Evaluation of saliva and nasopharyngeal swab sampling for genomic detection of SARS-CoV-2 in children accessing a pediatric emergency department during the second pandemic wave

SARS-CoV-2 infection is mainly detected by multiplex real-time RT-PCR from upper respiratory specimens, which is considered the gold-standard technique for SARS-CoV-2 infection diagnosis. A nasopharyngeal (NP) swab represents the clinical sample of choice, but NP swabbing can be uncomfortable to the patients, especially for pediatric-age participants, requires trained healthcare personnel, and may generate an aerosol, increasing the intrinsic exposure risk of healthcare workers. The objective of this study was to compare paired NP and saliva samples (SS) collected from pediatric patients to evaluate whether the saliva collection procedure may be considered a valuable alternative to the classical NP swab (NPS) sampling in children. In this study, we describe a SARS-CoV-2 multiplex real-time RT-PCR protocol for SS, comparing the results with the paired NPS specimens from 256 pediatric patients (mean age 4.24 ± 4.40 years) admitted to the hospital emergency room of Azienda Ospedaliera Universitaria Integrata (AOUI), Verona, and randomly enrolled between September 2020 and December 2020. The saliva sampling demonstrated consistent results when compared to NPS use. The SARS-CoV-2 genome was detected in 16 out of 256 (6.25%) NP samples, among which 13 (5.07%) were positive even when paired SS were analyzed. Moreover, SARS-CoV-2-negative NPS and SS were consistent, and the overall concordances between NPS and SS were detected in 253 out of 256 samples (98.83%). Our results suggest that saliva samples may be considered a valuable alternative to NPS for SARS-CoV-2 direct diagnosis with multiplex real-time RT-PCR in pediatric patients

Introductory Chapter: Malaria in 2022 – Promises and Unmet Needs

In this chapter were described: biological cycle of Plasmodia and diagnostic assays for malaria: microscopic examination, Rapid Diagnostic Test (RDT) based on antigen, Indirect Fluorescent Antibody Test (IFA test), and also, recently introduced and accepted, the nucleic-acid detection method by PCR or LAMP technologie

Different patterns of HIV-1 DNA after therapy discontinuation

Background: By persisting in infected cells for a long period of time, proviral HIV-1 DNA can represent an alternative viral marker to RNA viral load during the follow-up of HIV-1 infected individuals. In the present study sequential blood samples of 10 patients under antiretroviral treatment from 1997 with two NRTIs, who refused to continue any antiviral regimen, were analyzed for 16-24 weeks to study the possible relationship between DNA and RNA viral load. Methods: The amount of proviral DNA was quantified by SYBR green real-time PCR in peripheral blood mononuclear cells from a selected group of ten patients with different levels of plasmatic viremia (RNA viral load). Results: Variable levels of proviral DNA were found without any significant correlation between proviral load and plasma HIV-1 RNA levels. Results obtained showed an increase or a rebound in viral DNA in most patients, suggesting that the absence of therapy reflects an increase and/or a persistence of cells containing viral DNA. Conclusion: Even though plasma HIV RNA levels remain the basic parameter to monitor the intensity of viral replication, the results obtained seem to indicate that DNA levels could represent an adjunct prognostic marker in monitoring HIV-1 infected subjects

Prevalence of and short-term changes in conjunctival manifestations among patients with SARS-CoV-2 infection

[no abstract available

PI and OPG/RANKL levels in human osteoblast cells

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Oral microbiota in oropharyngeal cancers: friend or foe?

Oral microbiome is a complex population of micro-organisms, which by cross-talking with the local immune system, plays a major role in the immune homeostasis of the oral cavity, further contributing in the physiology of the gastro-intestinal microbiota. Understanding their involvement in the onset and pathogenesis of oropharyngeal cancers is paramount, despite very few reports deal with the fundamental role exerted by oral microbiota disorders, such as dysbiosis and impairment in the oral microbiome composition as causative factors in the development of oropharyngeal tumors. Current research, via metabolomic or meta-transcriptomic analyses, is wondering how this complex microbial population regulates the immune homeostasis in oral and pharyngeal mucosa and whether changes in bacterial composition may give insights on the role of oral microbiome in the development of oropharyngeal tumors, so to prevent their occurrence

Analysis of HIV-1 drug resistant mutations by line probe assay and direct sequencing in a cohort of therapy naive HIV-1 infected Italian patients

BACKGROUND: The routine determination of drug resistance in newly HIV-1 infected individuals documents a potential increase in the transmission of drug-resistant variants. Plasma samples from twenty seven therapy naive HIV-1 infected Italian patients were analyzed by the line probe assay (LIPA) and the TruGene HIV-1 assay for the detection of mutations conferring resistance to HIV-1. RESULTS: Both tests disclosed amino-acid substitutions associated with resistance in a variable number of patients. In particular, two mutations (K70R and V118I), detectable by LIPA and by sequencing analysis respectively, revealed resistance to NRTIs in two plasma samples. At least three mutations conferring resistance to NNRTIs, not detectable by commercial LIPA, able to reveal mutations associated only with nucleoside reverse transcriptase analogues, were disclosed by viral sequence analysis. Moreover, most samples showed mutations correlated with resistance to protease inhibitors. Remarkably, a key mutation, like V82A (found as a mixture), and some "indeterminate" results (9 samples), due the absence of signal on the lines corresponding to a specific probe, was revealed only by LIPA, while a variable number of secondary mutations was detectable only by TruGene HIV-1 assay. CONCLUSION: Even if further studies are necessary to establish the impact of different tests on the evaluation of drug-resistant strains transmission, LIPA might be useful in a wide population analysis, where bulk results are needed in a short time, while sequencing analysis, able to detect mutations conferring resistance to both NRTIs and NNRTIs, might be considered a more complete assay, albeit more expensive and more technically complex

Serum antibody reactivity to human intracisternal A-type particle retrovirus proteins in systemic sclerosis patients.

Serum antibodies against human intracisternal A-typeparticle (HIAP) endogenous retrovirus have been foundto be associated with various autoimmune pathologies.To evaluate the presence of serum antibody reactivityto HIAP proteins in systemic sclerosis, a Western blotanalysis was performed on sera from 42 patients withsystemic sclerosis, in comparison with 18 sera frompatients with primary biliary cirrhosis and 52 healthysubjects. A positive Western blot was found in 55.5% ofserum samples from patients with primary biliarycirrhosis and in 66.0% of patients with systemic sclerosis.None of the 52 healthy subjects showed positive results.Although this difference may be attributable either to anautoimmune response to antigenically related cellularproteins or to a specific antibody response to HIAPproteins expressed as an incidental consequence ofattendant pathological processes, the high prevalence ofantibodies against HIAP proteins demonstrated inpatients with systemic sclerosis may be considered ahallmark of this disease.(Accepted November 10, 2003.)Acta Derm Venereol 2004; 84: 177–180.Dr Michelangelo La Placa, Department of Clinicaland Experimental Medicine, Section of Dermatology,Via Massarenti, 1, 40138 Bologna, Italy. E-mail:[email protected] sclerosis (SSc) is a connective tissue diseasecharacterized by excessive deposition of collagen inthe skin and various internal organs, and by vascularabnormalities (1). SSc is considered to be an auto-immune disease. Although both cellular microchimer-ism (2) and molecular mimicry of some commoninfectious agents, such as cytomegalovirus and parvo-virus B19 (3), have been implicated in its pathogenesis,the aetiology of SSc remains unknown.Several publications have described the presence ofretroviral sequences associated with virions, producedby cells of patients with autoimmune diseases. In recentreviews (4, 5), these viruses, identified as human endo-genous retroviruses (HERVs), have been associatedwith Sjo¨grens's syndrome, type 1 or insulin-dependentdiabetes mellitus, multiple sclerosis, rheumatoid arthri-tis, congenital heart block, systemic lupus erythemato-sus and SSc. Serum antibodies specific for humanintracisternal A-type particles (HIAP), a HERV recog-nized by monoclonal antibody against HIV-1 p24capsid protein (6), have been found in primary biliarycirrhosis (PBC) (7), familial erosive arthritis (8) andsome patients with SSc, systemic lupus erythematosus,Still's disease and idiopathic T-lymphocytopenia (9,10).To further investigate serum antibody reactivity toHIAP proteins in SSc, we performed a Western blotanalysis of a substantial number of sera from SScpatients, in comparison with sera from PBC patientsand healthy subjects.MATERIALS AND METHOD

TiO2-Ag-NP adhesive photocatalytic films able to disinfect living indoor spaces with a straightforward approach

TiO2-Ag doped nanoparticulate (TiO2-Ag-NP) adhesive photocatalytic films were used to assess the ability in dropping down the burden of indoor microbial particles. The application of an easy-to use photocatalytic adhesive film to cleanse indoor living spaces from microbial pollution, represents a novelty in the field of photocatalytic devices. Reduction was attained by photocatalysis in selected spaces, usually with overcrowding (≥ 3 individuals) in the common working daily hours, and upon indoor microclimate monitoring. TiO2-Ag doped nanoparticulate (TiO2-Ag-NP) adhesive photocatalytic films were applied within five types of living spaces, including schools and job places. The microbial pollution was assessed at time 0 (far from routine clean, ≥ 9 h) and throughout 2-4 weeks following the photocatalyst application by relative light unit (RLU) luminometry and microbial indirect assessment (colony forming units per cubic meter, CFU/m3). TiO2-Ag-NP photocatalyst reduced RLU and CFU/m3 by rates higher than 70% leading to RLU ≤ 20 and microbial presence ≤ 35 CFU/m3. The described TiO2-Ag-NP is able to reduce microbial pollution to the lowest RLU threshold (≤ 20) within 60 min in open daylight in a standardized test room of 100 m2. The correlation between RLU and CFU/m3 was positive (r = 0.5545, p < 0.05), assessing that the microbial reduction of indoor areas by the TiO2-Ag-NP adhesive film was real. Titania photocatalysts represent promising tools to ensure air cleaning and sanitization in living indoor microclimates with a low cost, feasible and straightforward approach. This approach represents an easy to handle, cost effective, feasible and efficacious approach to reduce microbial pollution in indoor spaces, by simply attaching a TiO2-Ag-NP adhesive film on the wall