scholarly journals EXoO-Tn: Tag-n-Map the Tn Antigen in the Human Proteome

2019 ◽  
Author(s):  
Weiming Yang ◽  
Minghui Ao ◽  
Angellina Song ◽  
Yuanwei Xu ◽  
Hui Zhang
Keyword(s):  
Solid Tumors ◽  
Clinical Studies ◽  
Large Scale ◽  
Human Proteome ◽  
Jurkat Cells ◽  
Tn Antigen ◽  
Novel Technology ◽  
Glycosylation Sites ◽  
Unique Mass ◽  
New Vaccines

AbstractTn antigen (Tn), a single N-acetylgalactosamine (GalNAc) monosaccharide attached to protein Ser/Thr residues, is found on most solid tumors yet rarely detected in adult tissues, featuring it one of the most distinctive signatures of cancers. Although it is prevalent in cancers, Tn-glycosylation sites are not entirely clear owing to the lack of suitable technology. Knowing the Tn-glycosylation sites will spur the development of new vaccines, diagnostics, and therapeutics of cancers. Here, we report a novel technology named EXoO-Tn for large-scale mapping of Tn-glycosylation sites. EXoO-Tn utilizes glycosyltransferase C1GalT1 and isotopically-labeled UDP-Gal(13C6) to tag and convert Tn to Gal(13C6)-Tn, which has a unique mass being distinguishable to other glycans. This exquisite Gal(13C6)-Tn structure is recognized by OpeRATOR that specifically cleaves N-termini of the Gal(13C6)-Tn-occupied Ser/Thr residues to yield site-containing glycopeptides. The use of EXoO-Tn mapped 947 Tn-glycosylation sites from 480 glycoproteins in Jurkat cells. Given the importance of Tn in diseases, EXoO-Tn is anticipated to have broad utility in clinical studies.

Methods for the Quantification of Salivary Cortisol and of a-amylase in Biosensors and Portable Devices

2018 ◽  
Vol 68 (12) ◽  
pp. 2857-2859
Author(s):  
Cristina Mihaela Ghiciuc ◽  
Andreea Silvana Szalontay ◽  
Luminita Radulescu ◽  
Sebastian Cozma ◽  
Catalina Elena Lupusoru ◽  
...  
Keyword(s):  
Real Time ◽  
Medical Practice ◽  
Salivary Cortisol ◽  
Clinical Studies ◽  
Large Scale ◽  
Psychological Research ◽  
Portable Devices ◽  
Salivary Amylase ◽  
Specificity And Sensitivity

There is an increasing interest in the analysis of salivary biomarkers for medical practice. The objective of this article was to identify the specificity and sensitivity of quantification methods used in biosensors or portable devices for the determination of salivary cortisol and salivary a-amylase. There are no biosensors and portable devices for salivary amylase and cortisol that are used on a large scale in clinical studies. These devices would be useful in assessing more real-time psychological research in the future.

A large-scale targeted proteomics assay resource based on an in vitro human proteome

2016 ◽  
Vol 14 (3) ◽  
pp. 251-258 ◽  
Author(s):  
Masaki Matsumoto ◽  
Fumiko Matsuzaki ◽  
Kiyotaka Oshikawa ◽  
Naoki Goshima ◽  
Masatoshi Mori ◽  
...  
Keyword(s):  
Large Scale ◽  
Human Proteome ◽  
Targeted Proteomics

Electric Toothbrushes–For Whom are They Designed?

2002 ◽  
Vol 16 (1) ◽  
pp. 6-8 ◽  
Author(s):  
Sebastian Ciancio
Keyword(s):  
Clinical Studies ◽  
Large Scale ◽  
Economic Status ◽  
New Technology ◽  
Ease Of Use ◽  
Special Populations ◽  
Gingival Health

Powered toothbrushes were first introduced on a large scale in the early 1960s. However, because of a clear lack of superiority compared with manual brushes, and problems with mechanical breakdowns, their sales decreased significantly. However, recommendation for their use continued in special populations with dexterity and cognition problems. The 1990s ushered in an era of new technology, and studies began to suggest superiority of some powered brushes, particularly those using oscillating-rotating or counter-rotational actions. Some studies have shown interproximal cleansing abilities superior to those of manual brushes and yielding results similar to those achieved with the use of a manual brush and floss. Both controlled and open-labeled studies have suggested that electric brushes improve gingival health with patients who routinely used manual brushes prior to using these new powered brushes, and safety has been clearly established. In recommending powered toothbrushes, practitioners should familiarize themselves with the products available, with the clinical studies supporting their benefits compared with manual brushes, their safety and ease of use, and the patient's economic status.

Further clinical studies of combination chemotherapy using cyclophosphamide, vincristine, methotrexate and 5-flourouracil in solid tumors

1971 ◽  
Vol 261 (2) ◽  
pp. 73-78 ◽  
Author(s):  
CHARLES A. COLTMAN ◽  
GUILFORD M. DUDLEY ◽  
MONTAGUE LANE ◽  
JOHN J. COSTANZI ◽  
ARTHUR HAUT ◽  
...  
Keyword(s):  
Solid Tumors ◽  
Combination Chemotherapy ◽  
Clinical Studies

scholarly journals Reduction of Ambiguity in Phosphorylation-site Localization in Large-scale Phosphopeptide Profiling by Data Filter using Unique Mass Class Information

2014 ◽  
Vol 35 (3) ◽  
pp. 845-850 ◽  
Author(s):  
Inamul Hasan Madar ◽  
Seunghoon Back ◽  
Dong-Gi Mun ◽  
Hokeun Kim ◽  
Jae Hun Jung ◽  
...  
Keyword(s):  
Large Scale ◽  
Phosphorylation Site ◽  
Site Localization ◽  
Class Information ◽  
Unique Mass

scholarly journals Preclinical Activity of the MPS1 Inhibitor S81694 in Acute Lymphoblastic Leukemia (ALL)

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 2631-2631
Author(s):  
Anna Kaci ◽  
Emilie Adiceam ◽  
Melanie Dupont ◽  
Marine Garrido ◽  
Jeannig Berrou ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Line ◽  
Solid Tumors ◽  
Cell Lines ◽  
Small Molecule ◽  
Research Funding ◽  
Lymphoblastic Leukemia ◽  
Jurkat Cells ◽  
Cell Cycle Distribution ◽  
Monopolar Spindle

Introduction: The dual-specificity protein kinase, monopolar spindle 1 (Mps1) is one the main kinases of the spindle assembly checkpoint (SAC) critical for accurate segregation of sister chromatids during mitosis. A hallmark of cancer cells is chromosomal instability caused by deregulated cell cycle checkpoints and SAC dysfunction. Mps1 is known to be overexpressed in several solid tumors including triple negative breast cancer. Thus, Mps1 seems to be a promising target and small molecules targeting Mps1 entered clinical trials in solid tumors. ALL originates from malignant transformation of B-and T-lineage lymphoid precursors with a variety of genetic aberrations including chromosome translocations, mutations, and aneuploidies in genes responsible for cell cycle regulation and lymphoid cell development. While outcome is excellent for pediatric patients and younger adults, relapsed and refractory disease still remain a clinical challenge for elder patients. Here, we demonstrate for the first time preclinical efficacy of the small molecule Mps1 inhibitor (Mps1i) S81694 in T- and B- ALL cells including BCR-ABL1+-driven B-ALL. Materials and Methods: Expression of Mps1 was determined by RT-qPCR and WB in JURKAT, RS4-11 and BCR-ABL1+ cells (BV-173 and TOM-1). A small molecule Mps1i (S81694) was tested alone (0 to 1000nM) or in combination with imatinib, dasatinib, nilotinib and ponatinib in BCR-ABL1+ ALL cell lines. Cell viability and IC50 was assessed by MTS assays after exposure to Mps1i for 72h. In combination experiments, compounds were added simultaneously and relative cell numbers were determined at 72h with MTS assays and combination index (CI) values were calculated according to the Bliss model. Induction of apoptosis was evaluated by annexin-V exposure and PI incorporation at 72h with increasing doses of Mps1i. Cell-cycle distribution was determined by cytofluorometric analysis detecting nuclear propidium iodide (PI) intercalation at 48h. Phosphorylation of Mps1 was detected in synchronized (by nocodazole and MG-132) cells by immunofluorescence using an anti phospho-Mps1 antibody detecting Thr33/Ser37 residues. Time-lapse microscopy was used in cell lines in presence or absence of S81694 to determine mitosis duration. Bone marrow (BM) nucleated patient cells were obtained after informed consent and incubated in methylcellulose with cytokines with or without Mps1i for 2 weeks to determine colony growth. Results: Expression of Mps1 could be detected by RT-qPCR and at the protein level by WB in all cell lines (Figure 1A and B ). IC50 after Mps1i exposure alone was 126nM in JURKAT cells, 51nM in RS4-11 cells, 75nM in BV-173 cells and 83nM in TOM-1. Significant apoptosis as detected by phosphatidylserine exposure and PI incorporation in all cell lines with BCR-ABL1+ cell lines BV-173 and TOM-1 cells being the most sensitive (80% and 60% apoptotic cells respectively)(Figure 1C). Upon Mps1i exposure we observed targeted inhibition of Mps1 phosphorylation at Thr33/Ser37 residues indicating the specific on target effect of S81694 by inhibiting Mps1 autophosphorylation (Figure 1D and E). Cell cycle profile was generally lost after treatment with S81694 in all cell lines indicating aberrant 2n/4n distribution due to SAC abrogation (Figure 1F). Furthermore, we demonstrated that S81694 exposure accelerated significantly mitosis in BV-173 cell line from 36 minutes to 19 minutes indicating effective inhibition of SAC function (Figure 1G). Interestingly, S81694 induced significant apoptosis (70%) in the imatinib resistant BV173 cell line bearing the E255K-BCR-ABL1-mutation. Combination of S81694 with TKI imatinib, dasatinib and nilotinib (but not ponatinib) was strongly synergistic in BCR-ABL1+ cells (Figure 1H). Finally, we observed inhibition of colony formation in a patient with BCR-ABL1+ B-ALL after exposure to 100nM and 250nM S81694 (reduction of 85% and 100% respectively)(Figure 1I). Conclusion: Mps1i S81694 yields significant preclinical activity in T-and B-cell ALL including BCR-ABL1+ models. Interestingly S81694 was efficacious in a TKI resistant cell line. Disclosures Kaci: Institut de Recherches Internationales Servier (IRIS): Employment. Garrido:Institut de Recherches Internationales Servier (IRIS): Employment. Burbridge:Institut de Recherches Internationales Servier (IRIS): Employment. Dombret:AGIOS: Honoraria; CELGENE: Consultancy, Honoraria; Institut de Recherches Internationales Servier (IRIS): Research Funding. Braun:Institut de Recherches Internationales Servier (IRIS): Research Funding.

The ability to combine multiple mRNA antigens targeting multiple pathogens simultaneously, and the robust immune responses are confirmed in several clinical studies

2021 ◽  
Author(s):  
Moataz Dowaidar
Keyword(s):  
Immune Responses ◽  
Clinical Studies ◽  
Large Scale ◽  
Antigen Expression ◽  
Fine Tuning ◽  
Cold Chain ◽  
Disruptive Technology ◽  
Single Shot ◽  
Structural Components ◽  
Multiple Pathogens

In the last decade, great progress has been made on mRNA vaccines. MRNA vaccines that are well-tolerated and human immunogenic, stable and can be scaled up to hundreds of millions of doses have been produced with advancements in mRNA design, lipid nanoparticles (LNPs) composition and production techniques. The ability to combine multiple mRNA antigens in the same LNP, targeting multiple pathogens simultaneously, the lack of vector immunity, and the robust immune responses confirmed in several clinical studies make mRNA vaccines a disruptive technology that could change the development of vaccines in the coming years. Moreover, as mRNA was recently employed for large-scale vaccination applications, there is still plenty of room for refining and new advances.Ad-vector-based vaccines have also become promising immunization platforms. Ad vectors' structural components can be harnessed and modified for enhanced tropism, efficient transduction, and optimal antigen expression, and the structural components of Ad vaccine vectors can be harnessed and modified for enhanced tropism, effective transduction, and optimal antigen expression. Ad vectors can be readily created and mass-produced on a commercial basis, and their potency and stability make single-shot immunizations viable without using a frozen cold chain. Ad vectors' flexibility and promise for present and future vaccination applications is evidenced by their development against many illnesses.The use of biomaterials and engineering to improve vaccine delivery control has shown promise in boosting vaccination efficiency and fine-tuning the responses induced. Taken together, these vaccine science innovations have the potential to overcome many of the shortcomings in traditional vaccination technology, and they will almost probably play a crucial part in developing future known and novel disease vaccines.

scholarly journals High-Throughput Stool Metaproteomics: Method and Application to Human Specimens

mSystems ◽  
2020 ◽  
Vol 5 (3) ◽  
Author(s):  
Carlos G. Gonzalez ◽  
Hannah C. Wastyk ◽  
Madeline Topf ◽  
Christopher D. Gardner ◽  
Justin L. Sonnenburg ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
Clinical Studies ◽  
Large Scale ◽  
Dietary Intervention ◽  
Negative Impact ◽  
Mass Spectrometry Analysis ◽  
Careful Study ◽  
Stool Samples ◽  
Study Participants

ABSTRACT Stool-based proteomics is capable of significantly augmenting our understanding of host-gut microbe interactions. However, compared to competing technologies, such as metagenomics and 16S rRNA sequencing, it is underutilized due to its low throughput and the negative impact sample contaminants can have on highly sensitive mass spectrometry equipment. Here, we present a new stool proteomic processing pipeline that addresses these shortcomings in a highly reproducible and quantitative manner. Using this method, 290 samples from a dietary intervention study were processed in approximately 1.5 weeks, largely done by a single researcher. These data indicated a subtle but distinct monotonic increase in the number of significantly altered proteins between study participants on fiber- or fermented food-enriched diets. Lastly, we were able to classify study participants based on their diet-altered proteomic profiles and demonstrated that classification accuracies of up to 89% could be achieved by increasing the number of subjects considered. Taken together, this study represents the first high-throughput proteomic method for processing stool samples in a technically reproducible manner and has the potential to elevate stool-based proteomics as an essential tool for profiling host-gut microbiome interactions in a clinical setting. IMPORTANCE Widely available technologies based on DNA sequencing have been used to describe the kinds of microbes that might correlate with health and disease. However, mechanistic insights might be best achieved through careful study of the dynamic proteins at the interface between the foods we eat, our microbes, and ourselves. Mass spectrometry-based proteomics has the potential to revolutionize our understanding of this complex system, but its application to clinical studies has been hampered by low-throughput and laborious experimentation pipelines. In response, we developed SHT-Pro, the first high-throughput pipeline designed to rapidly handle large stool sample sets. With it, a single researcher can process over one hundred stool samples per week for mass spectrometry analysis, conservatively approximately 10× to 100× faster than previous methods, depending on whether isobaric labeling is used or not. Since SHT-Pro is fairly simple to implement using commercially available reagents, it should be easily adaptable to large-scale clinical studies.

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