<i>In-Vitro</i>Comparison of Antifungal Activity of Herbs (Darehald and Pomegranate) with Azoles

Aim: Candida, an opportunistic organism is one of the commonest causes of hospital acquired infections among fungi. Currently available antifungal drugs have numerous adverse effects and drug-drug interactions (DDIs) along with increase in resistance over the time. Therefore, it is highly emergent to consider alternative treatments for candidal infections, having fewer adverse effects and is cost-effective. The current in-vitro study is undertaken to assess and compare the antifungal effects of the herbs, Berberis aristata (B. aristata, Darehald/Darhald) and Punica granatum (P. granatum, Pomegranate) with fluconazole and voriconazole, based on culture and sensitivity of candidal isolates. Materials and Methods: Ethanolic extracts of herbs (Berberis aristata and Punica granatum) and concentrations were formulated as per standard procedure. 130 samples were obtained for the study from in and out patients reported in clinical subsets of Ziauddin Hospital, Karachi from March to May, 2018. Samples were collected and grown according to the standard procedures like, wet mount test and gram’s staining. Species were identified by CHROM agar candida and API 20 C AUX methods. Sensitivity tests were performed by Kirby Bauer’s disc diffusion method according to CLSI guide lines M-44 A2, 2009. Data analysis was done by one-way ANOVA to compare the antifungal activities of drugs and herbs. Results: Mean inhibitory zones of herbs, B. aristata and P. granatum were highly significant against clinical candidal isolates with respective p-values of 0.00 and 0.02. Both of the herbs, B. aristata and P. granatum were found to be more sensitive, 98.5% and 97.7% respectively in comparison to fluconazole showing 42.3% and voriconazole showing 29.2% sensitivity against candidal isolates. The most resistant candidal specie was C. tropicalis that showed resistance against both fluconazole and voriconazole, contrary to that, this specie was highly sensitive to both of the herbs, showing sensitivity of 100% respective for Darehald and Pomegranate. Conclusion: In comparison to azoles: Culture sensitivity of both herbs (B. aristata and P. granatum) displayed more sensitivity against candidal isolates of patients having non-invasive and invasive candidiasis. These herbs can be considered as substitute or alternative antifungal agents to the conventional antifungal therapy, particularly in cases of treating candidemia patients, which is a life threating condition.

Nitrogen-rich azoles as trifunctional electrolyte additives for high-performance lithium-sulfur battery

Rechargeable lithium-sulfur(Li-S)batteries are considered one of the most promising energy storage techniques owing to the high theoretical energy density.However,challenges still remain such as the shuttle effect of lithium polysulfides(LPSs)and the instability of lithium metal anode.Herein,we propose to use nitrogen-rich azoles,i.e.,triazole(Ta)and tetrazole(Tta),as trifunctional electrolyte additives for Li-S batteries.The azoles afford strong lithiophilicity for the chemisorption of LPSs.The density functional theory and experimental analysis verify the presence of Li bonds between the azoles and LPSs.The azoles can also interact with lithium salt in the electrolyte,leading to increase ionic conductivity and lithiumion transference number.Moreover,the azoles render particle-like lithium deposition on the lithium metal anode,leading to superlong cycling of a Li symmetric cell.The Li-S batteries with Ta and Tta exhibit the initial discharge capacity of 1425.5 and 1322.2 m Ah g^(-1),respectively,at 0.2 C rate,and promising cycling stability.They also enable enhanced cycling performance of a Li-organosulfide battery.

Contribution to a Comparative Study of the Corrosion Inhibiting Effect of Some Azoles, to Protect the Cu70-30Ni Alloy in Aerated NaCl 3% in Presence of Ammonia

Some azoles were tested such as 3-amino-1,2,4-triazole (ATA), 3-4’-bitriazole -1,2,4 (BiTA)and 2-Mercaptobenzimidazole (MBI) against Cu-30Ni alloy corrosion in 3%NaCl polluted by ammonia using potentiodynamic measurements and electrochemical impedance spectroscopy and non-electrochemical techniques (scanning Electron Microscopy (SEM)) studied surface morphology has been used to characterize electrode surface. This study permitted to follow the evolution of the inhibitory effect of some azoles, on Cu-30Ni alloy corrosion in 3%NaCl polluted by ammonia and indicate that the tested inhibitors act as a good mixed-type inhibitor retarding the anodic and cathodic reactions. An increase of the inhibitors concentration leads to a decrease of corrosion rate and inhibition efficiency increase.

Alkylene-functionality in bridged and fused nitrogen-rich poly-cyclic energetic materials:Synthesis,structural diversity and energetic properties

From the standpoint of chemical structures,the organic backbones of energetic materials can be classified into aromatic rings,nonaromatic rings,and open chains.Although the category of aromatic energetic compounds exhibits several advantages in the regulation of energetic properties,the nonaromatic heterocycles,assembling nitramino explosophores with simple alkyl bridges,still have prevailed in benchmark materials.The methylene bridge plays a pivotal role in the constructions of the classic nonaromatic heterocycle-based energetic compounds,e.g.,hexahydro-1,3,5-trinitro-1,3,5-triazine(RDX)and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine(HMX),whereas ethylene bridge is the core moiety of state-of-the-art explosive 2,4,6,8,10,12-hexanitro-2,4,6,8,10,12-hexaazaisowurtzitane(CL-20).In this context,it is of great interest to employ simple and practical bridges to assemble aromatic and nonaromatic nitrogen-rich heterocycles,thereby expanding the structural diversity of energetic materials,e.g.,bridged and fused nitrogen-rich poly-heterocycles.Furthermore,alkyl-bridged poly-heterocycles highlight the potential for the open chain type of energetic materials.In this review,the development of alkyl bridges in linking nitrogen-rich heterocycles is presented,and the perspective of the newly constructed energetic backbones is summarized for the future design of advanced energetic materials.

Cul/8-Hydroxyquinalidine Promoted N-Arylation of Indole and Azoles

An efficient catalytic system of CuI/8-hydroxyquinalidine was developed for the coupling of aryl iodides and indole as well as some azoles. The reaction could be carried out at 90 ~C under the condition of relatively low cata- lyst loading, affording various N-arylindoles and N-aryl azoles in good yields. The functionalized and hindered aryl iodides were suitable substrates for this transformation.

Iron（lll）-Catalyzed Direct N-Alkylation of Azoles via Oxidative Transformation of sp3 C H Bonds under Solvent-Free Conditions

A new approach to synthesize N-alkylation of azoles by iron（III）-catalyst under solvent-free conditions was de- veloped. The method is broad in scope and highly efficient.

Azole selenourea disrupted the midgut and caused malformed development of Plutella xylostella

Chemical insecticides targeting the digestive system of diamondback moth(DBM),Plutella xylostella,have not been developed.The discovery of an insecticide with novel mode of action is a challenge for the control of DBM.In this study,a class of selenium-and difluoromethyl-modified azoles(fluoroazole selenoureas,FASU)were designed and synthesized for the control of DBM.Of these azoles,compound B4 showed the highest insecticidal activity against DBM.The LC50of third-and second-instar larvae reached 32.3 and 4.6μg mL^(–1),respectively.The midgut tissue of larvae was severely disrupted,and the larval intestinal tissue was dotted with unique red spots after treatment with compound B4.Compound B4 led to disintegration of the peritrophic matrix,swelling of the midgut epithelium,fracture of the microvilli,and extensive leakage of cellular debris in the midgut lumen.Surviving larvae grew very slowly,and the larval duration was significantly prolonged after exposure to compound B4 at sublethal doses(LC10,LC25and LC50).Furthermore,the pupation rate,emergence rate and pupae weight were significantly decreased.Compound B4 also induced abnormal pupae,causing adults to be trapped in the cocoon or failure to fly due to twisted wings.These results demonstrated that FASU could reduce the population of DBM in sublethal doses.FASU is the first synthetic insecticidal lead compound that has been shown to disrupt the midgut tissue of the larvae of DBM,and its mode of action totally differs from that of commercial chemical insecticides.

微生物来源的Azole类抗真菌活性增强剂——FKI-0076A、B和C的研究

FKI- 0 0 76 A、B和 C是从真菌培养液中筛选 Azole类抗真菌活性增强剂过程中获得的 ,每 6 mm纸片 10μg FKI- 0 0 76 A、B或 1.0μg FKI- 0 0 76 C就显示增强活性。真菌 Talaromyces sp.FKI- 0 0 76培养 5 d后 ,经有机溶媒提取 ,常压 ODS柱层析和制备 HPL C柱层析分离 ,得到 FKI- 0 0 76 A、B和 C纯品。又经过测定其HR- MS、UV、IR、1 H- NMR、1 3C- NMR、DEPT、1 H- 1 H COSY、HMQC和 HMBC等光谱数据。将 FKI- 0 0 76 A的结构定为 BK 2 2 3- A,FKI- 0 0 76 B的结构定为 vermistatin,FKI- 0 0 76 C的结构定为 deoxyfunicone。

Efficacy of voriconazole on leishmaniasis by Leishmania major: An in vitro and in vivo study

Objective: To appraise the activity of voriconazole against Leishmania major(L. major) in vitro and its effectiveness on wound regeneration in cutaneous leishmaniasis in BALB/c mice Methods: The IC_(50) of voriconazole against promastigotes and intra-macrophage amastigotes of L. major was investigated in vitro. The in vivo study was performed by treating the L. major infected BALB/c mice. When the wounds appeared in the base of tail, treatment was started by administration of 30 mg/kg voriconazole for 28 consecutive days orally. Results: The IC_(50) of voriconazole against promastigotes and intra-macrophage amastigotes were 0.74 and 0.89μM, respectively. Voriconazole decreased lipid peroxidation and IL-6 level. Histopathologica findings indicated accelerated healing in the voriconazole treated group compared to other groups. Conclusions: Our results demonstrate that voriconazole can be an option in treating the cutaneous leishmaniasis by L. major.

ERG11 mutations associated with azole resistance in Candida albicans isolates from vulvovaginal candidosis patients

Objective: To investigate the azole susceptibility of Candida albicans(C.albicans)from vulvovaginal candidosis patients and to analyze the relationship between ERG11 gene mutations in these isolates and azole resistance.Methods: Three hundred and two clinical isolates of Candida species were collected.Azole susceptibility was tested in vitro in microdilution studies. The ERG11 genes of 17 isolates of C. albicans(2 susceptibles, 5 dose-dependent resistants and 10 resistants) were amplified and sequenced.Results: Of the 302 isolates collected, 70.2% were C. albicans, of which 8.5%, 3.8% and4.2% were resistant to fluconazole, itraconazole and voriconazole, respectively. In total,27 missense mutations were detected in ERG11 genes from resistant/susceptible dosedependent isolates. Among them, Y132 H, A114 S, and Y257 H substitutions were most prevalent and were known to cause fluconazole resistance. G464 S and F72 S also have been proved to cause fluconazole resistance. Two novel substitutions(T285A, S457P) in hotspot regions were identified.Conclusions: Twenty seven mutations in the ERG11 gene were identified in azoleresistant C. albicans isolates, which indicated a possible relation with the increase in resistance to azole drugs and the recurrence of vulvovaginal candidosis. The relationship of two novel substitutions(T285A, S457P) with fluconazole resistance needs to be further verified by site-directed mutagenesis.

Arylated analogues of cypronazole: fungicidal effect and activity on human fibroblasts. Docking analysis and molecular dynamics simulations

Triadimefon(TDM)and cyproconazole(CPZ)are two triazoles widely used as fungicides.Several azoles were synthe-sised starting from commercial TDM and CPZ.The compounds were evaluated against phytopathogenic filamentous fungi,including Aspergillus fumigatus(AF),A.niger(AN),A.ustus(AU),A.japonicus(AJ),A.terreus(AT),Fusarium oxyspo-rum and Botrytis cinerea isolated from grapevine in the province of San Juan,Argentina.Three of the synthesised compounds(1-(Biphenyl-4-yloxy)-3,3-dimethyl-1-(1H-1,2,4-triazol-1-yl)butan-2-one,1;2-(Biphenyl-4-yl)-3-cyclopropyl-1-(1H-1,2,4-triazol-1-yl)butan-2-ol,3;3-Cyclopropyl-2-(4’-fluorobiphenyl-4-yl)-1-(1H-1,2,4-triazol1-yl)butan-2-ol,4)presented remarkable in vitro fungicidal properties,with better effects than TDM and CPZ on some of the target fungi.Cytotoxicity was assessed using human lung fibroblasts MRC5.Derivative 1,with IC50 values of 389.4μM,was less toxic towards MRC-5 human lung fibroblasts than commercial TDM(248.5μM)and CPZ(267.4μM).Docking analysis and molecular dynamics simulations suggest that the compounds present the same interaction in the binding pocket of the CYP51B enzyme and with the same amino acids as CPZ.The derivatives investigated could be considered broad-spectrum but with some selectivity towards imperfect fungi.

Azole derivatives embedded in montmorillonite clay nanocarriers as corrosion inhibitors of mild steel

Azole derivatives such as 2-mercaptobenzothiazole(MBT) and 2-mercaptobenzimidazole(MBI) were introduced as corrosion inhibitors into the interlayer space of sodium montmorillonite clay(Na+-MMT). The corrosion protection behavior of mild steel in solutions containing MBT, MBI, MMT + MBT, MMT + MBI, Na^+-MMT, and NaCl(3.5 wt%) was evaluated using polarization and electrochemical impedance spectroscopy(EIS). Also, the release of penetrated species into the medium from the clay nanocarriers was evaluated using ultraviolet-visible(UV-Vis) spectroscopy. Small-angle X-ray scattering(SAXS) confirmed the insertion of MBT and MBI into the inner space of the clay layers and the interaction between two organic and inorganic phases. Scanning electron microscopy(SEM) was used to assess the morphology of the surface of the steel samples after the samples had been immersed for 24 h in the extraction solution. The corrosion protection in the solutions with clay nanocarriers containing MBT and MBI was better than that in solutions without MMT. The UV-Vis results showed that the release of MBI species from Na+-MMT nanocarriers in neutral pH was far lower than that of MBT species.

Synthesis of a Photolabeling Probe for the Study of Antiviral Mechanism of Ribavirin

Ribavirin has been used in urgency to treat SARS patients recently. In order to study its antiviral mechanism by photolabeling approach, we have synthesized and characterized 5-azido-1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxiamide 1 as a photolabeling probe of ribavirin. The azidotriazole nucleoside showed rapid and clean photochemical reaction, suggesting that l is a promising probe to study the antiviral mechanism of ribarivin by photolabeling.

Cardiac Toxicity of Azole Antifungals

Cardiac toxicity is an uncommon side effect of anti-fungal therapy. Until the recent reports of itraconazole (ITZ) associated cardiac failure, amphotericin was the antifungal most frequently reported with arrhythmias. We evaluated the cardiac effect of azole antifungals, specifically ITZ, and possible mechanisms of toxicity. Ex vivo live-heart studies were performed utilizing Sprague Dawley rats. Short exposure (<5 minutes), random crossover, dose ranging studies were performed with each pharmacologic agent. ITZ focused trials also included dose ranging utilizing a non-crossover design. The only azole found to have significant toxicity was ITZ. At ITZ ~ ED25 (2 - 2.5 ug/mL) exposures, contractility decreased by 22.2% ± 15.7% and amplitude of left ventricular pressure decreased by 11% ± 0.17%. Electron micrograph and alterations in mitochondrial respiration suggest mitochondrial toxicity as an underlying mechanism. In conclusion, ITZ was associated with reductions in contractility, possibly secondary to mitochondrial dysfunction and dilated cardiomyopathy.

Basic Research – Significance of Detection and Clinical Impact of <i>Candida albicans</i>in Non-Immunosuppressed Patients

Background: The clinical significance of the detection of Candida albicans on mucous membranes of the respiratory or intestinal tract from patients in intensive care units is still not finally clarified. Many patients reveal colonization, although, despite increased risk, there are only a few invasive infections detectable. Therefore, antimycotic therapy in this setting is strongly discouraged. In reality, however, many patients receive antimycotics as a pre-emptive therapy. To elucidate this point, a literature research was performed. Results: In the light of new results on the pathogenicity of C.albicans, the recommendation not to treat should be discussed anew. Without becoming invasive, C.albicans influences the immune system negatively in an anti-inflammatory sense (Th2) by means of at least two distinct mechanisms [action on toll like receptors (TLR), production of farnesol], which will be discussed. Conclusion: It is believed that patients in the phase of CARS or MARS can be further endangered by concomitant colonization of mucous membranes by C.albicans, i.e., in the sense of an anti-inflammatory immune response. Treatment with azole preparations, like fluconazole, which interacts with ergosterol synthesis in this phase of the disease, may trigger an additional effect on the patient, through increase of farnesol concentration by way of a negative feedback. Results of animal experiments on the immune system and concomitant therapeutic consequences indicate the need for verification through clinical trials.

Occurrence and risk assessment of azole fungicides during the urban water cycle:A year-long study along the Yangtze River,China

Azole fungicides(AFs)play an important role in the prevention and treatment of fungal diseases in agricultural crops.However,limited studies are addressing the fate and ecological risk of AFs in the urban water cycle at a large watershed scale.To address this gap,we investigated the spatiotemporal distribution and ecological risk of twenty AFs in the lower reaches of the Yangtze River across four seasons.Carbendazim(CBA),tebuconazole(TBA),tricyclazole(TCA),and propiconazole(PPA)were found to be the dominant compounds.Their highest concentrations were measured in January(188.3 ng/L),and November(2197.1 ng/L),July(162.0 ng/L),and November(1801.9 ng/L),respectively.The comparison between wastewater treatment plants(WWTPs)effluents and surface water suggested that industrial WWTPs are major sources of AFs in the Yangtze River.In particular,TBA and PPA were found to be the most recalcitrant AFs in industrial WWTPs,while difenoconazole(DFA)was found to be the most potent pollutant in municipal WWTPs,with an average removal rate of less than60%.The average risk quotient(RQ)for the entire AFs was 6.45 in the fall,which was higher than in January(0.98),April(0.61),and July(0.40).This indicates that AFs in surface water posed higher environmental risks during the dry season.Additionally,the exposure risk of AFs via drinking water for sensitive populations deserves more attention.This study provides benchmark data on the occurrence of AFs in the lower reaches of the Yangtze River,and offers suggestions for better reduction of AFs.

Synthesis, Crystal Structure and Catalytic Activity of a Hexa-μ-chloro-tetrakis-(1-triphenylmethyltriazole)-μ4-oxo-tetracopper(Ⅱ)

A new Cu(Ⅱ) complex,[Cu_(4)Cl_(6)O(1-TrTz)_(4)]where 1-TrTz is 1-trityl-1H-[1,2,4]triazole,was synthesized by the reaction of 1-trityl-1H-[1,2,4]triazole with two hydrated cupric chlorides,and its structure was characterized by X-ray single-crystal diffraction.[Cu4Cl6O(1-TrTz)4]is of monoclinic system,space group P21/c,a=12.5024(13),b=26.400(3),c=28.588(3)A,β=112.807(4)°,V=8698.1(16)A^(3),Z=4,ρcalc=1.411 g/cm^(3),μ=1.265 mm^(–1),F(000)=3772,the final R=0.1293 and w R=0.3644 for 55774 observed reflections (I>2s(I)),R(all data)=0.1972,w R(all data)=0.3989,completeness to theta of 25.242 is 99.9%and GOF=1.103.In the structure of[Cu_(4)Cl_(6)O(1-TrTz)_(4)],the central O atom is tetrahedral and is coordinated with four Cu atoms.The distance between copper and oxygen bond is 1.909(6)A.In addition,the catalytic property of this complex was investigated and it could effectively catalyze the Ullmann coupling reaction of various (hetero)aryl chlorides with azoles.

Alcohol dehydrogenase I expression correlates with CDR1, CDR2 and FLU1 expression in Candida albicans from patients with vulvovaginal candidiasis

Background The most critical mechanism governing drug resistance in Candida albicans （C. albicans） involves efflux pumps, the functionality of which largely depends on energy metabolism. Alcohol dehydrogenase I （ADH1） plays an important role in intracellular energy metabolism. The aim of this study was to explore the relationship between ADH1 and drug resistance in C. albicans. Methods Twenty clinical C. albicans samples isolated from individual patients diagnosed with vulvovaginal candidiasis, and two C. albicans strains obtained from a single parental source （the fluconazole （FLC）-sensitive strain CA-1s and the FLC-resistant strain CA-16R） were included in our study. In accordance with the Clinical and Laboratory Standards Institute （CLSI） M27-A3 guidelines, we used the microdilution method to examine the FLC minimum inhibitory concentrations （MICs） and real-time reverse transcription polymerase chain reaction （RT-PCR） to measure the mRNA expression levels of ADH1 and the azole resistance genes CDR1, CDR2, MDR1, FLU1 and ERG11 in all the isolates. Results A highly significant positive correlation between the mRNA levels of ADH1 and the MICs （r =0.921, P=0.000）, as well as positive correlations between the mRNA level of ADH1 and those of CDR1, CDR2 and FLU1 （rs of 0.704, 0.772 and 0.779, respectively, P 〈0.01）, were observed in the 20 clinical C. albicans samples. The relative expression of ADH1 was upregulated 10.63- to 17.61-fold in all of the drug-resistant isolates. No correlations were found between the mRNA levels of ADH1 and those of MDR1 or ERG11 （P 〉0.05）. The mRNA levels of the examined drug resistance genes were higher in the CA-16R strain than in CA-1s, and the mRNA levels of ADH1 in CA-16R were 11.64-fold higher than those in CA-1s （P 〈0.05）. Conclusions These results suggest that high levels of ADH1 transcription are implicated in FLC resistance in C. albicans and that the mRNA expression levels of ADH1 are positively correlated with those of CDR1, CDR2 and FLU1.

Two natural molecules preferentially inhibit azole-resistant Candida albicans with MDR1 hyperactivation

Antifungal drug resistance is a significant clinical problem, and antifungal agents that can evade resistance are urgently needed. In infective niches, resistant organisms often co-existed with sensitive ones, or a subpopulation of antibiotic-susceptible organisms may evolve into resistant ones during antibiotic treatment and eventually dominate the whole population. In this study, we established a co-culture assay in which an azole-resistant Candida albicans strain was mixed with a susceptible strain labeled with green fluorescent protein to mimic in vivo conditions and screen for antifungal drugs. Fluconazole was used as a positive control to verify the validity of this co-culture assay. Five natural molecules exhibited antifungal activity against both susceptible and resistant C. albicans. Two of these compounds, retigeric acid B(RAB) and riccardin D(RD), preferentially inhibited C. albicans strains in which the efflux pump MDR1 was activated. This selectivity was attributed to greater intracellular accumulation of the drugs in the resistant strains. Changes in sterol and lipid compositions were observed in the resistant strains compared to the susceptible strain, and might increase cell permeability to RAB and RD. In addition, RAB and RD interfered with the sterol pathway, further aggregating the decrease in ergosterol in the sterol synthesis pathway in the MDR1-activated strains. Our findings here provide an alternative for combating resistant pathogenic fungi.

Biodegradation of typical azole fungicides in activated sludge under aerobic conditions

Widespread use of azole fungicides and low removal efficiency in wastewater treatment plants(WWTPs) have led to the elevated concentration of azole fungicides in receiving environment. However, there was limited research about the removal mechanism of azole fungicides in the biological treatment of WWTPs. Imidazole fungicide climbazole and triazole fungicide fluconazole were selected to investigate the biodegradation mechanism of azole fungicides in activated sludge under aerobic conditions. Climbazole was found to be adsorbed to solid sludge and resulted in quick biodegradation. The degradation of climbazole in the aerobic activated sludge system was fitted well by the first-order kinetic model with a half-life of 5.3 days, while fluconazole tended to stay in liquid and had only about 30% of loss within 77 days incubation. Ten biotransformation products of climbazole were identified by high resolution mass spectrometry using suspect and non-target screening method. But no biodegradation products of fluconazole were identified due to its limited removal. The possible biodegradation pathways for climbazole were proposed based on the products identification and pathway prediction system, and involves oxidative dehalogenation, side chain oxidation and azole ring loss. The findings from this study suggest that it should be a concern for the persistence of fluconazole in the environment.