Proteomic characterization of a second-generation version of the BCGΔBCG1419c vaccine candidate by means of Electrospray-ionization quadrupole time-of-flight mass spectrometry

2020 ◽  
Author(s):  
Jesús Bernardino Velázquez-Fernández ◽  
Gustavo Henrique Martins Ferreira Souza ◽  
Jacobo Rodríguez-Campos ◽  
Michel de Jesús Aceves-Sánchez ◽  
Jorge Bravo-Madrigal ◽  
...  
Keyword(s):  
Second Generation ◽  
Vaccine Candidate ◽  
World Health ◽  
Label Free ◽  
Wild Type ◽  
Flight Mass Spectrometry ◽  
Proteomic Approach ◽  
Antigenic Proteins ◽  
Health Organization

Abstract Tuberculosis (TB) is the most important infectious disease worldwide, based on the number of new cases and deaths reported by the World Health Organization. Several vaccine candidates against TB have been characterized at the preclinical and clinical levels. The BCGΔBCG1419c vaccine candidate, which lacks the BCG1419c gene that encodes for a c-di-GMP phosphodiesterase, provides improved efficacy against chronic TB, reactivation from latent-like infection, and against TB in the presence of type 2 diabetes in murine models. We previously reported that compared to wild type BCG, BCGΔBCG1419c changed several proteins. Here, using a label-free proteomic approach, we confirmed that a novel, second-generation version of BCGΔBCG1419c maintains changes in antigenic proteins already reported, including differences in secreted proteins, and also found that it modifies its production of proteins involved in redox and nitrogen/protein metabolism, compared with wild type BCG. This work contributes to the proteomic characterization of a novel vaccine candidate that is effective against chronic TB.

scholarly journals Two Live Attenuated Vaccines against Recent Low–and Highly Pathogenic H7N9 Influenza Viruses Are Safe and Immunogenic in Ferrets

Vaccines ◽  
2018 ◽  
Vol 6 (4) ◽  
pp. 74 ◽  
Author(s):  
Larisa Rudenko ◽  
Irina Kiseleva ◽  
Elena Krutikova ◽  
Ekaterina Stepanova ◽  
Irina Isakova-Sivak ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Phase I ◽  
Reverse Genetics ◽  
Vaccine Candidate ◽  
Clinical Signs ◽  
Influenza Viruses ◽  
Phase I Clinical Trial ◽  
World Health ◽  
Histological Data ◽  
Health Organization

Influenza H7N9 virus is a potentially pandemic subtype to which most people are immunologically naïve. To be better prepared for the potential occurrence of an H7N9 pandemic, in 2017 the World Health Organization recommended developing candidate vaccine viruses from two new H7N9 viruses, A/Guangdong/17SF003/2016 (A/GD) and A/Hong Kong/125/2017 (A/HK). This report describes the development of live attenuated influenza vaccine (LAIV) candidates against A/GD and A/HK viruses and study of their safety and immunogenicity in the ferret model in order to choose the most promising one for a phase I clinical trial. The A/HK-based vaccine candidate (A/17/HK) was developed by classical reassortment in eggs. The A/GD-based vaccine candidate (A/17/GD) was generated by reverse genetics. Ferrets were vaccinated with two doses of LAIV or phosphate-buffered saline. Both H7N9 LAIVs tested were safe for ferrets, as shown by absence of clinical signs, and by virological and histological data; they were immunogenic after a single vaccination. These results provide a compelling argument for further testing of these vaccines in volunteers. Since the A/HK virus represents the cluster that has caused the majority of human cases, and because the A/HK-based LAIV candidate was developed by classical reassortment, this is the preferred candidate for a phase I clinical trial.

scholarly journals Diffuse Gliomas for Nonneuropathologists: The New Integrated Molecular Diagnostics

2018 ◽  
Vol 142 (7) ◽  
pp. 804-814 ◽  
Author(s):  
Sunhee C. Lee
Keyword(s):  
Krebs Cycle ◽  
Diagnostic Algorithm ◽  
World Health ◽  
Wild Type ◽  
Targeted Next Generation Sequencing ◽  
Idh Mutations ◽  
Diffuse Gliomas ◽  
Conventional Histology ◽  
Krebs Cycle Enzymes ◽  
Health Organization

Diffuse gliomas comprise the bulk of “brain cancer” in adults. The recent update to the 4th edition of the World Health Organization's classification of tumors of the central nervous system reflects an unprecedented change in the landscape of the diagnosis and management of diffuse gliomas that will affect all those involved in the management and care of patients. Of the recently discovered gene alterations, mutations in the Krebs cycle enzymes isocitrate dehydrogenases (IDHs) 1 and 2 have fundamentally changed the way the gliomas are understood and classified. Incorporating information on a few genetic parameters (IDH, ATRX and/or p53, and chromosome 1p19q codeletion), a relatively straightforward diagnostic algorithm has been generated with robust and reproducible results that correlate with patients' survival far better than relying on conventional histology alone. Evidence also supports the conclusion that the vast majority of diffuse gliomas without IDH mutations (IDH–wild-type astrocytomas) behave like IDH–wild-type glioblastomas (“molecular GBM”). Together, these changes reflect a big shift in the practice of diagnostic neuropathology in which tumor risk stratification aligns better with molecular information than histology/grading. The purpose of this review is to provide the readers with a brief synopsis of the changes in the 2016 World Health Organization update with an emphasis on diffuse gliomas and to summarize key gene abnormalities on which these classifications are based. Practical points involved in day-to-day diagnostic workup are also discussed, along with a comparison of the various diagnostic tests, including immunohistochemistry, with an emphasis on targeted next-generation sequencing panel technology as a future universal approach.

scholarly journals Microbiological and Physicochemical Characterization of Hospital Effluents before and after Treatment with Two Types of Sawdust

2019 ◽  
Vol 2019 ◽  
pp. 1-10 ◽  
Author(s):  
Rabea Elmountassir ◽  
Bahia Bennani ◽  
Youssef Miyah ◽  
Ahlam Fegousse ◽  
Ghita El Mouhri ◽  
...  
Keyword(s):  
Physicochemical Characterization ◽  
Oxygen Demand ◽  
Fecal Coliforms ◽  
World Health ◽  
Hospital Effluents ◽  
Before And After ◽  
Health Organization ◽  
Filter Bed ◽  
Microbiological Analyses

Physicochemical and microbiological analyses of liquid hospital effluents have demonstrated that they are loaded with organic and inorganic pollutants then discharged into the sewerage networks without treatment. The aim of this study is to suggest an effective solution for their treatment. Column filtration is an adequate method to reduce the pollutant load which makes it possible to have a rate of abatement of 97% and 79% by filtering the pollutant material using sawdust of catia and red sawdust, respectively, with a filter bed height equal to 13 cm. Physicochemical parameters such as chemical oxygen demand, biological oxygen demand, nitrate, ammonia, phosphorus, electrical conductivity and the bacteriological parameters like fecal coliforms, Streptococci, and Staphylococci have been measured. The analysis of heavy metals displays compliance with the World Health Organization standards. The red sawdust and catia sawdust have been characterized by scanning electron microscopy and Fourier-transform infrared spectroscopy.

scholarly journals Proteomic Analysis of Mucopolysaccharidosis IIIB Mouse Brain

Biomolecules ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 355 ◽  
Author(s):  
Valeria De Pasquale ◽  
Michele Costanzo ◽  
Rosa Siciliano ◽  
Maria Mazzeo ◽  
Valeria Pistorio ◽  
...  
Keyword(s):  
Mouse Brain ◽  
Clinical Manifestations ◽  
Label Free ◽  
Wild Type ◽  
Proteomic Approach ◽  
Cytoskeletal Regulation ◽  
Proteome Profile ◽  
Subsequent Storage ◽  
And Control ◽  
Differentially Abundant Proteins

Mucopolysaccharidosis IIIB (MPS IIIB) is an inherited metabolic disease due to deficiency of α-N-Acetylglucosaminidase (NAGLU) enzyme with subsequent storage of undegraded heparan sulfate (HS). The main clinical manifestations of the disease are profound intellectual disability and neurodegeneration. A label-free quantitative proteomic approach was applied to compare the proteome profile of brains from MPS IIIB and control mice to identify altered neuropathological pathways of MPS IIIB. Proteins were identified through a bottom up analysis and 130 were significantly under-represented and 74 over-represented in MPS IIIB mouse brains compared to wild type (WT). Multiple bioinformatic analyses allowed to identify three major clusters of the differentially abundant proteins: proteins involved in cytoskeletal regulation, synaptic vesicle trafficking, and energy metabolism. The proteome profile of NAGLU−/− mouse brain could pave the way for further studies aimed at identifying novel therapeutic targets for the MPS IIIB. Data are available via ProteomeXchange with the identifier PXD017363.

scholarly journals Characterization of Streptococcus thermophilus Host Range Phage Mutants

2006 ◽  
Vol 72 (4) ◽  
pp. 3036-3041 ◽  
Author(s):  
Martin Duplessis ◽  
Céline M. Lévesque ◽  
Sylvain Moineau
Keyword(s):  
Host Range ◽  
Point Mutations ◽  
Streptococcus Thermophilus ◽  
Structural Proteins ◽  
Laser Desorption Ionization ◽  
Wild Type ◽  
Ionization Time ◽  
Host Interactions ◽  
Flight Mass Spectrometry

ABSTRACT To investigate phage-host interactions in Streptococcus thermophilus, a phage-resistant derivative (SMQ-301R) was obtained by challenging a Tn917 library of phage-sensitive strain S. thermophilus SMQ-301 with virulent phage DT1. Mutants of phages DT1 and MD2 capable of infecting SMQ-301 and SMQ-301R were isolated at a frequency of 10−6. Four host range phage mutants were analyzed further and compared to the two wild-type phages. Altogether, three genes (orf15, orf17, and orf18) contained point mutations leading to amino acid substitutions and were responsible for the expanded host range. These three proteins were also identified in both phages by N-terminal sequencing and/or matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. The results suggest that at least three phage structural proteins may be involved in phage-host interactions in S. thermophilus.

scholarly journals Insights into SARS-CoV-2, the Coronavirus Underlying COVID-19: Recent Genomic Data and the Development of Reverse Genetics Systems

2020 ◽  
Vol 101 (10) ◽  
pp. 1021-1024
Author(s):  
Severino Jefferson Ribeiro da Silva ◽  
Renata Pessôa Germano Mendes ◽  
Caroline Targino Alves da Silva ◽  
Alessio Lorusso ◽  
Alain Kohl ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Reverse Genetics ◽  
World Health ◽  
Close Relative ◽  
Sequence Information ◽  
Recombinant Viruses ◽  
Protein Functions ◽  
Genomic Studies ◽  
Health Organization

The emergence and rapid worldwide spread of a novel pandemic of acute respiratory disease – eventually named coronavirus disease 2019 (COVID-19) by the World Health Organization (WHO) – across the human population has raised great concerns. It prompted a mobilization around the globe to study the underlying pathogen, a close relative of severe acute respiratory syndrome coronavirus (SARS-CoV) called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Numerous genome sequences of SARS-CoV-2 are now available and in-depth analyses are advancing. These will allow detailed characterization of sequence and protein functions, including comparative studies. Care should be taken when inferring function from sequence information alone, and reverse genetics systems can be used to unequivocally identify key features. For example, the molecular markers of virulence, host range and transmissibility of SARS-CoV-2 can be compared to those of related viruses in order to shed light on the biology of this emerging pathogen. Here, we summarize some recent insights from genomic studies and strategies for reverse genetics systems to generate recombinant viruses, which will be useful to investigate viral genome properties and evolution.

scholarly journals Mosquito Acetylcholinesterase as a Target for Novel Phenyl-Substituted Carbamates

2019 ◽  
Vol 16 (9) ◽  
pp. 1500
Author(s):  
James M. Mutunga ◽  
Ming Ma ◽  
Qiao-Hong Chen ◽  
Joshua A. Hartsel ◽  
Dawn M. Wong ◽  
...  
Keyword(s):  
Anopheles Gambiae ◽  
Parent Compound ◽  
World Health ◽  
Contact Toxicity ◽  
Ache Inhibitors ◽  
Mosquitocidal Activity ◽  
Health Organization ◽  
Reduced Activity

New insecticides are needed for control of disease-vectoring mosquitoes and this research evaluates the activity of new carbamate acetylcholinesterase (AChE) inhibitors. Biochemical and toxicological characterization of carbamates based on the parent structure of terbam, 3-tert-butylphenyl methylcarbamate, was performed. In vitro enzyme inhibition selectivity (Anopheles gambiae versus human) was assessed by the Ellman assay, as well as the lethality to whole insects by the World Health Organization (WHO) paper contact assay. Bromination at the phenyl C6 position increased inhibitory potency to both AChEs, whereas a 6-iodo substituent led to loss of potency, and both halogenations caused a significant reduction of mosquitocidal activity. Similarly, installation of a hexyl substituent at C6 drastically reduced inhibition of AgAChE, but showed a smaller reduction in the inhibition of hAChE. A series of 4-carboxamido analogs of the parent compound gave reduced activity against AgAChE and generally showed more activity against hAChE than AgAChE. Replacement of the 3-t-buyl group with CF3 resulted in poor anticholinesterase activity, but this compound did have measurable mosquitocidal activity. A series of methyl- and fluoro- analogs of 3-trialkylsilyl compounds were also synthesized, but unfortunately resulted in disappointing activity. Finally, a series of sulfenylated proinsecticides showed poor paper contact toxicity, but one of them had topical activity against adult female Anopheles gambiae. Overall, the analogs prepared here contributed to a better understanding of carbamate structure–activity relationships (SAR), but no new significant leads were generated.

scholarly journals A34 Molecular characterization of Zika virus in Cuba

2019 ◽  
Vol 5 (Supplement_1) ◽  
Author(s):  
P A Martínez ◽  
G Díaz ◽  
E M Castillo ◽  
M Álvarez ◽  
E Santana ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Latin American ◽  
Zika Virus ◽  
Evolutionary Dynamics ◽  
Western Hemisphere ◽  
World Health ◽  
Published Data ◽  
Reference Laboratory ◽  
Health Organization

Abstract Until now, three genotypes of Zika virus (ZIKV) have been detected (two African lineages and one Asian lineage). After the declaration of Public Health Emergency of International Concern issued by The Pan American Health Organization and the World Health Organization authors from some Latin American countries have identified the Asian genotype as the lineage responsible for the Zika epidemic in the western hemisphere. However, data from the Caribbean are sparse, and there is no published data regarding the genotypes that produced isolated outbreaks in Cuba. Aiming to realize the molecular characterization of ZIKV in Cuba, we will sequence by next-generation sequencing the full genome of the ZIKV identified in samples from Cuban patients of different provinces in which ZIKV produced outbreaks. All samples required for this study have been collected during the molecular surveillance of Arboviral diseases conducted at the National Reference Laboratory at Pedro Kourí Tropical Medicine Institute. Viral RNA will be purified from urine and serum samples collected from patients with confirmed ZIKV infection by real time PCR. Using evolutionary dynamics studies, we will map the spread of a virus or of particular variants in time and space in order to understand how frequently ZIKV has been introduced into Cuba. Moreover, we will evaluate the amino acid diversity of each ZIKV proteins. Further, we will evaluate the population dynamics of ZIKV in samples from patients with varying clinical outcomes. The results will allow us to characterize the ZIKV genome and its evolution into the Cuban population that would also have impact for vaccine development, diagnosis, and pathogenesis studies.

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