Human Laryngeal Epidermoid Carcinoma Cells Research Articles

Effectiveness of plant-based hand sanitizer incorporating Quercus infectoria gall extract.

Quercus infectoria (Qi), a traditional herbal plant with broad-spectrum of activities on multidrug-resistant bacteria has been develop as hand sanitizer applications. Antimicrobial activity was evaluated using agar-well diffusion and broth microdilution method. Bactericidal activity was determined following the European Standard 1276 antibacterial suspension test. Neutralization assay was performed to assess anti-respiratory syncytial virus. Safety, stability, and skin permeation of Qi hand gel was investigated. Qi hand sanitizer gel inhibited microorganisms ranging from 99.9-99.999% against Enterococcus faecalis, Staphylococcus aureus, methicillin-resistant S. aureus, S. epidermidis, S. pseudintermedius, S. saprophyticus, Streptococcus pyogenes, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Candida albicans. A significant reduction in main human dermatophytes including Microsporum canis, Microsporum gypseum, and Talaromyces marneffei of approximately 50% was observed (p<0.05). Qi hand sanitizer gel inactivated more than 99% viral particles entering human laryngeal epidermoid carcinoma cells in a dose-dependent manner. Scanning electron micrographs further illustrated that Qi hand sanitizer gel disrupted microbial cell membrane after 1-min contact time resulting in cell death. Qi hand sanitizer gel delivered emollient compounds through simulated human skin layers and showed no cytotoxicity on fibroblast cells. Moreover, Qi hand sanitizer gel demonstrated stability under extreme conditions. Qi hand sanitizer gel was able to inhibit various microorganisms including bacteria, dermatophytes, and virus.

Cytotoxic effects of water and ethanolic extracts of Turkish propolis on human laryngeal epidermoid carcinoma cell lines

Propolis is a natural resinous substance collected by bees from various types of trees and plants and has antibacterial, antiviral and antitumoral features depending on its antioxidant properties. Major aim is to investigate cytotoxic effect of Turkish propolis on human laryngeal epidermoid carcinoma (HEp-2) cells. HEp-2 cells/well were loaded on RTCA(real time cell analysis) system and the cell index was followed up during 48 hours. Water extract of Turkish propolis (WEP) of 250-10.000 μg/mL concentrations and ethanolic extracts of Turkish propolis (EEP) of 10-2.400 μg/mL concentrations were treated with HEp-2 cells and followed by RTCA system. The cell indexes and IC50 values were determined. HEp-2 cells were incubated with WEP and EEP. Proliferation was followed by flow cytometric DNA cycle analysis. WEP and EEP were found to be cytotoxic to HEp-2 cells. When WEP and EEP were incubated with HEp-2 cells during 72 hours, the highest antiproliferative effect was seen by interfering DNA cycles. Turkish propolis extracts were found to be cytotoxic and antiproliferative to HEp- 2 cells in the present study, therefore, it was concluded that it may fall within chemotherapy or target therapies for larynx cancers.

SRNA23, a novel small RNA, regulates to the pathogenesis of Streptococcus suis serotype 2

ABSTRACT Streptococcus suis serotype 2 (S. suis 2) is an important ubiquitous zoonotic pathogen. To date, regulatory factors and their implication in S. suis pathogenesis are not fully understood. Small non-coding RNAs (sRNAs) have been proven to function as important regulatory factors in bacterial pathogenesis and stress adaptation. Here, we identified a differentially downregulated S. suis 05ZYH33 sRNA after iron starvation by RNA-seq, which we named sRNA23. The presence of sRNA23 was further confirmed by RACE and Northern blot. Expression of sRNA23 was significantly altered under different environmental stresses such as nutritional starvation, osmotic pressure, oxidative stress, and lysozymal exposure. A sRNA23-deleted mutant exhibited relatively shorter streptococcal chains and weakened biofilm-forming ability. The mutation also resulted in decreased adherence of the S. suis 05ZYH33 to human laryngeal epidermoid carcinoma (HEp-2) cells, increased sensitivity to phagocytosis by RAW264.7 macrophages, and significantly reduced hemolytic activity. Furthermore, we observed that a sRNA23-deleted mutant had a low survival rate in pig whole blood and attenuated virulence in a mouse model. Moreover, based on RNA pull-down and electrophoretic mobility shift assay, we found that sRNA23 can directly bind to two proteins involved in adhesion and biofilm formation, namely, moonlighting protein FBA (fructose diphosphate aldolase) and rplB (50S ribosomal protein L2), respectively. Collectively, sRNA23 enhances S. suis 2 pathogenicity and the binding between sRNA23 and FBA/rplB might play an essential role in the adherence and biofilm-forming ability of S. suis 2. Abbreviation sRNA: small noncoding RNA; FBA: fructose diphosphate aldolase; rplB: 50S ribosomal protein L2; RACE: rapid amplification of cDNA ends; EMSA: electrophoretic mobility shift assay; THB: Todd-Hewitt broth; FBS: fetal bovine serum; BIP: 2,2ʹ-Bipyridine

Antiviral iridoid glycosides from Clerodendrum myricoides

The methanol root extract of Clerodendrum myricoides (Hochst.) Vatke afforded two new (1, 2) and two known (3, 4) iridoid glycosides. The structures of the isolated compounds were established based on NMR, IR, UV and MS data analyses. The crude extract and the isolated constituents were assayed for antiviral activity against the human respiratory syncytial virus (RSV) in human laryngeal epidermoid carcinoma (HEp-2) cells. The crude extract inhibited RSV infectivity at EC50 = 0.21 μg/ml, while it showed cytotoxicity against HEp-2 cells with CC50 = 9 μg/ml. Compound 2 showed 43.2% virus inhibition at 100 μM, while compounds 1 as well as 3 and 4 had only weak antiviral and cytotoxic activities.

The otc gene of Streptococcus suis plays an important role in biofilm formation, adhesion, and virulence in a murine model

Streptococcus suis (S. suis) is an emerging zoonotic pathogen that can cause meningitis, arthritis, pneumonia, and sepsis. It poses a serious threat to the swine industry and public health worldwide. Ornithine carbamoyltransferase (OTC) is involved in the arginine deiminase system. OTC, which is a widely distributed enzyme in microorganisms, mammals, and higher plants, catalyzes the conversion of ornithine to citrulline. The present study showed that the otc gene plays an important role in the pathogenesis of S. suis infections. The ability of an otc-deficient mutant (Δotc) to form a biofilm was significantly reduced compared to the wild-type (WT) strain, as determined by crystal violet staining. Confocal laser scanning microscopy and scanning electron microscopy observations showed that the weakening of biofilm formation by the Δotc strain is related to a decrease in the extracellular matrix. In addition, compared to the WT strain, the Δotc strain had a reduced capacity to adhere to human laryngeal epidermoid carcinoma (HEp-2) cells compared to the WT strain. A real-time PCR analysis showed that the expression of adhesion-related genes by the Δotc strain was also lower than that of the WT strain. The virulence of the Δotc strain was significantly lower than that of the WT strain in a murine infection model. In addition, a histological analysis showed that the pathogenicity of the Δotc strain was lower than that of the WT strain, causing only slight inflammatory lesions in lung, liver, spleen, and kidney tissues. No significant differences were observed between the complemented mutant (CΔotc) and WT strains with respect to biofilm formation, adhesion, gene expression, and virulence. The present study provided evidence that the otc gene plays a pivotal role in the regulation of S. suis adhesion and biofilm formation. It also suggested that the otc gene is indirectly involved in the pathogenesis of S. suis serotype 2 infections.

Disruption of Rho GTPase Prenylation by Statins Inhibits Respiratory Syncytial Virus

INTRODUCTIONRespiratory syncytial virus (RSV) is a leading cause of acute lower respiratory tract infections necessitating hospitalization in the pediatric population. Despite the significant healthcare burden associated with RSV, few options are available to prevent or treat infection. To address the lack of therapeutic strategies for RSV, we developed a high content screening platform to identify small molecule inhibitors of the virus. Following a screen of 2400 compounds, statins were found to significantly reduce RSV infection rates. Here we show that statin‐mediated disruption of isoprenoid biosynthesis and Rho GTPase prenylation blocks late events within the life cycle of RSV, including protein trafficking and viral assembly.METHODSTo screen for inhibitors of RSV, human laryngeal epidermoid carcinoma (HEp‐2) cells were infected with recombinant RSV engineered to express green fluorescent protein (GFP). Automated image analysis algorithms were used to quantify infection rate by measuring the proportion of GFP‐fluorescent cells. Statin‐induced changes to the life cycle of RSV were determined by immunofluorescence microscopy and temporal quantification of viral fusion and nucleoproteins. Effects on protein prenylation by statins and RSV were evaluated via click chemistry using azide‐modified isoprenoids with a biotin‐alkyne detection reagent. Rho GTPase activity was quantified by Western blot following affinity purification of active, GTP‐bound RhoA or Rac1.RESULTSInhibition of RSV by statins was dose‐dependent with an effective concentration (EC50) of 1 μM. Viral events occurring &gt;12 hours post‐infection, including protein trafficking and assembly, were inhibited by statins, whereas early events, such as viral entry, were unaffected. Viral trafficking and assembly are thought to be mediated by Rho GTPases, a group of prenylated proteins which modulate dynamics of the actin cytoskeleton. We found that RhoA‐specific inhibition by Rhosin (50 μM) moderately reduced RSV infection rate (26 ± 6% reduction), but Rac1‐specific inhibition by EHT1864 (20 μM) elicited a profound effect (90 ± 2% reduction). Critically, RSV infection promoted Rac1 prenylation and activity, while statin treatment blocked Rac1 prenylation and reduced active, GTP‐bound protein by 2.5‐fold.CONCLUSIONSOur results demonstrate the importance of protein prenylation to RSV infection and perturbation of this process by statins. Inhibition of Rac1 prenylation and activity by statins serves to block viral protein trafficking and assembly. Based on our findings, we propose repurposing statins as a treatment for RSV as these drugs potently inhibit the virus, are US Food and Drug Administration approved and have favorable clinical safety profiles.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Oxidative Response and Micronucleus Centromere Assay in HEp-2 Cells Exposed to Fungicide Iprodione.

The fungicide agents are a key component in the fruits and vegetables production. The Iprodione residues are one of the pesticide more frequently found in food products. The available data about the cytotoxicity of iprodione and its metabolites are scarce and do not allow characterization of its genotoxic potential and define the risk assessment.The human larynx epidermoid carcinoma cell line (HEp-2) has been shown to be sensitive to the toxic effects of xenobiotics of different origin and have been often used in citotoxicity and genotoxicity studies. The purpose of this paper is to evaluate the induction of genotoxicity and the role of oxidative stress in HEp-2cell line by exposure to the IP. The MTT test for viability resulted in CL50 85.86 (77.05-95.68) μg/mL of Iprodione. On the basis of this result, we proceeded to expose the cells to the sublethal concentrations (below the CL50) during 24 h to analyze the mitotic index and nuclear division index in order to determine the subcytotoxic concentrations of IP which the genotoxicity was evaluated. The subcytotoxic concentrations of 7, 17, and 25 μg/mL IP induced aneugenic effects as micronuclei centromere positive whereas 17 μg/mL was a threshold for centromere negative micronuclei induction in HEp-2 cells. The abnormal mitosis was induced for exposition of Hep-2 cells to the three concentrations. According to the result obtained, citotoxicity and genotoxicity oxidative stress studies were performed in 1.5, 7.0, and 25 μg/mL of IP. The results showed that the GSH intracellular content, the SOD activity and the levels of oxidative damage of the proteins were affected lead to redox imbalance. The decreased in the SOD activity and protein oxidation were in according to the result obtained to genotoxicity, suggesting that different biological targets could be affected.

Mechanisms underlying the cytotoxic effect of propolis on human laryngeal epidermoid carcinoma cells

Propolis has been used as a traditional remedy for centuries because of its beneficial effects, including anticancer properties. The aim of this study was to compare the cytotoxic mechanism of Cuban red propolis (CP) and Brazilian green propolis (BP) on human laryngeal carcinoma (HEp-2) cells. Cell viability, leakage of lactate dehydrogenase, fluorescence staining, mitochondrial membrane potential (ΔΨm) and the expression of pro/anti-apoptotic genes were assessed. Cell viability and cytotoxic assays suggested a dose-dependent effect of CP and BP extracts with a possible association of intracellular reactive oxygen species production and decreased ΔΨm. Both samples induced apoptosis via activation of TP53, CASP3, BAX, P21 signalling, and downregulation of BCL2 and BCL-XL. CP exerted a higher cytotoxic effect than BP extract. Our findings suggest further investigation of the main components of each propolis sample, what may lead to the development of strategies for the treatment of laryngeal cancer.

Mechanisms involved in the cytotoxic action of Brazilian propolis and caffeic acid against HEp-2 cells and modulation of P-glycoprotein activity

The effects of propolis and phenolic compounds (caffeic acid - Caf; dihydrocinnamic acid - Cin; p-coumaric acid - Cou) in the same quantity found in our propolis sample were investigated on human laryngeal epidermoid carcinoma (HEp-2) cells. Cell viability, apoptosis/necrosis and cell cycle arrest, P53 and CASPASE-3 gene expression, generation of reactive oxygen species (ROS) and the ability of propolis to induce doxorubicin (DOX) efflux using a P-glycoprotein (P-gp) inhibitor (verapamil) were assayed. Propolis exerted a cytotoxic effect on HEp-2 cells, whereas isolated compounds had no effect on cell viability. Higher concentrations were tested and Caf induced late apoptosis or necrosis in HEp-2 cells, while propolis induced apoptosis, both probably due to ROS generation. P53 expression was downregulated by propolis but not by Caf. CASPASE-3 expression was correlated with induction of both early and late apoptosis, with both propolis and Caf alone upregulating its expression. Propolis induced cell cycle arrest at G2/M phase and Caf at S phase. Propolis but not Caf may act as a P-gp inhibitor by modulating P-gp activity and inhibiting DOX efflux. Propolis exerted cytotoxic effects on HEp-2 cells, and the mechanisms are discussed, showing its potential as an antitumour drug.

Combinatorial effects of geopropolis produced by Melipona fasciculata Smith with anticancer drugs against human laryngeal epidermoid carcinoma (HEp-2) cells.

The identification of natural products exerting a combined effect with therapeutic agents could be an alternative for cancer treatment, reducing the concentration of the drugs and side effects. Geopropolis (Geo) is produced by some stingless bees from a mixture of vegetable resins, gland secretions of the bees and soil. It has been used popularly as an antiseptic agent and to treat respiratory diseases and dermatosis. To determine whether Geo enhances the anticancer effect of carboplatin, methotrexate and doxorubicin (DOX), human laryngeal epidermoid carcinoma (HEp-2) cells were treated with Geo alone or in combination with each drug. Cell growth, cytotoxicity and apoptosis were evaluated using 3-(4,5-dimethyl thiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release, and flow cytometry. Scratch assay was used to analyze cell migration and transmission electron microscopy to observe morphologic alterations. The influence of Geo on drug resistance was also investigated assessing P-glycoprotein (P-gp) action. Geo inhibited cell proliferation and migration. The combination Geo+DOX led to the highest cytotoxic activity and induced apoptosis, leading to loss of membrane integrity. Geo had no effect on P-gp-mediated efflux of DOX. Data indicate that Geo combined with DOX could be a potential clinical chemotherapeutic approach for laryngeal cancer treatment.

Synthesis and antitumor activity of some new pyrazolo[3,4-d]pyrimidine and pyrazolo[3,4-b]pyridine derivatives

A new series of pyrazolo[3,4-d]pyrimidine-3-carbonitrile and pyrazolo[3,4-b]pyridine-3-carbonitrile derivatives was synthesized by the reaction of 5-amino-1-tosyl-1H-pyrazole-3,4-dicarbonitrile as a key starting material with various electrophilic and nucleophilic reagents. All the newly synthesized compounds were structurally confirmed by various modern analytical methods (IR, 1H NMR and MS). All the title compounds have been evaluated for their potential cytotoxicity against human laryngeal epidermoid carcinoma cells (Hep2). Among the screened compounds, 3 and 4 exhibited the highest significant effect.

Antitumor Activity of Extracts from Medicinal Basidiomycetes Mushrooms.

Aqueous extracts from the vegetative submerged mycelia of cultivated Basidiomycetes Ganoderma lucidum, Lentinus edodes, and Grifola frondosa, as well as from the fruiting bodies of G. lucidum, were found to have antitumor activity. The antitumor effect of the mycelial extracts from all 3 fungal species was ascertained in vivo in rats with implanted kidney cancer. Dystrophic changes in tumor cells and tumor necrosis (up to 90%) were noted. In vitro cytotoxicity studies of the A549 human lung adenocarcinoma cell line and HEp-2 human laryngeal epidermoid carcinoma cells showed that the extracts from the G. lucidum fruiting bodies and from the L. edodes vegetative mycelium were the most effective. The animals' immune systems were activated, and the fungal extracts displayed no toxicity when administered orally.

Ocular biocompatibility of polyquaternium 10 gel: functional and morphological results

This paper deals with the characterization study of topical and intraocular biocompatibility and toxicity of cationic hydroxyethylcellulose Polyquaternium 10 (PQ10). It also evaluates the rheological properties of gels. The cytotoxicity assays were done in two cell lines: HEp-2 and VERO (human larynx epidermoid carcinoma cell and African green monkey kidney cells respectively). For the in vivo study, New Zealand albino rabbits were used. The in vitro cytotoxic activity of PQ10 shows no statistically significant differences in relation to the control of hydroxypropylmethylcellulose (HPMC) in any of the cell lines used in this study. Similarly, the signs of inflammation observed after treatment showed no significant difference between the groups of animals treated with the polymer compared to the control group. Normal histological characteristics were seen in both groups with no histological inflammatory reaction. After 1 month of the intracameral application of 2% PQ10 (treatment group) or 0.3% HPMC (control group), electroretinograms showed similar levels of a- and b-waves latencies and amplitude. In summary, PQ10 gel was well tolerated in these experiments, with proper monitoring, it could stand as a new alternative in the development of ophthalmic viscosurgical devices.

The chemical composition and pharmacological activities of geopropolis produced by Melipona fasciculata Smith in northeast Brazil

Aim: Geopropolis is produced by stingless bees from resinous materials of plants, adding salivary secretions, wax, soil, and has been used in folk medicine for the treatment of respiratory diseases and dermatoses. Therefore, it is important to study its antibacterial, antitumor and immunomodulatory properties to confirm the ethnopharmacological applications. This work aimed to evaluate its chemical composition, and its antimicrobial, anti-tumoral and immunomodulatory activities. Methods: Geopropolis composition was investigated using GC-MS analysis. Antibacterial tests were performed to determine the minimum inhibitory concentration. HEp-2 cells viability was determined by the reduction of MTT, and cytokine production by human monocytes was determined by ELISA. Results: The major constituents of geopropolis compounds were carbohydrates and their derivatives, triterpenes, anacardic acid, alkylresorcinols, and sugar alcohols. Geopropolis alone showed no antibacterial activity against Staphylococcus aureus and Escherichia coli, but its combination with chloramphenicol exerted a greater action against S. aureus than chloramphenicol alone. Geopropolis exhibited a cytostatic action toward human laryngeal epidermoid carcinoma cells and stimulated tumor necrosis factor alpha and interleukin-10 production by human monocytes, showing an activator profile for human monocytes. Conclusion: The synergistic effect of geopropolis and chloramphenicol deserves further investigation due to bacterial resistance to antibiotics. Geopropolis also displayed antitumoral and immunomodulatory activity, and its biological properties may be due to triterpenes - one of its major chemical constituents.

Low concentrations of trichosanthin induce apoptosis and cell cycle arrest via c-Jun N-terminal protein kinase/mitogen-activated protein kinase activation.

Trichosanthin (TCS) is a type I ribosome--inactivating protein, which inhibits cell viability in human epithelial type 2 (HEp-2) and AMC-HN-8 human laryngeal epidermoid carcinoma cells. Although TCS is a potential chemotherapeutic agent, its mechanism of action remains to be elucidated. In the present study, HEp-2 and AMC-HN-8 cells were treated with different concentrations of TCS combined with or without cisplatin. After 5 days of successive treatment, different experimental groups were detected using a cell counting kit-8 and the collected supernatants were analyzed using a lactate dehydrogenase kit. Flow cytometric assays were performed to detect apoptosis and cell cycle arrest in the HEp-2 and AMC-HN-8 cells, reverse transcription quantitative polymerase chain reaction was performed to detect the levels of p27, p21WAF and western blot analysis was performed to detect changes in c-Jun N-terminal protein kinase (JNK)/phosphorylated (phospho)-JNK, p38/phospho-p38, extracellular signal-regulated kinase (ERK)/phospho-ERK, caspase-3 and caspase-9 in the HEp-2 and AMC-HN-8 cancer cells. TCS significantly inhibited the cell viability of the HEp-2 and AMC-HN-8 cells, independently of necrosis. TCS induced apoptosis and increased the percentage of HEp-2 and AMC-HN-8 cells in the S-phase of the cell cycle. In addition, the JNK/mitogen-activated protein kinase (MAPK) pathway was activated by TCS in the HEp-2 and AMC-HN-8 cells. Low concentrations of TCS also induced apoptosis and S-phase cell cycle arrest in the HEp-2 and AMC-HN-8 cells. The antitumor effects of TCS may be associated with JNK/MAPK activation.

Anti-Viral Activity of Water Extract of Paeonia lactiflora Pallas Against Human Respiratory Syncytial Virus in Human Respiratory Tract Cell Lines

Paeonia lactiflora Pallas (P. lactiflora, Ranunculaceae) is a common ingredient of Sheng-Ma-Ge-Gen-Tang (SMGGT; Shoma-kakkon-to) and Ge-Gen-Tang (GGT; kakkon-to). SMGGT and GGT are different prescriptions of traditional Chinese medicine with different ingredients designed for airway symptoms. Both SMGGT and GGT have anti-viral activity against human respiratory syncytial virus (HRSV). Therefore, P. lactiflora was hypothesized to be the effective ingredient of both SMGGT and GGT against HRSV. However, P. lactiflora does not have any proven antiviral activity. This study used both human upper (Human larynx epidermoid carcinoma cell line, HEp-2) and lower (human lung carcinoma cell line, A549) respiratory tract cells to test the hypothesis that a hot water extract of P. lactiflora could effectively inhibit plaque formation induced by HRSV infection. The ability of P. lactiflora to stimulate anti-viral cytokines was evaluated by enzyme-linked immunosorbent assay (ELISA). The results showed that P. lactiflora was time-dependently and dose-dependently effective against HRSV in HEp-2 and A549 cells, particularly supplemented before viral inoculation (p < 0.0001). 10 μg/ml P. lactiflora had a comparable anti-HRSV activity with 10 μg/ml ribavirin, a broad-spectrum antiviral agent. P. lactiflora was dose-dependently effective against viral attachment (p < 0.0001), with a better effect on A549 cells (p < 0.0001). P. lactiflora was time-dependently (p < 0.0001) and dose-dependently (p < 0.0001) effective against viral penetration. Moreover, P. lactiflora stimulated IFN-β secretion without any effect on TNF-α secretion. Therefore, P. lactiflora could be beneficial at preventing HRSV infection by inhibiting viral attachment, internalization, and stimulating IFN secretion.

Isolation and cytotoxic activity of some secoiridoids from Gentiana olivieri

The genus Gentiana (Gentianaceae) is represented by 12 species in Turkish Flora with important biological effects such as antipyretic, stomachic, antidepressant, anticonvulsant, hepatoprotective and antidiabetic activities. Iridoids, secoiridoids, C-glycosylflavones and xanthones are considered to be the most promising groups of compounds responsible for the pharmacological activities of the Gentiana species. In this study swertiamarin, sweroside and gentiopicroside were isolated from Gentiana olivieri and their cytotoxic activities were evaluated against HEP-2 (human larynx epidermoid carcinoma) cell lines based on MTT assay for growth inhibition.

Antiviral Furanosesquiterpenes from Commiphora erythraea

The crude methanolic extract obtained from C. erythraea resin was chromatographed on silica gel with solvent of increasing polarity. The extract and fractions were evaluated for cytotoxicity and antiviral activity [parainfluenza type 3 virus (PIV3)] by plaque forming units (PFU) reduction assay using HEp-2 cells (human larynx epidermoid carcinoma cell line). From the active fraction, five compounds were isolated and tested. Only two of these showed anti-PIV3 activity with a selectivity index (SI) of 66.6 and 17.5, respectively. Both the compounds are furanosesquiterpenoids.

Isolation of Coxsackievirus A24 variant from patients with hemorrhagic conjunctivitis in Cuba, 2008–2009

BackgroundAn outbreak of acute hemorrhagic conjunctivitis occurred in Cuba in 2008 and 2009. ObjectiveTo determinate the etiological agent associated with the Cuban outbreaks of acute hemorrhagic conjunctivitis during 2008 and 2009. Study designConjunctival swabs and/or faecal samples from 382 patients with clinical diagnosis suggestive of acute hemorrhagic conjunctivitis were subject to viral culture in HEp-2 human laryngeal epidermoid carcinoma cells. Positive samples were identified by a specific Coxsackievirus A24 variant PCR and the 3C protease region of 16 isolates was sequenced for phylogenetic analysis. ResultsEnterovirus cytopathic effect was observed in 138 cases (36%). A higher percent of CA24v was recovered from faecal samples, 19 out of 45 cases (42.2%), than from conjunctival swabs, 127 out of 355 samples (35.8%). All isolates were identified as Coxsackievirus A24 variant. Phylogenetic analysis revealed that 2008 and 2009 Cuban outbreaks were caused by the same virus strains and that isolates were closely related to those from Taiwan (2006–2007), China (2007–2008) and Singapore (2005) with a bootstrap value of 71%. ConclusionsOutbreaks of acute hemorrhagic conjunctivitis occurred in Cuba in 2008 and 2009 were caused by Coxsackievirus A24 variant. The faecal-oral route is another mode of transmission of CA24v in the acute hemorrhagic conjunctivitis outbreaks. Phylogenetic analysis of Cuban CA24v strains involved in an acute hemorrhagic conjunctivitis outbreak in 2008 and 2009 confirms a new introduction of the CA24 variant into the Americas from South-east Asia.

Evaluation of antioxidative, protective effect against H2O2 induced cytotoxicity, and cytotoxic activities of three different Quercus species

Quercus species are used as antidiarrheic, for the treatment of hemorrhoid, oral and anal mucosa inflammation. These tree species have been of interest to researchers because of their usage in folk medicine, consumption as food, beverage and especially usage of oak woods for construction in wine barrels. The DPPH, SO and NO radical scavenging activities, protective effect against H2O2 induced cytotoxicity as well as their cytotoxic activity against Hep-2 human larynx epidermoid carcinoma cell line of the MeOH and water extracts of the barks of Quercus cerris var. cerris, Quercusmacranthera subsp. syspirensis and Quercus aucheri were investigated for the first time. Total phenolic content of the extracts was also evaluated by Folin–Ciocalteu method. Results demonstrated that the extracts showed strong radical scavenging activity comparable to those of standard compounds. Extracts also showed good protective effect against H2O2 induced cytotoxicity on human erythrocytes comparing to ascorbic acid. On the other hand, while each extract showed dose dependent cytotoxic activity, MeOH extract of Q.macranthera subsp. syspirensis showed the strongest cytotoxicity against the tested cell line. Taken together, the results showed that Quercus species may be a promising alternative to synthetic substances as natural compound with high antioxidant and antiproliferative activities.