bluuebunny/crossref_metadata_2025_split huggingface dataset:
PAX5‐Induced Expression of Endogenous Mucolipin‐2 (MCOLN2) Gene in Human Glial and Neuronal Cell Lines: A Potential Gene Complementation Therapy Approach for Mucolipidosis IV
Authors: Rosas Lauren , Cuajungco Math P. 
Published: June 2021

The loss of function in the Transient Receptor Potential Mucolipin‐1 (TRPML1) protein, encoded by the Mucolipin‐1 (MCOLN1) gene, is known to cause the neurodegenerative disease Mucolipidosis type IV (MLIV). Characteristics of MLIV are motor problems, cognitive dysfunction, achlorhydria, cataracts, and blindness caused by retinal cell death. The Ashkenazi Jew population is most afflicted by the disease, and MLIV has no known cure. The Mucolipin ion channels consist of TRPML1, ‐2, and ‐3 proteins, which are encoded by MCOLN1, ‐2, and ‐3 genes, respectively. They are known to serve as non‐selective cation channels located within endosomes and lysosomes and play a role in endosome‐lysosome fusion. Previous research indicates that TRPML1 and TRPML2 proteins share a high degree of homology, and we hypothesize that TRPML2, which has no known association to clinical disease, could substitute for the loss of functional TRPML1 in MLIV, resulting in a rescue of the disease phenotype. In order to study the potential therapeutic effects of TRPML2 in MLIV, we must first show that it is expressed in the brain, one of the tissues most affected by MLIV. We have previously discovered PAX5 as the transcriptional activator for MCOLN2 and have shown that MCOLN2 expression could be heterologously induced in both human neuroglioma (H4) cells and human embryonic kidney 293 (HEK‐293) cell lines. Previous reports demonstrated that MCOLN2 has tissue‐specific expression with little to no detectable transcripts in brain tissue. However, our preliminary research has detected endogenous levels of MCOLN2 transcripts in human H4 and neuroblastoma (SH‐SY5Y) cells. Moreover, to induce the expression of endogenous MCOLN2, we transduced SH‐SY5Y cells with adenovirus and lentivirus vectors containing the PAX5 cDNA. PAX5 serves as a B cell‐specific activator protein with tissue‐specific expression pattern as well. Standard reverse‐transcription polymerase chain reaction (RT‐PCR) and real‐time quantitative RT‐PCR both revealed MCOLN2 expression in SH‐SY5Y cells at 48 and 72 hours post‐transduction. We show that PAX5 transcriptionally activates and increases endogenous MCOLN2 expression in neuronal cells that are central to studying MLIV. Future studies include In‐Cell Western assay with an anti‐TRPML2 antibody and transcriptome analysis of PAX5‐treated cells to determine global gene expression. These results and future studies will further explore the therapeutic potential of gene complementation to rescue the cellular phenotype in MLIV disease.Support or Funding InformationThis work was partly funded by NIH R15 NS101594.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Use of the Phytochemical Neem as a Component of Combination Treatment Against Pseudomonas aeruginosa Biofilm
Authors: Bertrand Blake Patrick , Heuertz Rita M. 
Published: June 2021

According to the NIH, 80% of chronic infections are caused by biofilm‐associated bacteria. Biofilm communities contribute to antimicrobic resistance by providing a physical extracellular barrier and exhibit metabolic diversity within bacterial subpopulations. Clinically, combination antibiotic strategies are used to provide high dosage treatments to successfully combat biofilm‐associated infections. Microscopically, biofilms of Pseudomonas aeruginosa have been visualized as mushroom‐like structures with two distinct subpopulations: a high‐metabolic “mushroom cap” outer population and a low‐metabolic “stalk” inner population. Further, the inner subpopulation consists of persister cells that are not killed by many therapies and contribute to chronicity of biofilm‐associated infections. It has been proposed that complete eradication of biofilm is achieved by targeting specific biofilm populations with antimicrobic combination of colistin and ciprofloxacin that target inner and outer subpopulations of biofilm respectively. However, colistin is sparingly used clinically because of high renal toxicity. Use of phytochemical alternatives instead of antibiotics is under assessment for treatment of biofilm‐associated infections. Extract from Azadirachta indica, or neem tree, is reported to have antimicrobic and antibiofilm effects on P. aeruginosa with low human toxicity. For these reasons, P. aeruginosa was grown (24 h) and microplates washed to remove non‐adherent cells. Remaining biofilm and cells were treated overnight with neem, colistin, ciprofloxacin or dual combinations. Finally, biofilm was removed from the substratum by scraping and aspiration, and bacterial viability was determined by colony forming unit (CFU/mL) enumeration. Results indicated that neem and ciprofloxacin in combination had an inhibitory effect (≤103 CFU/ml) similar to colistin and ciprofloxacin combination (≤102 CFU/ml) compared to growth and vehicle controls (both of which were 1012 CFU/ml). Colistin and neem combination showed no combined effect (107 CFU/ml) different from single treatment effects (108 and 1010 CFU/ml, respectively). These results suggest that neem extract targets the inner, low‐metabolic subpopulation of bacteria in biofilms and, for this reason, neem has potential to be a non‐toxic alternative to colistin for use therapeutically. When colistin or neem (that target the inner, low‐metabolic bacterial subpopulation) are used in combination with ciprofloxacin (that targets the outer, high‐metabolic subpopulation), the entirety of the biofilm may be targeted and prevent chronicity in biofilm‐associated infections.Support or Funding InformationFinancial support for BPB was received from the DeNardo Education and Research Foundation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Subcellular Localization of The Enzymes In Serine Biosynthesis
Authors: Nance Bailey , Kyoung Minjoung , An Songon 
Published: June 2021

Serine biosynthesis uses 3‐phosphoglycerate, an intermediate of glycolysis, to catalyze the first reaction by phosphoglycerate dehydrogenase (PHGDH). This reaction produces 3‐phosphohydroxypyruvate, which eventually becomes serine through two additional steps. Upregulation of serine biosynthesis has been characterized as a hallmark of cancer cell metabolism, as serine is an essential metabolite that provides precursors for various building blocks supporting cancer cell growth and proliferation. We hypothesize that the enzymes involved in serine biosynthesis may associate with each other in the cytoplasm of human cancer cells. To test this hypothesis, we use molecular cloning techniques to tag human PHGDH and others with a fluorescent protein. We will then transfect the plasmids into human cancer cells to examine the localization of PHGDH and others using fluorescent live‐cell imaging. Ultimately, we will contribute to our understanding of the spatiotemporal regulation of serine biosynthesis in human cancer cells.Support or Funding InformationThis work was supported by UMBC Start‐up fund to S.A. This investigation was also sponsored in part by NIH/NIGMS MARC U*STAR T34 08663 National Research Service Award to B.N.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Targeting Glycolytic Metabolism in Cancer
Authors: Telang Sucheta , Trent John , Chesney Jason , Mojesky Aubrey 
Published: June 2021

Tumors exhibit high glucose uptake and glycolysis even in the presence of oxygen (i.e. the Warburg effect) and this metabolic phenotype serves to satisfy their increased requirement for energy and biosynthetic intermediates. Tumor glucose metabolism is stimulated in part by fructose‐2,6‐bisphosphate (F26BP), which is the product of a family of 6‐phosphofructo‐2‐kinase/fructose‐2,6‐bisphosphatases (PFKFB1–4) and activates a key rate‐limiting enzyme in glycolysis, 6‐phosphofructo‐1‐kinase (PFK‐1). We have found that the PFKFB4 family member is over‐expressed in human tumors and highly induced by HIF‐1α and that silencing PFKFB4 expression markedly reduced F26BP, glucose metabolism, cell survival and the growth of xenograft tumors in vivo. Using virtual screening, we recently discovered a novel small molecule inhibitor of PFKFB4, 5MPN, that selectively inhibits recombinant PFKFB4 activity, and decreases glucose metabolism, proliferation and tumor growth and have now identified a 5MPN derivative that is significantly more effective at decreasing glucose metabolism and proliferation in cancer cells and markedly reduces the growth of established tumors in vivo, without systemic toxicity. Since our previous studies have determined the PFKFB3 family member to be a dominant source of F26BP in cancer cells and additionally have demonstrated co‐expression of the PFKFB4 and PFKFB3 enzymes in multiple patient‐derived cancers, we compared the effects of silencing PFKFB4 and PFKFB3 in human cancer cell lines in vitro. We unexpectedly found that PFKFB4 expression is increased by PFKFB3 silencing, suggesting that PFKFB4 may compensate for decreased PFKFB3 expression and activity. Importantly, an inhibitor of PFKFB3 (PFK158) is currently in Phase I trials and compensation by PFKFB4 may limit the clinical efficacy of this and other PFKFB3 inhibitors. We then examined the effects of simultaneous silencing of PFKFB4 and PFKFB3 on cancer cell lines and observed that co‐knockdown led to a significant decrease in cell viability and glycolysis and a near‐complete abrogation of anchorage independent growth in vitro and, additionally, found that simultaneous small molecule inhibition of the PFKFB4 and PFKFB3 enzymes in vitro resulted in a synergistic decrease in cell viability. Taken together, our data indicate that targeting PFKFB4 may be a valid therapeutic option against cancer and strongly support the further development of PFKFB4 small molecule inhibitors as well as the co‐targeting of PFKFB4 and PFKFB3 as potential strategies for the treatment of cancer.Support or Funding InformationNIH REACH Program Kentucky Lung Cancer Research ProgramThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of dietary different energy sources on the growth performance, amino acid profile, blood profile, intestinal morphology and digestive enzyme in weaned piglets
Authors: Lv Dinghong , Xiong Xia , Yang Huansheng , Wang Meiwei , Liu Yanhong , Yin Yulong 
Published: June 2021

The low feed intake immediately after weaning is responsible for insufficient energy supply in weaned piglets. We investigated the effects of different dietary energy source on growth performance, serum amino acid profiles, biochemical parameters and immunoglobulin level, antioxidant capacity, and intestinal morphology and enzyme activity in weaned piglets. Forty‐two 21‐day‐old weaned piglets were randomly assigned to one of three treatments. They were fed isocaloric diets supplemented with three energy sources (glucose, soy oil, and glutamine) with two phases (14 and 28 d). Compared with glutamine group, piglets in the glucose and soy oil groups had greater (P < 0.05) average daily gain, average daily feed intake and gain:feed ratio from d 0 to 14, and gain:feed ratio for the overall period. Pigs fed with glutamine had greater (P < 0.05) serum glucose and creatinine on d 14 and high‐density lipoprotein on d 28, compared with pigs in the other two groups. The blood urea nitrogen of pigs was lower (P < 0.05) in soy oil group than in glucose and glutamine groups on d 28. Supplementation of glutamine or soy oil increased (P < 0.05) serum IgA on d 28, while supplementation of glutamine increased (P < 0.05) IgG and IgM on d 14 and 28. No differences were observed in serum amino acids among 3 treatments on d 14, except glycine and tyrosine. The serum concentrations of histidine, serine, threonine, proline, and cysteine in glutamine group was highest (P < 0.05), while the content of glycine and lysine in soy oil group was highest (P < 0.05) in three groups on d 28. Using glucose or soy oil as energy source increased (P < 0.05) serum total superoxide dismutase (T‐SOD) on d 14 and 28, but decreased (P < 0.05) serum malondialdehyde on d 28 compared with glutamine. On d 14, T‐SOD concentration and total antioxidant capacity (T‐AOC) in the duodenum and ileum of pigs supplemented with glutamine were higher (P < 0.05) than those in soy oil group; however, the concentration of T‐AOC in jejunum was lowest (P < 0.05) in the glutamine group on d 14 and 28. Results from the present experiment indicated that weaned pigs supplemented with glucose or soy oil have better growth performance likely due to their enhanced feed intake and better systemic antioxidant capacity, whereas glutamine supplements may increase immunity and intestinal oxidative capacity by modulating nutrition metabolism.Support or Funding InformationThe present study was jointly supported by the National Natural Science Foundation of China (31572420; 31330075)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of Omega‐3 Supplementation on the Lipid Profile and Adipose Indices in Hispanics with Type 2 Diabetes Mellitus
Authors: Jehi Toni , Mota Christy , Beeson Lawrence , Firek Anthony , Cordero‐MacIntyre Zaida , De Leon Marino 
Published: June 2021

BackgroundDiabetes mellitus (DM) is considered a serious affliction that highly augments the risk for serious health complications. Evidence is suggesting a role of n‐3 polyunsaturated fatty acids (PUFAs) on improving the lipid profile and adipose indices in patients with type 2 DM (DM2). However, this association hasn't been probed much in underserved populations. Thus, the objective of this study was to investigate the effect of the diabetes education program, En Balance Plus, on lipid profiles and adipose indices among Hispanic adults with DM2, using supplemental fish oil.MethodsSubjects (n=40) consumed a daily intake of 2000 milligrams of omega 3 fish oil in capsule form for the whole 3‐month study. At baseline and after the intervention, comprehensive assessments were conducted to evaluate the health status, dietary habits and total and regional body compositions of the subjects. During analysis, non‐parametric tests were utilized to conduct the comparisons.ResultsThere was a significant decrease in median glucose levels (−4.5 mg/dL, p=0.021), hemoglobin A1c (−0.1%; p = 0.001), trunk fat (−1%; p = 0.0134), and total fat (−0.50%; p = 0.0452). As for the changes in cholesterol levels, the only significant differences were observed in LDL(reduced by 5 mg/dL; p= 0.011)ConclusionThe omega‐3 supplementation for Hispanic patients in the En Balance Plus program contributed to significant attenuation in LDL cholesterol, an improvement in glycemic control, and a decrease in trunk and total fat. Further investigations should be conducted to assess the long term effects of this intervention.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of FSH‐SP on inhibition winkle formation, melaningenesis, and promoting skin hydration in UVB‐irradiated HRM‐2 hairless mice
Authors: Lee Dasom , Lee Minhee , Park Soo‐Jeng , Yun Jeong‐Moon , Kim Dakyung , Lee Minjae , Woo Young Ho , Lee Jeongmin 
Published: June 2021

Skin is important organ to protect against dangerous external factors including bacteria, chemicals, and UV irradiation. UVB is directly induced destruction in skin epidermis barrier by damaging DNA. Expose to UVB can lead to low skin hydration, winkle formation and melanogenesis. The aim of this study was to investigate the effect of FSH‐SP to protect skin barrier in HRM‐2 hairless mice induced UVB irradiation 150 mJ/cm2 three times a week for 6 weeks. The groups divided PL (FSH‐SP 100 mg/kg b.w.) and PH (FSH‐SP 200 mg/kg b.w.). We examined histopathological experiment about skin damage including changes in skin thickness, melanin pigmentation and winkle formation. Also, we experimented SOD, catalase in plasma and expression of mRNA about procollagens, HAS2 (hyaluronan synthesis 2), MMPs (matrix metalloproteinase) and MITF. The serum SOD and catalase activity were significantly increased PH. The result of procollagen, HAS2 and elastin expression was significantly increased in PH. The gene expression of MMPs (1 and 9) and MITF which is a key factor to synthesis melanin were significantly reduced in PH. The skin hydration exposed to back skin was increased in PH. The winkles, skin thickness and melanin pigment looked like decreased in PL and PH. In conclusion, FSH‐SP decreased the expression of MMPs and MITF increasing the activity of antioxidant enzymes, thereby protecting the skin moisturizing factors and inhibiting melanogenesis. It is believed that effect of the FSH‐SP was reduced to suppress the wrinkle formation, whitening effect and moisturizing.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Acetaminophen‐Induced Hepatotoxicity in Wistar Rats Treated with Annona Muricata Aqueous Stem Extract
Authors: Okpara Ifeoma Janet , Nosiri Chidi Ijeoma , chisom madukwe 
Published: June 2021

The central role of the liver in biochemical homeostasis cannot be overemphasized and its toxicity is owed greatly to its great involvement in xenobiotic metabolism. This study therefore evaluated the hepatoprotective effect of aqueous stem extract of Anonna muricata S. on Acetaminophen‐induced liver damage. Animals were divided into four groups of five rats. Group1‐ normal saline p.o, Groups 2–4 pretreated with 400mg/kg p.o Acetaminophen, then Groups 3 and 4 treated for 7days with 400mg/kg and 800mg/kg extract p.o respectively. The result revealed an elevated level of serum liver enzymes (ALT 32±1.00, AST 42±2.00) to the untreated Group II rats that received only 400 mg/kg acetaminophen, which at concurrent examination showed a significant decrease in these liver enzymes in Groups III (ALT 29±1.00 and AST 34.67±1.53) and IV (ALT 29±3.00 and AST 34±2.00). The increase seen in the treated groups were not dose dependent but were higher than the control. Group II when compared with the control (ALT 17±1.00 and AST 26±1.04) showed a significant (p≤0.05) increase. The ALP, total protein and bilirubin showed little or no effect in both treated and untreated. Acetaminophen overdose induces hepatotoxicity through its intermediate metabolite N‐acetyl‐p‐benzoquinone imine (NAPQI) which reacts with the natural antioxidant Glutathione reducing its concentration and generating more reactive oxygen species that damage the hepatic cells. These reductions in serum liver enzymes by the extract revealed that the aqueous stem extract of Anonna muricata S. has no toxic effect on the liver and could be hepatoprotective at the tested doses.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Wild yeast strains ferment galactose in whey permeate
Authors: Loughrin Molly R. , Nold Stephen , Delshadi Rana 
Published: June 2021

Whey permeate is a by‐product of cheese production that is rich in lactose but currently has little commercial value. Lactose in whey can act as a carbon source for ethanol production by yeast such as Kluyveromyces marxianus, but ethanol yield is typically low due to the inability of most strains to ferment galactose. In this study, we cultivated and characterized wild yeast strains that ferment galactose. Samples were collected from dairy operations and inoculated into enrichment cultures containing whey amended with galactose, then were streaked for isolation on plates to obtain isolates. Wild strains and K. marxianus were tested for their ethanol production efficiency and their ability to utilize different carbon sources. Of the 56 strains we obtained, 22 strains produced as much or more ethanol than K. marxianus and 37 strains fermented galactose as the sole carbon source. We are currently characterizing the optimal growth conditions and suitability for industrial ethanol production of these strains compared to a commercially available yeast. These findings may be useful in our efforts to convert a common dairy waste into ethanol, a useful fermentation product.Support or Funding InformationUniversity of Wisconsin ‐ StoutThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Group A Streptococcus Prevents Lysosomal Acidification in THP‐1 Macrophages By Inhibiting V‐ATPase Complex Fformation
Authors: Nishioka Scott , Snipper Joshua , Franscioni Henry , Yuen Isabelle , Gao Yu , Delahunty Claire , Yates John , Okumura Cheryl 
Published: June 2021

Group A Streptococcus (GAS) is a Gram‐positive bacterial pathogen responsible for approximately 750 million human‐specific infections worldwide every year, ranging in severity from mild pharyngitis to necrotizing fasciitis. To defend against invasive pathogens such as GAS, the innate immune system deploys macrophages to phagocytose and eradicate the bacterial pathogen via engulfment into the phagosome, fusion with lysosomes, phagosomal acidification, and activation of hydrolytic enzymes. We have demonstrated that GAS persists within macrophages despite trafficking normally through the phagocytic pathway. Because GAS is not capable of surviving highly acidic conditions, the data suggest that GAS instead prevents lysosomal acidification. Vacuolar‐ATPase (V‐ATPase), the proton pump on lysosomes responsible for generation of the proton gradient, is a complex of several proteins that must be properly assembled. Here, we investigated disruption of the formation of the V‐ATPase as a possible mechanism employed by GAS to promote intracellular survival. Immunofluorescence data suggest that membrane subunits fundamental to V‐ATPase assembly and function do not localize to the phagolysosomal membrane in GAS‐infected macrophages. Preliminary cell fractionation and mass spectrometry data also indicate that cytosolic components of the V‐ATPase are present in lower quantities in GAS‐infected phagosomes. We are in the process of immunoprecipitating the V‐ATPase complex from GAS‐infected macrophages to further confirm complex formation. The information gained through our investigations provide important insights on the specific mechanisms GAS has evolved to evade the innate immune response, which may contribute toward the development of more effective therapies.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.