Characterization of toxicological effects of complex nano‐sized metal particles using in vitro human cell and whole blood model systems

2021 ◽  
Author(s):  
Linda Elfsmark ◽  
Barbro Ekstrand‐Hammarström ◽  
Nina Forsgren ◽  
Christian Lejon ◽  
Lars Hägglund ◽  
...  
Keyword(s):  
Human Cell ◽  
Whole Blood ◽  
Model Systems ◽  
Metal Particles ◽  
Toxicological Effects

scholarly journals THU0080 PRECLINICAL CHARACTERIZATION OF TLL018, A NOVEL, HIGHLY POTENT AND SELECTIVE JAK1/TYK2 INHIBITOR FOR TREATING AUTOIMMUNE DISEASES

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 252.1-252
Author(s):  
X. Liu ◽  
F. Tan ◽  
C. Liang
Keyword(s):  
Bone Resorption ◽  
Autoimmune Diseases ◽  
Whole Blood ◽  
Significant Inhibition ◽  
Clinical Development ◽  
Therapeutic Window ◽  
Collagen Induced Arthritis

Background:Janus kinases (JAKs) are important regulators of intracellular responses triggered by many key proinflammatory cytokines and are clinically validated therapeutic targets for treating various autoimmune diseases. However, current approved JAK inhibitors failed to achieve maximal clinical benefit in part due to their unfavorable selectivity for individual JAKs such as JAK2 and/or JAK3, leading to dose-limiting toxicities or severe toxicities (e.g., thrombosis, anemia, immune suppression). Selective inhibition of JAK1 and/or TYK2 may minimize or avoid some of the toxicities and potentially offer a better therapeutic window for treating autoimmune diseases. No highly selective JAK1/TYK2 inhibitor has been reported to date.Objectives:Discovery of a highly selective JAK1/TYK2 inhibitor that maximally avoids JAK2 and JAK3 inhibition. We described preclinical characterization of a novel, highly potent and selective JAK1/TYK2 inhibitor TLL018 and its potential utility in treating autoimmune diseases such as rheumatoid arthritis (RA).Methods:Using predicting SAR, TLL018 was designed to achieve exquisite selectivity for both JAK1 and TYK2 while sparing JAK2, JAK3 and other human kinases. Its enzyme and cell activities, kinase selectivity, andin vivoefficacy were assessed in a battery of relevant enzyme, cell and whole blood assays, andin vivoarthritis animal models. Additional preclinical DMPK and toxicology studies were conducted to support its clinical development.Results:TLL018 is a highly potent and selective, orally bioavailable JAK1/TYK2 inhibitor against JAK1 (IC50= 4 nM) and TYK2 (IC50= 5 nM) as measured inin vitrokinase assays with ATP concentrations at individual Km. Its potency against JAK2 or JAK3 is greater than 1 µM. Profiling against a panel of over 350 human kinase showed that TLL018 is exclusively selective for JAK1 and TYK2, with ≥ 90-fold selectivity against all other kinases tested. TLL018 exhibited potent cellular activity for JAK1-mediated IL-6 signaling (IC50= 0.6 µM) with greater than 100-fold selectivity against JAK2-mediated cytokine (e.g., TPO) signaling in human whole blood-based assays.Oral administration of TLL018 demonstrated dose-dependent efficacy in commonly studied rat adjuvant-induced arthritis (rAIA) model and mouse collagen-induced arthritis (mCIA) model. Significant inhibition of inflammation, bone resorption, splenomegaly and body weight change was observed in adjuvant-induced disease in rats. In addition, significant inhibition of inflammation, cartilage destruction, bone resorption and histological signs was demonstrated in collagen-induced arthritis in mice. Noticeably, TLL018 exhibited significant anti-inflammation activity at doses that only blocked JAK1 and TYK2 and exerted little inhibition of JAK2 and JAK3.In support of clinical development of TLL018, preclinical ADME and PK studies and IND-enabling toxicology and safety pharmacology studies were completed, confirming that TLL018 possesses excellent ADME and PK properties, and exhibits a clean on-target safety profile.Conclusion:TLL018 is a highly potent and selective JAK1/TYK2 inhibitor that demonstrated excellent efficacy and tolerability in relevant mouse and rat arthritis models. The collective data of its preclinical pharmacology, PK and toxicology showed a favorable pharmaceutical profile, further supporting its development for treating autoimmune diseases including RA. Clinical evaluation of TLL018 is ongoing.Disclosure of Interests:Xiangdong Liu Shareholder of: I own shares of TLL Pharmaceutical LLC, Employee of: I am employed by TLL Pharmaceutical LLC, Fenlai Tan Shareholder of: I own shares of TLL Pharmaceutical LLC, Employee of: I am employed by TLL Pharmaceutical LLC, Chris Liang Shareholder of: I own shares of TLL Pharmaceutical LLC, Employee of: I am employed by TLL Pharmaceutical LLC

Characterization of microvesicles released from human whole blood after incubation with Bothrops jararaca venom: An in vitro preliminary study

Toxicon ◽  
2020 ◽  
Vol 177 ◽  
pp. S31
Author(s):  
Milene Cristina Menezes ◽  
DIlza Trevisan-Silva ◽  
Daniela Cajado-Carvalho ◽  
Solange Serrano
Keyword(s):  
Whole Blood ◽  
Human Whole Blood ◽  
Bothrops Jararaca ◽  
Preliminary Study

scholarly journals A new glycation product ‘norpronyl-lysine,’ and direct characterization of cross linking and other glycation adducts: NMR of model compounds and collagen

2014 ◽  
Vol 34 (2) ◽  
Author(s):  
Peter T. B. Bullock ◽  
David G. Reid ◽  
W. Ying Chow ◽  
Wendy P. W. Lau ◽  
Melinda J. Duer
Keyword(s):  
Model Systems ◽  
Cross Linking ◽  
Advanced Glycation ◽  
Model Compounds ◽  
Product Characterization ◽  
Glycation Product ◽  
Cross Links

NMR reveals numerous early and advanced glycation products, including a newly recognized ‘norpronyl-lysine,’ and cross links in solution, intact collagen and model systems. Solid state methods are directly applicable to in vitro and in vivo glycation pathway and product characterization.

scholarly journals Establishment and characterization of equine fibroblast cell lines transformed in vivo and in vitro by BPV-1: Model systems for equine sarcoids

Virology ◽  
2008 ◽  
Vol 373 (2) ◽  
pp. 352-361 ◽  
Author(s):  
Z.Q. Yuan ◽  
E.A. Gault ◽  
P. Gobeil ◽  
C. Nixon ◽  
M.S. Campo ◽  
...  
Keyword(s):  
Cell Lines ◽  
Fibroblast Cell ◽  
Model Systems ◽  
Fibroblast Cell Lines

scholarly journals ENHANCED FLUORESCENCE IMAGING WITH DMSO-MEDIATED OPTICAL CLEARING

2010 ◽  
Vol 03 (03) ◽  
pp. 153-158 ◽  
Author(s):  
SANJEEV KARMA ◽  
JAMES HOMAN ◽  
CHARLES STOIANOVICI ◽  
BERNARD CHOI
Keyword(s):  
Fluorescence Emission ◽  
Model Systems ◽  
In Vivo Studies ◽  
In Vivo Model ◽  
Fluorescence Signal ◽  
Optical Clearing ◽  
Treated Site

Recent studies have demonstrated that topical application of glycerol on intact skin does not affect its optical scattering properties. Investigators from our research group recently revisited the use of dimethyl sulfoxide (DMSO) as an agent with optical clearing potential. We address the use of optical clearing to enhance quantitation of subsurface fluorescence emission. We employed both in vitro and in vivo model systems to study the effect of topical DMSO application on fluorescence emission. Our in vitro experiments performed on a tissue-simulating phantom suggest that DMSO-mediated optical clearing enables enhanced characterization of subsurface fluorophores. With topical DMSO application, a marked increase in fluorescence emission was observed. After 30 min, the fluorescence signal at the DMSO-treated site was 9× greater than the contralateral saline-treated site. This ratio increased to 13× at 105 min after agent application. In summary, DMSO is an effective optical clearing agent for improved fluorescence emission quantitation and warrants further study in preclinical in vivo studies. Based on outcomes from previous clinical studies on the toxicity profile of DMSO, we postulate that clinical application of DMSO as an optical clearing agent, can be performed safely, although further study is warranted.

scholarly journals Studying the processes of free-radical oxidation in patients with purulent-inflammatory diseases of soft tissues

2015 ◽  
Vol 96 (3) ◽  
pp. 302-306
Author(s):  
T Z Zakiev ◽  
S R Tuysin ◽  
O V Galimov ◽  
A R Gil’fanov ◽  
R D Sagdeev
Keyword(s):  
Free Radical ◽  
Whole Blood ◽  
Soft Tissues ◽  
Local Treatment ◽  
Free Radical Oxidation ◽  
Model Systems ◽  
Radical Oxidation ◽  
Tissue Homogenates

Aim. To study the effects of combined dressings on the processes of free radical oxidation in patients with purulent wounds.Methods. The performance of luminol-dependent chemiluminescence of whole blood from 30 healthy donors and 122 patients with purulent wounds, as well as tissue chemiluminescence of festering wounds tissue homogenates were examined before and after the treatment. To assess the free radical oxidation in vitro, spontaneous and zymosan-induced whole blood chemiluminescence measurement, iron-induced chemiluminescence assessment of festering wounds tissue homogenates were performed. The distribution of chemiluminescence parameters compared to normal expected distribution was analyzed to group homogeneity on these criteria.Results. Group of donors was homogeneous in composition that allowed the calculation of mean values. By changing in whole blood chemiluminescence of patients with soft tissue festering wounds, they were allocated to two groups, in which a marked increase or decline of the investigated parameters was seen. Increased luminol-dependent blood chemiluminescence indicates excessive free radicals generation by phagocytes and is characteristic of acute inflammation. Decreased intensity of chemiluminescence was observed in the blood of patients who showed reduced functional activity of phagocytes, which, together with the clinical features indicate a long-term smoldering inflammation. The comparative assessment of «Poliderm», «Voskopran», «Polysorb», «Levomekol» medications, used for local treatment of suppurative wounds, influence on free radical oxidation in model systems in vitro, was performed.Conclusion. There is a triple increase in chemiluminescence intensity in the acute phase of the inflammatory process, marking increased production of reactive oxygen radicals with microbicide activity); in long-term smoldering processes, chemiluminescence decreases by half, indicating reduced efficiency of protective mechanisms

scholarly journals Physical, chemical, and toxicological characterization of fibrillated forms of cellulose using an in vitro gastrointestinal digestion and co-culture model

2020 ◽  
Vol 9 (3) ◽  
pp. 290-301 ◽  
Author(s):  
Sahar H Pradhan ◽  
Marina R Mulenos ◽  
London R Steele ◽  
Matthew Gibb ◽  
James D Ede ◽  
...  
Keyword(s):  
Food Safety ◽  
Chemical Characterization ◽  
Toxicological Effects ◽  
Simulated Digestion ◽  
Physical Chemical ◽  
Culture Model ◽  
The Usa ◽  
Physical And Chemical

Abstract Fibrillated cellulose is a next-generation material in development for a variety of applications, including use in food and food-contact materials. An alternative testing strategy including simulated digestion was developed to compare the physical, chemical, and biological characteristics of seven different types of fibrillated cellulose, following European Food Safety Authority guidance. Fibrillated forms were compared to a conventional form of cellulose which has been used in food for over 85 years and has Generally Recognized as safe regulatory status in the USA. The physical and chemical characterization of fibrillated celluloses demonstrate that these materials are similar physically and chemically, which composed of the same fundamental molecular structure and exhibit similar morphology, size, size distribution, surface charge, and low levels of impurities. Simulated gastrointestinal and lysosomal digestions demonstrate that these physical and chemical similarities remain following exposure to conditions that mimic the gastrointestinal tract or intracellular lysosomes. A toxicological investigation with an advanced intestinal co-culture model found that exposure to each of the fibrillated and conventional forms of cellulose, in either the pristine or digested form at 0.4% by weight, showed no adverse toxicological effects including cytotoxicity, barrier integrity, oxidative stress, or inflammation. The results demonstrate the physical, chemical, and biological similarities of these materials and provide substantive evidence to support their grouping and ability to read-across data as part of a food safety demonstration.

scholarly journals Isolation and characterization of novel primary cells from the human distal outflow pathway

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Uttio Roy Chowdhury ◽  
Cindy K. Bahler ◽  
Cheryl R. Hann ◽  
Bradley H. Holman ◽  
Michael P. Fautsch
Keyword(s):  
Trabecular Meshwork ◽  
Distal Region ◽  
Proximal Region ◽  
Cellular Growth ◽  
Model Systems ◽  
Type I ◽  
Isolation And Characterization ◽  
Outflow Pathway

AbstractOcular hypertension occurs due to increased resistance to aqueous humor removal through the conventional outflow pathway. Unlike the proximal region of the conventional outflow pathway, the distal region has not been well studied, mostly due to lack of model systems. Here we describe isolation and characterization of human primary vascular distal outflow pathway (VDOP) cells from the distal region of the conventional outflow pathway. Tissue from the distal region was isolated from human corneo-scleral rims, digested with collagenase type I (100 U/ml) and placed on gelatin coated plates to allow cellular growth in Dulbecco’s Modified Eagle’s Medium (low glucose) containing fetal bovine serum and antibiotic/antimycotic. VDOP cells showed consistent proliferation for up to 7 passages, retained endothelial-like nature of the parent tissues and showed a unique marker phenotype of Lectin+VEGFR2-CD34-NG2- that was distinct from neighboring trabecular meshwork (Lectin+VEGFR2-CD34-NG2+) and Schlemm’s canal (Lectin+VEGFR2+CD34+NG2+) cells. Dexamethasone treated VDOP cells did not express myocilin and did not form cross-linked actin networks, in contrast to trabecular meshwork cells. These data show that VDOP cells are unique to the distal outflow region and can be used as a viable in vitro model system to understand the biology of the distal outflow pathway and intraocular pressure regulation.

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