scholarly journals Repurposing ciclopirox as a pharmacological chaperone in a model of congenital erythropoietic porphyria

2018 ◽  
Vol 10 (459) ◽  
pp. eaat7467 ◽  
Author(s):  
Pedro Urquiza ◽  
Ana Laín ◽  
Arantza Sanz-Parra ◽  
Jorge Moreno ◽  
Ganeko Bernardo-Seisdedos ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
Ex Vivo ◽  
The Body ◽  
Missense Mutations ◽  
Allosteric Site ◽  
Cutaneous Lesions ◽  
Pharmacological Chaperone ◽  
Genetic Mouse Model ◽  
Congenital Erythropoietic Porphyria

Congenital erythropoietic porphyria is a rare autosomal recessive disease produced by deficient activity of uroporphyrinogen III synthase, the fourth enzyme in the heme biosynthetic pathway. The disease affects many organs, can be life-threatening, and currently lacks curative treatments. Inherited mutations most commonly reduce the enzyme’s stability, altering its homeostasis and ultimately blunting intracellular heme production. This results in uroporphyrin by-product accumulation in the body, aggravating associated pathological symptoms such as skin photosensitivity and disfiguring phototoxic cutaneous lesions. We demonstrated that the synthetic marketed antifungal ciclopirox binds to the enzyme, stabilizing it. Ciclopirox targeted the enzyme at an allosteric site distant from the active center and did not affect the enzyme’s catalytic role. The drug restored enzymatic activity in vitro and ex vivo and was able to alleviate most clinical symptoms of congenital erythropoietic porphyria in a genetic mouse model of the disease at subtoxic concentrations. Our findings establish a possible line of therapeutic intervention against congenital erythropoietic porphyria, which is potentially applicable to most of deleterious missense mutations causing this devastating disease.

The Role of Polyphenols in the Modulation of Plasma Non-Enzymatic Antioxidant Capacity (NEAC)

2012 ◽  
Vol 82 (3) ◽  
pp. 228-232 ◽  
Author(s):  
Mauro Serafini ◽  
Giuseppa Morabito
Keyword(s):  
Antioxidant Capacity ◽  
Dietary Intervention ◽  
Ex Vivo ◽  
The Body ◽  
Dietary Polyphenols ◽  
Enzymatic Antioxidant ◽  
Endogenous Antioxidant

Dietary polyphenols have been shown to scavenge free radicals, modulating cellular redox transcription factors in different in vitro and ex vivo models. Dietary intervention studies have shown that consumption of plant foods modulates plasma Non-Enzymatic Antioxidant Capacity (NEAC), a biomarker of the endogenous antioxidant network, in human subjects. However, the identification of the molecules responsible for this effect are yet to be obtained and evidences of an antioxidant in vivo action of polyphenols are conflicting. There is a clear discrepancy between polyphenols (PP) concentration in body fluids and the extent of increase of plasma NEAC. The low degree of absorption and the extensive metabolism of PP within the body have raised questions about their contribution to the endogenous antioxidant network. This work will discuss the role of polyphenols from galenic preparation, food extracts, and selected dietary sources as modulators of plasma NEAC in humans.

scholarly journals Current Challenges in Volatile Organic Compounds Analysis as Potential Biomarkers of Cancer

2015 ◽  
Vol 2015 ◽  
pp. 1-16 ◽  
Author(s):  
Kamila Schmidt ◽  
Ian Podmore
Keyword(s):  
Volatile Organic Compounds ◽  
Organic Compounds ◽  
Clinical Symptoms ◽  
Ex Vivo ◽  
Critical Evaluation ◽  
Detection Techniques ◽  
Volatile Organic ◽  
Potential Biomarkers

An early diagnosis and appropriate treatment are crucial in reducing mortality among people suffering from cancer. There is a lack of characteristic early clinical symptoms in most forms of cancer, which highlights the importance of investigating new methods for its early detection. One of the most promising methods is the analysis of volatile organic compounds (VOCs). VOCs are a diverse group of carbon-based chemicals that are present in exhaled breath and biofluids and may be collected from the headspace of these matrices. Different patterns of VOCs have been correlated with various diseases, cancer among them. Studies have also shown that cancer cells in vitro produce or consume specific VOCs that can serve as potential biomarkers that differentiate them from noncancerous cells. This review identifies the current challenges in the investigation of VOCs as potential cancer biomarkers, by the critical evaluation of available matrices for the in vivo and in vitro approaches in this field and by comparison of the main extraction and detection techniques that have been applied to date in this area of study. It also summarises complementary in vivo, ex vivo, and in vitro studies conducted to date in order to try to identify volatile biomarkers of cancer.

scholarly journals Extracellular vesicles engineered with valency-controlled DNA nanostructures deliver CRISPR/Cas9 system for gene therapy

2020 ◽  
Vol 48 (16) ◽  
pp. 8870-8882 ◽  
Author(s):  
Jialang Zhuang ◽  
Jizhou Tan ◽  
Chenglin Wu ◽  
Jie Zhang ◽  
Ting Liu ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Ex Vivo ◽  
Primary Liver Cancer ◽  
The Body ◽  
Dna Aptamer ◽  
Great Promise ◽  
Tumor Growth Inhibition ◽  
Specific Cell ◽  
Dna Nanostructures

Abstract Extracellular vesicles (EVs) hold great promise for transporting CRISPR–Cas9 RNA-guided endonucleases (RNP) throughout the body. However, the cell-selective delivery of EVs is still a challenge. Here, we designed valency-controlled tetrahedral DNA nanostructures (TDNs) conjugated with DNA aptamer, and loaded the valency-controlled TDNs on EV surface via cholesterol anchoring for specific cell targeting. The targeting efficacy of different ratios of aptamer/cholesterol from 1:3 to 3:1 in TDNs on decorating EVs was investigated. TDNs with one aptamer and three cholesterol anchors (TDN1) efficiently facilitated the tumor-specific accumulation of the EVs in cultured HepG2 cells and human primary liver cancer-derived organoids, as well as xenograft tumor models. The intracellular delivery of RNP by TDN1-EVs successfully realized its subsequent genome editing, leading to the downregulation of GFP or WNT10B in specific cells. This system was ultimately applied to reduce the protein expression of WNT10B, which presented remarkable tumor growth inhibition in vitro, ex vivo and in vivo, and could be extended to other therapeutic targets. The present study provides a platform for the directional display of aptamer on surface labeling and the EVs-based Cas9 delivery, which provides a meaningful idea for future cell-selective gene editing.

scholarly journals Platelet generation from circulating megakaryocytes is triggered in the lung vasculature

2021 ◽  
Author(s):  
Xiaojuan Zhao ◽  
Dominic Alibhai ◽  
Tony G. Walsh ◽  
Nathalie Tarassova ◽  
Semra Z. Birol ◽  
...  
Keyword(s):  
Blood Cells ◽  
Ex Vivo ◽  
Critical Role ◽  
The Body ◽  
Mouse Lung ◽  
Generation Process ◽  
Large Size ◽  
Large Numbers ◽  
Microfluidic Chamber

Platelets, small hemostatic blood cells, are derived from megakaryocytes, although the generation process is not clear. Only small numbers of platelets have been produced in systems outside the body, where bone marrow and lung are proposed as sites of platelet generation. Here we show that perfusion of megakaryocytes ex vivo through the mouse lung vasculature generates very large numbers of platelets, up to 3,000 per megakaryocyte. Despite their large size, megakaryocytes were able repeatedly to passage through the lung vasculature, leading to enucleation and fragmentation to generate platelets intravascularly. Using the ex vivo lung and a novel in vitro microfluidic chamber we determined the contributions of oxygenation, ventilation and endothelial cell health to platelet generation, and showed a critical role for the actin regulator TPM4.

PIR-B expressing CD8+ T cells exhibit features of Tc1 and Tc17 in SKG mice

Rheumatology ◽  
2019 ◽  
Vol 58 (12) ◽  
pp. 2325-2329 ◽  
Author(s):  
Kathrin Rothe ◽  
Dagmar Quandt ◽  
Gabriele Köhler ◽  
Simon Jasinski-Bergner ◽  
Barbara Seliger ◽  
...  
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Clinical Symptoms ◽  
Ex Vivo ◽  
Peripheral Tolerance ◽  
Autoimmune Arthritis ◽  
Specific Pathogen ◽  
Inflammatory Phenotype

Abstract Objective In autoimmune arthritis, TCR signalling is attenuated by peripheral tolerance mechanisms. We have described previously a population of inhibitory receptor LIR-1 expressing autoreactive CD8+ T cells in rheumatoid arthritis. Here, we investigated the role of CD8+ T cells in murine autoimmune arthritis by analysing their expression of the mouse orthologue of LIR-1, PIR-B. Methods Frequencies of PIR-B+CD8+ T cells were determined in the SKG arthritis model. The phenotype of those cells was determined ex vivo by FACS and functionality was investigated by means of cytokine production and cytolytic potential upon activation in vitro. Results SKG mice, under non-SPF (specific pathogen-free) conditions with clinical symptoms of arthritis, were found to harbour significantly increased frequencies of PIR-B+CD8+ T cells. Those cells showed a pro-inflammatory phenotype with preferential production of IL-17 and IFN-γ. The frequency of those cells correlated inversely with the arthritis score, indicating that they might represent autoreactive, but functionally inhibited, CD8+ T cells. Conclusion PIR-B+CD8+ T cells from SKG mice show a cytotoxic and pro-inflammatory phenotype. Inhibition of CD8+ T cell autoreactivity by PIR-B/LIR-1 receptor signalling might be a counter-regulatory mechanism to curb autoreactivity and arthritis.

scholarly journals Iodine-124 PET quantification of organ-specific delivery and expression of NIS-encoding RNA

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Matthias Miederer ◽  
Stefanie Pektor ◽  
Isabelle Miederer ◽  
Nicole Bausbacher ◽  
Isabell Sofia Keil ◽  
...  
Keyword(s):  
Cell Lines ◽  
Ex Vivo ◽  
Small Animal ◽  
Protein Translation ◽  
The Body ◽  
Small Animal Pet ◽  
Animal Pet ◽  
Organ Specific

Abstract Background RNA-based vaccination strategies tailoring immune response to specific reactions have become an important pillar for a broad range of applications. Recently, the use of lipid-based nanoparticles opened the possibility to deliver RNA to specific sites within the body, overcoming the limitation of rapid degradation in the bloodstream. Here, we have investigated whether small animal PET/MRI can be employed to image the biodistribution of RNA-encoded protein. For this purpose, a reporter RNA coding for the sodium-iodide-symporter (NIS) was in vitro transcribed in cell lines and evaluated for expression. RNA-lipoplex nanoparticles were then assembled by complexing RNA with liposomes at different charge ratios, and functional NIS protein translation was imaged and quantified in vivo and ex vivo by Iodine-124 PET upon intravenous administration in mice. Results NIS expression was detected on the membrane of two cell lines as early as 6 h after transfection and gradually decreased over 48 h. In vivo and ex vivo PET/MRI of anionic spleen-targeting or cationic lung-targeting NIS-RNA lipoplexes revealed a visually detectable rapid increase of Iodine-124 uptake in the spleen or lung compared to control-RNA-lipoplexes, respectively, with minimal background in other organs except from thyroid, stomach and salivary gland. Conclusions The strong organ selectivity and high target-to-background acquisition of NIS-RNA lipoplexes indicate the feasibility of small animal PET/MRI to quantify organ-specific delivery of RNA.

scholarly journals The fate of methylmercury through the formation of bismethylmercury sulfide as an intermediate in mice

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yumi Abiko ◽  
Yusuke Katayama ◽  
Wenyang Zhao ◽  
Sawako Horai ◽  
Kenji Sakurai ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Intraperitoneal Administration ◽  
Distal Colon ◽  
The Body ◽  
Gas Chromatography Mass Spectrometry ◽  
Dependent Manner ◽  
Free System ◽  
A Cell

AbstractA previous study by our group indicated that methylmercury (MeHg) is biotransformed to bismethylmercury sulfide [(MeHg)2S)] by interaction with reactive sulfur species (RSS) produced in the body. In the present study, we explored the transformation of MeHg to (MeHg)2S in the gut and the subsequent fate of (MeHg)2S in vitro and in vivo. An ex vivo experiment suggested the possibility of the extracellular transformation of MeHg to (MeHg)2S in the distal colon, and accordingly, the MeHg sulfur adduct was detected in the intestinal contents and feces of mice administered MeHg, suggesting that (MeHg)2S is formed through reactions between MeHg and RSS in the gut. In a cell-free system, we found that (MeHg)2S undergoes degradation in a time-dependent manner, resulting in the formation of mercury sulfide and dimethylmercury (DMeHg), as determined by X-ray diffraction and gas chromatography/mass spectrometry, respectively. We also identified DMeHg in the expiration after the intraperitoneal administration of (MeHg)2S to mice. Thus, our present study identified a new fate of MeHg through (MeHg)2S as an intermediate, which leads to conversion of volatile DMeHg in the body.

scholarly journals A Systematic Review of Clinical Studies on Mucocutaneous Manifestations of COVID-19: Virus-Related and Drug-Related

2021 ◽  
Author(s):  
Elham Behrangi ◽  
Mohammadreza Ghassemi ◽  
Afsaneh Sadeghzadeh-Bazargan ◽  
Masoumeh Roohaninasab ◽  
Niloufar Najar Nobari ◽  
...  
Keyword(s):  
Systematic Review ◽  
Clinical Symptoms ◽  
Skin Lesions ◽  
The Body ◽  
Inclusion Criteria ◽  
Cutaneous Lesions ◽  
Drug Induced ◽  
Skin Biopsies ◽  
Dermatologic Manifestations ◽  
Primary Skin Lesions

Coronavirus could affect almost any part of the body including the skin. In this systematic review, the primary skin lesions resulting from the direct activity of the virus or the medications used for treatment and the changes in the behavior of the virus regarding the occurrence of these symptoms over time were assessed. PubMed/MEDLINE, Embase, PsycINFO, TRIP Cochrane, Cochrane Skin were searched for all published articles from February 19 to July 1, 2020, which met the inclusion criteria. Thirty-six related articles were extracted. Twenty-eight studies reported virus-related mucocutaneous eruptions and 8 articles, the drug-reactions. Data of 583 patients were included. Skin lesions of COVID-19 could be caused by both the virus itself or the influence of drugs used for the treatment. Morbilliform rashes, urticaria, and acral-vasculopathic cutaneous lesions were at the forefront of primary COVID-dependent skin lesions with no significant change during time, Also, Hydroxychloroquine, lopinavir/ritonavir, paracetamol, and antibiotics were reported as the main causes of drug-induced rashes. Since dermatologic manifestations may occur prior or simultaneously/after other COVID clinical symptoms, so they may helpful in patients’ early diagnosis or prediction of internal organ involvements via histopathologic evaluations of skin biopsies especially about vasculopathic and vasculitic, respectively.

scholarly journals Modulating Mesenchymal Stem Cell Behavior Using Human Hair Keratin-Coated Surfaces

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Pietradewi Hartrianti ◽  
Ling Ling ◽  
Lyn Mei Ming Goh ◽  
Kok Seng Amos Ow ◽  
Rebekah Margaret Samsonraj ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Human Hair ◽  
Ex Vivo ◽  
The Body ◽  
Animal Origin ◽  
Homogeneous Distribution ◽  
Tissue Culture Polystyrene ◽  
Coated Surfaces ◽  
Hair Keratins

Human mesenchymal stem cells (hMSCs) have shown great potential for therapeutic purposes. However, the low frequencies of hMSCs in the body and difficulties in expanding their numbersin vitrohave limited their clinical use. In order to develop an alternative strategy for the expansion of hMSCsin vitro, we coated tissue culture polystyrene with keratins extracted from human hair and studied the behavior of cells from 2 donors on these surfaces. The coating resulted in a homogeneous distribution of nanosized keratin globules possessing significant hydrophilicity. Results from cell attachment assays demonstrated that keratin-coated surfaces were able to moderate donor-to-donor variability when compared with noncoated tissue culture polystyrene. STRO-1 expression was either sustained or enhanced on hMSCs cultured on keratin-coated surfaces. This translated into significant increases in the colony-forming efficiencies of both hMSC populations, when the cells were serially passaged. Human hair keratins are abundant and might constitute a feasible replacement for other biomaterials that are of animal origin. In addition, our results suggest that hair keratins may be effective in moderating the microenvironment sufficiently to enrich hMSCs with high colony-forming efficiencyex vivo, for clinical applications.

Artificial Super Para Magnetic Nano APC for Active and Adoptive Immunotherapy.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3888-3888
Author(s):  
Hugo De La Pena ◽  
J. Alejandro Madrigal ◽  
M. Bencsik ◽  
Gareth W.V. Cave ◽  
A. Selman ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Adoptive Immunotherapy ◽  
Ex Vivo ◽  
The Body ◽  
Cell Manipulation ◽  
Active Immunotherapy ◽  
Immune Gene

Abstract T cells are probably one of the most pivotal cell types in the human adaptive immune system. They have the capability to eradicate primary, metastatic, relapsed tumours and can ameliorate otherwise fatal viral infections. Not surprisingly therefore, activation and expansion of T cells has become one of the main focuses for immunotherapy and immune gene therapy. Sufficient T cells numbers however, are required to deliver a significant clinical impact to patients, and rapid reproducible expansion of viable T cells still remains one of the main challenges for significant improvement. One of the main concerns with adoptive immunotherapy is that it relies on one critical factor: ex-vivo cell manipulation; the problem with this is that the longer the in-vitro T cell culture, the shorter the in-vivo T cell survival after infusion. In-vivo artificial expansion systems for active immunotherapy would clearly circumvent this problem. Therefore ideally a flexible system should be constructed in order to performed both adoptive and/or active immunotherapy depending on the patients requirements. Currently there is no comprehensive artificial Antigen Presenting Cell system (aAPC) for both effective ex-vivo and in-vivo antigen specific T cell expansion. In order to address this, using nanotechnology, we have constructed a nano sized super-para-magnetic artificial targeted and traceable in-vivo APC system by coating liposomes (approved for human use) with an optimised number of MHC Class I / peptide complexes and a specific selected range of ligands for adhesion (anti LFA1), early activation (anti CD28, anti CD27), late activation (anti 4-1BB) and survival (anti CD40L) T cell receptors in the form of Fab antibody regions. We have constructed targeted liposomes (immuno-liposomes), which are also traceable in-vivo via fluorescent and Magnetic Resonance Imaging (MRI). Ex-vivo (human) and in-vivo (animal) models have been investigated showing firstly that these super-para-magnetic immuno-liposomes circulate the body safely and facilitate their own focusing to specific organs, tumour sites or body areas by applying external magnetic attraction. Secondly, in a viral (CMV) antigen specific model and measured by relevant and irrelevant tetramers, the system is capable of activating and expanding antigen specific T cells at greater levels (200 fold) than standard methods from CMV positive (memory) individuals. The system has also been able to accomplish a small successful level of T cell priming from naive CMV negative individuals. The T cells are phenotypicaly relevant and fully functional in terms of degranulation and cytokine production when specifically challenged. As mechanisms of action, we have demonstrated that the system functions directly on T cells as micro APCs and also semi-directly on the surface of natural APCs following a similar exosomes kinetics. The system is generated in less that 48 hr. Once the aAPC is created and it remains viable and stable for 7 days minimum. We have established optimal conditions for an efficient artificial APC, which embodies a superior and controllable approach and platform with enormous potential for cancer nanotechnology and T cell mediated immunotherapy.

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