Monkeypox (Mpox) virus isolation and ultrastructural characterisation from a Brazilian human sample case

BACKGROUND According to the last 2023 Monkeypox (Mpox) Outbreak Global Map from the Centres for Disease Control and Prevention (CDC), more than 100 countries with no Mpox infection report cases. Brazil stands out in this group and is the second country with the highest number of cases in the last outbreak. OBJECTIVE To contribute to knowledge of the virus infection effects in a cellular model, which is important for diagnosis infections not yet included in a provider´s differential diagnosis and for developing viral inhibition strategies. METHODS We describe a virus isolation protocol for a human clinical sample from a patient from Brazil, the viral growth in a cell model through plaque forming units (PFU) assay, reverse transcriptase polymerase chain reaction (RT-PCR) and transmission electron microscopy (TEM). FINDINGS We follow the viral isolation in Vero cell culture from a Mpox positive clinically diagnosed sample and show the infection effects on cellular structures using a TEM. MAIN CONCLUSIONS Understanding the impact of viral growth on cellular structures and its replication kinetics may offer better strategies for the development of new drugs with antiviral properties.

Municipal solid waste landfill leachate induced cytotoxicity in root tips of Vicia faba: Environmental Risk posed by non-engineered landfill

Landfills are considered the main option for dumping of municipal solid waste (MSW) all over the world, but these landfills are mostly non-engineered. The decomposition of solid waste in the landfill and rainwater penetration into the decomposing waste produces leachate that contains dissolved organic and inorganic compounds, heavy metals, suspended particles, and hazardous substances. Leachate migration in the environment may pose serious health risks to organisms exposed. Hence, the present study explored the cytotoxic potential of landfill leachate collected in different seasons from the Okhla landfill site, Delhi, India. Cytotoxicity of leachate samples was evaluatedby cell apoptosis and ultrastructural observation based on Transmission Electron Microscopy (TEM) of the cells of root tips of vicia faba seedlings treated with the leachates collected in summer, winter and monsoon in a time and dose dependent manner. Leachate collected in all the three seasons induced apoptosis in cells of root tips of vicia faba that increased in a time and dose dependent manner when compared to control. The apoptosis was highest in the samples treated with leachate collected in the summer season, followed by winter and monsoon. It was further confirmed with TEM images that there was induction of apoptotic-like morphological changes in the root cells treated with landfill leachate when compared with the control. The present study indicates that municipal solid waste leachate is very toxic and it should be treated before disposing it to the environment.

Electron Microscopy of the Separated Outer Tegument of the Sparganum and Its Antigenicity

The author reported previously on separation of the outer tegument of the spargana (plerocercoids of Spirometra mansoni) using high concentration of urea solution. To determine which layer of the tegument is separated by this method, an electron microscopic analysis has been processed in this study. It was confirmed that the basement layer of the tegument is separated from the parenchyme of the sparganum. In addition, the antigenicity of the separated outer tegument against the human sparganosis patient sera was evaluated. Numerous antigenic proteins, including 16 and 55 kDa proteins, were noticed in the separated tegument; however, there were no diagnostic 31/36 kDa molecules in this tegument. The molecules reactive with the patient sera in the tegument are to be characterized in future studies.

Ultrastructure observation of biological living skin equivalent / 实用口腔医学杂志

Objective:To observe the ultrastructure of biological living skin equivalent in vitro.Methods:Keratinocytes and fibroblasts were sourced from the back skin of legally aborted human foetus.Fibroblasts were seeded into bovine type I collagen gel and cultured for 3 days.Keratinocytes were seeded on the surface of collagen gel and cultured for another 2 days,then the equivalent skin was exposed to air-liquid interface to form a protective cornified layer.Histological structure and ultrastructure of the equivalent skin were observed by light microscope and transmission electron microscope.Results:The morphogy of equivalent skin of full thickness consisted of epithlium and dermis.Epithelium was made up of stratum basale,stratum spinosum,stratum granulosum and stratum corneum.lntercellular bridge existed within the cells of different layers.Some ultrastructures in epithelium such as desmosome,Lamelle,tonofibrils and lipid droplet were found in the equivalent skin.Conclusion:Tissue-engineered skin may have the features of natural skin.