bluuebunny/crossref_metadata_2025_split huggingface dataset:
Library of Recombinant Heparan Sulfates from Mammalian Cell Culture: Paving the Way to Bioengineered Heparin
Authors: Thacker Bryan , Lee Gloria , Scott Max , Groth Tobin , Thorne Kristen , Sharfstein Susan , Glass Charles 
Published: June 2021

Heparan sulfate is a polysulfated, unbranched polysaccharide that plays a role in numerous developmental, physiological and pathophysiological processes via its interaction with hundreds of heparan sulfate‐binding proteins. Heparin is a highly sulfated variant of heparan sulfate, used routinely in the clinic as an anticoagulant. Both molecules share a common biosynthetic pathway that includes numerous glycosyltransferases, sulfotransferases and an epimerase. Researchers in the field typically use heparan sulfate purified from porcine intestines or bovine kidney for experimentation. This material suffers from contamination, batch‐to‐batch variation and fails to reproduce the diversity of heparan sulfate structures found in nature. Heparin used in the clinic is also purified from pig intestines with variable compositions, risks of contamination and supply deficits. TEGA Therapeutics is developing a library of heparan sulfate products derived from cell culture. Genetic engineering gives rise to a diversity of heparan sulfate compositions. Each type of heparan sulfate is characterized with respect to disaccharide composition, chain length and protein binding affinity. This work paves the way for development of recombinant heparin bioengineered in cultured cells with a secure supply chain and the potential for enhanced properties and reduced side effects.Support or Funding InformationThis work is supported by NSF STTR Phase I grant 1622959 to C.G and S.S.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ER Stress marker GRP78 is not expressed on ER−/PR−/Her2− human breast cancer cell surface or secreted
Authors: Banerjee Dipak K. , Serrano Negron Jesus E. , Ruiz Andrea Rivera , Baksi Krishna 
Published: June 2021

GRP‐78 (a Mr 78kDa calcium dependent glucose binding protein) located in ER lumen. It functions as ER chaperone and translocates proteins for glycosylation at the asparagine residue present in the sequon Asn‐X‐Ser/Thr (i.e., N‐linked glycosylation). This important biochemical event is essential for glycoprotein folding and function. When targeted in ER−/PR−/HER2+ breast tumor in athymic nude mice with the N‐glycosylation inhibitor tunicamycin, the cancer growth was reduced ~55% in three weeks. Paraffin sections of the tumor exhibited reduced N‐glycan level, and reduced angiogenesis but increased GRP‐78 expression in microvasculature as well as in the tumor tissue (J. Biol. Chem. 286, 29127–29138, 2011). This supported the presence of ER stress. We have now evaluated the effect of tunicamycin on the proliferation of metastatic ER−/PR−/HER2− (MDA‐MB‐231) and non‐metastatic ER+ (MCF‐7) human breast cancer cells. Tunicamycin inhibited proliferation of both cells types in a time and dose‐dependent manner. Interestingly, GRP‐78 expression (protein and mRNA) was higher in tunicamycin (1.0 μg/ml) treated MDA‐MB‐231 and in MCF‐7 cells and supported by both Western blotting and qPCR. Since GRP‐78 is an ER stress marker, so we have followed its intracellular localization by immunofluorescence microscopy after treating the cancer cells with tunicamycin as well as after subjecting them to various stress conditions. We have used unfixed cells and stained with either FITC‐conjugated Concanavalin A (Con A), or Texas‐red conjugated wheat germ agglutinin (WGA) or with anti‐GRP‐78 antibody. There were expression of N‐glycans but not GRP‐78. GRP‐78 however became detectable after permeabilization of cells with a brief exposure to ice‐cold methanol. GRP‐78 was not detected in the conditioned media of the cancer cell but a high level of MMP‐1 did. We have therefore, concluded that GRP‐78 is neither expressed on the outer‐leaflet of the (ER−/PR−/HER2−) human breast cancer cells nor it is secreted into the culture media during tunicamycin‐induced ER stress. This is against the current dogma that GRP‐78 expression on the tumor cell surface or its secretion interferes with the cancer therapeutic(s) and make them as tumor promoters and not tumor suppressors. Our study, thus, suggests strongly that anti‐angiogenic and anti‐tumorigenic action of tunicamycin can be modeled to develop next generation cancer therapy, i.e., glycotherapy for treating breast tumor and all other sold tumors.Support or Funding InformationSupported in part by the university funds (DKB), NIH‐NIDDK STEP‐UP Program (AR) and NIH/NIMHD G12MD007583 (KB)..This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Using neurogenic explants to assess the role of the stem cell niche structure fractones in adult neurogenesis and pathology
Authors: Wulff Alina Sophia , Davis Harry , Mercier Frederic 
Published: June 2021

Neurogenesis occurs throughout adulthood in restricted zones of the adult brain. Growth factors influence neural stem cell proliferation and differentiation. However, the mechanisms orchestrating growth factors to regulate neurogenesis are not well known. In the adult brain, neural stem cells and their immediate progeny reside in a nurturing stem cell niche environment. These niches are located in the ventricular/sub ventricular zone (VZ/SVZ) of the lateral ventricles. In the SVZ, stem cells are directly contacted by extracellular matrix (ECM) structures that we have named fractones. Using in vivo experiments, we have previously shown that fractones contain heparan sulfate proteoglycans (HSPG) that bind, concentrate and dispatch growth factors to the target stem cells to ultimately regulate stem cell fate. Here, an ex vivo model was devised to expand our ability to study fractones, stem cells, and growth factor interactions in culture in a near‐physiological context. This model consists of brain tissue pieces of the neurogenic zone, termed neurogenic explant. Neurogenic explants survive for months in a simple culture medium. They contain neural stem cells, their immediate progeny, ependymocytes, fractones, all forming the VZ/SVZ, plus the adjacent superficial part of the caudate nucleus. Using mini‐incubator and fluorescence microscopy, we demonstrate that we can visualize fractones, cell proliferation and growth factor binding on the living neurogenic explants. The ease in altering the ECM of explants and assessing the effects on neurogenic stem cells will allow us to understand the role of fractones in neurogenesis. By using explants originating from animal models, we will also be able to understand the relationship between disrupted adult neurogenesis and neural pathology. We anticipate that we can determine from the neurogenic explant model which HSPGs are responsible for the binding of individual growth factors. This will provide insight into the possibility of manipulating HSPG expression in explants to ultimately alter the cell‐fractone interactions responsible for the progression of disease.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of botulinum toxin type A combined with exercise on the functional recovery after spinal cord injury
Authors: Jin Yunho , Sumsuzzman Dewan Mohammed , Hong Yonggeun 
Published: June 2021

Spinal cord injury (SCI) is a result of a traumatic damage of the vertebral column. This injury interrupts the signal conduction from upper motor neuron to lower motor neuron and spasticity may occur. Spasticity is common motor dysfunction induced from upper motor neuron lesions. Botulinum toxin type A (BTA) can inhibit muscle contraction as well as acetylcholine release by without affection to acetylcholine synthesis. Treadmill exercise (Ex) is known to reduce body and fat mass resulting improvement of neuroplasticity. Therefore, the purpose of this study was to evaluate the effect of BTA combined with treadmill exercise after SCI. Experimental animals were divided into three groups; SCI, SCI+BTA, SCI+BTA+Ex. BTA was injected into the gastrocnemius of SCI+BTA and SCI+BTA+Ex rats. And, SCI+BTA+Ex rats were assigned to treadmill exercise. After 14 postoperative day, all animals were sacrificed to perform further analyses. SCI+BTA+Ex group showed the greatest decrease of body weight and gastrocnemius mass. Also, BTA‐injected groups underwent significantly decreased muscle degradation compared to non‐injected group. Similarly, BTA treated muscle showed increased levels of autophagy markers. Especially, both beclin‐1 and LC3 were increased in the gastrocnemius of SCI+BTA+Ex group. Interestingly, this group showed the highest motor functioning scores. In addition, by light microscopy, several vacuoles were observed in the muscle fibers of SCI group rats, indicating delayed functional recovery. In this study, BTA is thought to have slight spasticity‐alleviating effect, and this effect is extended by combination of treadmill exercise.Support or Funding InformationFunds: NRF‐2017R1A2A2A01067169, KGM4611714This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Histopathological effect of some medicinal plants on kidney and liver functions in rats.
Authors: ADEYEMO ADENIKE CHRISTY , ADENIYI ADEDAYO FRANCIS 
Published: June 2021

Three medicinal plants which are Aframomum melegueta, Ricinus communis and Helianthus annuus are botanicals used in traditional medicine and they are known to possess insecticidal properties. This study aimed to determine the toxic effect these plant materials leave powders on kidney and liver function which was evaluated through histological examination of tissues from both organs from rats fed with the formulated diets. Forty two (42) male Wistar rats were randomly divided into seven groups (six each) and were fed daily with 1 and 10% of the three plant materials included in their diet for 30 days while the control group fed with basal diet without supplementation. At the end of 30 days, the rats were sacrificed to remove the kidney and liver for toxicity evaluation.Support or Funding InformationNONEThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of non‐esterified fatty acids on the expression and secretion of angiopoietin‐like protein 4 and fibroblast growth factor 21 in calf hepatocytes cultured in vitro
Authors: ZHU Xiaoyan , WANG Jianguo , LIU Lei 
Published: June 2021

IntroductionNegative energy balance (NEB) is considered as the pathological basis of energy metabolic disorders, such as fatty liver and ketosis, in periparturient dairy cows. An elevated serum non‐esterified fatty acids (NEFA) concentration is one of the most important indicators of NEB. Angiopoietin‐like protein 4 (ANGPTL4) and fibroblast growth factor 21 (FGF21) have been identified as hepatokines, which involved in regulation of metabolic adaptations. In dairy cows, most circulating ANGPTL4 and FGF21 originate from the liver. However, the direct effects of NEFA on the expression and secretion of ANGPTL4 and FGF21 in bovine hepatocytes are not entirely clear. The purpose of the present study was to investigate the effects of different NEFA concentrations on the expression and secretion of these two hepatokines in calf hepatocytes cultured in vitro.MethodsBovine hepatocytes were cultured using the collagenase perfusion method. After culture for 72 h, the hepatocytes were treated with NEFA at final concentrations of 0 (control), 0.6, 1.2, 1.8 and 2.4 mmol/L. After 24 h of continuous culture, the mRNA and protein expression levels of ANGPTL4 and FGF21 in the hepatocytes were determined by real‐time PCR and Western blot, respectively. ANGPTL4 and FGF21 secretions in the supernatant were determined by enzyme‐linked immunosorbent assay (ELISA).ResultsThe results showed similar levels of ANGPTL4 expression and secretion following treatment with 0.6 and 1.2 mmol/L NEFA, which were higher than those observed for the controls (P<0.05). ANGPTL4 expression and secretion were also similar at 1.8 and 2.4 mmol/L NEFA and significantly higher than those observed for controls (P<0.01). The expression and secretion of FGF21 at 0.6 mmol/L NEFA‐treated hepatocytes was higher than that of the control group (P<0.05). The FGF21 expression and secretion were similar at 1.2, 1.8 and 2.4 mmol/L NEFA‐treated hepatocytes and significantly higher than those observed for controls (P<0.01).ConclusionThe results of this study indicate that high concentrations of NEFA significantly increase the expression and secretion of ANGPTL4 and FGF21 in bovine hepatocytes cultured in vitro in a dose‐dependent manner. Specifically, NEFA promotes the expression and secretion of ANGPTL4 and FGF21, which may worsen fat mobilization under NEB conditions and thereby lead to the pathological processes of energy metabolism disorders in dairy cows in the peripartum period. However, further studies are needed to elucidate the underlying mechanisms.Support or Funding InformationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Pilot Study to Examine the Perspectives of Pharmacy Students towards Adding a Nutrition Course in Pharmacy Degree Curriculum
Authors: Syed‐Abdul Majid Mufaqam , Kabir Syed Sadath , Soni Dhwani Satishkumar , Barnes Jeremey T. , Faber Tony A. , Timlin Maureen T. 
Published: June 2021

Today's pharmacists are likely to encounter questions about nutritional products sold in the pharmacy. This is due, in part, to the increased number of pharmacies in grocery stores and the availability of pharmacists. However, many pharmacists report that they lack nutrition knowledge and believe the best time to educate pharmacists about nutrition is during pharmacy school.PURPOSEThe purpose of this study was to determine if pharmacy students realize the importance of nutrition education during pharmacy school and if a background in nutrition education or pharmacy‐related experience influences student's responses.METHODSBoth male and female students from nine different pharmacy schools from southern part of India [IND, n=225, age: 18–34y, male: n=74 (32.9%), female: 151 (67.1%)] and two pharmacy schools (Alabama and Illinois) from the United States [USA, n=95, age: 18–45y, male: n=23 (24.2%), female: 72 (75.8%)] participated in this study.RESULTSThe response rate from IND was 97.0% (seven students did not complete survey) and USA was 100% (no incomplete surveys). When the students were asked about their nutrition education, 77.3% of students from IND and 55.8% students from the USA reported never taking a nutrition course. Students who took a nutrition course had to take it as an elective. While 37.8% students from IND and 93.7% students from USA had pharmacy‐related experience, 6.7% of students from IND and 3.2% students from USA said they were very confident in handling nutrition‐related questions. Regardless of previous nutrition education and pharmacy‐related experience, 88.0% of students from IND and 70.5% of students from the USA, believed that nutrition education is important and should be incorporated in the pharmacy school curriculum. Among those who took a nutrition course, 81.8% of students from IND and 82.9% from USA believed that a nutrition course should be incorporated into the pharmacy school curriculum. When pharmacy‐related experience was considered, 92.9% from IND and 73.3% from USA also believed that a nutrition course should be incorporated into the pharmacy school curriculum.CONCLUSIONResults from this study suggest that a nutrition course should be incorporated into the pharmacy school curriculum, regardless of past nutrition education or pharmacy‐related experience. A limitation of this study was that only students from the southern part of the India and two pharmacy schools in the United States were surveyed. Additional data is needed from other pharmacy schools in order to generalize the results of this study and influence curriculum changes for pharmacy schools.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A redesigned pharmacology series increases students' satisfaction and is associated with improved performance in therapeutics
Authors: Brandl Katharina , Schneid Stephen , Tsunoda Shirley , Awdishu Linda 
Published: June 2021

ObjectiveIn this study we describe the redesign, implementation and evaluation of a three‐quarter pharmacology course within a Pharm.D. curriculum.MethodsThis retrospective study examined the pre and post implementation of a redesigned three‐course pharmacology curriculum for second‐year pharmacy students. Course evaluations and examination scores were analyzed from four different cohorts of students (Class of 2014 to Class of 2017).Qualitative analysis of the course evaluations identified key areas of weakness to help the new course director implement changes to the entire pharmacology curriculum starting with the Class of 2014. Descriptive statistics, qualitative analysis, and correlation analysis were used to evaluate outcomes of the new course.ResultsQualitative analysis of 155 second‐year pharmacy student evaluations from the Class of 2014 identified lack of integration and coordination within the three‐course pharmacology sequence. Poor exam quality and the absence of active teaching methods were the most frequently described shortcomings of the pharmacology curriculum. In response to this analysis, exam question flaws were eliminated and active teaching modalities were introduced. Also, the new director took over more lectures to reduce the number of different guest lecturers teaching in the course. Immediately following these changes, the course evaluations dramatically improved and continued to improve over the following three years (One‐Way ANOVA with Bonferroni post‐test, p< 0.05). Significant correlations were identified between course evaluations and the number of lecturers taught by the new course director as well as the number of active teaching methods (Pearson Correlation coefficient, p<0.05). Students' performance in the years following the redesign correlated better with their performance in the third‐year Therapeutics course providing evidence for improved exam validity (Pearson Correlation coefficient, p<0.05).ConclusionA rigorous redesign of a large pharmacology course resulted in increased students' satisfaction and an improved association with exam performance in the Therapeutics course. This study highlights the importance of a well‐designed pharmacology course for continued success in core courses in the PharmD curriculum.Support or Funding InformationN/AThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development and Preclinical Testing of a Vaccine for 3,4‐Methylenedioxypyrovalerone ((R,S)‐MDPV) Substance Use Disorders
Authors: McClenahan Samantha Jo , Kormos Chad , Gunnell Melinda , Hambuchen Michael , Lamb Pamela , Carroll Ivy , Lewin Anita , Owens Samuel Michael 
Published: June 2021

Synthetic cathinones like 3,4‐methylenedioxypyrovalerone ((R,S)‐MDPV) are medically dangerous psychoactive drugs which at high doses produce psychosis, cardiovascular toxicity, and death. The goal for these studies was to develop and test a vaccine for treating ((R,S)‐MDPV) substance use disorders. A racemic MDPV conjugate vaccine was developed from a hapten structure capable of generating antibodies that bind with high affinity to (R,S)‐MDPV and other potent synthetic cathinones. We hypothesized that treatment with an optimized MDPV vaccine could produce high affinity antibodies against (R,S)‐MDPV capable of mitigating the pharmacological effects of (R,S)‐MDPV. An (R,S)‐MDPV‐like hapten was conjugated to different carrier proteins, mixed with Sigma Adjuvant System, and used to immunize male Sprague Dawley rats (n=5/group). The immunochemical characteristics of the rats' immune response was then evaluated. The KD value as determined by radioimmunoassay for (R,S)‐MDPV binding was 2.5 ± 1.9 nM, which demonstrated production of high affinity antibodies. The KD value for the individual (R)‐ and (S)‐MDPV binding was 0.71 ± 0.8 nM and 2.8 ± 2.3 nM, respectively. These similar KD values for the MDPV enantiomers indicates antibodies for both enantiomers contribute to the high binding of (R,S)‐MDPV. The rat antisera also showed similar high affinity binding of (R,S)‐α‐pyrrolidinovalerophenone (α‐PVP). No significant cross reactivity was detected with over‐the‐counter medications like (+)‐pseudoephedrine, neurotransmitters (dopamine, norepinephrine, epinephrine, and serotonin), or non‐cathinone drugs of abuse (cocaine, morphine, phencyclidine, (+)‐methamphetamine, (+)‐MDMA, and (−)‐MDMA). Moreover, the vaccination protocol produced high serum antibody titers, determined by rapid equilibrium dialysis, from 5 weeks after the start of immunizations until termination of the rats 3 months later. Finally, locomotor studies of control and vaccinated rats were conducted to test the vaccine's efficacy using a series of increasing doses of (R,S)‐MDPV (0.3–5.6 mg/kg, sc, 2 days apart). The immunized rats displayed significantly (p<0.05) lower MDPV‐induced locomotor activity and a significantly shorter duration of action compared to controls. In conclusion, active vaccination with an (R,S)‐MDPV conjugate vaccine produced very high affinity antibodies against (R,S)‐MDPV; and the vaccinated rats showed significantly reduced (R,S)‐MDPV‐induced locomotor effects compared to controls.Support or Funding InformationFunded by NIDA grants DA039195 and T32 GM106999This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.