scholarly journals Virucidal effect of povidone-iodine against SARS-CoV-2 in vitro

2021 ◽  
Vol 49 (12) ◽  
pp. 030006052110636
Author(s):  
Ying Wang ◽  
Yutong Wu ◽  
Qingjing Wang ◽  
Jiajie Zhu ◽  
Wen Shi ◽  
...  
Keyword(s):  
Culture Medium ◽  
Povidone Iodine ◽  
Cell Counting ◽  
Viral Inhibition ◽  
Cytotoxic Effects ◽  
Infected Cells ◽  
Virucidal Effect ◽  
Polymerase Chain ◽  
The Ideal

Objective To evaluate the antiviral activity of the oral disinfectant povidone-iodine (PVP-I) against severe acute respiratory syndrome-coronavirus-2 (SARS-CoV2) in vitro. Methods The cytotoxic effects of PVP-I were determined in Vero and Calu-3 cell lines using that by Cell Counting Kit-8 assay. Viral load in the cell culture medium above infected cells was quantitated using real-time polymerase chain reaction. The cytopathic effect (CPE) and viral infective rate were observed by immunofluorescence microscopy. Results PVP-I at a concentration >0.5 mg/ml in contact with SARS-CoV-2 for 30 s, 1 min, 2 min and 5 min showed up to 99% viral inhibition. For in vitro testing, upon exposure for 1 min, PVP-I showed a virucidal effect. PVP-I had no cytotoxic effects at the range of concentrations tested (0.125–1 mg/ml; CC50 > 2.75 mM) in Vero and Calu-3 cells. Conclusion These results demonstrate that the ideal contact time was 1 min and the optimal concentration was 1 mg/ml, which provides an experimental basis for the use of oral disinfectants in dental hospitals.

scholarly journals Effects of PI3K and FSH on steroidogenesis, viability and embryo development of the cumulus–oocyte complex after in vitro culture

Zygote ◽  
2017 ◽  
Vol 26 (1) ◽  
pp. 50-61 ◽  
Author(s):  
Danielle Kaiser de Souza ◽  
Loise Pedrosa Salles ◽  
Ricardo Camargo ◽  
Laura Vanessa Mourão Gulart ◽  
Suellen Costa e Silva ◽  
...  
Keyword(s):  
Embryo Development ◽  
Culture Medium ◽  
Cumulus Cells ◽  
Expression Levels ◽  
Nuclear Maturation ◽  
Cumulus Oocyte Complex ◽  
Polar Bodies ◽  
17Β Estradiol ◽  
Polymerase Chain

SummaryThe purpose of this study was to evaluate the effects of FSH and PI3K on the nuclear maturation, viability, steroidogenesis and embryo development of bovine cumulus–oocyte complexes (COCs). Oocyte maturation was achieved with MIV B, MIV B+100 µM LY294002, MIV B+10 ng/mL follicle stimulating hormone (FSH), or MIV B+10 ng/mL FSH+100 µM LY294002 treatments for 22–24 h. After the cultured COCs were denuded, oocytes were separated into those that extruded polar bodies (mature) and those that did not, and real-time polymerase chain reaction (PCR) for BAX, BCL2, LHR, FSHR, CYP11A1, CYP19A1 and HSD17B1 genes was performed. The culture medium was collected to determine the levels of 17β-estradiol (E2) and progesterone (P4). The trypan blue test was used to study COC viability, and embryo development was evaluated. FSH increased nuclear maturation and PI3K blocked the maturation but did not influence oocyte viability. BAX and BCL2 expression levels in the cumulus cells were only affected by FSH, and the BAX levels decreased after treatment with LY294002. FSH increased the levels of E2 and P4, however inhibition of PI3K decreased E2 levels. MIV B enhanced levels of LHR, FSHR, CYP11A1, CYP19A1 and HSD17B1, whereas LY294002 inhibited the expression levels of all genes. MIV B+FSH decreased the expression levels of all genes except CYP11A1. LY294002 did not demonstrate any effects in the presence of FSH. Embryo development was significantly decreased when the MIV B+FSH medium was used. In conclusion, FSH controls the steroidogenesis, viability and gene expression in COCs. PI3K plays essential roles in nuclear maturation, steroidogenesis and embryo development.

scholarly journals Long noncoding RNA WEE2-AS1 plays an oncogenic role in glioblastoma by functioning as a molecular sponge for microRNA-520f-3p

2020 ◽  
Author(s):  
Hengzhou Lin ◽  
Dahui Zuo ◽  
Jiabin He ◽  
Tao Ji ◽  
Jianzhong Wang ◽  
...  
Keyword(s):  
Long Noncoding Rna ◽  
Noncoding Rna ◽  
Molecular Mechanisms ◽  
Cell Counting ◽  
Migration And Invasion ◽  
Tumor Diameter ◽  
Polymerase Chain ◽  
Specificity Protein

The long noncoding RNA WEE2 antisense RNA 1 (WEE2-AS1) plays anoncogenic role in hepatocellular carcinoma and triple negative breast cancerprogression. In this study, we investigated the expression and roles of WEE2-AS1 inglioblastoma (GBM). Furthermore, the molecular mechanisms behind the oncogenicactions of WEE2-AS1 in GBM cells were explored in detail. WEE2-AS1 expressionwas detected using quantitative real-time polymerase chain reaction. The roles ofWEE2-AS1 in GBM cells were evaluated by the Cell Counting Kit-8 assay, flowcytometric analysis, and Transwell cell migration and invasion assays, and tumorxenograft experiments. WEE2-AS1 expression was evidently enhanced in GBM tissuesand cell lines compared with their normal counterparts. An increased level of WEE2-AS1 was correlated with the average tumor diameter, Karnofsky Performance Scalescore, and shorter overall survival among GBM patients. Functionally, depleted WEE2-AS1 attenuated GBM cell proliferation, migration, and invasion in vitro, promoted cellapoptosis, and impaired tumor growth in vivo. Mechanistically, WEE2-AS1 functionedas a molecular sponge for microRNA-520f-3p (miR-520f-3p) and consequentlyincreased specificity protein 1 (SP1) expression in GBM cells. A series of recoveryexperiments revealed that the inhibition of miR-520f-3p and upregulation of SP1 couldpartially abrogate the influences of WEE2-AS1 downregulation on GBM cells. Inconclusion, WEE2-AS1 can adsorb miR-520f-3p to increase endogenous SP1expression, thereby facilitating the malignancy of GBM. Therefore, targeting theWEE2-AS1-miR-520f-3p-SP1 pathway might be a promising therapy for themanagement of GBM in the future.

The Effect of Some Natural Cytotoxic Peptides on Tumor Cells

2018 ◽  
Vol 69 (3) ◽  
pp. 597-601
Author(s):  
Bogdan Mihail Diaconescu ◽  
Daniela Jitaru ◽  
Mihaiela Loredana Dragos ◽  
Magda Badescu ◽  
Teodor Stefanache ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Culture Medium ◽  
Actinomycin D ◽  
Tumor Cell Lines ◽  
Cytotoxic Effects ◽  
Cecropin A ◽  
Viability Study ◽  
Tumor Cell Culture ◽  
Natural Peptides

Many studies have highlighted the anti-tumor properties of some natural peptides known to have antimicrobial virtues. In this study, we evaluated the tumoricidal potential of dermaseptin, defensin, cecropin A and B on tumor cell lines: M14K (human mesothelioma). The viability study was performed using PE V Annexin and 7-AAD (7-amino-actinomycin D) (BD Pharmingen). This test was used to detect and measure apoptosis by flow cytometry technique. The experimental results of our study revealed that the cytotoxic effects of the four peptides depend on their concentration. In this in vitro experimental study, we found that the cytotoxic effect of the four cytotoxic peptides used depended on their concentration in the tumor cell culture medium, being significant at concentrations of 120mM and maintained at concentrations of 60�M. At 30�M concentrations these tumoricidal effects were insignificant. Of all the studied peptides, dermaseptin has the most powerful effect and the weakest effect b - defensin - 1.

Effects of testicular interstitial fluid on the proliferation of the mouse spermatogonial stem cells in vitro

Zygote ◽  
2013 ◽  
Vol 22 (3) ◽  
pp. 395-403 ◽  
Author(s):  
Peng Wang ◽  
Yi Zheng ◽  
Ying Li ◽  
Hua Shang ◽  
Guang-Xuan Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Interstitial Fluid ◽  
Culture Medium ◽  
Physiological Role ◽  
Spermatogonial Stem Cells ◽  
Proliferation Rate ◽  
Pcr Analysis ◽  
Polymerase Chain

SummarySpermatogenesis is a process in adult male mammals supported by spermatogonial stem cells (SSCs). The cultivation of SSCs has potential value, for example for the treatment of male infertility or spermatogonial transplantation. Testicular interstitial fluid was added to culture medium to a final concentration of 5, 10, 20, 30 or 40%, in order to investigate its effects on proliferation of mouse SSCs in vitro, Alkaline phosphatase (AKP) assay, reverse transcription polymerase chain reaction (RT-PCR) analysis and indirect immunofluorescence of cells were performed to identify SSCs, and the proliferation rate and diameters of the SSCs colonies were measured. The results showed that the optimal addition of testicular interstitial fluid to culture medium was 30%. When medium supplemented with 30% testicular interstitial fluid was used to culture mouse SSCs, the optimum proliferation rate and diameter of the cell colonies were 72.53% and 249 μm, respectively, after 8 days in culture, values that were significant higher than those found for other groups (P < 0.05). In conclusion, proliferation of mouse SSCs could be promoted significantly by supplementation of the culture medium with 30% testicular interstitial fluid. More research is needed to evaluate and understand the precise physiological role of testicular interstitial fluid during cultivation of SSCs.

scholarly journals Cytotoxic Effects on Gingival Mesenchymal Stromal Cells and Root Surface Modifications Induced by Some Local Antimicrobial Products Used in Periodontitis Treatment

Materials ◽  
2021 ◽  
Vol 14 (17) ◽  
pp. 5049
Author(s):  
Irina Lupșe ◽  
Emoke Pall ◽  
Lucian Barbu Tudoran ◽  
Adriana Elena Bulboacă ◽  
Andreea Ciurea ◽  
...  
Keyword(s):  
Null Hypothesis ◽  
Cell Shape ◽  
Culture Medium ◽  
Root Surface ◽  
Cell Counting ◽  
Cytotoxic Effects ◽  
Commercial Products ◽  
Proliferation Capacity ◽  
Dose Dependent ◽  
Root Surfaces

(1) Background: this study aims to test the cytotoxicity of three antimicrobial products used in periodontitis treatment on gingival mesenchymal stem cells (gMSCs) and their influence on root surfaces and gMSC adhesion. We tested the null hypothesis that the effects of the antimicrobials did not differ. (2) Methods: the commercial products based on sulphonic/sulphuric acids, sodium hypochlorite and silver nanoparticles, in five different concentrations, were added to culture medium for growing gMSCs. Cell proliferation capacity was tested using the Cell Counting Kit-8 (CCK8) and their viability was determined by succinate dehydrogenase activity (MTT) assay. Scanning electron microscopy evaluated the adhesion of gMSCs on root samples treated mechanically and with commercial products. (3) Results: the products induced a dose-dependent cytotoxicity in terms of reduced proliferation and viability of gMSCs, as well as cell shape modifications. Significant differences in CCK8 values between the different commercial products were observed. Based on proliferation tests, the null hypothesis was rejected. When MTT values of the three products were compared with each other, no significant differences were observed for any of the five concentrations (p = 0.065, p = 0.067, p = 0.172, p = 0.256, p = 0.060). (4) Conclusions: the three antimicrobials had a certain degree of cytotoxicity on gMSCs. gMSCs repopulated treated root surfaces.

scholarly journals Synthesis and Screening of Anti-HSV-1 Activity of Thioglucoside Derivatives of Natural Polyhydroxy-1,4-Naphthoquinones

2019 ◽  
Vol 14 (6) ◽  
pp. 1934578X1986067 ◽  
Author(s):  
Sergey G. Polonik ◽  
Natalia V. Krylova ◽  
Galina G. Kompanets ◽  
Olga V. Iunikhina ◽  
Yuri E. Sabutski
Keyword(s):  
Antiviral Activity ◽  
Radical Scavenging ◽  
Effective Inhibition ◽  
Infected Cells ◽  
Virucidal Effect ◽  
Simplex Virus ◽  
Derivatives Of ◽  
Hsv 1

Four 1,4-naphthoquinone dithioglucoside derivatives based on natural polyhydroxy-1,4-naphthoquinones were synthesized. These thioglucosides were screened for their antiradical and antiviral activity in vitro. Antiradical activity of tested compounds was determined by the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay. The anti-herpes simplex virus type 1 (anti-HSV-1) activity of thioglucosides was analyzed by the cytopathic effect inhibition assay and mode of antiviral action was determined by the addition of the tested compounds to uninfected cells, to the virus prior to infection, or to herpes-infected cells. Most effective inhibition of HSV-1 replication was observed at pretreatment of virus by the compounds (direct virucidal effect). The dithioglucoside conjugate with the single β-OH group and lipophilic ethyl substituent in naphthoquinone core showed the greatest antiviral activity.

scholarly journals Human GnRH-secreting cultured neurons express activin betaA subunit mRNA and secrete dimeric activin A

2000 ◽  
pp. 133-138 ◽  
Author(s):  
P Florio ◽  
GB Vannelli ◽  
S Luisi ◽  
T Barni ◽  
R Zonefrati ◽  
...  
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Neuronal Cells ◽  
Activin A ◽  
Human Model ◽  
Subunit Mrna ◽  
Antigenic Properties ◽  
Polymerase Chain ◽  
Different Doses

OBJECTIVE: To evaluate the expression of activin betaA-subunit mRNA and the secretion of activin A in/from cultured GnRH-secreting neuronal cells cloned from human olfactory epithelium (FNC-B4), which showed biochemical and antigenic properties of GnRH-secreting neurons. DESIGN: FNC-B4 cells were cultured in basal and conditioned media. METHODS: Reverse transcription-polymerase chain reaction (RTR-PCR) evaluated the expression of activin betaA-subunit mRNA. By using a specific ELISA, dimeric activin A concentrations were measured in culture media, in the absence or presence of carvone or forskolin and with different doses of progesterone, GnRH, and estradiol. RESULTS: RT-PCR experiments performed on total RNA isolated from FNC-B4 cells, using specific primers for the activin betaA gene, showed a 787bp DNA band corresponding to the betaA gene. FNC-B4 cells secreted activin A, and the highest accumulation in conditioned medium was achieved after 3h culture: the addition of forskolin, but not of carvone, was able to stimulate the release of activin A from cultured neuronal cells (P<0.01). When progesterone or GnRH was added, a significant accumulation of activin A was observed (P<0.01), while estradiol administration did not significantly affect activin A secretion. CONCLUSION: To date, this is the only study, in an in vitro human model reporting, that GnRH-secreting neuronal cells expressed activin betaA-subunit mRNA, and released dimeric activin A in culture medium. The expression and secretion of activin suggests that in these cells activin A might exert its action by autocrine/paracrine mechanisms.

scholarly journals Validation d'une méthode automatisée d'extraction d'acides nucléiques et d'analyse en RT-qPCR : exemple du virus de la fièvre catarrhale ovine

2009 ◽  
Vol 62 (2-4) ◽  
pp. 122
Author(s):  
Kris De Clerq ◽  
Yves Van der Stede ◽  
Franck Vandenbussche
Keyword(s):  
In Silico ◽  
Bluetongue Virus ◽  
Western Europe ◽  
Amplification Factor ◽  
Analytical Sensitivity ◽  
Pcr Efficiency ◽  
Field Samples ◽  
Polymerase Chain ◽  
The Ideal

The control of bluetongue virus (BTV) in Central-Western Europe is greatly complicated by the coexistence of several BTV sero­types. Rapid, sensitive and specific assays are therefore needed to identify correctly the currently circulating BTV serotypes in field samples. In the present study, four serotype-specific real-time reverse-transcription quantitative polymerase chain reac­tion assays (RT-qPCR) are described for the detection of BTV-1, 6, 8 and 11. The analytical sensitivity of BTV-1/S2, BTV-6/S2, BTV-8/S2 and BTV-11/S2 serotype-specific RT-qPCR assays is comparable to the earlier described serogroup-specific pan- BTV/S5 RT-qPCR assay. In silico and in vitro analyses indicated that none of the assays cross-reacted with viruses which were symptomatically or genetically related to BTV and only detected the intended BTV serotypes. All assays exhibited a linear range of at least 0.05-3.80 log10 TCID50/mL and a PCR-efficiency approaching the ideal amplification factor of two per PCR cycle. Both intra- and inter-run variations were low with a total coef­ficient of variation of 1-2% for clear positive samples, and below 10% for very weak positive samples. Finally, the performance of the described assays was compared with commercially available kits for the detection of BTV-1, 6 and 8. Three in-house assays gave the same diagnostic results (positive/negative) as the com­mercial assays and can thus be used interchangeably. Together with the earlier-described serogroup-specific pan-BTV/S5, the serotype-specific RT-qPCR assays form a flexible and properly validated set of tools to detect and differentiate the BTV sero­types currently circulating in Central-Western Europe.

scholarly journals Receptiveness of the stigma and in vitro germination of orange pollen, submitted to different temperatures

2004 ◽  
Vol 28 (5) ◽  
pp. 1087-1091 ◽  
Author(s):  
Leila Aparecida Salles Pio ◽  
José Darlan Ramos ◽  
Moacir Pasqual ◽  
Flavia Carvalho Santos ◽  
Keize Pereira Junqueira
Keyword(s):  
Culture Medium ◽  
Calcium Nitrate ◽  
Pollen Grain ◽  
Effect Of Temperature ◽  
In Vitro Germination ◽  
Stigma Receptivity ◽  
Open Flower ◽  
Different Temperatures ◽  
The Ideal

The study was carried out in order to evaluate the effect of temperature and in vitor stigma receptivity on regeneration of orange cultivar (Valência, Pêra and Natal) pollen. Two experiments were carried out, in the first the ideal temperature of germination was assessed. Pollen was obtained from flowers at the balloon stage and inoculated in culture medium with 10 g L-1 agar, 200 mg L-1 boric acid, 100 g L-1 sucrose, 800 mg L-1 calcium nitrate, pH adjusted to 6,5 and incubated in a BOD chamber at temperatures of 23, 24, 25, 26 and 27ºC during a 24-hour photoperiod. After 12 hours of incubation, the best temperature for pollen grain germination was 25ºC for all varieties. In a second experiment, in order to test the receptivity of the stigma, flowers were collected at different flowering stages: small bud, balloon and open flower. The stigmas were by immersion exposed to hydrogen peroxide (perioxidase reaction), 3% for 3 minutes. Through the technique of Zeisler (1938), better results were detected at the balloon stage with 80 to 100% receptivity, while for the open flower the receptivity presented maximum indexes.

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