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2022 ◽  
Vol 22 (1) ◽  
Author(s):  
Evelyn Rute Carneiro Maciel ◽  
Eduarda Helena Leandro Nascimento ◽  
Hugo Gaêta-Araujo ◽  
Maria Luiza dos Anjos Pontual ◽  
Andrea dos Anjos Pontual ◽  
...  

Abstract Background This study aimed to investigate the effect of automatic exposure compensation (AEC) of intraoral radiographic systems on the gray values of dental tissues in images acquired with or without high-density material in the exposed region using different exposure times and kilovoltages. The influence of the distance of the high-density material was also investigated. Methods Radiographs from the molar region of two mandibles were obtained using the RVG 6100 and the Express systems, operating at 60 and 70 kV and 0.06, 0.10, and 0.16 s. Subsequently, a titanium implant was inserted in the premolar’s socket and other images were acquired. Using the ImageJ software, two regions of interest were determined on the enamel, coronary dentine, root dentine, and pulp of the first and second molars to obtain their gray values. Results In the RVG 6100, the implant did not affect the gray values (p > 0.05); the increase in kV decreased it in all tissues (p < 0.05), and the exposure time affected only the root dentine and pulp. In the Express, only enamel and coronary dentine values changed (p < 0.05), decreasing with the implant presence and/or with the increase in exposure factors. The distance of the implant did not affect the results (p > 0.05). Conclusions AEC’s performance varies between the radiographic systems. Its effect on the gray values depends not only on the presence or absence of high-density material but also on the kV and exposure time used.


2021 ◽  
Author(s):  
Simon Ng ◽  
Alexander Brueckner ◽  
Soheila Bahmanjah ◽  
Qiaolin Deng ◽  
Jennifer Johnston ◽  
...  

STIP1 homology and U-Box containing protein 1 (STUB1) plays a key role in maintaining cell health during stress and aging. Recent evidence suggested STUB1 also helps regulate immunity with the potential of clearing malignant cells. Indeed, we and others have shown that STUB1 is a pivotal negative regulator of interferon gamma sensing – a process critical to the immunosurveillance of tumors and pathogens. Thus far, investigation of STUB1’s role relies mostly on genetic approaches as pharmacological inhibitors of this protein are lacking. Identification of a STUB1 tool compound is important as it would allow therapeutically relevant target validation in a broader sense. Accordingly, we leveraged phage display and computational modeling to identify and refine STUB1 binders. Screening of >10E9 macrocyclic peptides resulted in several conserved motifs as well as structurally diverse leads. Co-crystal structure of the peptide hit and STUB1 has enabled us to employ structure-based in silico design for further optimization. Of the modifications employed, replacing the hydrophilic solvent-exposed region of the macrocyclic peptides with a hydrophobic scaffold improved cellular permeability, while the binding conformation was maintained. Further substitution of the permeability-limiting terminal aspartic acid with a tetrazole bioisostere retained the binding to certain extent while improving permeability, suggesting a path forward. The current lead, although not optimal for cellular study, provides a valuable template for further development into selective tool compounds for STUB1 to enable target validation.


Author(s):  
Barbara Kubickova ◽  
Jörg A. Schenk ◽  
Franziska Ramm ◽  
Kornelija Markuškienė ◽  
Jochen Reetz ◽  
...  

Abstract To generate a hepatitis E virus (HEV) genotype 3 (HEV-3)–specific monoclonal antibody (mAb), the Escherichia coli–expressed carboxy-terminal part of its capsid protein was used to immunise BALB/c mice. The immunisation resulted in the induction of HEV-specific antibodies of high titre. The mAb G117-AA4 of IgG1 isotype was obtained showing a strong reactivity with the homologous E. coli, but also yeast-expressed capsid protein of HEV-3. The mAb strongly cross-reacted with ratHEV capsid protein derivatives produced in both expression systems and weaker with an E. coli–expressed batHEV capsid protein fragment. In addition, the mAb reacted with capsid protein derivatives of genotypes HEV-2 and HEV-4 and common vole hepatitis E virus (cvHEV), produced by the cell-free synthesis in Chinese hamster ovary (CHO) and Spodoptera frugiperda (Sf21) cell lysates. Western blot and line blot reactivity of the mAb with capsid protein derivatives of HEV-1 to HEV-4, cvHEV, ratHEV and batHEV suggested a linear epitope. Use of truncated derivatives of ratHEV capsid protein in ELISA, Western blot, and a Pepscan analysis allowed to map the epitope within a partially surface-exposed region with the amino acid sequence LYTSV. The mAb was also shown to bind to human patient–derived HEV-3 from infected cell culture and to hare HEV-3 and camel HEV-7 capsid proteins from transfected cells by immunofluorescence assay. The novel mAb may serve as a useful tool for further investigations on the pathogenesis of HEV infections and might be used for diagnostic purposes. Key points • The antibody showed cross-reactivity with capsid proteins of different hepeviruses. • The linear epitope of the antibody was mapped in a partially surface-exposed region. • The antibody detected native HEV-3 antigen in infected mammalian cells.


2021 ◽  
Vol 17 (3) ◽  
pp. e1009393
Author(s):  
Fei Yuan ◽  
Dandan Li ◽  
Changyao Li ◽  
Yanan Zhang ◽  
Hao Song ◽  
...  

Classical swine fever virus (CSFV) is an important pathogen in the swine industry. Virion attachment is mediated by envelope proteins Erns and E2, and E2 is indispensable. Using a pull-down assay with soluble E2 as the bait, we demonstrated that ADAM17, a disintegrin and metalloproteinase 17, is essential for CSFV entry. Loss of ADAM17 in a permissive cell line eliminated E2 binding and viral entry, but compensation with pig ADAM17 cDNA completely rescued these phenotypes. Similarly, ADAM17 silencing in primary porcine fibroblasts significantly impaired virus infection. In addition, human and mouse ADAM17, which is highly homologous to pig ADAM17, also mediated CSFV entry. The metalloproteinase domain of ADAM17 bound directly to E2 protein in a zinc-dependent manner. A surface exposed region within this domain was mapped and shown to be critical for CSFV entry. These findings clearly demonstrate that ADAM17 serves as an essential attachment factor for CSFV.


Micromachines ◽  
2020 ◽  
Vol 11 (8) ◽  
pp. 762
Author(s):  
Satoshi Yamaguchi ◽  
Yumi Takasaki ◽  
Shinya Yamahira ◽  
Teruyuki Nagamune

Photo-responsive cell attachment surfaces can simplify patterning and recovery of cells in microdevices for medicinal and pharmaceutical research. We developed a photo-responsive surface for controlling the attachment and release of adherent cells on a substrate under light-guidance. The surface comprises a poly(ethylene glycol) (PEG)-based photocleavable material that can conjugate with cell-adhesive peptides. Surface-bound peptides were released by photocleavage in the light-exposed region, where the cell attachment was subsequently suppressed by the exposed PEG. Simultaneously, cells selectively adhered to the peptide surface at the unexposed microscale region. After culture, the adhered and spread cells were released by exposure to a light with nontoxic dose level. Thus, the present surface can easily create both cell-adhesive and non-cell-adhesive regions on the substrate by single irradiation of the light pattern, and the adhered cells were selectively released from the light-exposed region on the cell micropattern without damage. This study shows that the photo-responsive surface can serve as a facile platform for the remote-control of patterning and recovery of adherent cells in microdevices.


2019 ◽  
Vol 7 (4) ◽  
pp. 696-697
Author(s):  
Massimiliano Scalvenzi ◽  
Alessia Villani ◽  
Caterina Mazzella ◽  
Gabriella Fabbrocini ◽  
Claudia Costa

Bowen's disease (BD), also known as squamous cell carcinoma in situ, is a type of non-melanocytic intraepidermal malignancy characterised by a slowly enlarging erythematous to pink, scaly patch or plaque with irregular and well-demarcated borders. These lesions are usually persistent and progressive; it has been estimated that in general population around 3% to 5% of Bowen's disease transform into invasive squamous cell carcinoma. This report describes our experience with cutaneous BD and assesses the differences found about age, sex and anatomical site. Bowen’s disease was seen more frequently in male patients rather than in female patients in contrast to what confirmed in literature - this difference is probably because being head-neck an exposed region, patients are more easily induced to autoexam and to consult the dermatologist.


Antibodies ◽  
2019 ◽  
Vol 8 (1) ◽  
pp. 15 ◽  
Author(s):  
Mohamed Alfaleh ◽  
Neetika Arora ◽  
Michael Yeh ◽  
Christopher de Bakker ◽  
Christopher Howard ◽  
...  

CD117 (c-Kit) is a tyrosine kinase receptor that is overexpressed in multiple dog tumors. There is 100% homology between the juxtamembrane domain of human and canine CD117, and many cancer-causing mutations occur in this region in both species. Thus, CD117 is an important target for cancer treatment in dogs and for comparative oncology studies. Currently, there is no monoclonal antibody (mAb) specifically designed to target the exposed region of canine CD117, although there exist some with species cross-reactivity. We panned a naïve phage display library to isolate antibodies against recombinant CD117 on whole cells. Several mAbs were isolated and were shown to bind recombinant canine CD117 at low- to sub-nanomolar affinity. Additionally, binding to native canine CD117 was confirmed by immunohistochemistry and by flow cytometry. Competitive binding assays also identified mAbs that competed with the CD117 receptor-specific ligand, the stem cell factor (SCF). These results show the ability of our cell-based biopanning strategy to isolate a panel of antibodies that have varied characteristics when used in different binding assays. These in vitro/ex vivo assessments suggest that some of the isolated mAbs might be promising candidates for targeting overexpressed CD117 in canine cancers for different useful applications.


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