No abstract available.
Paramyxoviridae Infections* ; Polymerase Chain Reaction* ; Reverse Transcription*

Laccase activity of Pleurotus ostreatus is significantly increased by the addition of apple pomace. Among various conditions, the best concentration of apple pomace and cultivation time for the production of laccase by P. ostreatus was 2.5% and 9 days, respectively. Reverse transcription polymerase chain reaction analyses of laccase isoenzyme genes, including pox1, pox3, pox4, poxc, poxa3, and poxa1b, revealed a clear effect of apple pomace on transcription induction. Our findings reveal that the use of apple pomace can be a model for the valuable addition of similar wastes and for the development of a solid-state fermenter and commercial production of oyster mushroom P. ostreatus.
Fungi* ; Laccase* ; Pleurotus* ; Polymerase Chain Reaction ; Reverse Transcription

Objective. To determine the diagnostic accuracy of self-collected snorted and spit saliva in detecting COVID-19 using RT-PCR (ssRT-PCR) and lateral flow antigen test (ssLFA) versus nasopharyngeal swab RT-PCR (npRT-PCR). Methods. One hundred ninety-seven symptomatic subjects for COVID-19 testing in a tertiary hospital underwent snort-spit saliva self-collection for RT-PCR and antigen testing and nasopharyngeal swab for RT-PCR as reference. Positivity rates, agreement, sensitivity, specificity, and likelihood ratios were estimated. Results. Estimated prevalence of COVID-19 using npRT-PCR was 9% (exact 95% CI of 5.5% - 14.1%). A higher positivity rate of 13% in the ssRT-PCR assay suggested possible higher viral RNA in the snort-spit samples. There was 92.9% agreement between ssRT-PCR and npRT-PCR (exact 95% CI of 88.4% to 96.1%; Cohen’s Kappa of 0.6435). If npRT-PCR will be assumed as reference standard, the estimated Sensitivity was 83.3% (exact 95% CI of 60.8% to 94.2%), Specificity 93.9% (exact 95% CI of 89.3% to 96.5%), Positive predictive value of 57.7% (exact 95% CI of 38.9% to 74.5%), Negative predictive value of 98.2% (exact 95% CI of 95% to 99.4%), positive likelihood ratio of 3.65 (95% CI of 7.37 to 24.9), negative likelihood ratio of 0.178 (95% CI of 0.063 to 0.499). There was 84.84% agreement (95% exact CI of 79.1% to 89.5%; Cohen’s Kappa of 0.2356) between ssLFAvs npRT-PCR, sensitivity of 38.9% (exact 95% CI of 20.3% to 61.4%), specificity of 89.4% (exact 95% CI of 84.1% to 93.1%), PPV of 26.9% (95% CI of 13.7% to 46.1%), NPV of 93.6% (exact 95% CI of 88.8% to 96.4%), LR+ of 3.67 (95% CI of 1.79 - 7.51), LR – of 0.68 (95% CI of 0.47 - 0.99). Conclusion. Our data showed that snort-spit saliva RT-PCR testing had acceptable diagnostic performance characteristics and can potentially be used as an alternative to the standard nasopharyngeal/oropharyngeal swab RT-PCR test for COVID-19 in certain situations. However, our data also showed that snort-spit saliva antigen testing using lateral flow assay did not offer acceptable performance.
Saliva ; SARS-CoV-2 ; Reverse Transcription ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUND: Kruppel-like factor 4 (KLF4) is a transcription factor that regulates a diverse array of cellular processes, including development, differentiation, proliferation, and apoptosis. Although its function in keratinocytes has been widely studied, its exact role in psoriasis has not been elucidated. OBJECTIVE: We designed this study to investigate epidermal expression levels of KLF4 and the change in KLF4 expression after treatment in patients with psoriasis. METHODS: We compared the expression levels of KLF4 in the basal, suprabasal, and superficial epidermal layers, in psoriatic lesional, non-lesional, and normal skin, using an immunoreactivity intensity distribution index (IRIDI). In addition, we measured the change in KLF4 expression on the basis of the IRIDI and by reverse transcription polymerase chain reaction (RT-PCR) analysis after treatment. RESULTS: The combined IRIDI scores in psoriatic lesional skin were significantly higher than the scores in both non-lesional and normal skin. The psoriatic epidermis, particularly the suprabasal layer, showed a significantly increased IRIDI score compared to that of non-lesional and normal skin, which was significantly decreased after treatment. RT-PCR analysis exhibited a slight increase in KLF4 mRNA expression level after treatment; however, this increase was not significant. CONCLUSION: These data indicate that KLF4 could regulate epidermal proliferation and differentiation. Moreover, we believe that KLF4 may play an important role in the physiological reaction to counteract abnormal differentiation and proliferation of keratinocytes.
Apoptosis ; Epidermis ; Humans ; Keratinocytes ; Polymerase Chain Reaction ; Psoriasis* ; Reverse Transcription ; RNA, Messenger ; Skin ; Transcription Factors

This study was purposed to investigate the expression of the growth-factor independence 1 (GFI1) in patients with leukemia and its clinical significance. Bone marrow mononuclear cells were obtained from 65 newly diagnosed leukemia patients including 24 acute myeloid leukemia (AML), 18 chronic myelogenous leukemia (CML), 6 acute lymphoblastic leukemia (ALL), 17 blast crisis of chronic myelogenous leukemia and 13 patients with iron deficiency anemia (IDA) were used as controls. The relative expression of gene gfi1 was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and taqman quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR). The results showed that gene expression of gene gfi1 in leukemia patients was obviously higher than that in controls and the difference was statistically significant (p < 0.01), in which the expression of gene gfi1 in newly diagnosed CML patients was higher than that in newly diagnosed AML, newly diagnosed ALL, CML-BCP patients and the difference was significant (p < 0.01). Expression of gene gfi1 in lymphocytic blast crisis of CML was higher than that in nonlymphocytic blast crisis of CML, and the difference was significant. It is concluded that gene gfi1 may play an important role in leukemia, especially in CML incidence and progression. The high level expression of gene gfi1 may be participate in the development of lymphoma.
DNA-Binding Proteins ; genetics ; Gene Expression ; Humans ; Leukemia ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics ; Transcription, Genetic ; genetics

BACKGROUND: Norovirus is a leading cause of epidemic and sporadic acute gastroenteritis worldwide. Because of the rapid transmission of the virus, early detection is important to prevent outbreak of norovirus infection. To evaluate the performance of a newly introduced rapid antigen test for detecting human norovirus in stool specimens, we compared it with the established ELISA test and real time quantitative reverse transcription PCR (qRT-PCR). METHODS: One hundred and eighty-four stool samples were analyzed by rapid antigen test (Denka-Seiken, Japan), ELISA (R-Biopharm, Germany), and qRT-PCR (R-Biopharm, Germany). Overall percent agreement, percent positive agreement (PPA), and percent negative agreement (NPA) of the rapid antigen test in comparison with ELISA and qRT-PCR were obtained. RESULTS: Positive rates of rapid antigen test, ELISA, and qRT-PCR were 44.0% (81/184), 51.6% (95/184), and 42.9% (79/184), respectively. Seventy samples (38.0%) showed all positive, and 86 samples (46.7%) showed all negative results by three methods. Overall percent agreement of three methods was 84.8% (156/184). Overall percent agreement, PPA, and NPA of the rapid antigen test in comparison with qRT-PCR were 89.1%, 88.6%, and 89.5%, respectively, and those of the rapid antigen test in comparison with ELISA were 90.2%, 83.2%, and 97.8%, respectively. Total procedure of the rapid antigen test was finished within 20 min. CONCLUSIONS: Rapid antigen test was easier and quicker to perform, and showed high agreement rates with ELISA and qRT-PCR. This test may be useful for rapid screening of norovirus infection.
Enzyme-Linked Immunosorbent Assay ; Gastroenteritis ; Humans ; Mass Screening ; Norovirus ; Polymerase Chain Reaction ; Reverse Transcription ; Viruses

OBJECTIVE: To overcome the limitations of the single gene transfer, the authors present the results of wild-type p16 and p53 combined genes transfer in vitro to the U251MG and U373MG cell lines using cationic liposome as a vector. METHODS: To compare the therapeutic effect of the combined p16 and p53 genes transfer with the single p16 and p53 gene transfer, full length of wild-type human p16 and p53 gene, and combined p16-p53 genes were transferred in vitro to the U251MG and U373MG cell lines using cationic liposome as a vector. As the U251MG and U373MG cell lines are devoid of p16 and p53 genes, the therapeutic effect of the three groups of gene transfer could be evaluated by the growth suppression or percentage of the viable cells. Reverse transcription polymerase chain reaction(RT-PCR), flow cytometry, and electron microscopy(EM) were used for evaluation of the growth suppression or apoptosis of the tumor cells. RESULTS: p16 gene, p53 gene and the combined p16-p53 genes were effectively transferred to the cell lines using cationic liposome as a vector resulting in dramatic decrease of the viable tumor cells in comparison to the control group(p=0.004). The cytotoxic effect of the gene transfer in the U251MG cell line was the most significant in the combined p16-p53 group. However, in the U373MG cell line p53 single gene transfer group showed more significant effect than the combined gene transfer group. Apoptosis was confirmed by EM in the combined p16-p53 genes group. The G1 phase arrest effect, confirmed by the flow cytometry was more prevalent in the p16 gene transfer group than the other groups. CONCLUSION: Cationic liposome-mediated transfer of combined p16-p53 genes to the human glioblastoma cell lines is proven effective. However, the therapeutic effect of the combined p16-p53 genes transfer was not consistently superior to the single p16 or p53 gene transfer.
Apoptosis ; Cell Line* ; Flow Cytometry ; G1 Phase ; Genes, p16 ; Genes, p53* ; Glioblastoma* ; Humans* ; Liposomes ; Reverse Transcription

PURPOSE: Dysregulation of apoptosis may attribute to development of cancer by abnormally prolonging cell viability with accumulation of transforming mutations. Survivin and HIAP (Human Inhibitors of Apoptosis)-1 were recently described as apoptosis inhibitors. Their pathogenic roles in gastric cancer are largely unknown. In the present study, we examined the expression of survivin and HIAP-1 in gastric cancer tissues and cell lines in order to elucidate the roles of survivin and HIAP-1 in the process of gastric carcinogenesis. MATENRIALS AND METHODS: Eight gastric cancer cell lines and five gastric cancer tissues were studied. The expression of survivin and HIAP-1 were evaluated by reverse transcription -polymerase chain reaction (RT-PCR), immunohistochemistry, and Western blot. RESULTS: Western blot and RT-PCR analysis revealed survivin and HIAP-1 expression in all gastric cancer cell lines. Increased expression of survivin and HIAP-1 were found in all cases of gastric cancer tissues compared to normal tissues by Western blot analysis. In immunohistochemical analysis tumor cells were stained with anti-survivin and anti-HIAP-1 antibodies. Cell cycle dependence of survivin expression was preserved in gastric cancer cell lines. CONCLUSION: The results indicate that increased expression of survivin and HIAP-1 genes may play an important role in gastric cancer.
Antibodies ; Apoptosis ; Blotting, Western ; Carcinogenesis ; Cell Cycle ; Cell Line ; Cell Survival ; Immunohistochemistry ; Reverse Transcription ; Stomach Neoplasms*

A retron is a bacterial retroelement that encodes an RNA gene and a reverse transcriptase (RT). The former, once transcribed, works as a template primer for reverse transcription by the latter. The resulting DNA is covalently linked to the upstream part of the RNA; this chimera is called multicopy single-stranded DNA (msDNA), which is extrachromosomal DNA found in many bacterial species. Based on the conserved features in the eight known msDNA sequences, we developed a detection method and applied it to scan National Center for Biotechnology Information (NCBI) RefSeq bacterial genome sequences. Among 16,844 bacterial sequences possessing a retron-type RT domain, we identified 48 unique types of msDNA. Currently, the biological role of msDNA is not well understood. Our work will be a useful tool in studying the distribution, evolution, and physiological role of msDNA.
Biotechnology ; Chimera ; DNA ; DNA, Single-Stranded* ; Genome, Bacterial* ; Retroelements ; Reverse Transcription ; RNA ; RNA-Directed DNA Polymerase

Here, we describe a uracil-DNA glycosylase (UNG)-treated reverse transcription loop-mediated isothermal amplification (uRT-LAMP) for the visual detection of all subtypes of avian influenza A virus (AIV). The uRT-LAMP assay can prevent unwanted amplification by carryover contamination of the previously amplified DNA, although the detection limit of the uRT-LAMP assay is 10-fold lower than that of the RT-LAMP without a UNG treatment. To the best of our knowledge, this is the first successful application of deoxyuridine triphosphate/UNG strategy in RT-LAMP for AIV detection, and the assay can be applied for the rapid, and reliable diagnosis of AIVs, even in contaminated samples.
Animals ; Deoxyuridine ; Diagnosis ; DNA ; Influenza in Birds* ; Limit of Detection ; Reverse Transcription* ; Uracil-DNA Glycosidase