scholarly journals Development and Characterisation of Antibody-Based Optical Imaging Probes for Inflammatory Bowel Disease

2021 ◽  
Vol 14 (9) ◽  
pp. 922
Author(s):  
Matthijs David Linssen ◽  
Wouter Tjerk Rudolph Hooghiemstra ◽  
Annelies Jorritsma-Smit ◽  
Derk Pieter Allersma ◽  
Gerard Dijkstra ◽  
...  
Keyword(s):  
Drug Distribution ◽  
Production Methods ◽  
Sds Page ◽  
Medicinal Treatment ◽  
Imaging Probes ◽  
Treatment Paradigm ◽  
Important Addition ◽  
Inflammatory Bowel ◽  
High Degree ◽  
Further Development

Monoclonal antibodies are an important addition to the medicinal treatment paradigm for IBD patients. While effective, these agents show a high degree of primary and secondary non-response, and methods to predict response are highly desired. Information on drug distribution at the target level is often lacking. Fluorescent endoscopic imaging using labelled antibody drugs may provide insight regarding drug distribution, target engagement and drug response, but these assessments require stable and functional fluorescently-conjugated probes. Infliximab, vedolizumab, adalimumab and ustekinumab were conjugated to IRDye 800CW, IRDye 680LT and ZW800-1. The resulting 12 tracer candidates were analysed and characterised on SE-HPLC, SDS-PAGE, iso-electric focussing (IEF) and ELISA in order to evaluate their feasibility as candidate clinical tracers for cGMP development. Major differences in the conjugation results could be seen for each conjugated drug. For Infliximab, 2 conjugates (800CW and 680LT) showed formation of aggregates, while conjugates of all drugs with ZW800-1 showed reduced fluorescent brightness, reduced purification yield and formation of fragments. All 6 of these candidates were considered unfeasible. From the remaining 6, ustekinumab-680LT showed reduced binding to IL23, and was therefore considered unfeasible. Out of 12 potential tracer candidates, 5 were considered feasible for further development: vedolizumab-800CW, vedolizumab-680LT, adalimumab-800CW, adalimumab-680LT and ustekinumab-800CW. Infliximab-680LT and ustekinumab-680LT failed to meet the standards for this panel, but may be rendered feasible if tracer production methods were further optimized.

METHODS FOR RESTORING WOLVES OF ROLLING PRODUCTION

2020 ◽  
Vol 4 (141) ◽  
pp. 114-122
Author(s):  
DAR’YA LEBEDEVA ◽  
◽  
ANNA KARPUNICHEVA
Keyword(s):  
Thermal Effects ◽  
Machine Building ◽  
Research Purpose ◽  
Technical Literature ◽  
Production Methods ◽  
Advantages And Disadvantages ◽  
Rolling Production ◽  
Machine Parts ◽  
The Stability ◽  
High Degree

Large forces and significant thermal effects are created on the rolls when rolling sheets. The higher the stability of the rolls, the less downtime during their rerolling and higher productivity. (Research purpose) The research purpose is in analyzing the ways of restoring rolls and choose the most appropriate method for restoring these parts. (Materials and methods) The article presents the analysis of the scientific and technical literature on the topic of rolling production, methods for restoring large-sized machine parts of machine-building and metallurgical industries that work in difficult conditions and are subject to a high degree of wear. Authors try to solve the problem by means of comparative and logical analysis based on theoretical and empirical methods of scientific research. (Results and discussion) The article presents two groups of methods for restoring rolled rolls: banding and surfacing the working layer of the roll. Authors have analyzed each method in terms of technology, equipment, and feasibility. The article presents the advantages and disadvantages of the methods under consideration. (Conclusions) The most acceptable way to restore parts with a high degree of wear is surfacing. It is most efficient to apply submerged surfacing using an additional hot additive. Such surfacing, despite some complication of the equipment design, allows to deposit the metal on the roll with low heat input and in most cases in one pass. Surfacing using an additional hot additive allows to increase the productivity of the process by up to 250 percent while reducing the penetration depth by 2-3 times and saving energy by up to 40 percent.

scholarly journals Applications of Additively Manufactured Tools in Abrasive Machining—A Literature Review

Materials ◽  
2021 ◽  
Vol 14 (5) ◽  
pp. 1318
Author(s):  
Mariusz Deja ◽  
Dawid Zieliński ◽  
Aini Zuhra Abdul Kadir ◽  
Siti Nur Humaira
Keyword(s):  
Rapid Tooling ◽  
Rapid Manufacturing ◽  
Abrasive Machining ◽  
Production Methods ◽  
Industrial Environment ◽  
Hard And Brittle Materials ◽  
3D Printed ◽  
Abrasive Tools ◽  
Machine Parts ◽  
Further Development

High requirements imposed by the competitive industrial environment determine the development directions of applied manufacturing methods. 3D printing technology, also known as additive manufacturing (AM), currently being one of the most dynamically developing production methods, is increasingly used in many different areas of industry. Nowadays, apart from the possibility of making prototypes of future products, AM is also used to produce fully functional machine parts, which is known as Rapid Manufacturing and also Rapid Tooling. Rapid Manufacturing refers to the ability of the software automation to rapidly accelerate the manufacturing process, while Rapid Tooling means that a tool is involved in order to accelerate the process. Abrasive processes are widely used in many industries, especially for machining hard and brittle materials such as advanced ceramics. This paper presents a review on advances and trends in contemporary abrasive machining related to the application of innovative 3D printed abrasive tools. Examples of abrasive tools made with the use of currently leading AM methods and their impact on the obtained machining results were indicated. The analyzed research works indicate the great potential and usefulness of the new constructions of the abrasive tools made by incremental technologies. Furthermore, the potential and limitations of currently used 3D printed abrasive tools, as well as the directions of their further development are indicated.

scholarly journals Enhanced Cognitive Walkthrough: Development of the Cognitive Walkthrough Method to Better Predict, Identify, and Present Usability Problems

2013 ◽  
Vol 2013 ◽  
pp. 1-17 ◽  
Author(s):  
Lars-Ola Bligård ◽  
Anna-Lisa Osvalder
Keyword(s):  
Medical Equipment ◽  
Development Process ◽  
Product Development Process ◽  
Infusion Pumps ◽  
Usability Problems ◽  
Cognitive Walkthrough ◽  
High Level ◽  
High Degree ◽  
Further Development ◽  
Potential Use

To avoid use errors when handling medical equipment, it is important to develop products with a high degree of usability. This can be achieved by performing usability evaluations in the product development process to detect and mitigate potential usability problems. A commonly used method is cognitive walkthrough (CW), but this method shows three weaknesses: poor high-level perspective, insufficient categorisation of detected usability problems, and difficulties in overviewing the analytical results. This paper presents a further development of CW with the aim of overcoming its weaknesses. The new method is called enhanced cognitive walkthrough (ECW). ECW is a proactive analytical method for analysis of potential usability problems. The ECW method has been employed to evaluate user interface designs of medical equipment such as home-care ventilators, infusion pumps, dialysis machines, and insulin pumps. The method has proved capable of identifying several potential use problems in designs.

High Degree of Practice Variability in Colonic Dysplasia Surveillance for Patients With Inflammatory Bowel Disease

2022 ◽  
Author(s):  
James D Lewis ◽  
Raymond K Cross ◽  
Millie Long ◽  
Siddharth Singh ◽  
Hans Herfarth ◽  
...  
Keyword(s):  
Inflammatory Bowel Disease ◽  
Bowel Disease ◽  
Practice Patterns ◽  
High Volume ◽  
Practice Variability ◽  
Inflammatory Bowel ◽  
Surveillance Practice ◽  
Virtual Chromoendoscopy ◽  
High Degree

Lay Summary Dysplasia surveillance practice varies widely among high-volume inflammatory bowel disease providers. We surveyed high-volume inflammatory bowel disease providers about practice patterns to detect dysplasia. Regular use of dye-based chromoendoscopy was reported by 20%, virtual chromoendoscopy by 27%, and random biopsies by 58%.

scholarly journals Analysis of a rare recombination event within the multigenic Hor 2 locus of barley (Hordeum vulgare L.)

1990 ◽  
Vol 55 (3) ◽  
pp. 171-176 ◽  
Author(s):  
Peter R. Shewry ◽  
Saroj Parmar ◽  
Julian Franklin ◽  
Shirley R. Burgess
Keyword(s):  
Hordeum Vulgare ◽  
Recombination Event ◽  
Rflp Analysis ◽  
Genomic Clone ◽  
Class I ◽  
Class Iii ◽  
Hordeum Vulgare L ◽  
Ancestral Gene ◽  
Sds Page ◽  
High Degree

SummaryA rare recombinant within the multigenic Hor 2 locus of barley was detected by SD-PAGE of hordein fractions from F2 grain from the cross Bomi × P12/3. Analysis of a homozygous F4 line by 2-D IEF/SDS-PAGE showed that recombination between the class I/II and class III subfamilies of genes had occurred, indicating that they are spatially separate within the Hor 2 locus. RFLP analysis using a B hordein-related cDNA clone confirmed that recombination had occurred, while similar analysis using a genomic clone related to γ-type hordein (encoded by the Hor 5/HrdF locus) indicated the order of the two subfamilies of genes on the short arm of chromosome 5, the class I/II genes being closer to the centromere. The results are consistent with the origin of the B hordein gene family from initial duplication of a single ancestral gene to give two genes which diverged to become the ancestors of the class I/II and class III subfamilies. Subsequent cycles of duplication and divergence have resulted in the present high degree of polymorphism.

scholarly journals Phosphatase Shp2 exacerbates intestinal inflammation by disrupting macrophage responsiveness to interleukin-10

2019 ◽  
Vol 216 (2) ◽  
pp. 337-349 ◽  
Author(s):  
Peng Xiao ◽  
Huilun Zhang ◽  
Yu Zhang ◽  
Mingzhu Zheng ◽  
Rongbei Liu ◽  
...  
Keyword(s):  
Intestinal Inflammation ◽  
Interleukin 10 ◽  
Blood Monocytes ◽  
Stat3 Signaling ◽  
Tnf Α ◽  
Mouse Macrophages ◽  
Inflammatory Bowel ◽  
Anti Inflammatory ◽  
Further Development

Inflammatory cytokines produced by activated macrophages largely contribute to the pathological signs of inflammatory bowel disease (IBD). Interleukin-10 (IL-10) is the predominant anti-inflammatory cytokine in the intestine, and its therapeutic efficacy for IBD has been clinically tested. Nevertheless, how the function of IL-10 is regulated in the intestinal microenvironment remains unknown, which largely hinders the further development of IL-10–based therapeutic strategies. Here, we found that the expression of phosphatase Shp2 was increased in colonic macrophages and blood monocytes from IBD patients compared with those from healthy controls. Shp2 deficiency in macrophages protects mice from colitis and colitis-driven colon cancer. Mechanistically, Shp2 disrupts IL-10–STAT3 signaling and its dependent anti-inflammatory response in human and mouse macrophages. Furthermore, a Shp2-inducing role of TNF-α is unveiled in our study. Collectively, our work identifies Shp2 as a detrimental factor for intestinal immune homeostasis and hopefully will be helpful in the future exploitation of IL-10 immunotherapy for IBD.

scholarly journals Characterization and purification of a novel dATP-binding protein in eukaryotes

1992 ◽  
Vol 287 (3) ◽  
pp. 871-879 ◽  
Author(s):  
A Dalton ◽  
D P Hornby ◽  
S P Langston ◽  
G M Blackburn
Keyword(s):  
Binding Protein ◽  
Active Form ◽  
Direct Interaction ◽  
Nuclear Fraction ◽  
Yeast Saccharomyces Cerevisiae ◽  
Naturally Occurring ◽  
Sds Page ◽  
Marked Preference ◽  
High Degree ◽  
Page Electrophoresis

We characterized and purified an acidic dATP-binding protein, which, in its active form, resides in the nuclear fraction of a range of cells from mammals (including pig liver) and baker's yeast (Saccharomyces cerevisiae). This protein exhibits a high degree of specificity for the deoxy form of the naturally occurring nucleoside triphosphates and shows a marked preference for the purine deoxynucleoside triphosphates dATP and dGTP. The protein cleaves the terminal phosphate of dATP and appears to retain the dADP moiety of the nucleotide in a reaction that is resistant to both SDS and 8 M-urea. Fractionation of the nuclear preparation followed by non-denaturing PAGE and SDS/PAGE electrophoresis was sufficient to produce pure protein. The occurrence of this activity in all nuclei tested suggests that it plays an important role in nuclear metabolism. The specificity of the enzyme for deoxynucleoside triphosphates further suggests a role for this enzyme in DNA replication or repair, but the acidity of the protein argues against a direct interaction with DNA, and, indeed, the catalytic activity is not modulated by the inclusion of DNA in a variety of physical forms.

Nasal Endoscopy and the Definition and Diagnosis of Chronic Rhinosinusitis

2002 ◽  
Vol 126 (6) ◽  
pp. 623-627 ◽  
Author(s):  
James A. Stankiewicz ◽  
James M. Chow
Keyword(s):  
Computed Tomography ◽  
Chronic Rhinosinusitis ◽  
Sensitivity And Specificity ◽  
Nasal Polyps ◽  
Ct Scanning ◽  
Current Treatment ◽  
Current Definition ◽  
Treatment Paradigm ◽  
Definition Of ◽  
High Degree

OBJECTIVES: Although endoscopy has been shown by a few authors to be a valuable tool for the diagnosis of chronic rhinosinusitis, its true role in the evaluation of the patient with chronic rhinosinusitis has not been elucidated. The current definition of chronic rhinosinusitis is a symptom-based definition, and objective testing such as endoscopy or computed tomography (CT) is not included. However, the current treatment paradigm for chronic rhinosinusitis is dependent on the definition for diagnosis. Patients are treated with 4 weeks of antibiotics and decongestant/antihistamines/steroids based on the definition. This study aims to evaluate in a prospective fashion the place of endoscopy in the diagnosis of chronic rhinosinusitis. STUDY DESIGN: A group of 78 patients meeting the definition of chronic rhinosinusitis were subjected to same-day endoscopy and CT scanning. RESULTS: Seventeen (22%) of 78 patients had positive endoscopic and CT results. There were 20 (26%) of 78 patients with negative endoscopic and positive CT results. Six (8%) patients had positive endoscopic and negative CT results, and 35 (45%) had negative endoscopic and negative CT results. Overall, 37 (47%) patients had positive CT results, and 41 (53%) patients had negative CT results. Endoscopy showing the presence of purulence, nasal polyps, or watery congested mucosa correlated well with CT results. Negative endoscopy correlated with CT results in 65% of patients. CONCLUSION: The use of endoscopy to corroborate the diagnosis in nonpolypoid or nonpurulent rhinosinusitis in previously unoperated patients is questioned. Patients who meet the subjective definition of chronic rhinosinusitis should have a high degree of sensitivity and specificity with endoscopy or CT. The fact this is not the case questions the accuracy of the definition and the treatment paradigm. SIGNIFICANCE: According to this study, positive endoscopic results correlated well with CT, and negative endoscopic results correlated in 71% of patients with negative CT results.

scholarly journals Activation of a phospholipase Cβ2 deletion mutant by limited proteolysis

1998 ◽  
Vol 330 (1) ◽  
pp. 461-468 ◽  
Author(s):  
Petra SCHNABEL ◽  
Montserrat CAMPS
Keyword(s):  
Deletion Mutant ◽  
Limited Proteolysis ◽  
Gel Filtration ◽  
Cell System ◽  
Heterotrimeric G Proteins ◽  
Sds Page ◽  
Catalytically Active ◽  
Heterodimeric Complex ◽  
High Degree ◽  
Hydrolysis Of

All phosphoinositide-specific phospholipases C (PLC) identified until today exhibit a high degree of similarity within two regions of 170 and 260 residues, respectively, which are designated regions X and Y. The PLCβ family, including four members designated PLCβ1, PLCβ2, PLCβ3 and PLCβ4, is regulated by heterotrimeric G proteins. In order to investigate structure-function relationships of PLCβ2, we expressed PLCβ2Δ, a deletion mutant of PLCβ2 which lacks most of the sequence downstream of region Y, in the baculovirus/insect cell system. The mutant was present in both soluble and particulate fractions of Sf9 cells and was demonstrated to be catalytically active and sensitive to βγ-subunits. Sf9 cytosol containing this mutant was subjected to limited proteolysis by trypsin and S. aureus protease V8, respectively. Immunochemical analysis revealed that both proteases cleaved the enzyme between the regions X and Y. Most interestingly, proteolytic cleavage at this site by both proteases stimulated the catalytic activity of PLC2β2Δ. The proteolytically activated enzyme was still sensitive to βγ-subunits and showed an unchanged concentration dependence on Ca2+. Gel filtration chromatography indicated that the fragments generated by cleavage between the regions X and Y were still connected and formed a functional heterodimeric complex. In order to visualize all fragments generated by protease V8, PLCβ2Δ was purified to homogeneity from Sf9 cytosol. Limited proteolysis of the purified enzyme by S. aureus protease V8 and subsequent SDS/PAGE and silver staining revealed that several cuts take place between the regions X and Y and that the resulting fragments remain intact. We hypothesize that the activating proteolytic cut induces a conformational change of the enzyme which might facilitate hydrolysis of the phospholipid substrate.

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