bluuebunny/crossref_metadata_2025 huggingface dataset:
Sensitization of Anti‐EGFR/HER2 Targeted Therapy Resistance in Breast Cancer Cells by MiRNA Manipulation
Authors: Vigo‐Morales Paula Rocío , Borrero‐Garcia Luis Daniel , Del Mar Maldonado Maria , Dharmawardhane Suranganie
Published: June 2021

The aim of this study is to explore the resistance mechanisms to anti‐epidermal growth factor receptor (EGFR) and anti‐human epidermal growth factor receptor (HER2) targeted therapies in breast cancer (BC). Previous studies have implicated the microRNA (miR)‐200 family in maintenance of the epithelial phenotype and sensitivity to EGFR targeted therapies and miR‐221/222 have been associated with epithelial to mesenchymal transition (EMT), tumor progression and therapy resistance. To investigate mechanisms of EGFR/HER2 therapy resistance in breast cancer, therapy resistant variants of the therapy sensitive SKBR3 human breast cancer cell line were created by exposure to different concentrations of the EGFR/HER2 inhibitors lapatinib and gefitinib. Characterization of therapy sensitive and resistant variants of SKBR3 demonstrated that miR‐200a is downregulated while miR‐221 and miR‐222 are overexpressed in the therapy resistant cells. Therefore, the current objective is to validate the role of these miRNAs in therapy resistance. The hypothesis is that re‐expression of miR‐200a and downregulation of miR‐221/222 will help overcome therapy resistance to EGFR/HER2 targeted therapy. Therefore, we re‐expressed miR‐200a ectopically in the therapy resistant and sensitive cells using a lentiviral vector containing miR‐200a, or a control vector. MiR‐221/222 inhibition was achieved by expressing anti‐miR‐221/222 or control inhibitors. MTT assays were used to determine the IC50s for cell viability. Both SKBR3 parental and lapatinib resistant variants with ectopic expression of miR‐200a were more sensitive to lapatinib treatment as shown by a decrease in the IC50 when treated with lapatinib. Therapy sensitive and resistant cell lines also demonstrated a decrease in cell viability when miR‐221 or miR‐222 was downregulated using miRNA inhibitors. In conclusion, overexpression of miR‐200a or downregulation of miR‐ 221/222 family reduces the malignancy of SKBR3 parental cell lines and sensitizes EGFR/HER2 therapy resistant cells. These results suggest that manipulation of miR‐200a, miR‐221 and miR‐222 can be used as therapeutic strategies in combination with HER2/EGFR therapy to reduce cancer malignancy and therapy resistance.

Support or Funding Information

This work was supported by grants from NIH/NIGMS SC3GM094824 to SD, RCMI Programs G12RR03051 to UPR MSC, MBRS Program RISE (R25GM061838) to LDB and MM, and Bright Cure Program (U54) University of Puerto Rico/MD Anderson Cancer Center to PRV.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antimetastatic potential of Atovaquone against triple negative breast cancer: Involvement of the integrin‐FAK‐Src pathway
Authors: Gupta Nehal , Srivastava Sanjay
Published: June 2021

Triple negative breast cancer (TNBC) is considered to be the most aggressive and malignant neoplasm and is highly metastatic in nature. Metastatic TNBC is difficult to treat and thus leads to poor patient survival. Death of about 90% breast cancer patients is due to metastasis of primary tumor to distant sites. In the current study, we investigated the anti‐metastatic potential of atovaquone, a protozoal drug prescribed for Pneumocystis pneumonia. We show that atovaquone induced apoptosis and reduced the survival of several aggressive metastatic TNBC cell lines including few metastatic patient derived cells. The IC50 of atovaquone was around 11–18 μM in the breast cancer cell lines tested. Additionally, atovaquone treatment significantly reduced the expression of integrinα6, integrin β4, FAK, Src, and Vimentin in MDA‐MB‐231, HCC1806 and 4T1 cells after 72 hours of treatment. In order to study the efficacy of atovaquone in preventing metastasis of breast tumor cells to brain and lungs, we performed three in vivo experiments. We demonstrate that oral administration of 50mg/kg of atovaquone suppressed MDA‐MB‐231 breast tumor growth by 90% in lungs in an intravenous metastatic tumor model. Further anti metastatic effect of atovaquone was determined by intracardiac injection of 4T1‐luc breast tumor cells into the left ventricle of mouse heart. Once breast cancer cells lodged in the brain, mice were treated with 50 mg/kg atovaquone for 15 days. Our results show that atovaquone treatment suppressed the growth of metastatic tumors in brain by 30%. In this model, we also observed 70% and 50% reduced metastasis of breast tumors in lungs and liver, respectively, in atovaquone treated group when compared with control. In intracranial model, the growth of HCC1806‐luc brain tumors in atovaquone treated mice was about 55% less than that of control. Angiogenesis is one of the hallmarks of cancer and plays crucial role in metastasis. Interestingly, we observed reduced levels of VEGF in the cells treated with atovaquone. In‐vivo angiogenesis was evaluated by performing matrigel plug assay in mice. Our results showed reduced angiogenesis in atovaquone treated mice group. Atovaquone treated tumors from these in‐vivo models exhibited reduced metastatic markers and increased apoptosis. Taken together, our results indicate the anti‐metastatic effects of atovaquone in‐vitro and in‐vivo in various breast tumor metastasis models by inhibiting angiogenesis and integrin signaling and provides the rationale for further clinical investigation of atovaquone for breast cancer metastasis.

Support or Funding Information

This work is supported in part by RO1 grant CA129038, awarded by National Cancer Institute, (NIH) to Sanjay K. Srivastava.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Determining the role of Pal in Escherichia coli sepsis
Authors: Gallardo Leslie , Bauer Morgan , Pannullo Nicole , Michel Lea Vacca
Published: June 2021

Sepsis, a life‐threatening clinical condition affecting more than 1.5 million Americans per year, is defined as an over‐exuberant immune response to infection. Currently, sepsis is the leading cause of death in U.S. hospitals, and the incidence of sepsis caused by Gram‐negative bacteria, such as Escherichia coli (E. coli), has been steadily increasing since the late 1990's. While the detailed mechanism of sepsis is not fully understood, several bacterial components are thought to contribute to the hyper‐inflammatory response in humans. Past studies suggest that one E. coli lipoprotein, Peptidoglycan‐Associated Lipoprotein (Pal), may contribute to inflammation and the pathogenesis of sepsis. This work describes our efforts to elucidate the role of Pal in E. coli sepsis and the effect of antibiotics in Pal release from E. coli using a mouse model of sepsis. Our preliminary results corroborate the hypothesis that Pal releases from E. coli and contributes to inflammation and sepsis.

Support or Funding Information

Rochester Institute of Technology

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of non‐C2 symmetric salen ligands as novel antimicrobial and antifungal agents
Authors: Allgood Cory W. , Wolyniak Michael J , Deifel Nicholas P
Published: June 2021

The importance of salen ligands in homogeneous catalysis and material chemistry/biochemistry has unambiguously been demonstrated in procedures that include the synthesis of C2‐symmetrical complexes that are used as catalysts for many organic conversions with industrial applications. Of late, research on salen ligands has turned to the potential for non‐C2 symmetric ligands to act asantimicrobials and antifungals. As drug‐resistant microbes become a larger threat to public health, novel compounds like non‐C2 symmetric salen ligands could be an invaluable resource for disease control.

Here, a practical synthesis of a non‐C2 symmetric salen ligand is described using a 1:1:1 molar ratio of phenylenediamine and two different salicylaldehyde's which can be important in testing catalytic properties along with antimicrobial activity. Specially, we have built a non‐C2 symmetric ligand using varying R groups and metal complexes and have characterized the resultant product using 13C‐ NMR, 1H‐NMR, infrared spectroscopy, and gas chromatography‐mass spectroscopy. Current research focuses on the use of metal ions such as iron and zinc in the formation of efficient and cost‐effective novel antimicrobial and antifungal compounds that can be tested using Kirby‐Bauer analyses.

Support or Funding Information

Hampden‐Sydney College Office of Undergraduate Research

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structure‐activity relationship of a novel Staphylococcus aureus biofilm formation inhibitor
Authors: Shah Ashna , Baker Nnamdi , Loving Danielle , Hensel Linda , Goode David
Published: June 2021

Ideal biofilm inhibitors would not affect the growth of bacteria, in general, but would be specific for inhibiting reception of the signals to initiate biofilm formation. In this manner, the inhibitor would not affect the growth of normal flora that are imperative for the health of the host. Compounds that resemble biofilm signaling molecules have the potential to competitively inhibit the receptors from receiving signals. Over 400 novel compounds were synthesized and tested by undergraduate students in a sophomore integrated biology and organic chemistry classroom setting. Compounds that demonstrated a statistically significant inhibition of biofilm formation and that had no bacteriostatic or bactericidal activity in one or more of five species of bacteria (Streptococcus mutans, Escherichia coli, Bacillus subtilus, Staphylococcus aureus, and Pseudomonas aeruginosa) in the hands of the undergraduates underwent further testing. Confirmed hits were further tested for a dose response. Compound Tyr‐69 was a promising hit compound with specific inhibition of Staphylococcus aureus biofilms. A structure‐activity analysis was pursued via synthesis and dose response assays.

Support or Funding Information

This work is supported by the Biology and Chemistry Departments of Mercer University as well as the Mercer University Quality Enhancement Plan: Research that Reaches Out.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Glycogen phosphorylase inhibition activates intrinsic apoptosis pathway and potentiates multi‐kinase inhibitors in liver cancer cells
Authors: Barot Shrikant , Abo‐Ali Ehab M , Zhou Daisy , Palaguachi Christian , Dukhande Vikas D
Published: June 2021

Hepatocellular carcinoma (HCC) is one of the leading cancers in terms of incidence and mortality. There are fewer chemotherapies with HCC with a low survival rate. One of the prominent features of cancer is metabolic reprogramming and thus metabolic proteins are viable targets for cancer treatment. Glycogen is a polymer of glucose, which acts as a reservoir of energy in mammalian cells. The levels of glycogen have been found to correlate with biological processes in cancer cells, however, the role of glycogen metabolism in carcinogenesis, cancer cell growth, metastasis, and chemoresistance is not completely understood. Thus, we studied the cellular processes involved in the inhibition of glycogen metabolism in HCC. Pharmacological inhibition of glycogen phosphorylase (GP) by CP‐91149, a rate‐limiting enzyme in glycogen catabolism, led to a decrease in HepG2 viability. GP inhibition induced the intrinsic apoptosis pathway causing HepG2 cell death. This process was accompanied by mitochondrial dysfunction and autophagic adaptations, whereas endoplasmic reticulum stress, necrosis, and necroptosis were not involved. In addition, GP inhibition potentiated the effects of the multi‐kinase inhibitors sorafenib and regorafenib, which are key drugs in advanced‐stage HCC therapy. Our study provides mechanistic insights into cell death by perturbations of glycogen metabolism and identifies GP inhibition as a potential cancer pharmacotherapy target.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Heme Interaction with the Pyruvate Dehydrogenase Complex: A Novel Strategy to Promote Hypoxic Survival
Authors: Lynch John Allan , Fukuda Yu , Krishnamurthy Partha , Wijaya Juwina , Wang Yao , Herras Andras , Cheepala Satish , Bao Ju , Nourse Amanda , Milasta Sandra , Neale Geoffrey , Zheng Jie , Green Douglas , Chi Hongbo , Schuetz John
Published: June 2021

Heme is an important regulator of biological processes, but an unresolved question remains, can heme regulate the switch between glycolysis and mitochondrial oxidation in mammalian cells? Here we show that ABCB6, a member of the ATP‐binding cassette (ABC) family and an activator of heme synthesis facilitates a heme dependent metabolic transition between mitochondrial oxidative phosphorylation and glycolytic metabolism. ABCB6 is upregulated by the Hif‐1α pathway and mimicking this, overexpression of ABCB6 primarily increased mitochondrial heme, reduced pyruvate‐fueled mitochondrial oxygen consumption and increased hypoxic survival. Suppression of heme synthesis abolished this hypoxic survival advantage. Deletion of ABCB6 increased mitochondrial oxygen consumption in the inherently hypoxic double‐negative thymocytes. We hypothesized that pyruvate dehydrogenase (PDH) activity was suppressed by heme and directly demonstrated that heme potently inhibited PDH activity. Hemin‐agarose affinity chromatography revealed that only one subunit of the multimeric PDH complex, PDHA1, is bound to heme. Mutagenesis of PDHA1 suggested that the heme binding site was distinct from the substrate binding site, a finding consistent with heme inhibiting PDH non‐competitively. Heme binding to PDHA1 resulted in its dissociation from the PDH complex. PDHA1 dissociation was reversible, as dilution of heme containing PDH promoted reformation of the complex, which paralleled restoration of PDH activity. These findings reveal a new relationship between heme production and the ability to metabolically switch from pyruvate oxidation in the mitochondria to its glycolytic conversion to lactate.

Support or Funding Information

This work was supported by NIH and by the ALSAC.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of Non‐Polyamine based Polyamine Transport Inhibitors for the Treatment of Pancreatic Ductal Adenocarcinoma
Authors: Moots Holly , Phanstiel Otto
Published: June 2021

The native polyamines (putrescine, spermidine and spermine) are low molecular weight, biogenic amines, which play key roles in transcription, translation, immunity and chromatin remodeling. Polyamine depletion results in inhibited cell growth and has been used to treat human cancers with mixed results. While inhibitors of polyamine biosynthesis, like difluoromethylornithine (DFMO), result in intracellular polyamine depletion and decreased cell growth, the effect is mostly cytostatic. Indeed, cancer cells when treated with DFMO often rescue themselves by upregulating polyamine import to replenish their depleted polyamine stores. In this regard, a combination therapy comprised of a polyamine synthesis inhibitor (e.g., DFMO) and a polyamine transport inhibitor (PTI) are required to provide sustained polyamine depletion and limit tumor growth.

Our group has previously developed polyamine‐based PTIs and this report describes our efforts to develop non‐polyamine based PTI agents in hopes of improving drug efficacy. These agents are predicated upon piperazine scaffolds and provide an opportunity to develop orally active PTIs to be given in combination with DFMO. The remarkable thing about this technology is that the DFMO+PTI therapy not only inhibits cancer cell growth, but also unleashes the immune system to attack the tumor. In this regard, this approach provides a novel ‘chemoimmunotherapy’ to help patients with cancer. Since pancreatic cancer is the third leading cause of cancer‐related death with few treatment options available, we have developed these new medicines in hopes of providing new approaches to attack these seemingly ‘immunologically‐quiet’ tumors. This report will describe the development of our synthetic efforts to optimize these PTI designs.

Support or Funding Information

2016 Florida Translational Research Program Award and the 2018 Bankhead Coley Cancer Research Program Award # 8BC05.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Aqueous Gladiolus psittacinus Bulb Extract Influences Antioxidant Enzymes Activities in Diabetic Rats
Authors: Oyetayo Folake Lucy , Oseni Olatunde Abass
Published: June 2021

Aqueous extract of the bulb of Gladiolus psittascinus a tropical plant used as adjunct in the preparation of a non‐alcoholic drink was evaluated for its antioxidant activities in diabetic albino rats. After a 14‐day acclimatization period, rats were divided into five treatment groups of four animals per group as follows: Group 1: Control (Uninduced), Group 2: Alloxan induced diabetic rats, Group 3: Alloxan induced rats treated with standard drug, Group 4: Un‐induced rats treated 100mg/kg body weight aqueous Gladilous psittascinus bulb extract, Group 5: Alloxan induced diabetic rats treated 100mg/kg body weight aqueous Gladilous psittascinus bulb extract. On the establishment of hyperglycemia, animals were sacrificed; plasma antioxidant enzyme activities and lipid peroxidation levels were estimated. Activities of all antioxidant enzymes evaluated increased significantly. Plasma superoxide dismutase activity significantly increased (p<0.05) in the diabetic untreated rats from13.76±2.50U/L (Group 2) to 45.76±5.40U/L in the diabetic aqueous Gladilous psittascinus bulb extract treated rats (Group 5). Furthermore, Catalase activity in the diabetic group (2.48±0.43U/L) significantly increased (p<0.05) to 10.49±1.98U/L in the diabetic aqueous Gladiolus psittascinus bulb extract treated rats (Group 5) and there was a significant decrease in lipid peroxidation levels in plasma from 13.23±3.14 μmolMDA/L (Group 2) to 6.28±1.05 μmolMDA/L (Group 5). The foregoing shows the potentials of aqueous Gladilous psittascinus bulb extract in enhancing the production of antioxidant enzymes required to scavenge reactive oxygen species produced in the diabetic state. Hence, the Gladilous psittascinus bulb holds promise for the food industry in the control of oxidative changes that may occur during food processing and diet therapy in the management of oxidative stress related disorders such as Diabetes mellitus.

Support or Funding Information

Tertiary Education Trust Fund (TETFUND) through Ekiti State University Ado‐Ekiti, Nigeria

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Glycerol Improves Skin Lesions in a Mouse Model of Psoriasis: Possible Mechanism
Authors: Bollag Wendy B , Choudhary Vivek , Uaratanawong Rawipan , Kaddour‐Djebbar Ismail , Custer Victoria , Cohen Elyssa , Zhang Catherine
Published: June 2021

Psoriasis is a common skin disorder characterized by hyperproliferation and abnormal differentiation of the major cells comprising the skin epidermis, keratinocytes, as well as by skin inflammation. We have previously shown a physical and functional association between the glycerol channel aquaporin‐3 (AQP3) and the enzyme phospholipase D2 (PLD2), which can convert the AQP3‐transported glycerol to phosphatidylglycerol (PG). Further, we have recently demonstrated an ability of PG to inhibit toll‐like receptor‐mediated innate immune system activation by antimicrobial peptides, which are upregulated in psoriasis and can act as danger‐associated molecular patterns to activate toll‐like receptors (TLRs). Glycerol has been reported anecdotally by some patients to improve psoriasis, and we hypothesized that it does so by its PLD2‐mediated conversion to PG. To test this idea, we first demonstrated the ability of topical glycerol to ameliorate inflammation and lesion development in the imiquimod‐induced mouse model of psoriasis. Glycerol reduced ear edema, improved the psoriasis area and severity index (PASI) score, a measure of the appearance and degree of skin lesions, and inhibited the imiquimod‐increased epidermal thickness and tumor necrosis factor‐alpha immunoreactivity. When the same experiment was performed in PLD2 knockout mice, preliminary analysis indicated that glycerol also ameliorated skin lesions; however, topical glycerol is known to function as an emollient, with the capacity to improve skin function and appearance. To overcome this topical glycerol effect, we repeated the experiments using oral glycerol (2% in the drinking water). In the wild‐type mice, oral glycerol again improved the inflammation and skin lesion development induced by imiquimod. On the other hand, there was no significant effect of oral glycerol on these parameters in PLD2 knockout mice. These results provide the first experimental evidence to support the beneficial effects of glycerol in psoriasis. Furthermore, our data suggest that conversion of glycerol to PG underlies in part the glycerol effect and imply that glycerol and/or PG might constitute an effective treatment for this common skin disease.

Support or Funding Information

Supported in part by a Research Career Scientist Award and VA Merit Award #CX001357 to WBB. The contents of this article do not represent the official views of the Department of Veterans Affairs or the United States Government.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hemodynamic Characteristics of Elderly Patients with High Blood Pressure
Authors: Hao Chunyan , Duan Hubin , Wang Suping
Published: June 2021

Objective

To study hemodynamic characteristics of elderly patients with high blood pressure.

Methods

The study selected 94 geriatric patients older than 60 years old in the First Hospital of Shanxi Medical University hospitalized from November 1, 2016 to May 1, 2017. The study included 52 males, 42 females. The basic information were collected including age, sex, weight, and height, and years of hypertension and the level of blood pressure were collected for all hypertensive patients. Non‐invasive hemodynamic monitor records Stroke Volume (SV), Stroke Volume Index (SI), Cardiac Output (CO), Cardiac Output Index (CI), System Vascular Resistance (SVR), Thoracic Fluid Content (TFC) ), Left Ventricular Stroke Work (LSW) and Left Ventricular Stroke Work Index (LSWI), as well as Vascular Elasticity and Contractility Variability.

Results

According to blood pressure level classification, there was no significant statistical difference among the general clinical data of all subjects such as age, height, weight, body mass index(P>0.05). According to blood pressure levels of hypertensive patients, the hemodynamic parameters had no obvious variability pattern. And with the age increase, SV, SI, CO, CI, vascular elasticity and cardiac contractility variability reduced, SVR increase; and TFC did not change significantly. In addition, LSW and LSWI in elderly hypertensive patients over 80 years of age changed significantly (P <0.05). According to the level of systolic blood pressure vs diastolic blood pressure, SV, SI, CO, CI, cardiac contractility and other indicators were significantly higher than the control group, and the rest of hemodynamic parameters were not statistically different. As for the high systolic blood pressure group, SV, SI, CO, CI and vascular elasticity are significantly decreased, but SVR, TFC are significantly increased. For both of the high systolic and diastolic blood pressure, LSW and LSWI are significantly increased (P <0.05).

Conclusion

The use of non‐invasive hemodynamic tests to obtain different hemodynamic parameters may provide a scientific basis for the diagnosis and personalized treatment of hypertensive patients.

Support or Funding Information

This study was supported by the National Natural Science Foundation Youth Fund (30600637), China Postdoctral Foundation(2014M561207), the Shanxi Province Basic Research Program Youth Science and Technology Research Fund (2010021034‐4, 201601D011119), the Shanxi Scholarship Council of China (2011‐096 and Key Projects No.4), Department of Human Resources and Social Security of Shanxi province, The science and technology research projects of health department of Shanxi Province(2016D011119), The Teaching Reform Program of Shanxi Higher Education, The Education Reform Project of Postgraduate Students in Shanxi Province. The Education Reform Project of Shanxi Medical University.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of the Actin‐binding Protein α‐Adducin for Desmosome Turnover and Keratinocyte Adhesive Function
Authors: Hiermaier Matthias , Wanuske Marie‐Therès , Vielmuth Franziska , Waschke Jens , Spindler Volker
Published: June 2021

Desmosomes are cell‐cell junctions which provide strong intercellular adhesion and couple the intermediate filament cytoskeleton to sites of cell‐cell contact. We have previously shown that the actin‐binding protein α‐adducin is required for desmosome‐mediated intercellular adhesion. We here tested in more detail the mechanisms underlying α‐adducin‐dependent regulation of desmosomal turnover and adhesion. Experiments were conducted in human keratinocytes with siRNA‐mediated knock‐down of α‐adducin, in knock‐out cell lines obtained with the CRISPR/Cas9 system and in murine knock‐out cells. Dispase‐based dissociation assays confirmed reduced intercellular binding strength in cells lacking α‐adducin. A more immature desmosome state in α‐adducin knock‐outs was indicated by reduced Ca2+‐insensitivity. α‐adducin knockout led to elevated levels of desmoglein (Dsg) 2 and Dsg3 in the membrane. In atomic force microscopy (AFM) single molecule experiments, an increased interaction probability of Dsg3 was detectable in α‐adducin knock‐out cells. This was accompanied by increased amounts of membrane tethers which is in line with a less rigid membrane structure and a role of α‐adducin in organizing the cortical actin cytoskeleton. Ca2+‐depletion/repletion experiments revealed enhanced fragmentation of Dsg3 staining in the membrane in α‐adducin knock‐out cells, indicating dysfunctional desmosome turnover. Indeed, life cell imaging experiments showed less directed intracellular transport of Dsg3‐containing vesicles to the cell membrane whereas fluorescence recovery after photobleaching (FRAP) studies and single molecule imaging demonstrated increased mobility of extradesmosomal Dsg3 in the cell membrane. Interestingly, despite their decreased intercellular cohesion, cells lacking α‐adducin showed slower cell migration in in vitro scratch and ex vivo skin punch biopsy assays. Together, our data indicate that α‐adducin contributes to intercellular adhesion by regulating Dsg3‐dependent adhesive functions, Dsg3 trafficking and desmosome stability.

Support or Funding Information

German Research Council SP1300/1‐3

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Clinicopathological Assessment of Broiler Chickens Challenged with Field Infectious Bursa Disease Virus after Oral Vaccination Using Phytogenic Mucoadhesive Gum
Authors: Adeniran Gani , Emipke Benjamin Obukowho , Jarikre Theophillus Aghogho , Ola Olawumi , Ohore Godspower , Adigun Oyindamola
Published: June 2021

The rising socio‐economic loss incurred from mortalities and disharmonized vaccine control programs in poultry production has necessitated the search of adjuvants especially of natural origin. More so, productivity levels of poultry in many African countries fall far below desirable levels because of the economic losses incurred from chick mortality due to infectious bursa disease (IBD). This study determine the level of immune response conferred by Infectious bursa disease vaccine using a mucoadhesive delivery system from a phytogenic origin using the clinic‐pathological response of broilers vaccinated oraly with IBD vaccine using Khaya senegalensis and Cedrela odorata as phytogenic adjuvants and later challenged with the IBD virus. A total of 260 chicks were successfully brooded and raised from day old. The birds were distributed into four groups A to D with an average of 60 birds per group in a complete randomized block design, as A (Gum + IBD Vaccine oral + Virus challenge), B (IBD Vaccine alone oral + Virus challenge), C (Gum alone oral + Virus challenge) and D (No‐Vaccine‐No‐Gum + Virus challenge). Three birds were euthanised per Group for Serum, tracheal and intestinal washings and lymphoid tissue harvest including haderian gland, thymus, lungs, spleen, ileum, cecal tonsils and bursa of Fabricius pre and post vaccinations and post infection during the 7 weeks experiment. Data was analysed using Graph pad prism 21 at α=0.05. All the birds were apparently healthy pre and post vaccinations. The birds exhibited signs of picking and soiling of the vents and feathers with projectile yellowish diarrhoea few birds post infection (B, D). The feathers were ruffled and mortality varied from 0 to 5 in the non‐vaccinated and gum alone vaccinated birds. Pathologically, there was oedematous, hyperplastic to haemorrhagic changes in the bursa, congestive to hyperplastic changes in the spleen with severity in group D and haemorrhages of breast and pectoral muscles in some of the birds that died post infection. No recumbency was recorded in the birds given gum+IBD vaccine and challenged with live virus. Prophylactic application of IBDV vaccines and the Khaya senegalensis gum as phytogenic adjuvants enhanced the chicks immune response and provided protection against IBD.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular and histopathological diagnosis of Fowlpox virus in Chickens in Nigeria
Authors: Adesina Rofiat , Ogunleye Seto , Alaka Olugbenga , Fagbohun Olusegun
Published: June 2021

Fowlpox is a relatively slow developing disease characterized by discrete, proliferative lesions on the skin of toes, legs or head, and/or mucous membranes of the mouth and upper respiratory tract of birds. Several diseases of birds such as infectious coryza, infectious laryngotracheitis, Mareks, and vitamin A deficiency can be confused with fowlpox, therefore there is need for an accurate and reliable method of diagnosis. Polymerase Chain Reaction (PCR) and histopathology can be used to arrive at the confirmatory diagnosis of the disease. Consequently, fifty samples from suspected cases of fowlpox from backyard poultry in Ibadan, Nigeria were collected, and PCR was used to detect and characterize the open reading frame 168 (39 kDa core protein) gene of the virus. Also, lesions from the eyelids excised were and histopathology was carried out. Thirty‐one out of 50 samples (62%) tested positive for FPV. Histopathology revealed the presence of dermal hyperplasia and Bollinger bodies which are confirmatory of fowlpox. PCR amplification of 39 kDa core protein gene and histopathology provide a reliable diagnosis of fowlpox.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Connective Tissue Dysplasia is a Possible Predictor of Esophageal Variceal Bleeding in Liver Cirrhosis
Authors: Vasylyev Dmytro Volodymyrovych
Published: June 2021

The aim of this work is to assess the connective tissue skeleton of esophageal varices in patients with liver cirrhosis for the study of morphological predictors of variceal bleeding.

Material for morphological research were the fragments of the esophageal wall from the lower third of esophagus taken at autopsy (samples of 10 cases in each group). Four groups of samples were: cirrhotic patients who died due to initial variceal bleeding (group 3), due to recurrent variceal bleeding (group 4). The control (comparing group) consists of fragments of the intact wall of the lower third of the esophagus in patients who died from other pathologies (group 1), and dead from liver cirrhosis, who hadn't pronounced esophageal varices, whose death was not associated with bleeding (group 2).

Immunohistochemistry was performed on paraffin sections with a thickness of 5–6 μm using an indirect Coons method. The collagens were identified by monoclonal antibodies (MA) to collagen I, III, IV types (Chemicon International Inc., Daco Cytomation, USA). According to the original technique, it was determined the intensity of the fluorescence of preparation treated with MA and expressed in arbitrary units of fluorescence (AUF).

In cirrhotic patients with variceal bleeding revealed the predominance of immature collagen type III (instead of dominated collagen type I) in the composition of the connective tissue of the mucous membrane and submucosal layer of the esophagus, increased intensity of collagen type III in the vascular wall and perivascular space, decrease of fluorescence intensity of collagen type IV in the basal membranes of veins (tables 1, 2). Abnormalities of collagen formation in the basal membranes of the esophageal veins, as well as the replacement of the elastic and muscular elements of the venous walls with immature connective tissue can lead to thinning and rupture of the vein wall.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of P‐glycoprotein Inhibition on Norbuprenorphine‐induced Neonatal Abstinence Syndrome (NAS)
Authors: Salazar Paloma , Higgins William T , Brents Lisa K
Published: June 2021

Treating pregnant, opioid‐addicted women with the opioid buprenorphine improves mother‐child outcomes, but can cause neonatal abstinence syndrome (NAS), a potentially life‐threatening withdrawal syndrome newborns often develop following chronic prenatal exposure to opioids. NAS severity is independent of maternal buprenorphine dose, suggesting that inter‐individual variance in the pharmacokinetics of buprenorphine may influence NAS. We previously showed that prenatal exposure to the major active metabolite of buprenorphine, norbuprenorphine (NorBUP), can induce NAS. NorBUP is a substrate of the highly polymorphic placental efflux transporter P‐glycoprotein, which transports its substrates from fetal to maternal circulation. We tested the hypothesis that inhibiting P‐glycoprotein increases NorBUP‐induced NAS severity. We administered a subthreshold dose of NorBUP or vehicle to pregnant rats from gestation day (GD) 9 until post‐delivery and treated them twice daily with the p‐glycoprotein inhibitor Elacridar or vehicle on GD18‐21. After delivery, pups were observed for precipitated withdrawal signs following administration of naltrexone or saline. NorBUP and Elacridar together, but not individually, increased naltrexone‐precipitated withdrawal signs, providing the first evidence that P‐glycoprotein activity can substantially modulate the effects of NorBUP on the fetus and may therefore contribute to the uncoupling of maternal buprenorphine dose and NAS severity.

Support or Funding Information

The project described was supported by a Summer Undergraduate Research Fellowship from the American Society for Pharmacology and Experimental Therapeutics, the UAMS Fellow‐to‐Faculty Award (Arkansas Biomedical Institute, Tobacco Settlement Funds), a UAMS Research Scholar Pilot Grant Award in Child Health, and the Translational Research Institute (TRI), grants 1U54TR001629‐01A1 and KL2TR000063 through the National Center for Advancing Translational Sciences of the National Institutes of Health (NIH). The content is solely the responsibility of the authors and does not necessarily represent the official views of the funding agencies.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
c‐Fos Responses Produced by the Central Microinjection of Salvinorin A in Conscious Rats
Authors: Ali Aboozar , Franklin Cynthia , Rangel Yolanda , Gottlieb Helmut
Published: June 2021

Intracerebroventricular (ICV) injection of the kappa opioid agonists produces a marked diuresis in rats via inhibition of vasopressin secretion. Salvinorin A (SalA) is a plant‐derived high affinity agonist for the kappa opioid receptor (KOR). Previous studies have proposed that SalA has similar potency and efficacy as other synthetic kappa agonists in mediating the CNS effects of KOR activation. The present study examines the expression of the proto‐oncogene c‐fos in several forebrain regions of rats treated with SalA as compared to either acetonitrile or saline vehicle treated animals. SalA is a highly lipophilic drug and past studies utilized drug dissolving vehicles (e.g., DMSO) that produced significant cardiovascular and renal effects. In this study, we dissolved SalA in acetonitrile/saline (1:9 dilution) to minimize solvent viscosity and cardiovascular effects.

Methods

Animals were habituated to a metabolic cage and injected with either SalA, acetonitrile or saline. Urine samples were collected during two 15 min control periods and six 15 min periods beginning 15 min after injection into the lateral ventricle. After 90 minutes, animals were perfused with a phosphate buffer and PFA. Brains were removed and cut on a cryostat (40 μm thick). Brain sections were processed for c‐fos using a commercially available antibody. Sections were also double labeled with oxytocin to define anatomically specific forebrain regions.

Results

SalA decreased c‐fos expression in the parvocellular PVN, magnocellular PVN, supraoptic (SON) and median pre‐optic (MnPO) nucleus relative to the acetonitrile group. However, only in the parvocellular PVN was the decrease in c‐fos expression statistically significant (p<0.001). In contrast, there was no difference in c‐fos expression between SalA and saline vehicle treated rats, with the exception of the MnPO in which c‐fos expression was markedly increased (P<0.001).

Conclusion

The ability of kappa opioid receptor agonists to increase diuresis and decrease osmolality may be associated with changes in c‐fos expression in the PVN and lamina terminalis. Previous studies in our lab, demonstrated a small increase in urine excretion rate following ICV administration of SalA and is consistent with the present findings in which there is a concomitant decrease in c‐fos expression in the magnocellular PVN and SON forebrain regions in SalA treated animals. However, the decrease was not a prominent as observed with U50488H, a synthetic KOR agonist. These results support the findings that the diuretic effects of SalA are not as potent as synthetic kappa opioid agonists.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Female rats eating high fat chow acquire methamphetamine self‐administration faster than rats eating standard chow
Authors: Galindo Kayla I , Beltran Nina M , Serna Paloma , Ramos Jeremiah , Serafine Katherine M
Published: June 2021

Eating a diet that is high in fat contributes to increased risk of chronic diseases and mortality, even in the absence of obesity. Eating a high fat diet can also impact the same brain reward pathways that are targeted by drugs of abuse. Preclinical studies have demonstrated that rats eating high fat chow are more sensitive to the unconditioned behavioral effects of drugs that act on dopamine systems (e.g., cocaine). However, the majority of evidence demonstrating that diet can impact drug sensitivity has investigated only male subjects, despite probable sex differences. To test the hypothesis that eating high fat chow increases sensitivity of rats to the positive reinforcing effects of methamphetamine (0.01–1.78 mg/kg/infusion), female Sprague‐Dawley rats (n = 30) were fed either standard (17% kcal from fat) or high fat chow (60% kcal from fat) for 6 weeks prior to self‐administration procedures. Following i.v. catheter implantation, rats were trained to respond for 0.1 mg/kg/infusion methamphetamine under a Fixed Ratio 1 schedule of reinforcement. Rats eating high fat chow met acquisition criteria faster (14 +/−2 days) than rats eating standard chow (25 +/− 4 days). Preliminary (n = 5–8/group) dose‐response curve assessments under a Fixed Ratio 5 schedule of reinforcement revealed that rats eating high fat chow respond less for methamphetamine than rats eating standard chow, at least at the training dose (0.1 mg/kg/infusion). These results suggest that rats eating high fat chow are more sensitive to the reinforcing effects of methamphetamine than rats eating standard chow. Future work will examine the relative reinforcing effectiveness of methamphetamine in rats eating different diets using a progressive ratio schedule of reinforcement. These data expand previous research and add to the growing literature on the effects of diet on drug sensitivity.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison between the effects of propofol and midazolam on pentylenetetrazole kindled convulsions in rats
Authors: Hasan Zuheir A , Abdul Razzak Rima L
Published: June 2021

Propofol, a short acting intravenous anesthetic which is wildly used for induction and maintenance of anesthesia. In addition, propofol has been successfully used in the management of status epileptics. However, sporadic reports of the occurrence of seizures or seizure like phenomena in epileptic and non‐epileptic patients continue to appear. The purpose of the present study is to examine the effects of intravenous (IV) administration of various doses of propofol and midazolam on pentylentetrazole (PTZ) kindled convulsions in rat.

Experiments were conducted in adult females Sprague–Dawley rats (230–250 g). PTZ kindling was induced by intraperitoneal (IP) injection of a subconvulsivant dose of PTZ (35 mg/kg) every alternate day. After each injection of PTZ rats were placed in plexy glass cages and were monitored for 30 minutes for the occurrence of behavioral seizures. Severity of behavioral seizures and seizure score was rated according to Racine scale (Racin, 1972). An animal was considered fully‐kindled when it has had stage 4–5 seizure score on two consecutive trials. To examine the effects of propofol on PTZ kindled convulsions, fully kindled rats (8–10 animals per group) were injected with various doses of propofol (2.5, 5, 10 mg/Kg) prior to PTZ challenge. The drug was administered IV as a single dose through the dorsal tail vein via a 25G butterfly scalp vein cannula. Control animals received an equal volume of saline. Few minutes following drugs or saline injection, rats were challenged with a subconvulsant dose of PTZ (35 mg/Kg) and seizure score for individual animals was recorded. To compare the effects of midazolam on PTZ kindled seizures with these of propofol, the same animal groups were administered various doses of midazolam (2.5, 5, 10 mg/Kg). A period of a two weeks washout was allowed prior to midazolam treatment.

The result of our study shows that both anesthetic agents suppressed PTZ kindled seizures in a dose dependent manner indicating that both agents protects against PTZ kindled seizures. However, the effects of midazolam were more profound. Dose response analysis of percent inhibition of kindled seizures shows that the ED50 for midazolam was 1.85 mg/Kg with 95 % CI of (1.71–2.05), whereas the estimated ED50 of propofol was 5.36 mg/kg with 95 % CI of (4.85–5.96).

The data of the present study shows that propofol is an effective anticonvulsant against PTZ kindled seizures and provides further support of the anticonvulsant properties reported in different experimental seizures models.

Support or Funding Information

The research was supported by a grant from Arabian Gulf University.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterizing a Novel RGS14:MyosinV/VI Interaction in Brain
Authors: Montanez‐Miranda Carolina , Evans Paul R. , Gerber Kyle J. , Seyfried Nicholas T. , Hepler John R.
Published: June 2021

RGS14 is a complex multifunctional signaling protein that integrates G protein, Ras/ERK and Ca++/CaM/CaMKII signaling pathways. The protein is composed of an RGS domain that binds active Gαi/o, two tandem Ras/Rap binding domains (R1, R2) which bind active H‐Ras‐GTP and Rap2A‐GTP to regulate MAP kinase signaling, a G protein regulatory (GPR) motif that binds inactive Gαi1/3, and a Ca++/CaM binding site. RGS14 is highly expressed in the hippocampus, the brain center for learning and memory, which is composed of areas CA3, CA2, and CA1. Schaffer collateral (SC) projections from CA3 to CA1 display long term potentiation (LTP), a stable potentiation in post‐synaptic strength in response to brief periods of synaptic stimulation such as during learning in animals. Within the hippocampus RGS14 is highly expressed in postsynaptic dendrites and spines of area CA2, where SC inputs from CA3 to CA2 neurons show a surprising resistance to LTP. CA2 has been shown to be important for social learning and memory, and we have shown that RGS14 is a natural suppressor of LTP and synaptic plasticity in this area. That is, loss of RGS14 in CA2 (RGS14‐KO) restores synaptic plasticity (LTP) in CA2 and markedly enhances hippocampal learning. However, it remains unknown how RGS14 interacts with other signaling proteins to regulate synaptic plasticity. In proteomic studies, RGS14 recovered from mouse brain by immunoprecipitation was subjected to mass spectrometry and proteomic analysis, showing its association with a high molecular‐weight protein complex including actin‐based myosin motor proteins (Myosin V, Myosin VI and Myosin II) and their regulatory proteins, among others. MyoV is involved in activity‐dependent insertion of AMPA glutamate receptor subunits (GluR1, GluR2) and NMDA receptors, regulation of calcium and proper positioning of calcium intracellular stores. MyoVI is enriched at the postsynaptic density where it regulates AMPA receptor recycling. These myosin classes have crucial roles in regulating synaptic plasticity and LTP. Follow up studies have shown RGS14 in complex with MyoV and MyoVI in whole brain lysates. However, it remains unclear how RGS14 interacts with MyoV/VI to regulate synaptic plasticity. Ongoing studies will determine if RGS14 binds directly or indirectly to MyoV/VI and how that is regulated, as well as elucidate the physiological function of this interaction, thereby providing insight into the functions of both RGS14 and MyoV/VI in their roles in modulating synaptic plasticity and hippocampal‐dependent learning and memory.

Support or Funding Information

This research was supported by grants NIH/NINDS to JRH (5R01NS37112 and 1R21NS074975), and in part by the Emory Neuroscience NINDS Core Facilities and NIH/NINDS under award number P30NS055077.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.