bluuebunny/crossref_metadata_2025_split huggingface dataset:
Assessing the Role of Ndc80 Kinetochore Complex in Accurate Chromosome Segregation
Authors: Rodriguez‐Lopez Xavier Luis , Amin Mohammed , Wang Yu , Yao Yiyi , Varma Dileep 
Published: June 2021

Mitosis is required to grow and develop from a single cell to a multicellular organism. However, this process can often lead to cancer when its regulation is lost. The Highly Express in Cancer 1 (Hec1) protein is one of the four subunits of the Nuclear Division Cycle (Ndc80) complex present in mitosis. The function of this complex is required for robust kinetochore‐microtubule (kMT) attachment and is regulated by Aurora B Kinase phosphorylation of Hec1. Given that Hec1 protein binds directly to spindle microtubules, it certainly plays a critical role in chromosome alignment and segregation. To assess how Hec1 leads to defects in cell division cycle in human cells and induce cancer, we expressed a non‐phosphorylatable version of Hec1 (Hec1‐9A) in MDA‐MB‐436 (breast cancer) cells. We observed how mitosis was modified by the Hec1‐9A mutant by employing immunofluorescence microscopy to mark the chromosomes and the mitotic spindle in our Hec1‐9A positive and control cells. The Hec1‐9A satisfied the genome surveillance mechanism, thus, allowing cells that presented erroneous chromosome alignment and segregation to exit mitosis. Further studies will be aimed to investigate whether spindle checkpoint is defective at erroneous kMT attachments and whether Hec1 plays a role in other steps of mitosis.Support or Funding InformationFunded by NIH‐NIGMS #2R25GM096955Funded by NIH #R00CA178188This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
FLG loss of function mutations R501X and R2447X in Puerto Rico: Association of genetic ancestry with Atopic Dermatitis and Ichthyosis Vulgaris
Authors: Ramirez Aponte Edwin G , Martinez Cruzado Juan C 
Published: June 2021

In Puerto Rico, we do not have many studies of Loss of function (LoF) mutations but using the data from the 1000 Genome Project we identified several polymorphisms that predicted to eliminate protein function. In this research, we work with two single nucleotide polymorphisms that produce premature stop codons in the FLG gene. They are known as R501X and R2447X. Both are associated to skin conditions such as ichthyosis vulgaris and atopic dermatitis; they also have been associated to asthma and allergies. The 1000 Genome Project data is based on the analysis of 104 Puerto Ricans only, and we wanted to test if the frequencies in it were accurate for the Puerto Rican population. We genotyped 736 individuals for the mutation R501X and 292 for R2447X using real time PCR. From the 736 individuals, we found 21 heterozygotes for the SNP R501X, this mutation in the FLG gene having an allele frequency of 0.0142. This allele frequency compared to the one previously reported, shows a difference of 0.0114 suggesting that the mutation in Puerto Rico is not rare. This mutation has been identified mainly in 3 geographical regions of Puerto Rico with high European ancestry. We started working with the mutation R2447X and genotyped 292 individuals; and 3 heterozygotes were found. From this samples the mutation shows an allelic frequency of 0.005; and it appears to be localized in the east region of the island. Now, we expect to genotype the same individuals for both mutations and compare their distribution, prevalence and relation between both.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gene Expression of DNA Repair Proteins in Colon Cancer Tissue
Authors: Odufuwa Kuburat Temitope , Elder Rhoderick , Povey Andrew 
Published: June 2021

Cancer is a disease that arises from the disruption of cellular and genetic functions, either intrinsically or extrinsically. DNA repair genes play a critical role in maintaining the integrity of the genome and the dysregulation of the key DNA damage repair genes has been implicated in the development, progression and chemotherapeutic resistance of different cancers. Colorectal cancer (or colorectal adenocarcinoma) is one of the most common malignant neoplasms and a leading cause of cancer‐related deaths worldwide. This work will compare the level of gene expression of the DNA repair genes NEIL1, NEIL2, NEIL3, ERCC1, MLH1, OGG1, and NTHL1 with the housekeeping gene GAPDH in colon tumour tissue and matched normal colon tissue. Various molecular biology tools and techniques were employed in this research, including RNA extraction and purification, complementary DNA (cDNA) synthesis and qRT‐PCR analysis. Gene expression levels were determined using the comparative Ct method. For the twelve tumour tissues investigated, there is a reproducible and characteristic pattern of expression of the genes, with trends of expression observed between the Neil group of genes; Neil3 and Ercc1, and between Neil3, Ercc1 and Mlh1. The results showed that across the tumour tissues, the overexpression of one of the Neil genes marks the suppression of the others. Similarly, the results further demonstrated that there is a special relationship between Neil3 and Ercc1, with both genes antagonistic to each other. The expression patterns of these genes are indicators of the level of genetic instability and could serve as biomarkers for cancer diagnosis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Unstructured Proteins in Biological Structures: The Case of Human Teeth from a Protein Chemist's Perspective
Authors: Sharma Vaibhav , Srinivasan Alagiri , Roychoudhury Ajoy , Nikolajeff Fredrik , Kumar Saroj 
Published: June 2021

Proteins are the biomacromolecules that work downstream to genes in every living system. It is these protein molecules that are responsible for a plethora of functions in our body, ranging from building skeleton tissues to transmitting signal from one place to others. There was a period when biologists have the view that for a protein to function, the structure is of supreme importance. Nevertheless, this structure‐function paradigm is fairly obeyed by most of the proteins, if not all of them. Last two decades have witnessed the increased reports of proteins that do not have a stable three‐dimensional structure but are very much indispensable for the cellular and biological activities. These unstructured proteins are now known as “Intrinsically disordered proteins” (IDPs). Interestingly, these IDPs function not only in isolation (e.g. transcription factors) but helps in the formation of hard tissues like teeth, bone, and mollusks shell, by governing a process called biomineralization. Biomineralization is the process by which living organisms produce minerals, often to harden or stiffen existing tissues. Such tissues are called mineralized tissues.We have isolated total protein extracts from a healthy molar, premolar, canine and incisor teeth. We studied their protein secondary structural characteristics using various biophysical techniques. We have also investigated the effects of these protein extracts on mineralization of calcium phosphate through in vitro biomineralization assay followed by size‐based measurement analysis via Nanoparticle tracking analysis (NTA). The biophysical studies confirmed the presence of IDPs in the majority within these protein extracts. Moreover, these extracts were shown to elevate calcium phosphate biomineralization in a time‐dependent manner. This is the first time the effect of the whole protein extract was taken into account which in our view more closely imitates the in vivo process of tooth biomineralization. Furthermore, we also evaluated the differential proteome profile of the individual type of human teeth through Mass spectroscopy‐based method. The comparative qualitative and quantitative data of proteins from these different human teeth were obtained. The implication of these results will be discussed.Support or Funding InformationThere was no support or funding from any organizationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ordered and Disordered Segments of Amyloid‐β Drive Sequential Steps of the Toxic Pathway
Authors: Maiti Sudipta , Maity Barun Kumar , Das Anand Kant , Dey Simli , Moorthi Ullhas Kaarthi , Kaur Amandeep , Dey Arpan , Surendran Dayana , Pandit Rucha , Kallianpur Mamata , Chandra Bappaditya , Chandrakesan Murlidharan , Arumugam Senthil 
Published: June 2021

While the roles of intrinsically disordered protein domains in driving many interactions are increasingly well‐appreciated, the mechanism of toxicity of disease‐causing disordered proteins remains poorly understood. A prime example is Alzheimer's disease (AD) associated amyloid beta (Aβ). Aβ oligomers are highly toxic partially structured peptide assemblies with a distinct ordered region (residues ~10–40) and a shorter disordered region (residues ~1–9). Here, we investigate the role of this disordered domain and its relation to the ordered domain in the manifestation of toxicity through a set of Aβ fragments and stereo‐isomers designed for this purpose. We have measured their effects on lipid membranes and cultured neurons, probing their toxicity, intracellular distributions, and specific molecular interactions using the techniques of confocal imaging, lattice light sheet imaging, fluorescence lifetime imaging, and fluorescence correlation spectroscopy (FCS). Remarkably, we find that neither part ‐ Aβ10–40 or Aβ1–9, is toxic by itself. The ordered part (Aβ10–40) is the major determinant of how Aβ attaches to lipid bilayers, enters neuronal cells, and localizes primarily in the late endosomal compartments. However, once Aβ enters the cell, it is the disordered part (only when it is connected to the rest of the peptide) which has a strong and stereospecific interaction with an unknown cellular component, as demonstrated by distinct changes in the fluorescence lifetime of a fluorophore attached to the N‐terminal. This interaction appears to commit Aβ to the toxic pathway. Our findings correlate well with Aβ sites of familial AD mutations, a significant fraction of which cluster in the disordered region. We conclude that while the ordered region dictates attachment and cellular entry, the key to toxicity lies in the ordered part presenting the disordered part for a specific cellular interaction.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Protective Effect of DAS and DADS Against Doxorubicin Induced Toxicity
Authors: Taylor LaShaundra , Miles Jana , Webster Camille , Hudson Alicia , Soliman Karam , Flores‐Rozas Hernan , Darling‐Reed Selina 
Published: June 2021

PurposeCancer survival rates are increasing but the mechanisms involved in the adverse effects of cancer therapy such as doxorubicin are still being investigated along with treatment for these effects. Doxorubicin, an established chemotherapeutic agent used for various cancers, has limited use due to cardiotoxicity. The objective of this study, is to determine how garlic constituents, Diallyl sulfide (DAS) and Diallyl disulfide (DADS) may decrease the adverse effects of doxorubicin exposure.With a S. cerevisiae deletion library, a genome wide screen was conducted to detect mutants sensitive to doxorubicin exposure. A mutant deleted for the gene enconding for Mac1, a copper sensing transcription factor, was found to be highly sensitive. In S. cerevisiae the MAC1 gene is activated upon copper deficiency, resulting in the induction of copper transporters. Other doxorubicin sensitive strains included the sod1 mutant and its copper chaperone ccs1 mutant, both of which are associated with MAC1. The sensitivity of the mutant strains to doxorubicin, was in part due to the drug's capacity to generate reactive oxygen species (ROS). To test if garlic constituents, DAS and DADS could reduce the sensitivity of the mutants to ROS, the viability of the sensitive yeast strains were determined by exposing them to doxorubicin in the presence or absence of DAS and DADS.MethodsYeast survival assays were completed to determine the sensitivity of wt, mac1, ccs1, and sod1 strains to doxorubicin, menadione and etoposide, co‐treated with DAS and DADS. Statistical significance was set at p<0.05.Resultsmac1 shows sensitivity to doxorubicin and etoposide but not menadione. Menadione contains a quinone ring like doxorubicin, and acts through the induction of reactive oxygen species. Etoposide, a chemotherapeutic agent acts only through inhibition of DNA topoisomerase II. The sensitivity of mac1 to both etoposide and doxorubicin, shows that doxorubicin's sensitivity to mac1 is through its action on topoisomerase II. Unlike DAS, DADS was able to decrease the sensitivity of mac1 to doxorubicin and etoposide. Menadione has greater toxicity to ccs1 and sod1 than doxorubicin and etoposide, with DAS and DADS both increasing the survival of all chemicals to ccs1 and sod1.ConclusionGarlic constituents, das and dads may decrease the adverse effects of doxorubicin exposure, improving the quality of life of patients expose to doxorubicin.Support or Funding InformationResearch supported by National Institutes of Health (NIH) under Award Number G12MD007582This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Macromolecular Crowding on Alcohol Dehydrogenase and Citrate Synthase Kinetics
Authors: Slade Kristin , Chung Charmaine , Jackson Jasmine 
Published: June 2021

Most of our understanding of enzyme kinetics is based on experiments conducted in dilute solution. However, these conditions do not accurately represent the crowded cellular environment, which contains high concentrations (300–400g/L) of various macromolecules such as proteins, carbohydrates and ribosomes. This crowding reduces the volume of solvent available for other molecules in the solution and has previously been shown to impact the behavior of enzymes. To study these potential consequences, the Michaelis‐Menten kinetics of yeast alcohol dehydrogenase (YADH) and citrate synthase (CS) were monitored under crowded conditions. Spectroscopic assays were performed in the presence and absence of high concentrations of dextran or glucose, the small‐molecule counterpart. This study also explored the effects of mixed crowding by combining equal ratios of different sizes of dextran. Overall, the effects of crowding depend on the size and concentration of the crowding agent. Furthermore, the results demonstrated that, in addition to merely excluded volume effects, chemical interactions from crowders must also be considered.Support or Funding InformationCamille and Henry Drefyus Foundation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Examination of specific binding activity of packaging sequence RNAs to the SARS‐CoV nucleocapsid by using cell‐based in vivo assay for RNA‐Protein interaction
Authors: Woo Jiwon , Cho Yong‐Eun 
Published: June 2021

Severe acute respiratory syndrome coronavirus (SARS‐CoV) is the etiological agent of SARS, an emerging disease characterized by atypical pneumonia. The SARS‐CoV nucleocapsid protein serves multiple functions in viral replication, transcription and the formation of viral genome complex. Coronaviruses specifically package genomic RNA into assembled virions, little is known about the specific interaction between Nucleocapsid protein and packaging RNA sequence in previously. Herein, we exploited a cell‐based translation inhibition assay to elucidate the protein‐RNA interaction in living cells. Our results demonstrated that the packaging signal is pivotal in the specific interaction of the nucleocapsid and viral genomic RNA for incorporation into virions. We identified that the specific conserved RNA sequence of the nsp12 encoding vRNA of SARS‐CoV, and conserved RNAs of other corona viruses are major determinant sequences for the N‐protein interaction, respectively. In addition, using deletion mapping of the N‐protein, we also found that the specific in vivo RNA binding activity of the Nucleocapsid protein of the SARS‐CoV requires both the N‐terminal and C‐terminal domain. These results provide the first evidence for a specific interaction of the SARS‐CoV Nucleocapsid protein with the its viral packaging RNA sequence in vivo that will be useful as a valuable research tool for further detailed studies and providing new targets for antiviral therapies.Support or Funding InformationThis work was supported by the Brain Korea 21 PLUS Project for Medical Science, Yonsei University. In addition, this work was supported by a grant from the National Research Foundation of Korea (NRF‐2017R1D1A1B03030315).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Assessing anti‐cancer potential of newly synthesized staurosporine analogs
Authors: Hurysz Brianna , McFadden Marissa , Pelkey Erin , Mowery Patricia 
Published: June 2021

The discovery of staurosporine in 1977 and subsequent finding that it was a potent inhibitor of protein kinase C suggested that it might have anti‐cancer potential. Subsequently staurosporine was found to be a pan‐kinase inhibitor, making it to toxic for drug development, however, derivatives have been found with increased specificity and have been tested in clinical trials. The structure of staurosporine can be divided into three parts: a pyran ring that bridges the indole nitrogen atoms, an indolocarbazole central core, and a fused pyrrol‐2‐one. We focus on the synthesis and biological activity of the pyrrolinone group and bis‐indole motifs. Using a cytotoxicity assays we have analyzed structure‐function activity of our derivatives and endeavor to synthetic new targets with increased potency. We recently reported on the synthesis and biological activity of a 1st generation of analogs with low micromolar potency. Our findings suggested that biologically effective compounds require: a) an indole cis to the carbonyl oxygen atom on the 3‐pyrrolin‐2‐one central core ring, b) a flexible, rather than a planar, fused analog, c) aryl/heteroaryl groups at both the C3 and C4 position of the 3‐pyrrolin‐2‐one core, and d) multiple methoxy groups on the C4 ring. Building upon this work, we now report on a 2nd generation of compounds with nanomolar cytotoxic activity.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evaluation of Ant‐Lion Larvae Extract As Alpha‐Glucosidase and Alpha‐Amylase Inhibitor
Authors: Adeyanju Muinat Moronke , OLANIYI Omolayo Omobolawa , RASAK Olashile Fatimat , ADEOSUN Oyewale Oluwapelumi , ASHIDI Joseph Senu , OBUOTOR Efere Martins 
Published: June 2021

Diabetes mellitus is a serious condition with potentially devastating complications that affects all age groups worldwide with the percentage of affected people increasing at an alarming rate. Inhibition of alpha‐amylase and alpha‐glucosidase are the most important strategies in controlling the postprandial blood glucose and invariably for the treatment of disorders in carbohydrate uptake, such as diabetes and obesity. There had been several efforts geared towards sourcing anti‐diabetic therapeutics, the most studied being plants and have been reported to effectively do this but quite a number of them are toxic to humans. Thus we investigated alternative means of managing the ailment using ant lion larvae extract. In this study, alpha‐amylase and alpha‐glucosidase inhibitory potentials of the ant‐lion larvae extract were investigated. About 0.50 g of live ant‐lion larvae samples were hand‐picked from the soil within the school premises and homogenized in 25 ml of 100 mM phosphate buffer at pH 6.0. The protein concentration of the extract was determined while alpha‐glucosidase and alpha‐amylase inhibitory activities were determined by calculating the percentage inhibition and the inhibitory concentrations at 50 percent (IC50) of the crude extract. The values were compared to that of acarbose, a standard inhibitor and the mode of inhibition determined. The protein concentration of the extract was 5.256 ± 0.062 mg/ml with an IC50 value of 13.329 ±1.739 mg/ml for alpha‐glucosidase and 0.710±0.040 mg/ml for alpha‐amylase inhibitory activities while acarbose gave lower IC50 values of 0.620±0.090 mg/ml and 0.399±0.004 mg/ml for alpha‐glucosidase and alpha‐amylase respectively. The extract was noted to exhibit uncompetitive inhibition for both enzymes, mode of action attractive for drug design. It can therefore be concluded that the crude ant‐lion larvae extract could be more potent if purified and therapeutically exploited in the management of Diabetes mellitus or other related diseases.Support or Funding InformationSelf SupportedThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
SP101, a novel synthetic compound displays survivin suppression, apoptosis and tumor inhibition in both the EGFR‐wild type and ‐T790M of non‐small cell lung cancer
Authors: Chao Jui‐I 
Published: June 2021

Gefitinib is a tyrosine kinase inhibitor of epidermal growth factor receptor (EGFR‐TKI) which is currently used therapy in non‐small cell lung cancer (NSCLC) patients; however, drug resistance of cancer patients treatment is a critical problem. Here, we show a novel synthetic compound, dodecyl‐4‐(4‐(3‐(4‐(3‐chloro‐4‐fluorophenylamino)‐7‐methoxyquinazolin‐6‐yloxy)propyl)piperazin‐1‐yl)‐4‐oxobutanoate derivative from gefitinib, which is called SP101 that attenuated drug resistance in both the EGFR‐wild type and ‐T790M of non‐small cell lung cancer. SP101 inhibited EGFR kinase activity in the gefitinib‐resistant lung cancer cells. SP101 induced cancer cell death and apoptosis in both the A549 (EGFR‐wild type) and H1975 (EGFR‐T790M) lung cancer cells. SP101 inhibited anti‐apoptotic Survivin protein levels but conversely induced active caspase 3 proteins for apoptosis induction. The ectopic expression of survivin by a survivin‐expressed vector decreased the SP101‐induced cell death. The cell death was higher after treatment with SP101 than that gefitinib in the drug‐resistant H1975 cells. SP101 inhibited tumor growth and substantially reduced survivin protein but conversely elicited active caspase 3 in the xenograft human H1975 lung tumors of nude mice. Moreover, SP101 markedly inhibited the patient‐derived xenografted patient's lung tumors in nude mice. We demonstrate a novel compound SP101 that can reduce the drug resistance of the EGFR mutation at T790M as well as the EGFR wild type in lung cancer cells.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.