We describe the first application of stapled peptides in human platelets. Bim BH3 stapled peptides are used to overcome the limitations of traditional methods and uncover a new role for Bim in ...platelet activation.
Platelets are blood cells with numerous crucial pathophysiological roles in hemostasis, cardiovascular thrombotic events and cancer metastasis. Platelet activation requires the engagement of intracellular signalling pathways that involve protein–protein interactions (PPIs). A better understanding of these pathways is therefore crucial for the development of selective anti-platelet drugs. New strategies for studying PPIs in human platelets are required to overcome limitations associated with conventional platelet research methods. For example, small molecule inhibitors can lack selectivity and are often difficult to design and synthesise. Additionally, development of transgenic animal models is costly and time-consuming and conventional recombinant techniques are ineffective due to the lack of a nucleus in platelets. Herein, we describe the generation of a library of novel, functionalised stapled peptides and their first application in the investigation of platelet PPIs. Moreover, the use of platelet-permeable stapled Bim BH3 peptides confirms the part of Bim in phosphatidyl-serine (PS) exposure and reveals a role for the Bim protein in platelet activatory processes. Our work demonstrates that functionalised stapled peptides are a complementary alternative to conventional platelet research methods, and could make a significant contribution to the understanding of platelet signalling pathways and hence to the development of anti-platelet drugs.
Infants with extremely low birth weights (≤1000 g) have low plasma and tissue concentrations of vitamin A,
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and vitamin A deficiency may predispose these infants to chronic lung disease.
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A ...meta-analysis
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of clinical trials of vitamin A supplementation for preterm infants
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revealed a 17 percent increase in the rate of survival without chronic lung disease, which approached statistical significance. The current relevance of these trials is limited by changes in the vitamin A concentration in infant formulas and by the use of antenatal glucocorticoid and postnatal surfactant therapy. Moreover, in a pilot study
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we found that many infants . . .
Placentas from gestational diabetes mellitus (GDM) patients undergo significant metabolic and immunologic adaptations due to hyperglycemia, which results in an exacerbated synthesis of ...proinflammatory cytokines and an increased risk for infections. Insulin or metformin are clinically indicated for the treatment of GDM; however, there is limited information about the immunomodulatory activity of these drugs in the human placenta, especially in the context of maternal infections. Our objective was to study the role of insulin and metformin in the placental inflammatory response and innate defense against common etiopathological agents of pregnancy bacterial infections, such as E. coli and S. agalactiae, in a hyperglycemic environment. Term placental explants were cultivated with glucose (10 and 50 mM), insulin (50-500 nM) or metformin (125-500 µM) for 48 h, and then they were challenged with live bacteria (1 × 10sup.5 CFU/mL). We evaluated the inflammatory cytokine secretion, beta defensins production, bacterial count and bacterial tissue invasiveness after 4-8 h of infection. Our results showed that a GDM-associated hyperglycemic environment induced an inflammatory response and a decreased beta defensins synthesis unable to restrain bacterial infection. Notably, both insulin and metformin exerted anti-inflammatory effects under hyperglycemic infectious and non-infectious scenarios. Moreover, both drugs fortified placental barrier defenses, resulting in reduced E. coli counts, as well as decreased S. agalactiae and E. coli invasiveness of placental villous trees. Remarkably, the double challenge of high glucose and infection provoked a pathogen-specific attenuated placental inflammatory response in the hyperglycemic condition, mainly denoted by reduced TNF-α and IL-6 secretion after S. agalactiae infection and by IL-1β after E. coli infection. Altogether, these results suggest that metabolically uncontrolled GDM mothers develop diverse immune placental alterations, which may help to explain their increased vulnerability to bacterial pathogens.
The original version of this Article contained an error in the spelling of the author Manuela Gago-Dominguez, which was incorrectly given as Manuela G. Dominguez. This has now been corrected in both ...the PDF and HTML versions of the Article.
Abstract
Fanconi anemia (FA), a model syndrome of genome instability, is caused by a deficiency in DNA interstrand crosslink (ICL) repair resulting in chromosome breakage. The FA repair pathway ...comprises at least 22 FANC proteins including BRCA1 and BRCA2 and protects against carcinogenic endogenous and exogenous aldehydes. Individuals with FA are hundreds to thousands-fold more likely to develop head and neck (HNSCC), esophageal and anogenital squamous cell carcinomas (SCCs) with a median onset age of 31 years. The aggressive nature of these tumors and poor patient tolerance of platinum and radiation-based therapy have been associated with short survival in FA. Molecular studies of SCCs from individuals with FA (FA SCCs) have been limited, and it is unclear how they relate to sporadic HNSCCs primarily driven by tobacco and alcohol exposure or human papillomavirus (HPV) infection. Here, by sequencing FA SCCs, we demonstrate that the primary genomic signature of FA-deficiency is the presence of a high number of structural variants (SVs). SVs are enriched for small deletions, unbalanced translocations, and fold-back inversions that arise in the context of TP53 loss. The SV breakpoints preferentially localize to early replicating regions, common fragile sites, tandem repeats, and SINE elements. SVs are often connected forming complex rearrangements. Resultant genomic instability underlies elevated copy number alteration (CNA) rates of key HNSCC-associated genes, including PIK3CA, MYC, CSMD1, PTPRD, YAP1, MXD4, and EGFR. In contrast to sporadic HNSCC, we find no evidence of HPV infection in FA HNSCC, although positive cases were identified in gynecologic tumors. A murine allograft model of FA pathway-deficient SCC was enriched in SVs, exhibited dramatic tumor growth advantage, more rapid epithelial-to-mesenchymal transition, and enhanced autonomous inflammatory signaling when compared to an FA pathway-proficient model. In light of the protective role of the FA pathway against SV formation uncovered here, and recent findings of FA pathway insufficiency in the setting of increased formaldehyde load resulting in hematopoietic stem cell failure and carcinogenesis, we propose that high copy-number instability in sporadic HNSCC may result from functional overload of the FA pathway by endogenous and exogenous DNA crosslinking agents. Our work lays the foundation for improved FA patient treatment and demonstrates that FA SCC is a powerful model to study tumorigenesis resulting from DNA crosslinking damage.
Citation Format: Andrew L. Webster, Mathijs A. Sanders, Krupa Patel, Ralf Dietrich, Raymond J. Noonan, Francis P. Lach, Ryan R. White, Audrey M. Goldfarb, Kevin Hadi, Matthew M. Edwards, Frank X. Donovan, Moonjung Jung, Sunandini Sridhar, Olivier Fedrigo, Huasong Tian, Joel Rosiene, Thomas Heineman, Jennifer Kennedy, Lorenzo Bean, Rasim O. Rosti, Rebecca Tryon, Ashlyn-Maree Gonzalez, Allana Rosenberg, Ji-Dung Luo, Thomas Carrol, Eunike Velleuer, Jeff C. Rastatter, Susanne I. Wells, Jordi Surrallés, Grover Bagby, Margaret L. MacMillan, John E. Wagner, Maria Cancio, Farid Boulad, Theresa Scognamiglio, Roger Vaughan, Amnon Koren, Marcin Imielinski, Settara Chandrasekharappa, Arleen D. Auerbach, Bhuvanesh Singh, David Kutler, Peter J. Campbell, Agata Smogorzewska. Fanconi anemia pathway deficiency drives copy number variation in squamous cell carcinoma abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6196.
The Mouse Lymphoma Assay (MLA) Workgroup of the International Workshop on Genotoxicity Testing (IWGT), comprised of experts from Japan, Europe, and the United States, met on August 29, 2003, in ...Aberdeen, Scotland, United Kingdom. This meeting of the MLA Workgroup was devoted to reaching a consensus on the appropriate approach to data evaluation and on acceptance criteria for both the positive and negative/vehicle controls. The Workgroup reached consensus on the acceptance criteria for both the agar and microwell versions of the MLA. Recommendations include acceptable ranges for mutant frequency, cloning efficiency, and suspension growth of the negative/vehicle controls and on criteria to define an acceptable positive control response. The recommendation for the determination of a positive/negative test chemical response includes both the requirement that the response exceeds a defined value the global evaluation factor (GEF) and that there also be a positive dose–response (evaluated by an appropriate statistical method). Environ. Mol. Mutagen., 2006. Published 2005 Wiley‐Liss, Inc.
The purpose of this study was to investigate trunk and knee strength in practitioners of hard-style martial arts. An additional objective was to examine reaction times in these participants by ...measuring simple reaction times (SRT), choice reaction times (CRT) and movement times (MT). Thirteen high-level martial artists and twelve sedentary participants were tested under isokinetic and isometric conditions on an isokinetic dynamometer. Response and movement times were also measured in response to simple and choice auditory cues. Results indicated that the martial arts group generated a greater body-weight adjusted peak torque with both legs at all speeds during isokinetic extension and flexion, and in isometric extension but not flexion. In isokinetic and isometric trunk flexion and extension, martial artists tended to have higher peak torques than controls, but they were not significantly different (p > 0.05). During the SRT and CRT tasks the martial artists were no quicker in lifting their hand off a button in response to the stimulus reaction time (RT) but were significantly faster in moving to press another button movement time (MT). In conclusion, the results reveal that training in a martial art increases the strength of both the flexors and extensors of the leg. Furthermore, they have faster movement times to auditory stimuli. These results are consistent with the physical aspects of the martial arts. Key PointsMartial artists undertaking hard-style martial arts have greater strength in their knee flexor and extensor muscles as tested under isokinetic testing. Under isometric testing conditions they have stronger knee extensors only.The trunk musculature is generally higher under both conditions of testing in the martial artists, although not significantly.The total reaction times of the martial artists to an auditory stimulus were significantly faster than the control participants. When analysed further it was revealed that the decrease in reaction time was due to the movement time component of the total reaction time.The training involved for the practice of the hard-style martial arts increases the strength of muscles involved in kicking. This increased strength is not seen in the trunk muscles. Furthermore, martial artists have a faster response time; the cause of which appears to be only the faster movement time.
En esta investigación bibliográfica se observa una gran variedad de especies responsables de precipitar a cuatro metales de interés (Cu, Pb, Zn y Fe). En la mayoría de las investigaciones no ...solamente precipitan estos metales, sino que otros elementos que están presentes en cada investigación. Los artículos aquí mencionados tienen una relación directa con el efluente proveniente de la operación unitaria de flotación. Aportan conocimiento acerca del proceso de sulfato-reducción, comprendiendo el mecanismo mediante microorganismos con características específicas, especialmente su versatilidad pues se desarrollan en diferentes ecosistemas. Se observa que varias especies, como Desulfobacter o Desulfovibrio son comunes pues tienen condiciones relativamente sencillas para desarrollarse. Los MSR son eficientes para reducir la acidez del agua (de la operación unitaria de flotación de una mina, de cocinas, de corrientes marinas, etc.). También lo son para precipitar diferentes elementos pues no requieren de algún agente externo salvo en contadas ocasiones donde debe actuar un catalizador. Hay investigaciones sobre nutrientes que deben adicionarse para incrementar su actividad. Los artículos revisados reconocieron las variables a controlar para obtener buenos resultados en la remoción de metales y su impacto en el ambiente. Es de vital importancia el desarrollo de proyectos que tomen un sistema natural como la degradación anaerobia para alcanzar un punto en el cual la tecnología y el ambiente puedan convivir logrando bienes de consumo sin causar daños irreparables a la naturaleza.
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