Quality assurance of postharvest grapes against Botrytis cinerea by terbinafine

Worldwide,fruit is an indispensable treasure house of nutrition for human beings,occupying a vital position of human diet.Postharvest fruit storage requires efficient antifungal agents to control Botrytis cinerea,which is a vital postharvest disease affecting fruit and leading to enormous losses.However,with the enormous abuse of existing antifungal drugs,the problem of drug-resistant fungi is imminent,making the controlling diseases caused by pathogenic fungi even more challenging.Drug repurposing is an efficient alternative method,we evaluated a well-known antifungal chemical,terbinafine,against the agricultural pathogen,B.cinerea in vitro,as a result,terbinafine showed strong anti-fungal activity.Furthermore,the in vivo antifungal activity of terbinafine was evaluated,the results showed that terbinafine could reduce the decay area on grapes.Terbinafine could disrupt the cell membrane integrity,increase cell membrane permeability,and eventual cell death of B.cinerea.In addition,terbinafine reduced decay incidence,and weight loss and maintained the soluble solids,titratable acidity,ascorbic acid,total phenolic,and malondialdehyde content during the storage period of grapes.Overall,terbinafine could be an antifungal preservative for postharvest table grapes fresh-keeping.

新型高效抗真菌药——Terbinafine

Terbinafine(商品名lamisil,下简称TBF)是近年来合成的新型广谱丙烯胺类抗真菌药。化学结构式如图所示,该药无论是口服和外用均具有高效杀真菌和抑制真菌的作用,是一个很有前途的药物。1988年在西班牙巴塞罗那召开的国际人和动物真菌学会第十届大会举行了TBF首次专题讨论会。现将主要内容综合如下。

Analytical methods for determination of terbinafine hydrochloride in pharmaceuticals and biological materials

Terbinafine is a new powerful antifungal agent indicated for both oral and topical treatment of myco- sessince. It is highly effective in the treatment of determatomycoses. The chemical and pharmaceutical analysis of the drug requires effective analytical methods for quality control and pharmacodynamic and pharmacokinetic studies. Ever since it was introduced as an effective antifungal agent, many methods have been developed and validated for its assay in pharmaceuticals and biological materials. This article reviews the various methods reported during the last 25 years.

Cutaneous Sporotrichosis Treated with Terbinafine

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Evaluation of the Permeation and Penetration of Two Formulations of Terbinafine Chlorhydrate Incorporated in Liposomes (Cream 1%) vs. a Conventional Formulation (Cream 1%), in an <i>in Vitro-ex Vivo</i>Model

In this trial, a topical formulation of a 1% liposome TH cream was compared to a conventional TH formulation. Permeation was evaluated through a synthetic membrane. The study provided information on the rate of diffusion and percentage of drug release and also compared the penetration dynamics in the layers of the stratum corneum of both formulations. Liposome TH cream was able to deliver the active ingredient to a larger number of SC layers, with a higher amount of drug, which is very promising for the treatment of superficial mycoses.

Effect of topical application of terbinafine on fungal keratitis

Background Fungal keratitis is a rare but serious corneal disease that may result in loss of vision. The poor prognosis might be due to limited treatment option. This study aimed to evaluate the clinical efficacy of 0.25% terbinafine eye drops comparing with 5% natamycin suspension on fungal keratitis. Methods A retrospective clinical trial was performed on 90 patients presenting with direct smear and/or culture positive fungal keratitis at Beijing Tongren Hospital, Beijing, China from January 2006 to May 2008. Corneal ulcers were categorized as mild or severe. Forty-five patients were treated with topical terbinafine and the next 45 cases received topical natamycin hourly. Results Filamentous fungi were found in corneal scrapings among all 90 cases. Fungal cultures were positive in 64 patients （71%）. Species of Fusarium and Aspergillus were the principal isolates. Forty （89%） patients showed favorable response to terbinafine, while forty-two （93%） patients exhibited favorable response to natamycin （P 〉0.05）. The mean course of treatment was significantly showed in the terbinafine treatment group than natamycin group （（26.5±11.2） days versus （19.3±6.4） days; P 〈0.05）. In terbinafine group, twenty patients with ulcers smaller than 4 mm had favorable outcome, while 20 of 25 patients with ulcers more than 4 mm in diameter had favorable response （P 〈0.05）. Twenty-seven patients with depth of infiltration less than half of stroma thickness had favorable response to terbinafine, while 13 of 18 patients with depth of infiltration more than half of stroma responded to terbinafine. This difference was statistically significant （P 〈0.05）. Conclusions Our findings suggest that topical terbinafine is an effective antifungal drug for the management of filamentous mycotic keratitis, particularly in cases with smaller and shallower ulcers. Its mean duration of treatment was longer than natamycin.

Genome-wide expression profiling of the response to terbinafine in Candida albicans using a cDNA microarray analysis

Background Candida albicans is the most frequently seen opportunistic human fungal pathogen. Terbinafine is an allylamine antifungal agent that has been proven to have high clinical efficacy in the therapy of fungal infections, the mechanism of action of terbinafine involves the specific inhibition of fungal squalene epoxidase, resulting in ergosterol deficiency and accumulation of intracellular squalene. We used cDNA microarray analysis technology to monitor global expression profile changes of Candida albicans genes in response to terbinafine treatment, and we anticipated a panoramic view of the responses of Candida albicans cells to the representatives of allylamine antifungal agents at the molecular level in an effort to identify drug class-specific and mechanism-independent changes in gene expression.Methods Candida albicans strain ATCC 90028 was exposed to either medium alone or terbinafine at a concentration equivalent to the 1/2 minimal inhibitory concentrations （MICs, 4 mg/L） for 90 minutes. RNA was isolated and gene expression profiles were compared to identify the changes in the gene expression profile using a cDNA microarray analysis. Differential expression of 10 select genes detected by cDNA microarray analysis was confirmed by semi-quantitative reverse transcription-polymerase chain reaction （RT-PCR）. Results A total of 222 genes were found to be responsive to terbinafine, including 121 up-regulated genes and 101 down-regulated genes. These included genes encoding membrane transport proteins belonging to the members of the ATP-binding cassette （ABC） or major facilitator supeffamily （MFS; CDR1, AGP2, GAP6, PHO84, HOL3, FCY23, VCX1）, genes involved in stress response and detoxification （CDR1, AGP2, HOL3）, and gene involved in the ergosterol biosynthesis pathway （ERG12）. The results of semi-quantitative RT-PCR were consistent with that of the cDNA microarray analysis. Conclusions The up-regulation of the gene encoding the multidrug resistance efflux pump CDR1 may contribute to the terbinafine resistance in Candida albicans. However, the precise roles of other affected genes remain unclear, further studies of these genes and their respective products that play roles in the context of antifunqal resistance are warranted.

角鲨烯环氧酶通过积累骨化三醇和激活CYP24A1介导的MAPK信号通路促进结直肠癌细胞增殖

背景与目的结直肠癌(colorectal cancer,CRC)是一种高发的恶性肿瘤,其分子机制和靶向治疗有待进一步研究。代谢紊乱是癌症的重要特征。靶向调节肿瘤代谢途径已成为现代肿瘤治疗的重要方向。在本研究中,我们旨在研究一种新的促进CRC增殖的代谢酶及相关的分子机制。方法我们对人CRC组织样本进行了RNA测序和组织微阵列分析,来筛选CRC相关的新基因。角鲨烯环氧酶(squalene monooxygenase,SQLE)在CRC患者组织中高表达。通过检测CRC细胞的细胞活力、集落和类器官形成、细胞内胆固醇浓度和异种移植肿瘤生长,来明确SQLE在CRC进展过程中的调节功能和SQLE抑制剂的治疗效果。通过转录组和非靶向代谢组学联合分析,探讨SQLE发挥功能的分子机制。应用蛋白质免疫印迹和实时荧光定量PCR评估SQLE对MAPK信号通路的激活。结果在CRC患者中SQLE相关的胆固醇生物合成被上调,并且与预后不良相关。SQLE在体外和体内促进CRC生长。抑制SQLE可降低骨化三醇(维生素D3的活性形式)和CYP24A1的水平,使细胞内Ca2+浓度增加。抑制MAPK信号通路可抑制CRC细胞生长。SQLE抑制剂terbinafine可抑制CRC细胞增殖、类器官和异种移植肿瘤的生长。结论SQLE通过积累骨化三醇和激活CYP24A1介导的MAPK信号通路促进CRC生长,提示SQLE可作为潜在的CRC治疗靶点。

Comparison of the Effects of Three Different Anti-fungus Drugs on Candida Albicans of Murine Vaginal Mucosa

To compare the therapeutic effects of three different anti-fungal drugs （i.e., terbinafine, fluconazole and intraconazole） in the treatment of experimental vaginitis caused by Candida albicans （C. albicans） in mice, the fungal vaginitis model was established in female ICR mice by intravaginal inoculation of suspension of C. albicans after the animal had been pretreated with estradiol. Mice were divided at random into different groups and then respectively treated with terbinafine, fluconazole and intraconazole given by gastrogavage. The burden of the fungus in the vaginal lavage fluids in the mice of the different groups was measured dynamically at different time points after the beginning of the drug treatment. The fungal burdens in the vaginal lavage fluids taken at different time points from the mice treated with terbinafine were significantly higher than those taken at corresponding time points from mice treated with fluconazole or itraconazole （P〈0.01）. The fungal burdens in the vaginal lavage fluids taken from mice 1 week after the beginning of the treatment with terbinafine remained at a relatively high level. A dramatic drop in the fungal burden was noted in the vaginal lavage fluids taken on the 2nd day of the treatment from mice treated with itraconazole or fluconazole group and the fungal burden on the 3rd day of the treatment in these mice were at a very low level, suggesting that fluconazole or itraconazole were highly effective for the treatment. However, the difference in the therapeutic effect between the two drugs was not significant （P〉0.05）. Itraconazole or fluconazole, but not terbinafine, is very effective for the treatment of fungal vaginitis caused by C. albicans in mice.

<i>Rhizopus oryzae</i>Retro-Orbital Abscess: Comparison of Spores and Hyphae Antifungal Susceptibility and Clinical Outcome

Rhizopus is fungus of the order Mucorales, responsible for invasive diseases called mucormycosis. The correct identification of these fungi is one of the main questions raised in the literature since the diagnosis is not simple, being the necessary detection in biopsies of infected tissues and isolation of the microorganism for identification by morphological and molecular methods. In vitro susceptibility testing results are not fully clear since there are reports showing that in vitro resistance does not always mean therapeutic failure. This paper describes a case of rhino-orbito-cerebral mucormycosis in a patient with alcoholic chronic pancreatitis and secondary mellitus diabetes involving microbiological diagnostic, antifungal susceptibility tests for conidia and hypha and clinical correlation. The causative agent was identified by morphological characteristics and DNA sequencing. Minimal Inhibitory Concentration (MIC) for amphotericin B (AMB), itraconazole (ITZ) and terbinafine (TERB) was determined by broth microdilution method. Standardization of the dynamic growth evaluation patterns using the automated system BCT (BioCell-Tracer?) was obtained allowing to determining hyphae growth control and evaluating the effect of different concentrations of amphotericin B, itraconazole and terbinafine directly on growing hyphae. The fractional inhibitory concentration (FIC) for the combination of the two antifungals that the patient received at the same time, terbinafine and itraconazole, was also determined trying to correlate with clinical outcome. Strains isolated of retro-orbital abscess were identified as Rhizopus oryzae. MIC results (MIC) testing conidia showed high sensibility to amphotericin B and itraconazole in comparison to the obtained for terbinafine. The tests in the automated system BCT using hyphae showed sensibility to all tested antifungal agents and the FIC results for combination in vitro between terbinafine and itraconazole showed synergism consistent with the therapeutic success obtained for the patient who received initial treatment with amphotericin B followed for maintenance treatment with the combination itraconazole and

Terbinafin治疗真菌性指甲感染

真菌性指甲感染是一种由真菌引起常见指甲病.在美国,真菌性指甲感染约占该类病总数的50%.最近Pollak及其同事在他们的一份最新报告中指出terbinafine用于治疗该病有较好疗效并证实该药物不仅可以杀灭真菌性指甲感染,还可以预防72%的该病复发.

Depletion of squalene epoxidase in synergy with glutathione peroxidase 4 inhibitor RSL3 overcomes oxidative stress resistance in lung squamous cell carcinoma

Background:Lung squamous cell carcinoma(Lusc)lacks effective targeted therapies and has a poor prognosis.Disruption of squalene epoxidase(SQLE)has been implicated in metabolic disorders and cancer.However,the role of SQLE as a monooxygenase involved in oxidativestressremainsunclear.Methods:We analyzed the expression and prognosis of lung adenocarcinoma(LUAD)and LUSC samples from GEO and TCGA databases.The proliferative activity of the tumors after intervention of SQLE was verified by cell and animal experiments.JC-1 assay,flow cytometry,and Western blot were used to show changes in apoptosis after intervention of sQLE.Flow cytometry and fluorescence assay of ROs levels were used to indicate oxidative stress status.Results:We investigated the unique role of SQLE expression in the diagnosis and prognosis prediction of LUSC.Knockdown of SQLE or treatment with the SQLE inhibitor terbinafine can suppress the proliferation of LUsC cells by inducing apoptosis and reactive oxygen species accumulation.However,depletion of SQLE also results in the impairment of lipid peroxidation and ferroptosis resistance such as upregulation of glutathione peroxidase 4.Therefore,prevention of SQLE in synergy with glutathione peroxidase 4 inhibitor RSL3 effectively mitigates the proliferation and growth of LUSC.Conclusion:Our study indicates that the low expression of sQLE employs adaptive survival through regulating the balance of apoptosis and ferroptosis resistance.In future,the combinational therapy of targeting sQLE and ferroptosis could be a promising approach in treating LUSC.

Primary cutaneous aspergillosis due to Aspergillus flavus: a case report

Infections caused by opportunistic organisms which have been known asetiologic agents of disease become more and more frequent. Aspergillus spp. is one of the agents.Fungi of aspergillus genus are widely distributed in nature, particularly in the soil and in thedecomposed vegetation. They are frequent opportunist pathogens in immunocompromised patients. Themost frequent causative organisms that cause cutaneous aspergillosis are A. fumigatus and A. flavus.In this report, we present a case of primary cutaneous aspergillosis manifested by ulceration ofthe shank due to A. flavus. The patient had no deficiency of immunological status and severe diseaseassociated with fungal infection. Excellent response was shown to anti-fungal therapy.

Squalene epoxidase promotes colorectal cancer cell proliferation through accumulating calcitriol and activating CYP24A1-mediatedMAPK signaling

Background:Colorectal cancer(CRC)is one of the most malignant tumorswith high incidence,yet its molecular mechanism is not fully understood,hindering the development of targeted therapy.Metabolic abnormalities are a hallmark of cancer.Targeting dysregulated metabolic features has become an important direction for modern anticancer therapy.In this study,we aimed to identify a new metabolic enzyme that promotes proliferation of CRC and to examine the related molecular mechanisms.Methods:We performed RNA sequencing and tissue microarray analyses of human CRC samples to identify new genes involved in CRC.Squalene epoxidase(SQLE)was identified to be highly upregulated in CRC patients.The regulatory function of SQLE in CRC progression and the therapeutic effect of SQLE inhibitors were determined by measuring CRC cell viability,colony and organoid formation,intracellular cholesterol concentration and xenograft tumor growth.Themolecularmechanism of SQLE functionwas explored by combining transcriptome and untargeted metabolomics analysis.Western blotting and realtime PCR were used to assess MAPK signaling activation by SQLE.Results:SQLE-related control of cholesterol biosynthesis was highly upregulated in CRC patients and associated with poor prognosis.SQLE promoted CRC growth in vitro and in vivo.Inhibition of SQLE reduced the levels of calcitriol(active form of vitamin D3)and CYP24A1,followed by an increase in intracellular Ca2+concentration.Subsequently,MAPK signaling was suppressed,resulting in the inhibition of CRC cell growth.Consistently,terbinafine,an SQLE inhibitor,suppressed CRC cell proliferation and organoid and xenograft tumor growth.Conclusions:Our findings demonstrate that SQLE promotes CRC through the accumulation of calcitriol and stimulation of CYP24A1-mediated MAPK signaling,highlighting SQLE as a potential therapeutic target for CRC treatment.

Fungal Translocation Marker in People Living with HIV Needing Treatment for Onychomycosis: A Protocol for the Prospective Pilot Study

Increased microbial translocation and chronic immune activation are two critical problems for people living with HIV(PLWH)in the antiretroviral therapy(ART)era.Compared with numerous studies on bacterial microbiomic communities,there are only a limited number of studies focusing on fungal microbiomic composition and products in PLWH.This study protocol is used to evaluate the changes in bacterial and fungal microbiome populations induced by terbinafine treatment,which is an antifungal agent widely used amongst PLWH.Twenty-two PLWH on a stable ART regimen for more than six months,who require treatment for onychomycosis,will be recruited.The participants will be followed-up for a 12-week treatment period(oral terbinafine 250mg daily)and another 12-weeks of terbinafine discontinuation.Plasma and fecal samples will be collected before and after terbinafine treatment,and for 12weeks after the discontinuation of terbinafine.Plasma gut injury and microbial translocation biomarker assays,in addition to testing for gut microbiome composition,will be undertaken.With this pilot study,we will perform formal sample size calculations and test study feasibility for a possible full-scale study.