Biofilm inhibition by Cymbopogon citratus and Syzygium aromaticum essential oils in the strains of Candida albicans

Ethnopharmacological relevance

Oils of Cymbopogon citratus and Syzygium aromaticum have been used in traditional medicine to treat fungal infections of skin, mouth, urinary and vaginal tract in Asian countries particularly India and other developing countries.

Aim of the study

To evaluate essential oils of Cymbopogon citratus and Syzygium aromaticum for their anti-biofilm activity against strong biofilm forming strains of Candida albicans.

Materials and methods

XTT reduction assay, Time kill assays, light microscopy and scanning electron microscopy (SEM) were employed to determine the effect of test oils on the Candida albicans biofilms.

Results

Most of the Candida albicans strains tested displayed formation of moderate to strong biofilms. Preformed Candida biofilms showed ≥1024 times increased resistance to antifungal drugs, 2 times to Syzygium aromaticum, but no increased tolerance for Cymbopogon citratus. Test oils were more active against preformed biofilms compared to amphotericin B and fluconazole. At 0.5× MIC, Cymbopogon citratus followed by Syzygium aromaticum were most inhibitory against biofilm formation. Light and electron microscopic studies revealed the deformity of three dimensional structures of biofilms formed in the presence of sub-MICs of Cymbopogon citratus. The cell membranes appeared to be the target site of compounds in sessile cells as displayed by SEM observations.

Conclusion

Our data had demonstrated promising in vitro anti-biofilm activity by Cymbopogon citratus and Syzygium aromaticum and confirm the ethnopharmacological use of these oils in muco-cutaneous Candida infections. Furthermore, it suggests exploitation of these oils as new anti-biofilm products to deal with the problem of drug-resistance and recurrent infection associated with biofilm mode of growth of Candida spp.     

Differential gene expression of chemokines in KRAS and BRAF mutated colorectal cell lines:Role of cytokines

AIM:To study KRAS/BRAF mutations in colorectalcancer(CRC)that influences the efficacy of treatment.To develop strategies for overcoming combination of treatment.METHODS:Five colonic cell-lines were investigated:DLD-1 with KRAS(G13D)mutation,HT 29 and Colo205 with BRAF(V600E)mutation as well as the wild type(Wt)cell-lines Caco2 and Colo-320.DLD-1(KRAS),HT-29(BRAF)and Caco2(Wt)cell lines were treated with cytokines(TNFα50 ng,IL-1β1 ng and IFNγ50ng)and harvested at different time points(1-24 h).KRAS inhibition was performed by the siRNA-approach.Two colorectal cancer cells DLD-1 and Caco2 were used for KRAS inhibition.About 70%confluency were confirmed before transfection with small interferring RNA(siRNA)oligonucleotides.All the synthetic siRNA sequences were designed in our laboratory.Total RNA and protein was isolated from the cells for RT-PCR and Western blotting.Densitometry of the Western blotting was analyzed with the Image J software(NIH).Results are shown as mean±SD.RESULTS:RT-PCR analysis in non-stimulated cells showed a low basal expression of TNFαand IL-1βin the DLD-1 KRAS-mutated cell-line,compared to Caco2wild type.No detection was found for IL-6 and IFNγin any of the studied cell lines.In contrast,pro-angiogenic chemokines(CXCL1,CXCL8)showed a high constitutive expression in the mutated cell-lines DLD-1(KRAS),HT-29 and Colo205(BRAF),compared to wild type(Caco2).The anti-angiogenic chemokine(CXCL10)showed a high basal expression in wild-type,compared to mutated cell-lines.KRAS down-regulation by siRNA showed a significant decrease in CXCL1 and CXCL10gene expression in the DLD-1(KRAS)cell-line in comparison to wild type(Caco2)at 72 h after KRAS silencing.In contrast,the specific KRAS inhibition resulted in an up-regulation of CXCL1 and CXCL10.The results of our study show a higher expression of pro-angiogenic chemokines at basal level in mutated cell-lines,which was further increased by cytokine treatment.CONCLUSION:To summarize,basal chemokine gene expression for pro-angiogenic chemokines was high in mutated as compared to wild type cell-lines.This reflects the likely existence of a different microenvironment in tumours consistent of wild type or mutated cells.This may help to rationalize the choice of molecular targets for suitable therapeutic investigation in clinical studies.     

Genotoxicity Assessment of Amaranth and Allura Red Using Saccharomyces Cerevisiae

Amaranth (E123) and Allura red (E129), very important food azo dyes used in food, drug, paper, cosmetic and textile industries, were assessed for their genotoxic potential through comet assay in yeast cells. Comet assay was standardized by with different concentration of H₂O₂. Concentrations of Amaranth and Allura red were maintained in sorbitol buffer starting from 9.76 to 5,000 μg/mL and 1 × 10⁴ cells were incubated at two different incubation temperatures 28 and 37°C. Amaranth (E123) and Allura red (E129) were found to exhibit their genotoxic effect directly in Saccharomyces cerevisiae. No significant genotoxic activity was observed for Amaranth and Allura red at 28°C but at 37°C direct relation of Amaranth concentration with comet tail was significant and no positive relation was seen with time exposure factor. At 37°C the minimum concentration of Amaranth and Allura red at which significant DNA damage observed through comet assay was 1,250 μg/mL in 2nd h post exposure time. The results indicated that food colors should be carefully used in baking products as heavy concentration of food colors could affect the fermentation process of baking.     

Molecular characterization of Staphylococcus aureus isolated from children with adenoid hypertrophy: emergence of new spa types t7685 and t7692

Objective

Adenoids have been associated with the pathogenesis of acute, recurrent and chronic infectious diseases of the upper respiratory system and their hypertrophy is one of the most common causes of upper airway obstruction affecting children. In this study, the characteristics of Staphylococcus aureus isolates from patients who had undergone adenoidectomy were investigated via spa typing method.

Methods

A total of 113 children with adenoid hypertrophy who underwent adenoidectomy during September 2009 to November 2010, were included in the study. The isolates were identified to the species level as S. aureus using standard biochemical methods, following which the amplification and sequencing of the spa gene X region were carried out.

Results

S. aureus was found in the adenoid tissue of 26 (23%) patients. Out of the 26 S. aureus isolates, 5 (19%), 3 (11.5%) and 3 (11.5%) were resistant to tetracycline, erythromycin and oxacillin respectively. All the isolates were susceptible to vancomycin, rifampin, ciprofloxacin, gentamicin, mupirocin and quinupristin-dalfopristin and were typed using spa typing method. All the isolates were found to include 21 spa types, including two previously unreported types (t7685 and t7692). The most prevalent spa types were t7685 (11.5%), t230 (8%), t325 (8%) and t1149 (8%).

Conclusion

This study demonstrates that the prevalence rate of S. aureus in the adenoid tissue of the children assessed was 23%. An interesting point to note was the dominance of the spa type t7685 that has not been previously reported by other studies.     

Portal Hypertension and an Atypical Reactive Arthritis Like Presentation in a patient infected with Hepatitis C Virus Genotype 3

Background:Reactive arthritis (ReA) is defined as a peripheral arthritis lasting longer than 1 month, associated with urethritis, cervicitis, or diarrhea. The reported annual incidence of ReA is approximately 30-40 cases per 100,000 adults, occurring commonly in the age group of 16 and 35 years. It is known to be associated with gastrointestinal infections with Shigella, Salmonella, and Campylobacter species and other microorganisms, as well as with genitourinary infections (especially with Chlamydia trachomatis).Conclusions:Our case is unlike classical ReA because the patient is older, HLA B27 negative, and without florid urethritis. Admitted for fever and lower urinary tract symptoms, along with respiratory distress, the primary objective of the emergency doctors was to prevent the patient from progressing to organ failure. The diagnosis of underlying atypical/incomplete ReA could easily have been missed without adequate awareness, dermatological consultation, and a skin biopsy.     

Effects of silymarin supplementation on blood lipids: A systematic review and meta‐analysis of clinical trials

Dyslipidemia is a leading cause of endothelial dysfunction and cardiovascular disease. Several studies used silymarin as an herbal supplement in hyperlipidemic subjects. The aim of the present systematic review and meta‐analysis was to examine the effect of silymarin supplementation on blood lipids. PubMed, Scopus, Ovid (Cochrane library), ISI Web of Science, and Google Scholar were systematically searched until March 2018 to find intervention studies that examined the impact of silymarin supplementation on blood lipids in adults. Changes in blood lipids and potential sources of between‐study variation were extracted. We run a subgroup analysis to determine potential sources of inter‐study heterogeneity. Ten clinical trials fulfilled the eligibility criteria. Meta‐analysis indicated that silymarin supplementation in combination with other treatments (not silymarin alone) reduced total cholesterol (change: ?25.45 mg/dl; 95% confidence interval [CI] [?47.89, ?3.01 mg/dl]) and low‐density lipoprotein (change: ?28.25 mg/dl; 95% CI [?53.09, ?3.42 mg/dl]). Also, silymarin increased high‐density lipoprotein concentration (change: 4.82 mg/dl; 95% CI [2.01, 7.63 mg/dl]). Blood concentration of triglyceride was significantly after silymarin supplementation in comparison with controls (change: ?22.55 mg/dl; 95% CI [?44.32, ?0.78 mg/dl]). Present systematic review and meta‐analysis revealed that silymarin supplementation in combination with other treatments had a favorable effect on blood lipids.     

Association of OPRK1 gene polymorphisms with opioid dependence in addicted men undergoing methadone treatment in an Iranian population

Previous studies have shown significant associations between OPRK1 and susceptibility to opioid dependence and the relationships between libido dysfunction and insomnia among opium addicts who underwent methadone maintenance treatment. The authors investigated the single locus and haplotype association of rs997917, rs6985606, and rs6473797 with susceptibility to opioid addiction. Samples were selected among 202 healthy individuals and 202 opium addicts undergoing methadone maintenance treatment. Genomic DNA was extracted from the whole blood samples of all subjects through a salting out procedure. All three variants were genotyped in the studied subjects using Amplification Refractory Mutation System-Polymerase Chain Reaction (ARMS-PCR). The whole analysis process was performed using SNPAlyze and SPSS ver.20 software packages. According to the single locus analyses, rs997917 and rs6985606 represented significant associations with opium addiction under recessive (p = 0.0128) and co-dominant (p = 0.0001) inheritance models, respectively. The haplotypes C-T-C (Permutation p = 0.014) and C-T-T (Permutation p = 0.0002) were significantly associated with opioid dependence. Among methadone maintenance treatment individuals, rs997917 was significantly associated with insomnia in both allelic and genotypic levels (p = 0.0001 and p = 0.038, respectively). Furthermore, rs6985606 had the only significant association with the co-incidence of insomnia and libido dysfunction in the methadone maintenance treatment group (p = 0.038). The OPRK1 gene variants showed significant association with susceptibility to opioid dependence among Iranians.