bluuebunny/crossref_metadata_2025_split huggingface dataset:
Generation and partial characterization of cancer cell lines from Mexican breast cancer patients
Authors: Valencia María Lourdes , De Leon Roberto , Lüthy Isabel Alicia , Angulo Aracely , Ríos Omar , Velázquez Carlos Arturo 
Published: June 2021

Numerous cell models have been approached to study cancer biology. Some of those models are primary cell cultures, as well as, cancer cell lines derived from tumors. Several breast cancer cell lines are available for scientific research. Nevertheless, those cell lines do not represent all the cancer types and subtypes, neither their origin corresponds to most of the ethnic groups of the world. The main goal of this study is to generate and partially characterize cancer cell lines from tumor tissue obtained from Mexican patients with breast cancer, which can be used as models to study cancer biology and to characterize the phenotype and genotype of breast cancer affecting Mexican population. At present, four primary cell cultures (ID010, ID012, ID013 and ID014) have been established from primary tumor tissues representative of infiltrating ductal carcinomas, such as luminal B and HER‐enriched molecular subtypes. In addition, all the primary cell cultures showed the expression of cytokeratins, which suggests the presence of epithelial cells. The development and partial characterization of new breast cancer cell lines with different molecular subtypes, will allow us a better comprehension about the variations in morbidity, mortality and susceptibility to anti‐cancer drugs among different ethnicities.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mechanism of Fgf‐Mediated Lhx2 Upregulation
Authors: Shah Neera , Watson Billy , Pira Charmaine , Oberg Kerby 
Published: June 2021

IntroductionDuring development, fibroblast growth factors (FGFs) secreted from the apical ectodermal ridge (AER) direct limb outgrowth. FGFs also coordinate limb patterning through cross‐talk with other signaling molecules. The LIM homeobox transcription factor LHX2 is a suspected regulator of patterning cross‐talk, is expressed at the distal tip of the limb bud subjacent to the AER, and is a primary response target of FGF (Figure 1). However, the mechanism by which FGFs upregulate LHX2 is unknown. There are four major FGF signaling pathways through which regulation may occur: Jak/Stat, PLCγ, AKT, and Ras‐related (Figure 2). Based on the suspected roles of each signaling pathway, we hypothesize that FGF mediates LHX2 upregulation through the Ras‐related pathway.MethodsTo determine whether FGFs regulate LHX2 through the Ras‐related pathway, we injected the chemical inhibitor Salirasib (2.5 mM) into the anterior, posterior, and distal tips of the right limb of chick embryos at Hamburger‐Hamilton stage 21–22 (Figure 3). We also examined the potential function of the other Fgf pathways using pathway‐specific inhibitors. The embryos were harvested 24 hours after treatment and assessed for morphology. Whole mount in situ hybridization for LHX2 was then performed (Figure 4).ResultsPreliminary results indicate that inhibition of the Ras‐related pathway caused growth inhibition of the treated limbs in all embryos. In addition, LHX2 expression was markedly decreased in the these limbs. Treatment with a PLCγ‐specific inhibitor did not alter limb growth, while an AKT‐specific inhibitor impaired limb growth and LHX2 expression.ConclusionThese results suggest that the Ras‐related and AKT pathways control Fgf‐mediated LHX2 upregulation, whereas PLCγ does not. Further studies are needed to confirm these findings and to evaluate the Jak/Stat pathway.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Muscle Fiber‐type Composition of Primates in Relation to Brain Size
Authors: Omstead Kailey M , Muchlinski Magdalena N , Hemingway Holden W , Burrows Anne M 
Published: June 2021

There are many metabolic hypotheses for how humans can afford such large brains. The Expensive Tissue Hypothesis (ETH) addresses the metabolic cost of the brain in accordance to the costs of other metabolically “expensive” organs in the body. Skeletal muscle is composed of two main fiber types; type I (slow‐twitch) and type II (fast‐twitch). Type I fibers convert glucose to ATP using oxygen, while type II fibers rely primarily on anaerobic metabolic processes. When active, skeletal muscle is in direct competition with the brain for glucose and is an expensive tissue to maintain. Skeletal muscle acting as expensive tissue however, has not been addressed in context to the ETH. Based on these observations, we hypothesize that primates will have a decrease in type I fibers in accordance to their brain mass to account for the expensive tissue. For this preliminary study, our goal was to measure type I and type II fiber frequencies in various primates using immunohistochemistry to provide us with a muscle fiber composition profile. Two forelimb muscles (deltoid, pectoralis major) and two hindlimb muscles (gastrocnemius, soleus) were biopsied from 10 species of primates (N=11) for histological analysis. Formalin‐fixed paraffin embedded muscle samples were sectioned (12um) and immunostained using type I and type II Myosin heavy‐chain antibodies. Histological analysis of muscle morphology was visualized and quantified using light microscopy. Through muscle fiber‐typing, we found that primates appear to have fewer type I fibers than type II fibers. Results from this study have been implemented as part of a larger study (Muchlinski et al., 2018) to clarify the relationship between muscle mass and brain mass to better illustrate how muscle fiber composition could be used to address evolutionary questions in relation to energetics.Support or Funding InformationNational Society for Histotechnology (NSH)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Capillary density and succinate dehydrogenase in skeletal muscle on chronic kidney disease rats
Authors: Murakami Shinichiro , Sakita Masahiro , Huo Ming , Kondo Hiroyo , Hayashi Satoshi , Kameyama Kazuyoshi , Fujino Hidemi 
Published: June 2021

Background and PurposeKidney disease increases risk of blood vessel and capillary disease. In case of chronic kidney disease (CKD), these problems may happen slowly over a long period of time. The thing about heart in particular is well known. Chronic kidney disease may be caused by diabetes, high blood pressure and other disorders. Hypertension causes CKD and CKD causes hypertension. Anyone can get CKD at any age and any sex. However, some people are more easily to get kidney disease than others. That two main causes of CKD are diabetes and high blood pressure. Those diseases fail with blood vessel and capillary in various factors. In any case, the blood vessel system is connected with kidney disease deeply. We confirmed muscle capillary architecture on rat last time. We clarified muscle capillary density and mitochondrial activity this time.Materials/Methods12 Wistar rats (age; 16wks) were randomly divided into CKD group (n=6) and Sham operation (SHAM) group (n=6). Rats in CKD group suffered from a chronic kidney failure artificially. The details are as follows; 1. When 16wks old, under anesthesia with pentobarbital, we removed the two‐third left kidney surgically. 2. When 17wks old, under anesthesia with pentobarbital, we removed the right kidney surgically. 3. When 23wks old, we removed soleus muscle out. 4. Those muscles were frozen, were stained by SDH, AP and ATPase (pH4.2) method, and observed with light microscope (LM). In SHAM group, they were operated for at the same time, but the kidney was not removed surgically. The kidney was extracted to outside from rat and it was returned to rat without removing it surgically. The microscope image of SDH stained was analyzed in NIH Image software and it was compared. All data were presented as the means ± SEM. All statistical tests were done using an unpaired Student's – test. P<0.05 indicates a significant difference.ResultsOn 23 wks old CKD, body weight and soleus weight were decrease. Feeding volumes were same. Urinary albumin/urinary creatine value was increase. Data from measurement of the LM images by the NIH Image software of SDH, the mean SDH activity on CKD rats were significantly higher than that in the SHAM rats. Capillary density on CKD rat was lower than that in SHAM. However, muscle myosin heavy chain types of ratio on CKD were same to SHAM.ConclusionsThese results indicate that muscle requires oxygen, but shows that blood supply to muscle decreases on CKD.Support or Funding InformationThis study was supported by Grants‐in‐Aid for Scientific Research from the Japanese Ministry of Education, Culture, Sports, Science and Technology (Grant no. 17K01879)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Serum Brain‐derived neurotrophic factor promotes repair and regeneration of aged myelinated fibers via aerobic exercise
Authors: Sakita Masahiro , Murakami Shinichiro , Isobe Wataru , Kameyama Kazuyoshi , Hayashi Satoshi , Fujino Hidemi 
Published: June 2021

PurposeDegeneration of the distal peripheral nerve (DPN) with aging is associated with balance and gait disorders. Brain‐derived neurotrophic factor (BDNF) contributes to the repair and remodeling of myelinated fiber. However it is not obvious whether BDNF in the DPN increases with aerobic exercise (AE) in old age and promotes remodeling of MFs. We investigated morphological changes of MFs and BDNF expression levels of both serum and DPN via AE.MethodsEight male wistar rats (aged 95 weeks) were randomly divided into AE and control (Cont) groups. Rats in AE group performed running on the treadmill for 1 hour per day, 6 days per a week during 2 weeks. At 97 weeks of age, the bilateral tibial nerves of 2 groups were removed. To measure myelin thickness and axon diameter, semi‐thin sections of the unilateral specimens were stained with 0.5% Sudan Black B solution following double‐fixation. In addition, each expression level of serum and tissue BDNF in the contralateral specimens was analyzed with enzyme‐linked immunosorbent assay.ResultsMyelin thickness and axon diameter of AE group were significantly larger than those of Cont group (p<0.01). The serum BDNF level of AE group significantly decreased in comparison to that of Cont group (p<0.01). In contrast, BDNF level of AE group was significantly higher than that of Cont group (p<0.01).ConclusionIt was thought that serum BDNFs that were promoted by AE release into nerve fibers and contribute to the remodeling of myelinated fibers in aged DPNs.Support or Funding InformationThe present study was supported by a Grant‐in‐Aid for Scientific Research (C) from the Japan Society for the Promotion of Science (https://kaken.nii.ac.jp; 26350853).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 5‐Fluorouracil/Leucovorin Chemotherapy on Cognitive Function in Mice
Authors: Groves Thomas , Alexander Tyler , Anderson Julie , Kiffer Fred , Allen Antino 
Published: June 2021

5‐fluorouracil (5‐Fu) and leucovorin (LV) are often given in combination to treat colorectal cancer. 5‐Fu/LV prevents cell proliferation by inhibiting thymidylate synthase (TS), which is an enzyme that catalyzes the conversion of deoxyuridine monophosphate to deoxythymidine monophosphate. While 5‐Fu has been shown to cause cognitive impairments, the synergistic effect of 5‐Fu with LV has not been fully explored. The present investigation was designed to assess how the combination of 5‐Fu and LV affect cognition in a murine model. A total of 30 nine‐month‐old male mice were used in this study; 8 mice received 0.9 % saline injections, 6 mice received LV (90 mg/kg) injections, 8 mice received 5‐Fu (50 mg/kg) injections, and 8 mice received 5‐Fu and LV (50 mg/kg and 90 mg/kg) injections. One‐month post‐treatment, Y‐maze and Morris water maze (MWM) behavioral tasks were performed. Brains were then extracted, cryosectioned, and stained with CD68 and tomato lectin. All animals in the MWM task were able to locate the visible and hidden platform locations in the water maze; however, treatment effects were seen when spatial memory retention was assessed in the probe trials (no platform). A significant impairment in spatial memory retention was observed in the probe trial after the first day of hidden platform training (first probe trial) in animals that received LV, 5‐Fu, and 5‐Fu/LV. However, by day 5, animals that received 5‐Fu only and 5‐Fu/LV showed spatial memory retention in the probe trials, whereas the LV only mice were impaired. We are currently performing microscopy to quantify changes in the activated microglia and brain vasculature. In studies conducted by Mustafa et. al and El Betagy et. al, Lister‐hooded rats were given 5‐Fu and LV and performed the novel location recognition task. For both studies, rats that received 5‐Fu/LV failed to discriminate between the two object locations, suggesting a spatial memory deficit. While these studies are promising, there is still a need to determine how this drug combination affects other aspects of memory and brain function. The present data suggests that LV may exacerbate the cognitive deficits seen with 5‐Fu treatment, as mice that received LV treatment only remained impaired by day 5 of the MWM. Mice that received 5‐Fu and 5‐Fu/LV were also impaired but showed spatial memory retention by day 5.Support or Funding InformationThis work was supported by Pilot Grant under NIH P20 GM109005 (ARA).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of microRNAs associated with apoptosis in neurospheres of glioblastoma cell line subjected to treatment with ionizing radiation and temozolomide.
Authors: Schiavoni Vagner Sarraipo , Rodrigues Andressa Romualdo , de Assis Cirino Mucio Luiz , Neto Fermino Sanzes Lizarte , da Cunha Lourenço Luana Pretti , Peria Fernanda Maris , Bisson Gabriela Silva , Tiezzi Daniel Guimaraes , Tirapelli Luis Fernando , Junior Carlos Gilberto Carlotti , da Cunha Tirapelli Daniela Pretti 
Published: June 2021

IntroductionGlioblastomas (GBM) are the most common malignant primary tumors in the brain, and show high mortality rate. Despite the current advances in the therapy, the GBMs are extremely resistant to ionizing radiation and chemotherapy, and the number of relapse is high. Studies bring this eminent tumorigenic potential due to the presence of a subpopulation of neoplastic cells with characteristics of stem cells, called cancer stem cells (CSCs). In gliomas, the isolation of tumor stem cells have been done by antigenic markers and observing the culture conditions of normal neural stem cells in vitro. That is, the floating proliferation of tumor cells when cultured, are analogous to neurospheres derived from normal neural stem cells in defined culture conditions. It is believed that CSCs are responsible for the restoration of the tumor and the low efficiency of the treatment, as these cells demonstrate malignant properties as tumorigenesis, chemoresistance and radioresistance. The practical implication of this finding is that no current therapy is able to suppress or stop the proliferation of these cells. Several microRNAs have been linked to the development and proliferation of glioblastomas, associated with different molecular mechanisms, among them programmed cell death. Studies have shown that these microRNAs in brain tumors exhibit altered expression levels, one of the essential mechanisms for the regulation of tumor stem cells (CSCs).ObjectivesTo evaluate the effect of ionizing radiation and temozolomide, alone or associated, on the expression of miRNAs that act as tumor suppressors (miR‐15 and miR‐16) and as oncogene (miR‐21), and adhered cells in neurospheres in culture of glioblastoma cell line (U343‐MG).Materials and MethodsThe trypan blue was used to examine cell viability before and after treatment, and method of real time PCR to check the expression of microRNAs in time 0h, considered to be immediately after the treatments, and 48h after exposure to treatments.Results and ConclusionsThe presented microRNAs are differentially expressed when comparing the different treatment modalities and time. We also observed differences between adhered cells and neurospheres. The oncogene miR‐21 remained with higher expression on treatment with ionizing radiation in neurospheres compared with the group of adherent cells.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endurance exercise training protects against the upregulation of nitric oxide in the striatum of MPTP/probenecid mouse model of Parkinson's disease.
Authors: Bataineh Ziad Mohedeen 
Published: June 2021

Parkinson's disease (PD) is a neurodegenerative disorder, caused by the gradual loss of cells in substantia nigra. Nitric oxide (NO) plays an important role in a variety of signal transduction pathways that are crucial for maintaining the physiologic functions of nervous system. The aims of this study are: 1) To investigate the expression of the inducible form of NO (iNOS), and compare it to neuronal nitric oxide (nNOS) in the brain of a chronic mouse model of PD and 2) To study the effect of endurance exercise training on the expression of these markers.METHODMouse models of PD were obtained using 10 doses of 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) (25 mg/kg) and probenecid (250 mg/kg) over 5 weeks. Forty C57BL/6 albino mice were randomly divided into four groups: sedentary control (SC, N = 10), exercise control (EC, N = 10), sedentary PD (SPD, N = 10), exercise PD (EPD, N = 10). At the end of training program, nNOS and iNOS were evaluated in the striatum in all animal groups using immunohistochemistry.RESULTSnNOS showed significant increases in striatum (ST) of SPD mice compared to SC mice (P > 0.03). There was also decreased expression of nNOS in EC group compared to SC mice, but this decrease was not significant (P > 0.8). Exercise training significantly decreased the level of nNOS in the EPD compared to SPD, (P > 0.04). Although, iNOS expression followed almost the same trend as nNOS, but exercise training did not significantly decrease the expression of iNOS in both EC and EPD groups, P > 0.2 and 0.3 respectively.DISCUSSIONThe data from this study suggests that 4 weeks of treadmill exercise has a positive impact on the expression of nNOS and iNOS in the striatum of a PD model. This might clear in part the pathogenicity of the diseases and the positive impact of training on PD.Support or Funding InformationThis work was funded from the Deanship of Scientific Research at Jordan University of Science and TechnologyThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mechanistic Insight into Species‐specific Redox Partner Interactions in the Vitamin D Carbon‐24 Hydroxylase CYP24A1
Authors: Kumar Amit , Estrada D. Fernando 
Published: June 2021

The mitochondrial cytochrome P450 24A1 (CYP24A1) is responsible for side chain hydroxylation of vitamin D, the initial step in deactivation of the bioactive hormone. Mutations in CYP24A1 lead to incomplete clearance of active vitamin D and correlate with idiopathic infantile hypercalcemia, or elevated calcium in blood serum, and can lead to loss of appetite, nausea, vomiting, and failure to thrive in newborns. The regiospecific hydroxylation of the vitamin D side chain at either carbon‐23 (C23) or carbon‐24 (C24) relies in part on differences between species; while human isoforms are both C23 or C24 hydroxylases, other species are either C23 or C24 pure hydroxylases, despite an overall sequence identity of 80% between isoforms. Recent work from our group using solution NMR to evaluate the interaction between CYP24A1 and the redox accessory protein adrenodoxin (from bovine) suggests that species displaying distinct regioselectivity for vitamin D also form distinct P450‐redox partner complexes. Evidence of species‐specific complexes combined with the fact that mix‐species P450‐redox complexes are widely used to examine P450 function, highlights the need to examine this redox complex using proteins from the same species. Therefore, in this work we have used a combined approach of site‐directed mutagenesis, protein NMR, chemical cross‐linking, and vitamin D hydroxylation assays to evaluate the CYP24A1‐adrenodoxin complex from rat (a C24 hydroxylase). Differences in the 2D NMR spectra between bovine and rat adrenodoxin led to the complete re‐assignment of the protein backbone for rat adrenodoxin. Nonetheless, differential broadening of the NMR signal when titrated with rat CYP24A1 bound to 1α‐hydroxyvitamin D suggests the complex is mediated by surface charges. Dependence on surface charges was also verified by site‐directed mutagenesis of both proteins as well as zero‐length chemical cross‐linking using 1‐Ethyl‐3‐(3‐dimethylaminopropyl)carbodiimide. While preliminary, this works demonstrates the combined use of biophysical and biochemical approaches to evaluate nuanced differences in a P450‐redox complex that is critical to the maintenance of proper levels of bioactive vitamin D.Support or Funding InformationNIGMSR00GM112862 (DFE)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.