scholarly journals A recombinant SARS-CoV-2 RBD antigen expressed in insect cells elicits immunogenicity and confirms safety in animal models

2021 ◽  
Author(s):  
Ricardo Choque-Guevara ◽  
Astrid Poma-Acevedo ◽  
Ricardo Montesinos-Millan ◽  
Dora Rios-Matos ◽  
Kristel Gutierrez-Manchay ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Insect Cells ◽  
Present Report ◽  
Safety Evaluation ◽  
Etiologic Agent ◽  
Histopathological Analysis ◽  
Binding Assays ◽  
Immunization Strategies ◽  
Reducing Conditions ◽  
Liver And Kidney

COVID-19 pandemic has accelerated the development of vaccines against its etiologic agent, SARS-CoV-2. However, the emergence of new variants of the virus requires new immunization strategies in addition to the current vaccines approved for human administration. In the present report, the immunological and safety evaluation in mice and hamsters of a subunit vaccine based on the RBD sub-domain with two adjuvants of oil origin is described. The RBD protein was expressed in insect cells and purified by chromatography until >95% purity. The protein was shown to have the appropriate folding as determined by ELISA and flow cytometry binding assays to its receptor, as well as by its detection by hamster immune anti-S1 sera under non-reducing conditions. In immunization assays in mice and hamsters, the purified RBD formulated with adjuvants based on oil-water emulsifications and squalene was able to stimulate specific neutralizing antibodies and confirm the secretion of IFN-γ after stimulating spleen cells with the purified RBD. The vaccine candidate was shown to be safe, as demonstrated by the histopathological analysis in lungs, liver and kidney. These results demonstrate the potential of the purified RBD administered with adjuvants through an intramuscular route, to be evaluated in a challenge against SARS-CoV-2 and determine its ability to confer protection against infection.

Preparation and Biochemical Evaluation of Functionalized Multi-Walled Carbon Nanotubes with Punica granatum Extract

2019 ◽  
Vol 15 (1) ◽  
pp. 138-144 ◽  
Author(s):  
Ahmed A. Haroun ◽  
Abdel-Tawab H. Mossa ◽  
Samia M.M. Mohafrash
Keyword(s):  
Liver Enzymes ◽  
Histopathological Analysis ◽  
Pomegranate Peel ◽  
Liver And Kidney ◽  
Multi Walled Carbon Nanotubes ◽  
Pomegranate Peel Extract ◽  
Walled Carbon Nanotubes ◽  
Functionalized Mwcnts ◽  
Peel Extract

Background: Funcionalized multi-walled carbon nanotubes (ox-MWCNTs) were used for the preparation of therapeutic nanoparticles for delivery of some bioactive compounds. Consequently, this work deals with the preparation of grafted MWCNTs with n-vinyl caprolactam in the presence of pomegranate peel extract (P. granatum), titanium dioxide (TiO2) and/or silver nanoparticeles and their toxic effects on male mice using in vivo biological examination (liver and kidney dysfunction biomarkers) and the histopathological analysis. Methods: P. granatum extract was immobilized onto functionalized MWCNTs using simple adsorption technique. Moreover, The prepared materials were analyzed by Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM). In vivo examination using liver and kidney dysfunction biomarkers was investigated. In addition, the histopathological study was carried out. Results: The ox-MWCNTs induced significant elevation in the liver enzymes including AST, ALT and ALP relative to the control group. While, the treatment with P. granatum extract only did not induce any change in the liver and kidney biomarkers. In other words, P. granatum extract loaded onto functionalized MWCNTs showed low effects on liver enzymes and kidney function biomarkers in the treated mice in comparison with ox-MWCNTs and extract separately. Moreover, histopathological analysis revealed that the P. granatum extract functionalized MWCNTs exhibited normal renal tissue with no histopathological alteration. Conclusion: The grafted MWCNTs with n-vinyl caprolactam in the presence of pomegranate peel extract (P. granatum), titanium dioxide (TiO2) and/or silver nanoparticeles were successfully prepared. SEM-micrographs showed complete coating of MWCNTs fiber with the extract. The prepared materials resulted in no toxic effects and the histopathological findings were confirmed by inflammation of the liver and kidney tissues.

scholarly journals Biological Safety Evaluation and Surface Modification of Biocompatible Ti–15Zr–4Nb Alloy

Materials ◽  
2021 ◽  
Vol 14 (4) ◽  
pp. 731
Author(s):  
Yoshimitsu Okazaki ◽  
Shin-ichi Katsuda
Keyword(s):  
Dental Implant ◽  
Safety Evaluation ◽  
Histopathological Analysis ◽  
Negative Effects ◽  
Biological Safety ◽  
Artificial Hip Joint ◽  
Artificial Hip ◽  
Pullout Load ◽  
Al2o3 Particles ◽  
Zr Alloys

We performed biological safety evaluation tests of three Ti–Zr alloys under accelerated extraction condition. We also conducted histopathological analysis of long-term implantation of pure V, Al, Ni, Zr, Nb, and Ta metals as well as Ni–Ti and high-V-containing Ti–15V–3Al–3Sn alloys in rats. The effect of the dental implant (screw) shape on morphometrical parameters was investigated using rabbits. Moreover, we examined the maximum pullout properties of grit-blasted Ti–Zr alloys after their implantation in rabbits. The biological safety evaluation tests of three Ti–Zr alloys (Ti–15Zr–4Nb, Ti–15Zr–4Nb–1Ta, and Ti–15Zr–4Nb–4Ta) showed no adverse (negative) effects of either normal or accelerated extraction. No bone was formed around the pure V and Ni implants. The Al, Zr, Nb, and Ni–Ti implants were surrounded by new bone. The new bone formed around Ti–Ni and high-V-containing Ti alloys tended to be thinner than that formed around Ti–Zr and Ti–6Al–4V alloys. The rate of bone formation on the threaded portion in the Ti–15Zr–4Nb–4Ta dental implant was the same as that on a smooth surface. The maximum pullout loads of the grit- and shot-blasted Ti–Zr alloys increased linearly with implantation period in rabbits. The pullout load of grit-blasted Ti–Zr alloy rods was higher than that of shot-blasted ones. The surface roughness (Ra) and area ratio of residual Al2O3 particles of the Ti–15Zr–4Nb alloy surface grit-blasted with Al2O3 particles were the same as those of the grit-blasted Alloclassic stem surface. It was clarified that the grit-blasted Ti–15Zr–4Nb alloy could be used for artificial hip joint stems.

scholarly journals Evaluation of the Coating with TiO2 Nanoparticles as an Option for the Improvement of the Characteristics of NiTi Archwires: Histopathological, Cytotoxic, and Genotoxic Evidence

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Javier Morán-Martínez ◽  
Roberto Beltrán del Río-Parra ◽  
Nadia Denys Betancourt-Martínez ◽  
Rubén García-Garza ◽  
Joel Jiménez-Villarreal ◽  
...  
Keyword(s):  
Cell Viability ◽  
Blood Sample ◽  
Tio2 Nanoparticles ◽  
Niti Alloy ◽  
Renal Tissue ◽  
Male Rats ◽  
Histopathological Analysis ◽  
Liver And Kidney ◽  
The Times ◽  
Nuclear Alterations

For the EPD, different voltages and different times were used. Male rats were used in four groups (n=3) with different treatments. The blood sample was obtained for genotoxic analysis and liver and kidney organs were removed for histopathological analysis. The amount of NPs TiO2 deposited on the samples of the arches increases gradually in the times of 15 and 30 s. At all voltages, however, at 45, 60, 75, and 90 s, there is an increase up to 25 V. Cell viability in lymphocytes treated with TiO2 NPs did not cause genotoxicity. In the histopathological findings of hepatic and renal tissue, nuclear alterations and necrosis were observed. The objective of the study was to improve the physical and biocompatibility characteristics of the NiTi arches for which the EPD is used. The technique for the deposition of TiO2 NPs was used, where this technique could be used as an economical and versatile way to perform homogeneous depositions even on surfaces with the complexity of the NiTi alloy. As for genotoxicity and cytotoxicity, we continue to have controversial results.

scholarly journals Dermatophytosis due to Microsporum nanum infection in a canine

2017 ◽  
Vol 38 (1) ◽  
pp. 317
Author(s):  
Marilia Avila Valandro ◽  
João Paulo da Exaltação Pascon ◽  
Maria Lígia de Arruda Mistieri ◽  
Irina Lubeck
Keyword(s):  
Wild Boar ◽  
Clinical Features ◽  
Therapeutic Response ◽  
Systemic Treatment ◽  
Present Report ◽  
Etiologic Agent ◽  
Good Clinical Response ◽  
Circular Pattern ◽  
Source Of Infection ◽  
Wild Boar Hunting

Miscrosporum nanum is a dermatophyte found in swine that causes non-pruritic lesions with desquamation, alopecia, and circular characteristics. M. nanum infection in dogs is rare and poorly understood in terms of its epidemiological and clinical features, and its therapeutic response. The present report describes a case of dermatophytosis due to M. nanum in a Dogo Argentino breed of dog that was used for wild boar hunting. The dermatophytosis presented with hypotrichosis, erythema, and non-pruritic desquamation in the back of the neck and chest area. The dermatophytosis was responsive to systemic treatment with itraconazole and topical (miconazole 2%) for 60 days. Thus, we conclude that the practice of hunting wild boar should be considered as a possible source of infection of M. nanum in the reported dog. The M. nanum infection showed clinical features that were similar to the lesions observed in swine, except for the absence of the circular pattern, and showed a good clinical response to the therapy. Finally, M. nanum should be considered as an etiologic agent of dermatophytosis in dogs that in some manner have had direct contact with domestic or wild swine.

scholarly journals ASSESSMENT OF EXTRAPULMONARY TOXICITY INDUCED BY CARBON NANOMATERIALS FOLLOWING INTRA-TRACHEAL INSTILLATION IN RATS

2017 ◽  
Vol 10 (5) ◽  
pp. 82 ◽  
Author(s):  
Bhikku Angoth ◽  
Harikiran Lingabathula ◽  
Narsimha Reddy Yellu
Keyword(s):  
Carbon Nanomaterials ◽  
Intratracheal Instillation ◽  
Histopathological Analysis ◽  
Carbon Nanofibres ◽  
Multi Wall Carbon Nanotubes ◽  
Post Exposure ◽  
Liver And Kidney ◽  
Serum Biochemical ◽  
Commercial Applications ◽  
Dose Dependent

Objective: Carbon nanomaterials (CNMs) such as carbon nanofibres (CNFs), multi wall carbon nanotubes (MWCNTs) and carbon nanorods (CNRs) were found various industrial and commercial applications. The occupational exposure for these CNMs were also increased enormously. The present study evaluated the extrapulmonary toxicity induced by these CNMs. Methods: The extrapulmonary toxicity was assessed following intratracheal instillation of test CNMs in rats after 1 day, 1 week, 1 month and 3 months post exposure periods by using serum biochemical parameters such as alanine transaminase (ALT) and creatinine using diagnostic assay kits. Further, the histopathological analysis was performed for liver and kidneys of particle exposed rats.Results: The results have displayed that increased levels of serum ALT and creatinine were found after 1 day, 1 week and 1 month post exposure periods indicating liver and kidney toxicity respectively. This toxicity was further confirmed by the changes observed in the histopathological analysis of rat liver and kidneys.Conclusion: The CNFs, MWCNTs and CNRs able to translocate from the lungs into other extrapulmonary organs such as liver and kidney, and also cause dose dependent toxicity to them.

scholarly journals The Evaluation of Toxicity Induced by Psoraleae Fructus in Rats Using Untargeted Metabonomic Method Based on UPLC-Q-TOF/MS

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Yanyan Xu ◽  
Yiwei Zhao ◽  
Jiabin Xie ◽  
Xue Sheng ◽  
Yubo Li ◽  
...  
Keyword(s):  
Metabolic Pathways ◽  
Acid Metabolism ◽  
Safety Evaluation ◽  
Rat Plasma ◽  
Leguminous Plant ◽  
Control Group ◽  
Kidney Toxicity ◽  
Flight Mass Spectrometry ◽  
Liver And Kidney ◽  
Tof Ms

Psoraleae Fructus is the dry and mature fruit of leguminous plant Psoralea corylifolia L., with the activity of warming kidney and enhancing yang, warming spleen, and other effects. However, large doses can cause liver and kidney toxicity. Therefore, it is necessary to evaluate the toxicity of Psoraleae Fructus systematically. Although traditional biochemical indicators and pathological tests have been used to evaluate the safety of drug, these methods lack sensitivity and specificity, so a fast and sensitive analytical method is urgently needed. In this study, an ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) method was used to analyze the metabolic profiles of rat plasma. The changes of metabolites in plasma samples were detected by partial least squares-discriminant analysis (PLS-DA). Compared with the control group, after 7 days of administration, the pathological sections showed liver and kidney toxicity, and the metabolic trend was changed. Finally, 13 potential biomarkers related to the toxicity of Psoraleae Fructus were screened. The metabolic pathways involved were glycerol phospholipids metabolism, amino acid metabolism, energy metabolism, and so forth. The discovery of these biomarkers laid a foundation for better explaining the hepatotoxicity and nephrotoxicity of Psoraleae Fructus and provided a guarantee for its safety evaluation.

scholarly journals Susceptibility to Neutralization by Broadly Neutralizing Antibodies Generally Correlates with Infected Cell Binding for a Panel of Clade B HIV Reactivated from Latent Reservoirs

2018 ◽  
Vol 92 (23) ◽  
Author(s):  
Yanqin Ren ◽  
Maria Korom ◽  
Ronald Truong ◽  
Dora Chan ◽  
Szu-Han Huang ◽  
...  
Keyword(s):  
T Cells ◽  
Infected Cell ◽  
Neutralizing Antibodies ◽  
Binding Assays ◽  
Cell Binding ◽  
Broadly Neutralizing Antibodies ◽  
Clade B ◽  
Latently Infected ◽  
Living With Hiv ◽  
Selection Of

ABSTRACTEfforts to cure human immunodeficiency virus (HIV) infection are obstructed by reservoirs of latently infected CD4+T cells that can reestablish viremia. HIV-specific broadly neutralizing antibodies (bNAbs), defined by unusually wide neutralization breadths against globally diverse viruses, may contribute to the elimination of these reservoirs by binding to reactivated cells, thus targeting them for immune clearance. However, the relationship between neutralization of reservoir isolates and binding to corresponding infected primary CD4+T cells has not been determined. Thus, the extent to which neutralization breadths and potencies can be used to infer the corresponding parameters of infected cell binding is currently unknown. We assessed the breadths and potencies of bNAbs against 36 viruses reactivated from peripheral blood CD4+T cells from antiretroviral (ARV)-treated HIV-infected individuals by using paired neutralization and infected cell binding assays. Single-antibody breadths ranged from 0 to 64% for neutralization (80% inhibitory concentration [IC80] of ≤10 μg/ml) and from 0 to 89% for binding, with two-antibody combinations (results for antibody combinations are theoretical/predicted) reaching levels of 0 to 83% and 50 to 100%, respectively. Infected cell binding correlated with virus neutralization for 10 of 14 antibodies (e.g., for 3BNC117,r = 0.82 andP < 0.0001). Heterogeneity was observed, however, with a lack of significant correlation for 2G12, CAP256.VRC26.25, 2F5, and 4E10. Our results provide guidance on the selection of bNAbs for interventional cure studies, both by providing a direct assessment of intra- and interindividual variabilities in neutralization and infected cell binding in a novel cohort and by defining the relationships between these parameters for a panel of bNAbs.IMPORTANCEAlthough antiretroviral therapies have improved the lives of people who are living with HIV, they do not cure infection. Efforts are being directed towards harnessing the immune system to eliminate the virus that persists, potentially resulting in virus-free remission without medication. HIV-specific antibodies hold promise for such therapies owing to their ability to both prevent the infection of new cells (neutralization) and direct the killing of infected cells. We isolated 36 HIV strains from individuals whose virus was suppressed by medication and tested 14 different antibodies for neutralization of these viruses and for binding to cells infected with the same viruses (critical for engaging natural killer cells). For both neutralization and infected cell binding, we observed variation both between individuals and amongst different viruses within an individual. For most antibodies, neutralization activity correlated with infected cell binding. These data provide guidance on the selection of antibodies for clinical trials.

scholarly journals Cross reactivity of neutralizing antibodies to the encephalitic California Serogroup orthobunyaviruses varies by virus and genetic relatedness

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alyssa B. Evans ◽  
Karin E. Peterson
Keyword(s):  
Gold Standard ◽  
Neutralizing Antibodies ◽  
Viral Encephalitis ◽  
Genetic Relatedness ◽  
Cross Reactivity ◽  
Etiologic Agent ◽  
Snowshoe Hare ◽  
Neuroinvasive Disease ◽  
California Serogroup ◽  
High Degree

AbstractThe California Serogroup (CSG) of Orthobunyaviruses comprises several viruses capable of causing neuroinvasive disease in humans, including La Crosse (LACV), Snowshoe Hare (SSHV), Tahyna (TAHV), Jamestown Canyon (JCV), and Inkoo (INKV) viruses. Diagnosis of specific CSG viruses is complicated by the high degree of antibody cross-reactivity between them, with laboratory standards requiring a fourfold higher titer of neutralizating antibody (NAb) activity to positively identify the etiologic virus. To help elucidate NAb relationships between neuroinvasive CSG viruses, we directly compared the cross-reactivity of NAb between LACV, SSHV, TAHV, JCV, and INKV. Mice were inoculated with individual viruses and the NAb activity of plasma samples was compared by plaque reduction neutralization tests against all five viruses. Overall, the results from these studies show that the CSG viruses induced high levels of NAb against the inoculum virus, and differing amounts of cross-reactive NAb against heterologous viruses. LACV, SSHV, and INKV elicited the highest amount of cross-reactive NAb. Interestingly, a fourfold difference in NAb titer between the inoculum virus and the other CSG viruses was not always observed. Thus, NAb titers, which are the gold-standard for diagnosing the etiologic agent for viral encephalitis, may not clearly differentiate between different CSG viruses.

scholarly journals Microangiopathic Anemia of Acute Brucellosis – is it a True TTP?

2010 ◽  
Vol 2 (3) ◽  
pp. e2010031 ◽  
Author(s):  
Amir Kuperman
Keyword(s):  
Thrombotic Microangiopathy ◽  
Neutralizing Antibodies ◽  
Molecular Mechanisms ◽  
Brucella Melitensis ◽  
Present Report ◽  
Severe Disease ◽  
Acute Infection ◽  
High Dose ◽  
Microangiopathic Anemia ◽  
Work Up

Thrombotic thrombocytopenic purpura (TTP) is a severe disease, potentially fatal, if not diagnosed and treated promptly. TTP is clinically characterized by the pentad of thrombocytopenia, Coombs-negative hemolytic anemia, fever, renal abnormalities and neurological disturbances. Advances in recent years have delineated the molecular mechanisms of acquired and hereditary TTP.Many infectious organisms have been reported to be associated with TTP, especially mycoplasma, but only 6 cases of Brucella infection associated with thrombotic microangiopathy were reported.We describe a young woman who presented clinically with TTP following acute infection with both Brucella melitensis and Brucella abortus. The patient completely recovered after an aggressive therapy with plasmapharesis, high-dose corticosteroids and appropriate antimicrobial therapy.Since measurement of ADMTS13 activity and neutralizing antibodies is now available, and in none of the reported cases of brucellosis with thrombotic microangiopathy (including the present report) were tested, we recommend this work-up in future cases for better understanding of this rare association.   

scholarly journals Isolation of Potent CGRP Neutralizing Antibodies Using Four Simple Assays

2015 ◽  
Vol 21 (1) ◽  
pp. 24-34 ◽  
Author(s):  
Frances Neal ◽  
Joanne Arnold ◽  
Christine J. Rossant ◽  
Sadhana Podichetty ◽  
David Lowne ◽  
...  
Keyword(s):  
Functional Activity ◽  
Neutralizing Antibodies ◽  
Cross Reactivity ◽  
Affinity Maturation ◽  
Binding Assays ◽  
Time Resolved ◽  
Calcitonin Gene ◽  
Potent Vasodilator ◽  
Competition Assays ◽  
Species Cross

Calcitonin gene-related peptide (CGRP) is a small neuropeptide and a potent vasodilator that is widely associated with chronic pain and migraine. An antibody that inhibits CGRP function would be a potential therapeutic for treatment of these disorders. Here we describe the isolation of highly potent antibodies to CGRP from phage and ribosome display libraries and characterization of their epitope, species cross-reactivity, kinetics, and functional activity. Homogenous time-resolved fluorescence (HTRF) binding assays identified antibodies with the desired species cross-reactivity from naïve libraries, and HTRF epitope competition assays were used to characterize and group scFv by epitope. The functional inhibition of CGRP and species cross-reactivity of purified scFv and antibodies were subsequently confirmed using cAMP assays. We show that epitope competition assays could be used as a surrogate for functional cell-based assays during affinity maturation, in combination with scFv off-rate ranking by biolayer interferometry (BLI). This is the first time it has been shown that off-rate ranking can be predictive of functional activity for anti-CGRP antibodies. Here we demonstrate how, by using just four simple assays, diverse panels of antibodies to CGRP can be identified. These assay formats have potential utility in the identification of antibodies to other therapeutic targets.

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