bluuebunny/crossref_metadata_2025_split huggingface dataset:
Learned Modulation of Innate Odor‐Driven Behavior Requires the Orbitofrontal Cortex
Authors: Miyamoto Kiana , Victoriano Joelle , Kathrotia Mayuri , Root Cory M. 
Published: June 2021

Animals use a combination of innate and learned behaviors to survive. Innate behaviors do not require experience and drive stereotypical, hard‐wired responses to particular environmental cues, such as predator odor. However, learning provides a mechanism to integrate past experience and even modulate innate behaviors. We have created a behavioral paradigm to study the learned override of mouse aversion to the fox odor, TMT. The ability of the brain to suppress innate aversion in favor of a learned attraction is a form of executive control. What are the neural circuits involved in this behavior? Our previous work demonstrated that a third order olfactory brain region, the cortical amygdala, mediates innate responses to odor. We hypothesized that a higher order structure should be required to suppress innate aversion and reinforce a positive value. A likely candidate is the orbitalfrontal cortex (OFC), which receives olfactory input from piriform cortex and has outputs to the innate pathways and reward circuitry. The OFC is implicated in executive functions such as value reassignment. We have tested the role of the OFC in modulation of behavioral response to fox odor.We designed a two‐port decision making task to measure the innate response of mice to attractive or aversive odor stimuli. When odor is paired with an equal water reward at each port, water restricted mice prefer the port with the attractive rose odor, 2‐phenylethanol, to the TMT port. However decreasing the amount of water available at the rose port teaches mice to prefer the fox odor port. The attraction persists even in the absence of water. Thus, mice can learn to be attracted to predator odor that predicts reward.To test if the OFC is involved in this preference reassignment, we performed a loss of function experiment using optogenetic tools to suppress activity in the neurons of the OFC of trained animals. Bilateral expression of the light gated chloride pump, halorhodopsin, and optical stimulation in the OFC during an unrewarded test phase were sufficient to block the learned behavior and revert the animals to their innate behavioral response of avoiding aversive odor. These findings reveal that the orbitofrontal cortex is indeed required for the learned override of innate behavior. We next plan to record neural activity in the OFC as mice perform the learning task to reveal changes that occur in OFC to accommodate learning, and to map out the relevant downstream targets of OFC.Support or Funding InformationGrant #: 4R00DC014516‐03 from NIH National Institute on Deafness and Other Communication Disorders (NIDCD)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
RGS4 Regulates Neurite Outgrowth And Cell Proliferation Mediated by STAT5B Transcriptional Responses
Authors: Pallaki Paschalina , Serafimidis Ioannis , Papadimitriou Elsa , Papakonstantinou Maria Pagona , Thomaidou Dimitra , Gaitanou Maria , Georgoussi Zafiroula 
Published: June 2021

Regulators of G proteins signaling (RGS) serve several cellular functions, despite their initial role as GTPase activating proteins. Recent evidence suggests that RGS proteins interact with transcription factors to mediate transcriptional responses. RGS4, a member of the B/R4 family of RGS proteins, is a multitask protein highly expressed in developing neurons, that has been shown to be a key regulator of opioid receptor signaling (ORs) by confering selectivity for G protein coupling, inhibiting OR‐mediated ERK1,2 phosphorylation and accelerating receptor internalization (Leontiadis et al. 2009; Georgoussi et al. 2006). We have previously shown that δ‐OR forms a multiprotein signaling complex, consisting of Gi/Go proteins and the Signal Transducer and Activator of Transcription 5B (STAT5B) that leads to neurite outgrowth upon DSLET‐δ‐OR activation (Georganta et al. 2010;2013). Based on these findings and the fact that RGS4 directly interacts with STAT5B to negatively regulate STAT5B activation (Pallaki et al. 2017) we wondered whether RGS4‐STAT5B interplay could be involved in cellular mechanisms controlling neurogenesis.Primary neuronal cortical cultures from RGS4−/− mice, lacking a functional RGS4, exhibited differential neuronal sprouting compared to wild types, as assessed by an increased axonal length and branch number, upon DSLET administration. Additional studies in brain extracts from adult RGS4−/− mice demonstrated increased levels of phospho‐STAT5B while exhibited altered expression levels of STAT5B inducible genes Snx9 and Ptk2 which are both highly implicated in axonogenesis. This result indicates that RGS4 is implicated in neurite outgrowth via a STAT5B‐mediated transcriptional regulation. On the other hand, Neuro2A cells stably expressing RGS4 exhibited a significant reduction in the cell number and proliferation rate whereas isolated RGS4−/− neural stem cells exhibited elevated proliferative properties with a concomitant increase of the mRNA levels of the STAT5B anti‐apoptotic target genes, Bcl‐2 and Bcl‐xl. The same expression pattern was also confirmed in RGS4−/− mature adult brain tissues. Collectively, these studies demonstrate for the first time a novel selective role of RGS4 in STAT5B mediated‐transcriptional responses providing insights into the role of RGS4 involvement in neuronal development and synaptic signaling upon δ‐opioid administration.Support or Funding InformationThis work was supported by grants NORMOLIFE (LSHC‐CT037733) and the GSRT Excellence II, 3722 ‐”NO‐ALGOS” to Z.G. The facilities of the Hellenic Research Infrastructure OPENSCREEN‐GR were used for protein expression.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Developing an Intranasal Colonization Model for NTHi in Mice
Authors: O'Neil Meghan , LaClair Ciara , Zavorin Mark , Pryharski Karin , Khan Nadeem , Kaur Ravinder , Pichichero Michael , Michel Lea Vacca 
Published: June 2021

Nontypeable Haeomphilius influenzae (NTHi) is a Gram‐negative pathogenic bacterium that can live as a commensal in the nasopharynx or can be the cause of Acute Otitis Media (AOM). Although there are currently no approved vaccines in the US to protect against NTHi, several NTHi recombinant protein vaccines are currently being studied for their safety and efficacy of protection. To assess the effectiveness of these prototype vaccines, we had to first establish NTHi colonization and AOM infection models in mice. To establish colonization, C57BL/6J mice were intranasally (IN) infected with PR8 (a mouse‐adapted H1N1 virus) prior to IN infecting with NTHi. Several coinfection protocols were tested to optimize NTHi colonization. Bacterial densities were determined at days 3 and 7 post‐NTHi infection by harvesting blood, nasal washes, and ear washes from the infected mice. Using this coinfection strategy, we were successful in developing a robust NTHi nasal colonization model in mice.Support or Funding InformationRochester Institute of Technology Honors Program Summer Fellowship and Rochester General Hospital Research Institute Summer Scholars FundThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A comparison of a Natural and Synthetic Stilbenoid, Arachadin‐3, on a Rotavirus Infected Human Intestinal Cell Line
Authors: Napier‐Jameson Rebekah , Witcher Caleb M. , Wisdom Stormey B. , Strange Essence B. , Triggs Doran S. , Saade Luanna L. , Taylor Josephine , Ball Judith M. , Medina‐Bolivar Fabrecio , Parr Rebecca D. 
Published: June 2021

Rotavirus (RV) causes severe diarrhea in young children and immunocompromised individuals. Several effective vaccines have been developed to immunize children between 2 to 8 months of age, but there are no vaccines or antiviral therapeutics for RV‐infected immunocompromised patients of any age. Our laboratory has shown a decrease in the number of infectious RV particles produced in HT29.f8 cells with the addition of the stilbenoid, arachidin‐3 (A3). Additionally, the viral protein, NSP4, that is important for viral replication is decreased, suggesting an effect on virus replication. Also, ultrastructural changes were observed with TEM studies that imply a connection with the apoptosis and autophagy pathways which is supported by qRT PCR data. Recently, a synthetic A3 (sA3) has been developed. This study compared the effects of the natural A3 and sA3 on HT29.f8 cell viability, production of infectious RV particles, production of structural and nonstructural viral proteins, ultrastructural changes and the gene regulation of apoptosis and autophagy transcripts. From the data collected, the viability of the cells was not affected with the introduction of either natural or sA3, but the amount of infectious virus particles produced was significantly decreased by 50‐fold, respectively. The TEM images depicted changes in ultrastructures of RV‐infected HT29.f8 cells that appear to show signs of apoptosis while RV‐infected cells with the addition of either the natural or synthetic A3 showed a more normal appearance like the negative controls. The nucleus to cytoplasm ratios of TEM micrographs of RV‐infected cells was greater than 1.0 whereas the arachidin treated RV‐infected cells was about 0.5 which was like the control cells. To validate this data, whole cell staining of the nucleus and cytoplasm showed similar changes with the addition of both the A3 and sA3. qRT PCR analysis of transcripts for genes that are important in regulating the apoptosis and autophagy pathways regulations corroborated the TEM observation, and suggests a cross talk between the two pathways during an RV infection that is regulated with the addition of the arachidins. Our data suggests that both the natural A3 and sA3 have anti‐RV activity that implies a potential therapeutic application.Support or Funding InformationThis work was supported by: 1) The Faculty‐Student Collaborative Research Program of The Office of Research and Sponsored Programs at Stephen F. Austin State University and 2) The NSF‐EPSCoR (grant# EPS‐ 0701890; Center for Plant‐Powered Production‐P3).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
RASG12D causes more Proliferation than RASG12V in Drosophila Pancreatic Cancer Models
Authors: Bernard Faviola , Sonoshita Masahiro , Cagan Ross 
Published: June 2021

Pancreatic cancer is the fourth leading cause of death in the United States. Over 95% of pancreatic cancer patients have activating mutations in the KRAS oncogene, suggesting that these mutant KRAS forms play a critical role in development of the disease. The most common mutations in KRAS are G12D and G12V missense mutations, but little is known about functional differences between them. Therefore, our objective is to compare the outcomes between KRASG12D and KRASG12V mutations in pancreatic cancer models. Using the Gal4‐UAS system, which allows the expression of transgenes in a specific tissue, we developed pancreatic cancer models in the fruit fly Drosophila. The patched promoter‐driven Gal4 transcription factor targeted the Drosophila RasG12D or RasG12V transgene to the midline of the fly wing epithelium. Using green fluorescence protein (GFP) as a marker for Ras‐expressing cells, we found that RASG12D directed more aggressive expansion of the patched domain than wild type or RASG12V wing epithelia. Oral dosing of trametinib—a MEK inhibitor that blocks signaling downstream of RAS—decreased proliferation of RASG12D expressing cells. These results indicate that trametinib could be a useful drug against RAS‐positive cells for RASG12D mutations. Future experiments include understanding the mechanisms by which RASG12D and RASG12V direct different phenotypes, and further developing novel treatments based on trametinib.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antimycobacterial and macrophage apoptosis inducing effects of Psychotria capensis and Psychotria zombamonatana species
Authors: Aro Abimbola , Fonteh Pascaline , McGaw Lyndy Joy 
Published: June 2021

The causative agent of tuberculosis (TB), Mycobacterium tuberculosis (Mtb), has succeeded in infecting one‐third of the human race through inhibition or evasion of the innate and adaptive immunity. Mtb was responsible for 1.5 million deaths and the emergence of 9.6 million new cases of TB in 2014. Apoptosis is a cellular biology process that is closely involved in the interaction between Mtb and the phagocytes it infects such as macrophages, dendritic cells and neutrophils. It has been suggested that apoptosis of infected macrophages is one way in which the host deals with intracellular pathogens. To effectively combat these disease, new TB drugs with novel modes of action different from the presently administered drugs are urgently needed. Approximately 65% of antibacterial approved for use between 1981 and 2010 were natural products or natural product derivatives including currently employed TB drugs such as rifampicin and the aminoglycosides The acetone extracts of Psychotria capensis and Psychotria zombamontana have been reported to be active against non‐pathogenic mycobacteria This study aims to assess the antimycobacterial activity of P. capensis and P. zombamontana against pathogenic strains of mycobacteria and also to shed some light on elucidating the anti‐tuberculosis mechanism of the selected plant extracts.The antimycobacterial activities of the crude extracts of these two species belonging to the rubiaceae family were screened against Mycobacterium tuberculosis ATCC, Mycobacterium bovis ATCC and H37Rv strains to determine the minimum inhibitory concentration (MIC). THP‐1 cells were treated with varying concentrations (20, 50, 100 μg/mL) of each extract for 24 and 48 h. Cell apoptosis was measured by annexin V/propidium iodide apoptosis kits using flow cytometry. The concentration of IL‐12 in the supernatant of the cell culture was determined by using the human enzyme linked immunosorbent assay (ELISA) to assess the immune modulatory effect of the extracts.The acetone crude extracts of the plant species had an excellent antimycobactrial activity against the tested mycobacteria strains with a MIC values ranging from 78–156 μg/mL. P. capensis and P. zombamontana extracts induced apoptotic effects on THP‐1 cells following treatment in a time and dose‐dependent manner. Interestingly, the acetone extracts of P. capensis revealed a profound macrophage apoptosis induction (16%) and also enhanced the expression of IL‐12 in a dose dependent manner. Results obtained from this study suggest that the acetone extracts of P. capensis may contain promising lead compounds for anti‐TB therapy. There is on‐going work to isolate compounds and analogues responsible for the observed anti‐TB activity.Support or Funding InformationUniversity of Pretoria, South AfricaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Carnosic acid activates AMPK, inhibits Akt and inhibits H1299 human lung cancer cell survival
Authors: Nyforovskyy Dmytro , Moore Jessy , Tsiani Evangelina 
Published: June 2021

Non‐small cell lung cancer (NSCLC) represents 80% of all lung cancers and is characterized by low survival rates due to chemotherapy and radiation resistance. Thus, finding alternative treatments to reduce NSCLC proliferation and survival is of high importance. The survival pathways of Akt, the energy sensor AMP‐activated protein kinase (AMPK), and the apoptotic protein poly (ADP‐ribose) polymerase (PARP) are key modulators of cancer cell growth and survival. In previous studies, we found inhibition of NSCLC proliferation and survival by rosemary extract but the exact components responsible for the anticancer effects and the cellular mechanisms involved are not known. Rosemary extract contains many polyphenols including carnosic acid (CA). The objectives of the present study were to examine the effects of CA on NSCLC cell survival and apoptosis, and to investigate its effects on Akt, AMPK and PARP. The human NSCLC cell line H1299 was used. Clonogenic cell survival assays were performed to examine the effects of CA on cell survival, and immunoblotting with phospho‐specific antibodies was performed to examine signaling events. CA dose‐dependently inhibited H1299 cell survival. A significant inhibition of Akt phosphorylation/activation and enhanced AMPK phosphorylation/activation was seen with 25 and 50 mM CA while the total levels of each protein were not affected. PARP cleavage, an indicator of apoptosis, was enhanced by carnosic acid treatment. Our findings indicate that CA may have robust anticancer properties in NSCLC, and strongly support the need for further studies to investigate its role against lung cancer.Support or Funding InformationSupported by a Brock University Advancement Fund (BUAF) grant to ETThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of A Novel RON Targeted Antibody‐Drug Conjugates using Cysteine Bridging Technology for Potential Treatment of Pancreatic Cancer
Authors: suthe sreedhar reddy , Yao Hang‐Ping , trippier paul c. , wang ming‐hai 
Published: June 2021

Therapeutics targeting known oncoproteins have been applied for pancreatic cancer treatment but clinical outcomes are not promising. Hence, there is an urgent need to identify novel targets and develop effective drugs to improve pancreatic cancer therapeutic index. Antibody‐drug conjugates (ADC) represent a promising class of drugs for targeted cancer therapy. Here we developed a novel ADC targeting RON receptor tyrosine kinase for potential pancreatic cancer treatment. To this end, we have synthesized a bis‐alkylating linker (BL), attached to a lysosomal protease‐cleavable dipeptide with payload Monomethyl auristatin E (MMAE). The BL‐MMAE was then conjugated to Zt/g4 (anti‐RON mAb) through cysteine bridging technology to produce Zt/g4‐BL‐MMAE with a relatively homogeneous conjugation profile and an antibody to drug ratio of 1:4. Zt/g4‐BL‐MMAE showed significant improvement in drug conjugation homogeneity and serum stability over conventional ADCs prepared by maleimide‐based linkers. In pancreatic cancer cell lines overexpressing RON, Zt/g4‐BL‐MMAE specifically targeted RON‐expressing tumor cells and was effective in rapid induction of cell surface RON endocytosis. Functional analysis revealed that Zt/g4‐BL‐MMAE caused cell cycle arrest at G2/M phase, reduction of cell viability and subsequently resulted in massive cell death. The calculated IC50 is in the range of 1 to 2 μg/ml. Therapeutic studies using mouse xenograft models are currently underway to determine the efficacy of Zt/g4‐BL‐MMAE for pancreatic and other epithelial cancers. We conclude that Zt/g4‐BL‐MMAE is a novel anti‐RON ADC with excellent conjugation profile, serum stability, and selective cytotoxicity for pancreatic cancer cells. This work provides a pharmaceutical opportunity for evaluating potentials of RON‐targeted ADCs in pancreatic cancer treatment in the future.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
How cAMP Homeostasis is Controlled in Leishmania by Receptor Adenylate Cyclases and Acidocalcisomal Pyrophosphatases Aiding its Survival in Phagolysosomal Conditions?
Authors: BISWAS ARUNIMA , BHATTACHARYA ARIJIT , Vij Amit , Das Pijush K. 
Published: June 2021

Leishmania donovani while infecting macrophages is exposed to phagolysosome conditions (PC) (37 °C and pH 5.5) from the sandfly gut environment of 22 °C and pH 7.4. This shift in temperature and pH led to increased intracellular cAMP and cAMP‐mediated PKA activity in the parasites, which help in intra‐macrophage survival, a pre‐requisite for infectivity and transformation to amastigotes. Though cAMP and cAMP‐triggered signaling pathways seemed to be one of the important factors for parasite survival and infectivity, the mechanism of cAMP production and its stringent control is obscure. In the absence of typical orthologs for G‐proteins and G‐protein coupled receptors, the precise mechanisms for positive modulation of cAMP production during exposure to PC seemed an important aspect to be studied. Amongst various membrane bound isoforms of receptor adenylate cyclase, two promastigote‐stage specific membrane bound receptor adenylate cyclases (LdRAC‐A and LdRAC‐B), LdRAC‐A appeared to function as a major cAMP generator following PC exposure. Following PC exposure, pyrophosphate (PPi), an energy storage compound as well as a by‐product of cAMP biosynthesis by adenylate cyclase, was found to be decreased. This may be due to microtubule and microfilament‐driven translocation of acidocalcisomes near plasma membrane vicinity with concomitant increase of acidocalcisome membrane pyrophosphatase (LdV‐H+PPase) and acidocalcisomal soluble pyrophosphatase (LdVSP1). Episomal over‐expression and conditional silencing demonstrated regulatory role of V‐H+PPase on cAMP trigger and consequent induction of resistance to macrophage‐derived pro‐oxidants and parasite killing. Moreover, immunofluorescence analysis indicated towards a possible colocalization of LdV‐H+PPase and LdRAC‐A during PC exposure. The results suggested that translocation of acidocalcisome in membrane vicinity functions as a trigger for LdRAC‐A‐driven cAMP generation through depletion of PPi pool by LdV‐H+PPase. For the first time, one of the molecular mechanisms behind the positive modulation of cAMP by leishmanial adenylate cyclases in absence of Gproteins was deciphered by us.Support or Funding InformationDST‐INSPIRE, Department of Science and Technology (DST), Govt of India, DST‐PURSE, Department of Science and Technology (DST), Govt of India, UGC‐SAP, UGC, Govt of IndiaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.