TY - JOUR. T1 - Differential mRNA expression of prostaglandin receptor subtypes in macrophage activation. AU - Hubbard, Neil. AU - Lee, S. H.. AU - Lim, D.. AU - Erickson, Kent L. PY - 2001. Y1 - 2001. N2 - Assessing the regulation of macrophage receptors for prostaglandin (PGE2) is essential to understanding the control which that potent lipid mediator has in modulating macrophage activities. The purpose of this study was to assess the differential mRNA expression of PGE2 receptor subtypes (EP) during macrophage exposure to activating and transducing agents. RAW 264.7 macrophages constitutively expressed mRNA for EP2, EP3 and EP4 receptor subtypes. Messenger RNA for EP4 was expressed at a much higher level when compared to EP2 in unstimulated macrophages as assessed by kinetic quantitative RT-PCR. When macrophages were stimulated with LPS, EP2 mRNA levels were 12-fold higher when compared to unstimulated macrophages, while EP4 mRNA remained unchanged. Conversely, mRNA levels of both EP2 and EP4 ...
The substantial amount of RNA required for expression analysis is a limiting factor for the cDNA microarray technology in a number of potentially important applications. Two main approaches, signal amplification and global mRNA amplification, have been developed to overcome this obstacle. Signal amplification, such as dendrimer technology [1] and tyramide signal amplification (TSA) [2] aim to increase the fluorescent signal emitted per mRNA molecule. Global mRNA amplification has the purpose of increasing the number of available transcript equivalents for sufficient labeling from a limited starting amount. In current implementations, mRNA amplification techniques require less RNA than those based on signal amplification.. Van Gelder et al. [3] devised a multistep strategy to amplify mRNA from limited quantities of cDNA in studies of gene expression. Their method is commonly referred to in the literature as the Eberwine method. The general steps involve reverse transcription of mRNA with an oligo ...
The present study was designed to characterize the regulation of the type II corticosteroid receptor (GR) mRNA in two tissues involved in the control of the hypothalamic-pituitary-adrenal axis. We have used a solution hybridization/S1 nuclease protection assay to quantitate GR mRNA levels in the rat …
TY - JOUR. T1 - Human lymphocyte messenger RNA activity profiles in type I and type II diabetes. T2 - A tool for classification of metabolic disease. AU - Mariash, C. N.. AU - Burmeister, L. A.. PY - 1988/12/1. Y1 - 1988/12/1. N2 - We have previously used rat hepatic messenger ribonucleic acid (mRNA) activity profiles to categorize various pathophysiologic states. To test the hypothesis that similar techniques can be used to categorize disease states in humans, we examined the mRNA activity profiles by using in vitro translational assays of Ficoll-Hypaque-separated mononuclear cells obtained from six normal volunteers, six patients with type I diabetes, and five patients with type II diabetes as examples of different disease states. Translated proteins were labeled with sulfur 35-labeled methionine, separated by two-dimensional gel electrophoresis, and quantitated by videodensitometry of autoradiographs derived from the two-dimensional gels. Of approximately 160 quantitated mRNAs, the levels of ...
TY - JOUR. T1 - BCL2 protein expression parallels its mRNA level in normal and malignant B cells. AU - Shen, Yulei. AU - Iqbal, Javeed. AU - Huang, James Z.. AU - Zhou, Guimei. AU - Chan, Wing C.. PY - 2004/11/1. Y1 - 2004/11/1. N2 - The regulation of B-cell lymphoma 2 (BCL2) protein expression in germinal center (GC) B cells has been controversial. Previous reports have indicated post-transcriptional regulation plays a dominant role. However, a number of recent studies contradicted these reports. Using real-time polymerase chain reaction (PCR) and Standardized Reverse Transcriptase-PCR (StaRT-PCR), we measured the level of mRNA expression in GC, mantle zone (MNZ), and marginal zone (MGZ) cells from laser capture microdissection. Both quantitative RT-PCR measurements of microdissected GC cells from tonsils showed that GC cells had low expression of BCL2 transcripts commensurate with the low protein expression level. These results are in agreement with microarray studies on fluorescence-activated ...
Figure 3 Thermogenic program and β3-adrenergic receptor signaling is mediated by membrane-initiated ERα signaling. A: Immunoblot analysis of UCP1 levels in BAT and pWAT of ERα+/+, ERα−/−, WT, and KRRki/ki mice. Representative immunoblots and quantification are shown (n = 5-8 per group). #P , 0.01. B: qRT-PCR analysis of genes consistent with beige adipocytes in adipose tissues of ERα+/+, ERα−/−, WT, and KRRki/ki mice (n = 6-8). Relative mRNA expression levels are normalized to gapdh. *P , 0.05, #P , 0.01. C: Hematoxylin and eosin staining of pWAT of ERα+/+, ERα−/−, WT, and KRRki/ki mice. Scale bar indicates 100 µm. The graph depicts the quantification of mean cell area (n = 4). #P , 0.01. D: qRT-PCR analysis of genes consistent with beige adipocytes in NIH 3T3-L1 preadipocytes treated with vehicle (control), 100 nmol/L E2, or 2 µmol/L rosiglitazone for 72 h. Relative mRNA expression levels are normalized to gapdh. Data depict the results from three independent experiments. ...
Non-small cell lung tumor (NSCLC) may be the most common tumor as well as the leading reason behind death from tumor worldwide. in comparison to people that have low mRNA amounts (20.three months vs 34.three months, respectively; Log Rank Check, p?=?0.016), when contemplating all NSCLC levels which difference is buy 958772-66-2 a great deal larger when contemplating only sufferers with stage IV (15.9 months vs 31.three months, respectively; Log Rank Check, p?=?0.036). Furthermore, circulating Ang-2 mRNA amounts independently determine general survival, as well as the concordance (c) index evaluation showed that this is of the nomogram which has information relating to tumor stage, sufferers smoking position and circulating Ang-2 mRNA amounts present an elevated capacity to anticipate overall success in NSCLC sufferers (c-index 0.798). These outcomes claim that this nomogram could serve as a distinctive and practical device to determine prognosis in NSCLC, not really counting on the option of ...
RNA expression patterns of cancer-adjacent breast tissue could be to gauge future survival outcomes for women with estrogen receptor-positive breast cancer
Gene expression differs among individuals and populations and is thought to be a major determinant of phenotypic variation. Although variation and genetic loci responsible for RNA expression levels have been analysed extensively in human populations, our knowledge is limited regarding the differences in human protein abundance and the genetic basis for this difference. Variation in messenger RNA expression is not a perfect surrogate for protein expression because the latter is influenced by an array of post-transcriptional regulatory mechanisms, and, empirically, the correlation between protein and mRNA levels is generally modest. Here we used isobaric tag-based quantitative mass spectrometry to determine relative protein levels of 5,953 genes in lymphoblastoid cell lines from 95 diverse individuals genotyped in the HapMap Project. We found that protein levels are heritable molecular phenotypes that exhibit considerable variation between individuals, populations and sexes. Levels of specific sets of
Figure 3. Western blots of Cos7 cell extracts stained with CCM2 antibody (A and B). Cells were transfected with an expression vector encoding a full-length CCM2-GFP fusion protein ( ). Untransfected cells served as controls ( ). A, In transfected cells, the CCM2 antibody detects an 82-kDa protein (expected size for the fusion protein). No staining is observed in untransfected controls. B, Peptide competition eliminates staining, demonstrating the specificity of this antibody for the CCM2 protein. C, Multitissue Western blot reveals CCM2 protein expression in the brain, heart, lung, and kidney. - CCM2 expression parallels that of CCM1.
Gene transcription is a random process in single cells manifested by the observed distribution of mRNA copy numbers in homogeneous cell populations. A central question is to understand how mRNA distribution is modulated under environmental changes. In this work, we initiate a theoretical study on mRNA distribution dynamics for the stochastic transcription model that involves cross-talking signaling pathways to direct gene activation in response to external signals. We first express the distribution in mathematical dynamical formulas under both moderate and high transcriptional upregulations. In each scenario, our further numerical examples display an observed dynamical transition type among three distribution modes for stress genes in yeast. In particular, the intermediate bimodal stage sustains within a certain length of early time and lasts much longer than that generated by the single pathway. This shows the general and robust bimodal transcription regulated by the cross-talk of signaling pathways.
Principal Investigator:FUKUDA Takeshi, Project Period (FY):1991 - 1993, Research Category:Grant-in-Aid for General Scientific Research (C), Research Field:Respiratory organ internal medicine
gene, would facilitate examination of the role of this gene in the inheritance of human obesity. Northern blot analysis revealed that OB RNA is present at a high level in adipose tissue but at much lower levels in placenta and heart. OB RNA is undetectable in a wide range of other tissues. Comparative mapping of mouse and human DNA indicated that the ob gene is located within a region of mouse chromosome 6 that is homologous to a portion of human chromosome 7q. We mapped the human ...
Supplementary MaterialsSupplementary information 41467_2019_10135_MOESM1_ESM. reduces right center hypertrophy, restores the cardiac index, and order KOS953 decreases pulmonary vascular redecorating. These total results demonstrate that inhibition of CDKs by palbociclib could be a therapeutic strategy in PAH. and (Fig.?1f) and (and mRNA amounts could possibly be demonstrated, helping increased activity of the CDK-induced Rb-E2F pathway in HPASMCs from IPAH sufferers. To confirm the fact that predicted upsurge in activity of CDK2, CDK4, CDK6, and CDK9 is because order KOS953 of an enhanced appearance level under disease circumstances, real-time PCR analyses had been performed in isolated major HPASMCs 48?h after hunger (Supplementary Fig.?2aCh) and in homogenates of explanted individual lungs (Supplementary Fig.?2iCp). In HPASMCs (Supplementary Fig.?2aCompact disc), aswell such TM4SF4 as lung homogenates from IPAH sufferers (Supplementary Fig.?2iCl), increased and mRNA amounts were noted, whereas ...
The application of synthetic modified messenger RNA (mRNA) is a promising approach for the treatment of a variety of diseases and vaccination. In the past few years, different modifications of synthetic mRNA were applied to render the mRNA more stable and less immunogenic. However, the repeated application of synthetic mRNA still requires the suppression of immune activation to avoid cell death and to allow a sufficient production of exogenous proteins. Thus, the addition of type I interferon (IFN) inhibiting recombinant protein B18R is often required to avoid IFN response. In this study, the ability of B18R encoding mRNA to prevent the immune response of cells to the delivered synthetic mRNA was analyzed. The co-transfection of enhanced green fluorescent protein (eGFP) mRNA transfected fibroblasts with B18R encoding mRNA over 7-days resulted in comparable cell viability and eGFP protein expression as in the cells transfected with eGFP mRNA and incubated with B18R protein. Using qRT-PCR, significantly
Individual fractions of polysomes were isolated from yeast. Pulse labeling experiments in vivo show constant specific activity of messenger RNA in each polysome peak; this suggests a uniform density of ribosomes per unit length of messenger RNA. In the cell-free incorporating system, the amount of peptide per ribosome unit increased with the size of polysome.. ...
RNA binding proteins (RBPs) can regulate the stability and/or translatability of messengerRNAs (mRNAs) throughinteractions with their 30-untranslated regions. However, individual mRNAs may be regulated simultaneously or successivelyby more than one RBP, as well as by Argonaute (AGO)-bound miRNAs; the coordination of these various influenceson an individual mRNA is therefore complex and not well studied. In this report we examine the roles of two RBPs thatbind to AU-rich elements (ARE) - AUF1 and HuR - in the stability and translation of cyclin D1 (Ccnd1) mRNA in ratmyoblasts transiting the G phase of the cell cycle, and their interactions with miRNAs. Knockdown (KD) of AUF1 resultedin (1) transient upregulation of the mRNA level as well as an advancement of translation onset time (TOT) from 6 to 5 hpost-serum addition, (2) loss of miRNA loading on AGO1 and AGO2 and (3) reduction in the level of AGO-1 and AGO-2bound mRNA. In contrast, KD of HuR had no effect on the mRNA level, or on the AGO-mRNA ...
酵母细胞通过提高葡萄糖合成酶和糖酵解作用维持能量代谢的平衡。在人体细胞中我们也发现了同样的代谢调节途径,主要通过eIF3e蛋白与代谢相关的mRNA结合并促进这些与能量代谢相关的蛋白表达来维持代谢的平衡。在肿瘤形成的过程中,这种通过eIF3d和eIF3e调控的能量代谢机制是被打破的,因此该研究成果将有助于科学家通过靶向代谢通路研发治愈肿瘤的药物。该研究利用转录组学、蛋白组学和代谢组学的方法发现缺少了eIF3e和eIF3d的裂殖酵母细胞无法合成线粒体电子传递链相关的蛋白,从而导致呼吸功能阻断,内源性的氧化应激压力和细胞老化产生。 课题组硕士研究生苏丹为共同第一作者 。 Dieter A. ...
Shinde and Klein are not yet sure whether GSK-3s effect on RNA splicing explains its role in mood disorders. The effect of GSK-3 on messenger RNA in neuronal cells, with or without lithium, would need to be examined to determine this. The study underlines how investigations into the basic biological function of a drug target can lead in unexpected directions. [The GSK-3 phosphoproteome] is a really large data set, Shinde said. Its a resource for the field. The relevance to leukemia could be direct and something worthy of immediate study, Klein said. The role in psychiatric disorders is a major interest of the work, but the impact would be down the road, not immediate. ...
Effect of gefitinib on CYP mRNAs expression and EROD activity in NSCLC cell lines The baseline transcript levels of had been determined in both sensitive and resistant cell lines selleck chemical MK-0457 by RT PCR and information are summarized in Figure 4A. CYP1A1 and CYP1A2 had been expressed at important levels only in H322, H292 and Calu three cell lines, CYP2D6 was detected in all cell lines, whereas CYP3A4 was undetected. CYP3A5 was present at higher level only in A549 cells. The inducibility of individual CYP genes by gefitinib was then investigated along with the levels of CYP1A1, CYP1A2, CYP2D6 and CYP3A5 mRNAs have been assessed just after treating cells with the drug. After six h, significantly greater gene expression levels of CYP1A1 and CYP1A2 had been observed in all sensitive cell lines. By contrast no substantial modulation of gene expression was observed in resistant cell lines. As a way to evaluate no matter if modulation of the CYP1A1 transcript levels was connected with ...
Time-dependent induction of connected cells by overexpression of cdc5ΔN. (A) Strain KLY1083 expressing three copies of GAL1-EGFP-cdc5ΔN homogeneously induced
Our mRNA assays have exceptional sensitivity and specificity using short, LNA-enhanced primers and are optimized to eliminate nonspecific amplification
Our mRNA assays have exceptional sensitivity and specificity using short, LNA-enhanced primers and are optimized to eliminate nonspecific amplification
Alternative splicing (AS) is a post-transcriptional regulatory mechanism for gene expression regulation. Splicing decisions are affected by the combinatorial behavior of different splicing factors that bind to multiple binding sites in exons and introns.
Messenger RNA (mRNA) is a molecule of RNA encoding a chemical blueprint for a protein product. mRNA is transcribed from a DNA template, and carries coding information to the sites of protein synthesis: the ribosomes. Here, the nucleic acid polymer is translated into a polymer of amino acids: a protein. In mRNA as in DNA, genetic information is encoded in the sequence of nucleotides arranged into codons consisting of three bases each. Each codon encodes for a specific amino acid, except the stop codons that terminate protein synthesis. This process requires two other types of RNA: transfer RNA (tRNA) mediates recognition of the codon and provides the corresponding amino acid, while ribosomal RNA (rRNA) is the central component of the ribosomes protein manufacturing machinery.. ...
Background: Current methodology of gene expression analysis limits the possibilities of comparison between cells/tissues of organs in which cell size and/or number changes as a consequence of the study (e.g. starvation). A method relating the abundance of specific mRNA copies per cell may allow direct comparison or different organs and/or changing physiological conditions. Methods: With a number of selected genes, we analysed the relationship of the number of bases and the fluorescence recorded at a present level using cDNA standards. A lineal relationship was found between the final number of bases and the length of the transcript. The constants of this equation and those of the relationship between fluorescence and number of bases in cDNA were determined and a general equation linking the length of the transcript and the initial number of copies of mRNA was deduced for a given pre-established fluorescence setting. This allowed the calculation of the concentration of the corresponding mRNAs per ...
Fingerprint Dive into the research topics of Scleraxis messenger ribonucleic acid is expressed in C2C12 myoblasts and its level is down-regulated by bone morphogenetic protein-2 (BMP2). Together they form a unique fingerprint. ...
I would like to know what amounts of protein are necessary for pregnant women? I have been eating the Paleo Diet since you introduced me to it. This is my first
Im prepping for surgery right now with a liquid diet. What would you say is the minimum grams of protein to get a day? My doctor wants 100g but Ive been lucky to get 60g. Is this going to hurt me down the road?Thanks!
YV Subrahmanyam, S Yamaga, Y Prashar, HH Lee, NP Hoe, Y Kluger, M Gerstein, JD Goguen, PE Newburger, SM Weissman (2001). Blood 97: 2457-68 ...
Mouse monocytic Mm-A, Mm-P, Mm-S1, and Mm-S2 cells are sublines of mouse monocytic and immortalized Mm-1 cells derived from spontaneously differentiated, mouse myeloblastic M1 cells. Although these subline cells retain their monocytic characteristics in vitro, Mm-A and Mm-P cells are highly leukemogenic to syngeneic SL mice and athymic nude mice, whereas Mm-S1 and Mm-S2 cells are not or are only slightly leukemogenic. To better understand the molecular mechanisms of these levels of leukemogenicity, we investigated putative leukemogenesis-associated genes or oncogenes involved in the maintenance of growth, especially in vivo, by means of differential mRNA display. We isolated a fragment clone (15T01) from Mm-P cells. The mRNA probed with 15T01 was expressed at high levels in leukemogenic Mm-P and Mm-A cells but not in nonleukemogenic Mm-S1 and Mm-S2 cells. The gene corresponding to 15T01, named TRA1, was isolated from an Mm-P cDNA library. The longest open reading frame of the TRA1 clone predicts ...
Mouse monocytic Mm-A, Mm-P, Mm-S1, and Mm-S2 cells are sublines of mouse monocytic and immortalized Mm-1 cells derived from spontaneously differentiated, mouse myeloblastic M1 cells. Although these subline cells retain their monocytic characteristics in vitro, Mm-A and Mm-P cells are highly leukemogenic to syngeneic SL mice and athymic nude mice, whereas Mm-S1 and Mm-S2 cells are not or are only slightly leukemogenic. To better understand the molecular mechanisms of these levels of leukemogenicity, we investigated putative leukemogenesis-associated genes or oncogenes involved in the maintenance of growth, especially in vivo, by means of differential mRNA display. We isolated a fragment clone (15T01) from Mm-P cells. The mRNA probed with 15T01 was expressed at high levels in leukemogenic Mm-P and Mm-A cells but not in nonleukemogenic Mm-S1 and Mm-S2 cells. The gene corresponding to 15T01, named TRA1, was isolated from an Mm-P cDNA library. The longest open reading frame of the TRA1 clone predicts ...
For zooming please use the mouse wheel. The movie shows a sequence of slides with expression data at t=0h, t=5h, t=7h and t=9h after a shift of an aerobically grown culture to anaerobic conditions. Expression on protein level has been encoded in shades of blue with white (no expression at all) and blue (highest observed expression along the shown time line). Cell sizes encode the maximum protein amount over the monitored sampling points t0 to t9 ...
Data Availability StatementThe first study data used to aid the results of the scholarly research are included within this article. 5-aza-2′-deoxycytidine downregulated the methylation of NKX2.2 and retrieved its manifestation of mRNA and proteins amounts (p 0.05). No significant association was discovered Ntf5 between your NKX2.2 sex and methylation, age group, tumor differentiation, TNM stage, CEA, CA199, and fecal occult bloodstream (p 0.05). Kaplan-Meier evaluation indicated that NKX2.2 hypermethylation showed a tendency however, not statistical FK-506 (Tacrolimus) significance for predicting poor overall success in CRC individuals (p=0.33). NKX2.2 overexpression suppressed cell proliferation, colony formation, and inhibited tumor invasion and migration in CRC cells (both p 0.05). Conclusions: This research shows that NKX2.2 is a tumor suppressor in CRC because of hypermethylation. strong course=kwd-title Keywords: Colorectal tumor, Hypermethylation, NK homeobox 2.2, Epigenetic Intro ...
Cancer cells. (A) Relative expression levels of Nox1, 2, 3, 4, and 5 mRNAs in A549 cells were determined by real-time RT-PCR and are presented as mean delta Ct
The first and foremost point to be made here is that messenger RNA (mRNA) vaccines are not legally vaccines at all according to the CDC official definition. The FDA granted
Every cell in the human body contains DNA, Messenger RNA and Protein among many other components. The DNA stores instructions for what that cell is to actually…
For this article, RNA was isolated from mouse and rat brain followed by denaturing agarose gel electrophoresis and Northern blot analysis of isolated RNA. RT-PCR analysis was performed on total RNA from mouse and rat brain as well as rat poly(A)+ RNA followed by T-vector cloning of beta-actin RT-PCR product and colony screening for positive recombinants.
Alcyomics are looking for a Scientist, Grade 1 (with 1-3 years post doctoral experience and industry experience an advantage).. The work will primarily be to ensure client projects are completed efficiently and in a timely manner and will also involve research and development (R&D) within Alcyomics Ltd. The job entails working with a team of scientists within Alcyomics Ltd in the completion of client proposals as well as development of primary cell lines and differentiation of these lines to develop in vitro 3D skin models or 3D osteoarthritic (OA) joint models. Experience of tissue culture, immunohistochemistry, mRNA expression analysis and cytokine analysis will be required. Work will also involve sectioning and embedding of tissue, Skimune® assays and cytokine analysis. The job will also involve the Scientist contributing to other commercial activities.. Essential Requirements: -. Must have extensive tissue culture experience and tissue sectioning and staining expertise.. Working within a ...
J:60127 Pascolo S, Tsoukatou D, Mamalaki C, Identification of thymus specific and developmentally regulated genes by an improved version of the mRNA differential display technique. Dev Immunol. 1999;7(1):1-7 ...
Comments, concepts and statistics about Systematic discovery of structural elements governing stability of mammalian messenger RNAs..
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The technology we have available to us today in the lab is both a boon and a bafflement. Example: The screens we have for RNA expression in cells is so sensitive we can see tiny changes in RNA expression levels in healthy/diseased/drug treated/etc cells. YAY! More information! More observations! More new ideas for research!… Except,…. ...
DS was born at 36 weeks by EMCS, following a failed induction attempt. There were complications in the pregnancy that meant he was monitored closely
DNA is the informational basis from which living cells derive instructions for synthesizing proteins. Many of the resulting proteins are enzymes that catalyze biochemical reactions from which the cell derives energy or generates other molecules essential to its health and safety. The process normally occurs when the sequence of nucleotides in DNA is transcribed into a complementary, single strand of nucleotides known as messenger RNA, or mRNA. The mRNA provides the instructions by which other components in the cell synthesize proteins. Because not all genes are transcribed (or expressed) but all genes that are transcribed do so through mRNA, the presence of mRNA is an indicator that a gene from the cells DNA has been expressed. The DNA from which the mRNA is obtained is sometimes interspersed with oligonucleotide spacers that do not appear in the final mRNA. Because mRNA is used for the cells molecular machinery to generate the protein, the sequence of DNA (or gene) that corresponds to a ...
overexpressing GPX1 in endothelial cells is able to change the basal mRNA and protein BAX levels without affecting those of TP53 and BCL2 (useful to antiatherogenic therapies which use antioxidants with the aim of protecting the vascular wall against ...
Question 1 (1 point) Which of the following is a true statement about genes?Question 1 options: A) Genes are structures within chromosomes of each cell that contain deoxyribonucleic acid B) Genes are messenger ribonucleic acid (mRNA) C) Genes are cells that determine gender during fertilization D) Genes are antibodies that promote disease SaveQuestion 2 (1 point) Which of the following is the major cause of death from cancer?Question 2 options: A) Infection B) Hemorrhage C) Pain D) Metastasis SaveQuestion 3 (1 point) Which of the following statements is true with respect to the stages of cancer?Question 3 options: A) Stage 1 represents a poor prognosis B) Localized cancer is a stage 4 C) Benign tumors are stage 2 D) &
CureVac is the first company to successfully harness messenger RNA (mRNA) for medical purposes. The principle is promising: use natural mRNA as a data carrier to instruct the human body to produce its own proteins to fight a wide range of diseases. ...
Study Flashcards On FA page 199 G-protein linked 2nd messengers, major functions plus major grouping at Cram.com. Quickly memorize the terms, phrases and much more. Cram.com makes it easy to get the grade you want!
Last weekend I had the opportunity to learn from one of the most intelligent fitness minds in the industry. Aside from learning an abundant amount of