX‐2 cell growth and expression of HSC activation marker α‐smooth muscle actin, which was similar following luteolin treatment.

Conclusion

Luteolin treatment inhibited STAT3 signaling and its target genes. STAT3 inhibitors attenuate LX‐2 cell growth and activation. Luteolin may affect HSC activation through the STAT3 pathway.

Support or Funding Information

National Institutes of Health: T32‐GM8256
Cloning, Identification And Characterization of Two MyD88s during Anterior Regeneration in Aeolosoma viride
Authors: Chen Chiao Ping
Published: November 2023

A 2 mm long freshwater annelid, belonging to Aeolosomatidae, with an exceptional regenerative ability, called Aeolosoma viride, serves as an excellent model for regenerative research. Additional reasons are its small size and semi‐transparent color, which allow manageable culture and experimentation. A. viride can regenerate its amputated head with a functional brain within 5 days. The regeneration process includes several steps, starting with wound repair followed by blastema formation and tissue remodeling. In addition, the immune system and inflammatory responses associated with injury to prevent deadly infections have been implicated as critical modulators during wound healing and regeneration. The aim of our study is to unravel the involvement of immune responses in A. viride regeneration. The Toll‐like receptor (TLR) signaling pathway is one of many pathogen recognition mechanisms, which initiates by ligand binding to the receptors and then recruits myeloid differentiation factor 88 (MyD88) to serve as an adaptor molecule to activate the transcription factor NF‐κB. Two MyD88 sequences have been cloned from A. viride. Based on blasting results, one of the sequences has two regular functional domains of MyD88, namely Toll/IL‐1R (TIR) domain and death domain. However, the other one lacks the TIR domain, responsible for TLR binding. Furthermore, we identified and analyzed their gene expressions during the regenerative process. Real‐time PCR results of blastemal tissue showed different expression patterns of these two genes. One is down‐regulated during blastema formation, while the other is up‐regulated during wound healing. Therefore, we propose the two MyD88s may play different roles during regeneration in A. viride. Avi‐MyD88a and Avi‐MyD88i may activate or inactive the TLR pathway.
Streptococcal Collagen‐Like Protein 1 (Scl‐1) Protects Group A Streptococcus from Innate Immune Defenses
Authors: Whalen Ryan , Dohrmann Simon , Shimamoto Jacqueline , Etesami Neelou , Martin Gary , Nizet Victor , Okumura Cheryl
Published: November 2023

Group A Streptococcus (GAS) is a gram‐positive bacterium that ranks within the top 10 of human pathogens responsible for mortalities every year. The human innate immune system combats and kills GAS through the enlistment of macrophages and neutrophils. Neutrophils can kill bacteria intracellularly via the generation of reactive oxygen species and release of antimicrobial granule contents. These molecules also contribute to a specific mechanism of neutrophil death, resulting in the production of neutrophil extracellular traps (NETs), which can kill bacteria extracellularly. We have shown that the GAS virulence factor Streptococcal collagen‐like protein (Scl‐1) increases bacterial survival in NETs while simultaneously lowering the overall production of NETs. In this work, we are exploring the mechanism by which Scl‐1 prevents NET formation, specifically investigating whether Scl‐1 decreases the production, activity or availability of antimicrobial factors that eventually lead to the formation of NETs. While the release of granules were not significantly different between cells infected with strains expressing or lacking Scl‐1, the content of the granules was shown to be an important factor in Scl‐1 mediated GAS survival. The antimicrobial peptide LL‐37 is released by the granules and subsequently aids in direct bacterial killing and NET formation. We found that Scl‐1 mediates protection against LL‐37 mediated bacterial killing in vitro. Transmission electron microscopy (TEM) imaging of bacteria within phagocytic cells indicates that strains expressing Scl‐1 are more intact, further supporting the idea that Scl‐1 is protective against LL‐37. We are currently exploring whether there is a linkage between LL‐37 activity within the phagocytic cells and extracellular trap formation. Our results shape our understanding of not only Scl‐1 prevention of NET formation, but also the general mechanism of NET generation in phagocytic cells.
Effects of Sub‐inhibitory Concentrations of Hydrogen Peroxide and Fluoride on the Innate Immune Resistance of Streptococcus mutans
Authors: Peacock Stephanie , Okumura Cheryl
Published: November 2023

Streptococcus mutans is a gram‐positive bacterium that resides in biofilms on teeth and is thought to be a large contributor to the formation of dental caries. S. mutans grown in sucrose is able to adhere to teeth and form plaque deposits by producing glucans. Neutrophils and innate immune defense proteins such as lysozyme and antimicrobial peptides found in the mouth are not enough to combat oral pathogens such as S. mutans, especially when in biofilms, and therefore antiseptic compounds such as fluoride and hydrogen peroxide are utilized to maintain dental health. We found that while high concentrations of fluoride and peroxide inhibited the growth of S. mutans, bacteria grown in sub‐inhibitory concentrations of fluoride and peroxide were induced to form biofilms. Analogous research in our lab demonstrated that sub‐inhibitory concentrations of the antibiotic nafcillin weakened antibiotic resistant Staphylococcus aureus to allow innate immune cells to kill the bacteria more efficiently. However, our current results provide the possibility that fluoride and hydrogen peroxide may function in the opposite way for S. mutans by inducing a protective response. Given our results and the ways in which bacteria evolve resistance to survive antimicrobial compounds, it becomes advantageous to look at alternative means to antiseptics for fighting pathogens.
Effect of Opioids on Integrity of White Matter
Authors: Korneeva Nadejda L , Fan Ruping , Schrott Lisa M , Arnold Thomas , Graham Derrel
Published: November 2023

Oxycodone (6‐deoxy‐7,8‐dehydro‐14‐hydroxy‐3‐O‐methyl‐6‐oxomorphine) is a strong semi‐synthetic opioid. It is prescribed to treat different types of pain especially when other opioids are ineffective. Oxycodone has close structural similarity to morphine and heroin and binds not only m‐ but also k‐opioid receptors. Unfortunately, similar to other opioids, repetitive oxycodone administration has the potential to develop analgesic tolerance, withdrawal, and addiction. Recently, another concern regarding the effect of chronic or over‐dose opioid exposure has emerged: their effect on neuronal degeneration. Leukoencephalopathy or damage of white matter has been described as one of the effects that heroin abusers develop, and was recently documented in a patient after morphine and oxycodone administration. The exact mechanisms underlying the neurotoxic effect of opioids on the Central Nervous System are still not fully understood. To investigate the effect of opioid treatment on rat white matter we used an animal model in which female rats were gavaged with 15 mg/kg oxycodone every 24 hours for 30 days. In our study we observed that chronic oxycodone exposure induces oxidative/nitrosidative stresses in three brain areas in rats: nucleus accumbens, cortex and brain stem. We demonstrated activation of pro‐apoptotic machinery in rat brains after oxycodone treatment. Interestingly, activated caspase 3 was predominantly localized in areas containing white matter suggesting increased axonal damages after chronic oxycodone exposure. The increased level of a‐synuclein aggregates and amyloid precursor protein (APP) in oxycodone‐exposed cerebellum confirmed the axonal injury. We also demonstrated induction of pro‐apoptotic protein Bax by oxycodone, morphine and DAMGO in breast carcinoma cells MCF7, which are known to express the m‐opioid receptor.

Support or Funding Information

LMS was supported by Grant No. 2R01GM20818 from the National Institute of General Medical Sciences.
Antinociceptive effects of synthetic tetracycline compound and influence of Cx3cr1 in inflammatory pain
Authors: Guindon Josee , Bezboruah Christian , Blanton Henry , Sherfey Caitlyn , Marquardt Patrick C , Martinez Joseph M , Bergeson Susan E
Published: November 2023

Tetracycline compounds are broad‐spectrum antibiotics used against different bacterial infectious diseases. Synthetic tetracycline compounds have been developed and are able to reduce infection as well as inflammation. However, the effect on alleviation of pain, and more specifically reduction of inflammatory pain, has not been investigated. The goal of the study is to evaluate the effect of synthetic tetracycline compounds using an inflammatory pain model and mechanical and cold allodynia responses. The formalin test (10 μl at 2.5 % intraplantar) was use to evaluate pain threshold differences between male wild‐type C57BL/6 mice and male GFP Cx3cr1 +/− mice. First, our preliminary data demonstrate that male GFP Cx3cr1 +/− mice show lower pain threshold in both (acute and inflammatory) phases of the formalin test. Second, we evaluated the effect of a synthetic tetracycline compound on inflammatory pain and on mechanical and cold allodynia. Our data indicate that mechanical and cold allodynia remains unchanged after administration of a synthetic tetracycline compound meaning values are similar to baseline levels. However, synthetic tetracycline compound lower pain threshold in both phases of the formalin test from 4 hours up to 20 hours after its administration and showed antinociceptive effects to mechanical and cold allodynia following the formalin test (60 minutes after injection of formalin). Moreover, mechanical and cold allodynia responses return to baseline levels 2 hours after the formalin test. These results demonstrate potential antinociceptive properties of synthetic tetracycline compound with long lasting effect up to 20 hours after its administration. Further studies are needed to investigate the mechanism underlying this antinociceptive effect of synthetic tetracycline compounds. We will also evaluate potential stronger antinociceptive effect of the synthetic tetracycline compound in Cx3cr1 +/− male mice and evaluate if there is any gender specific differences. Antinociceptive properties of synthetic tetracycline compound could be a great alternative avenue to relieve inflammatory pain and improve alleviation of pain in patients.

Support or Funding Information

Funded by grant from NIAAA 021142 (SEB) and funded by Texas Tech University Health Sciences Center School of Medicine grant 121035 (JG).
Lysergic Acid Diethylamide (LSD) Modulates Circadian Activity Behavior And The Expression Level of Circadian Genes in Drosophila melanogaster in Both Sexes
Authors: Sherman Kelly Jean , Nichols Charles D.
Published: November 2023

Circadian rhythms modulate biochemical and molecular pathways in living systems. In humans an endogenous clock mechanism is synchronized by a daily light‐dark cycle: sun up and sun down. These external cues from the environment regulate the core‐clock through auto‐regulating circadian genes’ expression levels. These oscillating gene products modulate physiological, biochemical, and molecular pathways that maintain homeostasis of cells and the overall homeostasis of a living organism. Drosophila and other model organisms are utilized to study the input pathways that modulate the core clock and its regulation of circadian rhythms.

Our laboratory has previously used serotonin agonists to study behaviors in the fly, but the role of serotonin and its receptors in circadian behaviors have not been studied extensively. Therefore to study the impact of serotonin receptor linked pathways on circadian gene expression, we investigated the effects of the pharmacological agent lysergic acid diethylamide (LSD), a mixed serotonin receptor agonist, on circadian activity in the fly. In humans, studies have shown that LSD affects the sleep/wake circadian rhythms, but the mechanism is not known. Although, LSD has not been used to study circadian activity rhythms in the fly, previous work in the laboratory demonstrated that serotonin and its receptors play a role in modulating circadian related activity in the fly, We therefore hypothesize that LSD will alter clock gene expression levels and disrupt circadian activity in Drosophila.

The Drosophila Activity Monitor (DAM) was used to record circadian activity in entrain (light/dark) and free run (dark/dark) environments. The Capillary Feeding (CAFE) assay was simultaneously used to measure food/drug consumption. Five concentrations (0, 0.01, 0.1, 1.0, 3.0, and 10.0 mM) of LSD were administered to the flies for seven days in a light/dark environment, during which time the flies were monitored for activity levels. Our data show that LSD affects circadian activity in the fly in a dose‐dependent manner in both males and females, with a more robust effect in females. Changes were measured in the RNA levels of circadian and P450 genes in the fly heads between LSD‐treated and control flies. In free run experiments three doses of LSD (0, 1.0 and 3.0mM) were tested, and these data show that LSD also disrupts the endogenous circadian rhythm in a dose dependent manner in both males and females. These behavioral experiments and molecular studies suggest that LSD affects circadian activity, potentially through modulating expression of circadian genes downstream of serotonin receptor activation.
Structural Basis for Allosteric Coupling Between G Protein and the Agonist‐Binding Pocket in GPCRs
Authors: Mahoney Jacob , DeVree Brian , Velez‐Ruiz Gisselle , Rasmussen Soren , Kuszak Adam , Edwald Elin , Manglik Aashish , Masureel Matthieu , Du Yang , Matt Rachel , Pardon Els , Steyaert Jan , Kobilka Brian , Sunahara Roger
Published: November 2023

G protein‐coupled receptors (GPCRs) translate extracellular ligand binding into intracellular signaling responses via allosteric communication between two distant binding sites. For many GPCRs, allosteric communication between agonists and G proteins has been observed in the form of G protein‐mediated enhancement of agonist affinity. However, the mechanism by which G proteins allosterically regulate a receptor's orthosteric ligand binding site has remained unclear. Based on recent crystal structures of the β2‐adrenergic receptor (β2AR), we hypothesized that stabilization of an active receptor conformation by nucleotide‐free G protein leads to closing of the receptor's orthosteric site around a bound agonist, thus slowing agonist dissociation and enhancing affinity. Using purified receptors reconstituted into high‐density lipoprotein (rHDL) particles, we performed equilibrium and kinetic radioligand binding assays to characterize the allosteric effects of G protein and G protein‐mimetic nanobodies on the ligand‐binding properties of multiple GPCRs. We also investigated the contribution of individual residues within β2AR in mediating high‐affinity agonist binding. When β2AR was bound to nucleotide‐free Gs heterotrimer, we observed slowed association of the antagonist [3H]DHAP, which was reversible by GDP or GTPγS. A Gs‐mimetic nanobody (Nb80), like the nucleotide‐free G protein, slowed [3H]DHAP association but also slowed [3H]DHAP dissociation. However, this effect was not antagonist‐specific, as Nb80 also inhibited association of the full agonist [3H]formoterol, the partial agonist [3H]CGP‐12177, and the inverse agonist [3H]carvedilol. Mutation of tyrosine 308 to alanine in β2AR abolished the rate‐slowing effects of Nb80 on antagonist and agonist association. Together, these data suggest that transit into or out of the orthosteric site in the active β2AR conformation (stabilized by either Gs or Nb80) is restricted due to closing of the orthosteric site, and that Tyr308 of β2AR is an important residue for controlling access to the orthosteric site. Interestingly, the M2 muscarinic acetylcholine receptor and mu opioid receptor behaved similarly to β2AR when bound to nucleotide‐free G protein or G protein‐mimetic nanobodies, suggesting that G protein‐mediated closing of the orthosteric site may extend to GPCRs beyond β2AR and the amine receptor family.

Support or Funding Information

This work was supported by AHA 13PRE17110027 and NIH GM083118.
LPA1 Receptor Antagonists Exhibit Signaling Bias and Differential Efficacy in Models of Renal Fibrosis
Authors: Hazelwood Lisa A , Leys Laura , Lundgaard Greta , Putman C. Brent , McGaraughty Steve , Clapham Bruce , DeGoey David , Lee Lance
Published: November 2023

The endogenous phospholipid, lysophosphatidic acid (LPA), is known to mediate an array of physiologic processes, including proliferation and cell migration. In addition, LPA signaling is implicated in the pathophysiology of fibrotic disease and evokes a pro‐inflammatory response. The various effects of LPA are mediated by interaction with one of six cognate receptors, termed LPA1–6. In particular, many of the pro‐fibrotic and pro‐inflammatory effects of LPA are attributed to interaction with the LPA1 receptor. LPA1 receptor stimulation promotes signaling through Gq, Gi/o, G12/13 and arrestin. Notably, while LPA species 18:1 is equipotent at evoking each of these downstream signals, we identified antagonists that exhibit biased inhibition of the various pathways. To better understand the molecular pharmacology of LPA signaling in renal fibrosis and inflammation, we studied the signaling pathways activated by 18:1 LPA in various primary human renal cell types, including proximal tubule epithelial cells and renal fibroblasts. These studies enabled mapping of the complex LPA signaling network that drives key pro‐fibrotic and pro‐inflammatory endpoints. Finally, we will present efficacy data comparing full antagonists against antagonists with various biased profiles in the mouse UUO model of renal fibrosis. In total, these data demonstrate the complexity of the LPA‐LPA1 signaling events that drive renal fibrosis, and highlight the requisite antagonist profiles to reverse renal fibrosis endpoints in vivo.

Disclosures: All authors are employees of AbbVie. The design, study conduct, and financial support for this research were provided by AbbVie. AbbVie participated in the interpretation of data, review, and approval of the publication.

Support or Funding Information

All research was funded by AbbVie Inc.
Efficacy at the Mu Opioid Receptor: Insights from Orthosteric and Allosteric Ligands
Authors: Livingston Kathryn E , Mahoney Jacob , Manglik Aashish , Kobilka Brian , Sunahara Roger , Traynor John R
Published: November 2023

The mu opioid receptor (MOPr) represents one of the most pharmacologically targeted G protein‐coupled receptors (GPCRs) worldwide. Activation by an orthosteric agonist like morphine causes robust pain relief but also results in unwanted effects including respiratory depression, constipation, and addiction liability. However, these actions have different agonist efficacy requirements. Consequently an understanding of the factors governing efficacy is required. In addition, we have recently described a series of positive allosteric modulators (PAMs), exemplified by the thiazolidine BMS‐986122 that control the affinity and/or efficacy of orthosteric MOPr ligands. Here we report that BMS‐986187, a delta opioid receptor (DOPr) PAM, although structurally very distinct from BMS‐986122 acts as a MOPr‐PAM with increased allosteric efficacy compared to BMS‐986122, but with the same probe dependence and the same mechanism of action: by allosterically disrupting Na+ binding. Furthermore, competition experiments suggest that BMS‐986187 binds to the same site as BMS‐986122 indicating a somewhat conserved allosteric binding site on MOPr and DOPr. Utilizing purified MOPr reconstituted into high density lipoprotein (HDL) particles we have studied binding of the MOPr state‐sensitive sensor nanobody39 (Nb39) by interferometry to monitor formation of MOPr active state. Differences in orthosteric ligand efficacy were seen to correlate with different kinetics of Nb39 association and dissociation. The PAMs alone enhanced the on‐rates of Nb39 but differences were seen reflective of distinct allosteric efficacies. Finally, we show that cooperativity between agonist and BMS‐986187 is unchanged in the absence or presence of G proteins suggesting that the allosteric modulators alone are able to stabilize of an active state of MOPr.

Support or Funding Information

Funded by DA039997.
Investigation of a cyanobacterial collection from Curaçao for CNS activity
Authors: Hilton Edward , Staub Corinne , Kolber Benedict , Tidgewell Kevin
Published: November 2023

Cyanobacteria are photosynthetic prokaryotes found in diverse terrestrial and aquatic habitats around the world. Secondary metabolites produced by marine cyanobacteria are known to exhibit pharmacological activity and resemble endogenous ligands to mammalian receptors1. Our lab is interested in discovering novel compounds from marine cyanobacteria that have activity at targets involved in CNS disorders. This project focuses on the serotonin and peripheral benzodiazepine receptors, which are involved in pain, depression, and anxiety in the central nervous system. A marine cyanobacterium, field classified as a paintbrush Symploca, was collected off the coast of Santa Barbara Beach, Curaçao and was extracted and fractioned following standard lab protocol. Extracted fractions of compounds will be sent to the Psychoactive Drug Screening Program run by the Roth Lab at the University of North Carolina Chapel Hill. Fractions are screened against a panel of GPCRs, ion channels, and transporters, which are linked to CNS disorders. Active fractions and compounds will be tested in behavioral assays using mouse models of depression and anxiety, such as the forced swim test and elevated zero maze. Concurrently, active compounds in these fractions will undergo structure elucidation using 1D‐ and 2D‐NMR and LC‐MS. Phylogenetic analysis will be carried out using 16S rRNA sequencing to definitively determine the species of cyanobacteria. Results to date will be presented.

Support or Funding Information

R15AT008060 (NIH NCCIH)
Evaluation of the effect of CdTe quantum dots on cell viability
Authors: Rodríguez‐Lopez Anahi , Reyes‐Esparza Jorge , Agarwal Vivechana , Rodriguez‐Fragoso Lourdes
Published: November 2023

Evaluate the effect of CdTe quantum dots on cell viability of three cell line. Nanotechnology is the study, design, synthesis, manipulation and application of materials, devices and functional systems through control of matter at the nanoscale (1–100 nm). The quantum dots (QDs) are colloidal crystals semiconductor scales 1–10 nm, producing fluorescence of different colors depending on their size. Due to its several applications in areas such as medicine and the pharmaceutical, there has been a need to understand their interaction with biological systems. Parameters such as size, composition, surface charge, and functionalization can be modified to either enhance biocompatibility or reduce their toxic effects. In this work, the cell viability is evaluated in kidney (Hek293), cervical cancer (HeLa) and liver (HepG2) cells by thiazolyl blue tetrazolium bromide (MTT) technique. Cells were seeded in 96‐well plates in culture media respectively. These cells were treated with CdTe quantum dots of different sizes (3 and 10 nm) (pregunta : do you want to put three different sizes as we prepared in the lab or it is only 1 type from SA? ) at concentrations of 0.1, 1, 10, 100 and 1000 ng/ml for 24 hours. In order to determine the effect of the cell proliferation by MTT technique on HeLa cell line. These cells were treated with CdTe‐COOH QDs (by SIGMA ALDRICH) at the same concentrations as was described above, but also evaluated from one to seven days. The CdTe quantum dots (3 nm) were have no effect on reducing cell viability Hek293Q, HeLa and HepG2. We hypothesized that the cytotoxic effect of CdTe QDs may have a little correlation function its size, as indicated by recent studies which have emphasized on the composition of these QDs and the possibility of toxic effects. Although CdTe QDs of 3 and 10 nm caused no decrease in cell viability Hek293, HeLa and HepG2, except, compared to about 10 nm QDs where HeLa had cytotoxic effect from maximum concentrations of 1000 ng/ml. The quantum dots CdTe‐COOH at concentrations of 1 and 10 ng/ml had no effect on cell proliferation HeLa significantly, but there was an increase in cell proliferación of HeLa significant compared to the control group at concentrations of 0.1 and 100 ng/ml which increased by up to 500% of cell proliferation. Although, in the case of HeLa cells treated to concentration of 1000 ng/ml had no effect on cell proliferation, this result is consistent with that obtained in the cytotoxicity assay.
Bioengineered miR‐1291prodrugas sensitizing agent for the control of pancreatic carcinoma cell proliferation and tumor growth
Authors: Tu Meijuan , Li Meimei , Yu Aiming
Published: November 2023

MicroRNAs (miRs or miRNAs) are critical factors in the control of tumor initiation and progression via crosstalk with cancer signaling pathways. Our recent studies have demonstrated that miR‐1291 is able to sensitize pancreatic carcinoma cells to chemotherapy through downregulation of efflux transporter ABCC1/MRP1. We have also developed a novel biological method to produce large quantity ofmiR‐1291 prodrug for research and development. In this study, we aimed to assess the utility of bioengineered miR‐1291 prodrug for improving the efficacy of chemotherapy. Our data showed that miR‐1291 prodrug was readily processed to mature miR‐1291 in human carcinoma PANC‐1 cells and consequently reduced the protein levels of miR‐1291 targeted genes including efflux transporter ABCC1/MRP1. Furthermore, miR‐1291 prodrug significantly enhanced the sensitivity of ABCC1‐overexpressing PANC‐1 cells to doxorubicin, a substrate of ABCC1. In addition, in vivo‐jetPEI formulated miR‐1291 prodrug was distributed to PANC‐1 xenograft tumors in mouse models and enhanced the efficacy of chemotherapy. These results indicate that bioengineered miR‐1291prodrug may be utilized as sensitizing agent for the development of more effective cancer therapy.

Support or Funding Information

National Institutes of Health (1U01CA175315 to A.M.Y.)
A Dock‐Derived PEDF Mimic Targeting Laminin Receptor Downregulates VEGF Receptor
Authors: Umbaugh Charles S , Diaz‐Quiñones Adriana , Figueiredo Marxa
Published: November 2023

Pigment epithelium derived factor (PEDF) is regarded as an exceptionally potent inhibitor of angiogenesis. Our lab and others have used PEDF as a novel prostate cancer therapy, as PEDF inhibits tumor establishment, vascularization, and alters malignant signaling. Recently, PEDF was identified as a binding partner for the cancer associated protein Laminin Receptor (LAMR1). LAMR1 overexpression is common in a variety of cancers including prostate, breast, and colon. In an attempt to mimic the protein‐protein interaction between PEDF and LAMR1, we used a structure based drug design approach. After screening 14,000 compounds in silico against the crystal structure of LAMR1 and screening 7 compounds in vitro against five cell types, we identified a hit compound with low micromolar anti‐proliferative capacity against the prostate cancer cell line PC3. Our hit compound markedly decreased VEGFR at the mRNA level, a finding that has been associated with PEDF expression. In an unbiased redock, our hit compound bound a laminin binding site on the crystal structure of LAMR1. This binding appears to be facilitated by a hydrogen bond between the compound and HIS169 of LAMR1. Next, we will confirm compound activity via a fluorescent polarization assay and endothelial tube formation.

Support or Funding Information

We acknowledge support from the Basic Medical Sciences Department in the College Of Veterinary Medicine at Purdue University.
Discovering potent PTP4A3 phosphatase small molecule inhibitors for colorectal cancer
Authors: McQueeney Kelley E , Salamoun Joseph M , Handing Kasia , Boulton Adam , Sharlow Elizabeth R , Bushweller John H , Minor Wladek , Wipf Peter , Lazo John S
Published: November 2023

Although there has been significant progress in understanding colorectal cancer pathogenesis, the 5‐year patient survival rate in the United States of ~60% underlines the need for better treatment options when surgical resection and current therapies are not sufficient. In recent years, protein tyrosine phosphatase 4A3 (PTP4A3) has garnered significant interest and extensive validation as an anticancer target. The Oncomine database documents PTP4A3 is in the top 1% of the most highly expressed colorectal cancer genes with >650 tumor samples. Moreover, it has been noted that increased PTP4A3 expression correlates with increased tumor invasiveness and poor patient prognosis. We synthesized the thienopyridone JMS‐631‐050, which is the most potent published PTP4A3 small molecule inhibitor. Using 6,8‐difluoro‐4‐methylumbelliferyl phosphate as an artificial substrate, we found JMS‐631‐050 non‐competitively inhibited recombinant PTP4A3 in vitro phosphatase activity with an IC50 value of 138 nM. We have generated the first NMR spectra of PTP4A3 in complex with JMS‐631‐050 to more thoroughly characterize the nature of their interaction. Preliminary analysis of NMR data suggests that JMS‐631‐050 binds to a region of the phosphatase outside of the active site, which we hypothesize decreases protein flexibility and proximity of the general acid to the catalytic cysteine. This binding site is also consistent with molecular docking simulation studies. Recently, we discovered a new, more potent thienopyridone analog, JMS‐631‐053, which inhibited recombinant PTP4A3 in vitro phosphatase activity with an IC50 value of 8 nM and showed limited or no inhibition against other phosphatases at concentrations < 1 μM. Both JMS‐631‐050 and JMS‐631‐053, which computationally have drug‐like properties, reduced cellular migration of mouse colorectal tumor cells and prevented the formation of cohesive spheroids in an extracellular matrix phenocopying results observed with PTP4A3 null colorectal cancer cells. JMS‐631‐050 and JMS‐631‐053 should be valuable reagents for further validation of PTP4A3 as a cancer target as well as potential leads for drug discovery.

Support or Funding Information

NIH R21CA191944, the Farrow Fellowship, and NCI Cancer Center Support Grant P30 CA44579
Evidence that aspirin blocks platelet‐induced growth, invasive activity and epithelial‐mesenchymal‐transition (EMT) of mouse colon cancer cells
Authors: Lichtenberger Lenard M , Fang Dexing , Dial Elizabeth , Vijayan K. Vinod
Published: November 2023

Background

Aspirin consumption is associated with a significant reduction in the incidence and mortality of colorectal cancer (CRC). The evidence was sufficiently compelling to lead a subcommittee of the US Preventive Services Task Force (USPSTF) this year to recommend that individuals aged 50–69 years of age take low‐dose aspirin for chemoprevention of CRC. There is also evidence that platelet number is increased in late‐stage cancer patients and that platelets may stimulate cancer cell growth and EMT.

Objective

To determine if aspirin's chemopreventive action against CRC may be due to the drugs ability to inhibit platelet activation (via irreversible COX‐1 acetylation) and thereby block platelet‐ induced effects on CRC cell growth, invasive activity and EMT.

Methods

MC‐26 mouse colon cancer cells were incubated in cell culture on Transwell plates in the absence and presence of freshly isolated mouse platelets, at a cell : platelet ratio of 1:400 and 1:800, in medium containing aspirin from 0–1mM. Cell growth, invasive activity (translocation across the Matrigel‐coated membrane insert), and EMT (change in cell shape, and β‐catenin nuclear translocation) were assessed at different periods of culture. Medium Thromboxane B2 (TXB2) conc was measured by ELISA.

Results

Co‐culture of colon cancer cells with platelets induced a significant increase in MC‐26 cell number and invasive activity and an activation of EMT, as the cells transformed from an epithelial to a mesenchymal shape with the generation of intracellular and extracellular microfilaments and an increase in medium TXB2 conc. Aspirin at concentrations between 0.3–1.0 mM blocked platelet activation (as indicated by a >95% reduction in medium TXB2) and all of the above dysplastic platelet‐induced changes in the colon cancer cells in culture.

Conclusion

Aspirin's chemopreventive action against colon cancer may be dependent on the drug's ability to irreversibly block platelet activation and the ability of platelets to promote cancer cell growth, invasion and EMT.

Support or Funding Information

Supported by NIH grant R21 CA182798.
Anti‐Cancer Effects of a Proposed Sigma‐2 Receptor Agonist Structurally Similar to Tamoxifen
Authors: Adams Bryce R. , Stang Christopher R.T. , Carlston Leah , Faler Ashton D. , Thomas Colin D. , Schuler Brian R. , Eichel Amanda M. , Whited Tawna L. , Kinder David H. , Schneider Ryan A.
Published: November 2023

Sigma receptor agonists present a new area for anti‐cancer drug development. Agonists of the sigma‐2 receptor are known to induce tumor cell death, but the mechanism by which they work is clouded by associated sigma‐1 receptor binding. We developed a series of potential sigma‐2 receptor agonists that take their design motifs from the drug tamoxifen. Here we report the anti‐cancer activity of the first in that series, SP‐1. The design of SP‐1 utilizes portions of tamoxifen's structure with the elimination of phenyl and ethyl groups from the terminal ethylene bridge, removing key features required for estrogen receptor binding. We hypothesized that SP‐1 would function as a sigma‐2 receptor agonist and thereby elicit non‐caspase‐dependent cell death in several diverse human cancer cells lines (A375 melanoma, CRL‐1620 glioblastoma, and HT‐29 colon carcinoma). Each of the three cell lines was treated with increasing concentrations (1–100 μM) of SP‐1 for 48 hours, followed by quantification of relative cell number with the CyQUANT® NF assay. Results from these experiments suggested relatively potent GI50 values of 32 μM in A375 cells, 36 μM in CRL‐1620 cells, and 64 μM in HT‐29 cells. To assess potential mechanisms of cytotoxicity, we treated each of the cell lines with 40 or 80 μM of SP‐1 for 24 and 48 hours and then measured effector caspase activity using the Caspase‐Glo® 3/7 Assay. Significant effector caspase activity was observed in each of the three lines treated with 80 μM of SP‐1 at 24 hours. Effector caspase activity was not noted at 24 hours with 40 μM treatments. However, caspase 3/7 activity was slightly, but significantly, increased using 40 μM of SP‐1 at 48 hours. Collectively, these results suggest that the proposed sigma‐2 receptor agonist, SP‐1, elicits anti‐cancer activity in several human cancer cell lines. While it is evident that SP‐1 can induce effector caspase activity, the timing and doses required to detect this activity suggest that alternative cell death pathways may indeed be involved. These results are consistent with what would be expected from a sigma‐2 receptor agonist. Further studies are ongoing to investigate alternative death pathways, to determine sigma receptor binding, and to optimize the structural requirements necessary for activity.
Ex‐vivo response of Plasmodium falciparum to Dihydroartemisinin in an area of Ota South‐west Nigeria
Authors: Dokunmu Titilope M. , Akinbohun Adesola E , Rotimi Solomon Oladapo , Rotimi Oluwakemi A , Ogunlana Olubanke O.
Published: November 2023

Artemisinin drugs remain the mainstay of falciparum malaria control globally. Ring‐stage survival assay (RSA) is a simple new method that can be used for detecting artemisinin resistance, thus this study aims to determine ex‐vivo parasite susceptibility to dihydroartemisinin and its correlation with in‐vivo parasite response to artemisinin combinations in an endemic area of Nigeria. Using the simple and sensitive ring‐stage survival assay (RSA), 50 parasite isolates from patients with malaria were exposed to DHA concentrations for 6 hours and ring stage survival was determined by microscopy within 72hours of drug exposure. In‐vivo parasite clearance was compared with parasite survival in matched patient samples. Parasitemia cleared rapidly within 24hours (mean 22.4 ± 0.2h) in all subjects. There was a dose‐dependent growth inhibition in drug‐exposed plates, and ring‐stage growth inhibition correlated positively with parasite clearance rates in all samples (r = 0.97, p<0.0001). P. falciparum isolates from this endemic area show sensitivity to artemisinin drugs both in‐vivo and ex‐vivo. The RSA method has potential to detect declines in parasite responses to artemisinins and can be used for continuous monitoring of responses especially in areas where resistance has not emerged.
Advances and trend analysis on science and technology against antibiotic‐ resistant bacteria through scientometric approaches
Authors: Sohn Eunsoo
Published: November 2023

Over the past century, the overuse and misuse of antibiotics have lead to create antimicrobial‐resistant bacteria(namely, superbugs) such as methicillin‐resistant Staphylococcus aureus(MRSA) and vancomycin‐resistant Staphylococcus aureus(VRSA). Worldwide growing of antibiotic use and the dramatic rise of antibiotic‐resistant bacteria have recently become serious health and social issues. Many researches have been conducted to get free from the threats of antibiotic resistance. This review is focused research and development trend on S&T advances against antibiotic resistant bacteria through scientometric analysis. Especially some implications in the case of South Korea, which has much use of antibiotics were derived from R&D trend analysis of each country.
EVIDENCES OF ANTIHYPERTENSIVE POTENTIAL OF EXTRACT FROM SOLANUM CAPSICOIDES ALL. IN SPONTANEOUSLY HYPERTENSIVE RATS
Authors: Simões Letícia Oliveira , da Silva Ana Quênia Gomes , Conceição‐Filho Ginaldo , Ribeiro Thácio , Niero Rivaldo , Filho Valdir Cechinel , Silva Darizy Flavia
Published: November 2023

The aims of this study were to characterize the pharmacological activity of the crude extract of S. capsicoides All. (MeOH‐Sc) and its fractions, on the cardiovascular system in spontaneously hypertensive rats (SHR) and their normotensive controls Wistar Kyoto (WKY), and analyze the phytochemical extract of this plant drug. The HPLC indicated the presence of at least two main compounds previously isolated from this plant, Cilistadiol and Cilistol. Intravenous injections MeOH‐Sc (5, 10, 20 and 40 mg/kg) induced hypotension in both mouse strains in a dose‐dependent manner. The reduction was prominent in SHR compared to WKY (** p<0.01) in doses of 40 mg/kg. This hypotensive effect was associated with a tachycardia was significantly greater (* p< 0.05) in animals WKY compared to SHR. In animals receiving MeOH‐Sc orally for 21 days a significant antihypertensive effect was observed in comparison to control. In addition, treatment with captopril and MeOH‐Sc prevented the development of increased of cardiac mass. Significant increase in urine volume of animals treated with MeOH‐Sc (**p<0.001 and *p<0.05) in the fifth and fifteenth day of treatment was observed. Mesenteric artery rings from animals treated with MeOH‐Sc pre‐contracted with Phe, have suffered vasorelaxation by SNP (10‐13‐10‐4M) concentration‐dependent manner with increased potency of pharmacological significant manner (*** p< 0.001) compared to control. The concentration‐response curve of Phe in mesenteric rings of these animals showed a significant decrease (** p <0.01) the power to Phe compared to control. The cumulative addition of MeOH‐Sc (0,001–1000 μg/ml) in the isolated superior mesenteric artery of SHR and WKY rats, in the presence or absence of functional endothelium intact and rings precontracted with phenylephrine (Phe 1μM), induced a vasorelaxation concentration‐dependent. In vascular rings precontracted with depolarizing solution (KCl 80mM), we observed a significant decrease (*p<0.05 and ** p<0.01) of pharmacological potency induced by MeOH‐Sc in both SHR and WKY. Additionally, MeOH‐Sc reversed contractions induced by CaCl2 in depolarizing medium without nominally Ca2+ in both animal strains. When administered cumulatively in the tonic phase of the contraction induced by S‐(−)‐ Bay K 8644 (200 nM), MeOH‐Sc induced significant changes in the values Emax and CE50 besides demonstrating induce their vasorelaxant effect through inhibition of the influx Ca2+ by ROC and SOC. These results suggest that the MeOH‐Sc has hypotensive effect, vasorelaxant and antihypertensive effect potential, possibly due to an influence of the drug on the plant Ca2+ influx in rat vascular smooth muscle.

Support or Funding Information

Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)