Chronic obstructive pulmonary disease (COPD) is a progressive disease characterized by irreversible airflow limitation, airway inflammation and remodeling, and enlargement of alveolar spaces. COPD is in the top five leading causes of deaths worldwide and presents a high economic cost. However, there are some preventive measures to lower the risk of developing COPD. Low-level laser therapy (LLLT) is a new effective therapy, with very low cost and no side effects. So, our objective was to investigate if LLLT reduces pulmonary alterations in an experimental model of COPD. C57BL/6 mice were submitted to cigarette smoke for 75 days (2x/day). After 60 days to smoke exposure, the treated group was submitted to LLLT (diode laser, 660 nm, 30 mW, and 3 J/cm2) for 15 days and euthanized for morphologic and functional analysis of the lungs. Our results showed that LLLT significantly reduced the number of inflammatory cells and the proinflammatory cytokine secretion such as IL-1β, IL-6, and TNF-α in bronchoalveolar lavage fluid (BALF). We also observed that LLLT decreased collagen deposition as well as the expression of purinergic P2X7 receptor. On the other hand, LLLT increased the IL-10 release. Thus, LLLT can be pointed as a promising therapeutic approach for lung inflammatory diseases as COPD.
The gold standard for the laboratory diagnosis of COVID-19 is the reverse
transcription quantitative real-time polymerase chain reaction (RT-qPCR) assay,
which searches for SARS-CoV-2 target genes in nasopharyngeal/oropharyngeal
(NP/OP) samples, and its performance depends on the quantity and quality of the
RNA input. This study compared the performance and cost-effectiveness of three
different kits/reagents for RNA extraction used in COVID-19 diagnosis in Sao
Paulo, Brazil. A total of 300 NP/OP samples belonging to suspected cases of
COVID-19 stored in a biorepository were randomly selected, and RNA was extracted
using (i) automated extraction (Loccus, Extracta Kit FAST), (ii) manual
extraction (BioGene Kit, Bioclin, Quibasa), and (iii) quick extraction methods
(Lucigen, Quick DNA Extract Kit). Next, the samples were tested using RT-qPCR
for SARS-CoV-2 with the Allplex 2019-nCoV modified assay and the Charité-Berlin
protocol. All assays/kits were used according to the manufacturer’s
instructions. For the Allplex kit, the sensitivity in detecting SARS-CoV-2 with
previously extracted RNA by different procedures was 100.0% for Loccus, 100.0%
for BioGene and 91.9% for Quick. Using the Charité-Berlin protocol, the
sensitivities were 81.4% for Loccus, 81.2% for BioGene and 60.7% for Quick. The
least sensitive target gene and the gene most affected by RNA extraction
procedures was the RNA-dependent RNA polymerase gene (Charité-Berlin protocol).
No false-positive SARS-CoV-2 results were detected using RNA obtained from any
of the different protocols. In conclusion, Loccus and BioGene RNA extractions
were efficient for RT-qPCR assays, and although the BioGene procedure is less
expensive, Loccus is the best choice because it allows the rapid handling of
hundreds or thousands of samples, a desirable feature during pandemics. Although
less sensitive, the Quick extraction is useful during outbreaks coupled with the
Allplex amplification kit for SARS-CoV-2 diagnosis (κ = 0.925).
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