Search

Objective: Effective adjuvant therapeutic strategies are urgently needed to overcome MAPK inhibitor (MAPKi) resistance, which is one of the most common forms of resistance that has emerged in many types of cancers. Here, we aimed to systematically identify the genetic interactions underlying MAPKi resistance, and to further investigate the mechanisms that produce the genetic interactions that generate synergistic MAPKi resistance. Methods: We conducted a comprehensive pair-wise sgRNA-based high-throughput screening assay to identify synergistic interactions that sensitized cancer cells to MAPKi, and validated 3 genetic combinations through competitive growth, cell viability, and spheroid formation assays. We next conducted Kaplan-Meier survival analysis based on The Cancer Genome Atlas database and conducted immunohistochemistry to determine the clinical relevance of these synergistic combinations. We also investigated the MAPKi resistance mechanisms of these validated synergistic combinations by using co-immunoprecipitation, Western blot, qRTPCR, and immunofluorescence assays. Results: We constructed a systematic interaction network of MAPKi resistance and identified 3 novel synergistic combinations that effectively targeted MAPKi resistance (ITGB3 + IGF1R, ITGB3 + JNK, and HDGF + LGR5). We next analyzed their clinical relevance and the mechanisms by which they sensitized cancer cells to MAPKi exposure. Specifically, we discovered a novel protein complex, HDGF-LGR5, that adaptively responded to MAPKi to enhance cancer cell stemness, which was up- or downregulated by the inhibitors of ITGB3 + JNK or ITGB3 + IGF1R. Conclusions: Pair-wise sgRNA library screening provided systematic insights into elucidating MAPKi resistance in cancer cells. ITGB3- + IGF1R-targeting drugs (cilengitide + linsitinib) could be used as an effective therapy for suppressing the adaptive formation of the HDGF-LGR5 protein complex, which enhanced cancer stemness during MAPKi stress.

The aim of this study is to compare the azole synergy across an insect, Chironomus riparius, and a crustacean species, Daphnia magna. We use a combination of in vivo measurements of cytochrome P450 monooxygenase (CYP) biotransformation potential and toxicokinetic (TK) and toxicodynamic (TD) modeling to understand the mechanism behind the synergy of two azole fungicides: the imidazole prochloraz and the triazole propiconazole on the pyrethroid insecticide α-cypermethrin. For both species, the synergistic effect of prochloraz was well-described by its effect on in vivo CYP activity, which corresponded to the biotransformation rate of the TK model parameterized on the survival data of the mixture experiment. For propiconazole, however, there were 100-fold and 50-fold differences between the 50% effect concentration of in vivo CYP activity and the modeled biotransformation rate for C. riparius and D. magna, respectively. Propiconazole, therefore, seems to induce synergy through a mechanism that cannot be quantified solely by the CYP activity assay used in this study in either of the two species. We discuss the differences between prochloraz and propiconazole as synergists across the two species in the light of the type and time dynamics of affected biotransformation processes.

Background The European grapevine moth, Lobesia botrana (Denis and Schiffermüller) (Lepidoptera: Tortricidae) is currently the most damaging pest in many viticultural regions across the Mediterranean basin and elsewhere. Its feeding activity also enhances the development of secondary infections by Botrytis cinerea - and other fungi - to wine grapes. The gram-positive bacterium Bacillus thuringiensis (Bt) has been reported to partially control larval populations of L. botrana, while it requires to be digested by the insect to cause infection. Entomopathogenic fungi (EPF) are possibly capable of acting synergistically with Bt to increase its efficacy against insect pests. Results The hypothesis of synergy or antagonism between Bt and EPF for the control of L. botrana was tested in two bioassays: A) Insects fed on Bt diet and subsequently some groups were sprayed by conidia of Beauveria bassiana or Paecilomyces fumosoroseus, and B) Grapes were sprayed by Bt, or B. bassiana, or combination of the two, and then untreated insects were placed to feed on the grapes. In both bioassays, combination treatments performed better than single treatments, indicating additive action or synergy. The Bt and B. bassiana combination treatment (Bt diet for 30 h and then sprayed with conidia of B. bassiana) resulted in 91% larval mortality while the single Bt and B. bassiana treatments caused 28% and 34% mortality respectively. Such results indicated synergism. Combination treatment on grapes also caused significantly higher mortality on L. botrana larvae, compared to single treatments. The median lethal time (LT50) was estimated as 8.43 days for the single Bt treatment, 7.87 days for the single B. bassiana treatment and 6.3 days for the combination Bt + B. bassiana treatment. Conclusions Absence of antagonism as well as additive action or synergy were indicated by the results. Larval populations of the pest can be effectively controlled by using microbial biocontrol agents. Further research is needed to investigate the biotic and abiotic factors that affect interactions between insect hosts and entomopathogenic organisms. However, the entomopathogens used in the present study showed remarkable action and may be included parallelly in control strategies against vineyard pests.

Earlier in our laboratory, the role of various individual sperm impairing microorganisms on sperm parameters and female infertility has been elucidated at higher doses. As, multiple bacterial species tend to exert more pathogenic effect in comparison to single organism hence, present study was carried out to evaluate that if consortia of these sperm impairing organisms can lead to infertility in female mice at sub fertility dose. For this, impact of individual bacterial strains of Escherichia coli, Pseudomonas aeruginosa, Pseudomonas aeruginosa, Klebsiella pneumoniae and consortia of Escherichia coli and Pseudomonas aeruginosa, Escherichia coli and Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae was examined on the motility, viability of mouse spermatozoa and fertility outcome. The results showed that the individual bacterial strains of E. coli, S. marcescens and K. pneumoniae could led to immobilization of spermatozoa by agglutination and P. aeruginosa led to immobilization of spermatozoa without agglutination. Also, all of them led to 100% sperm death in 45 min of incubation. In case of consortia of bacterial strains, the results showed sperm agglutination in all the cases and they were able to induce 100% sperm death at 30 min of incubation time. Further, in vivo studies were carried out to evaluate the impact of individual bacterial strains and consortia of bacterial strains on the fertility outcome in female Balb/c mice. For this, female mice were administered intravaginally with 101 cfu/20µl of individual bacterial strains or consortia of strains for 10 consecutive days or PBS. The results showed that both individual bacterial strains and consortia of bacterial strains were able to efficiently colonize the mouse vagina. Further, control group receiving phosphate buffer saline and groups receiving individual bacterial strains showed all the pregnancy related changes viz. abdominal distension, string of pearls on palpation as well as delivery of pups on completion of gestation period and delivery of pups. The histological examination of reproductive organs viz. uterus and ovary, of the female mice receiving PBS or individual bacterial strains showed the formation of corpus luteum in the ovary and the formation of decidua’s in the uterus, indicative of pregnancy. However, mice receiving consortia of bacterial strains did not show any pregnancy related changes throughout the experiment. Thus, these results indicate that the presence of consortia of sperm impairing microorganisms in vaginal milieu is efficient in provoking infertility even at subfertility doses.

Botrytis cinerea, which causes gray mold disease in more than 200 plant species, is an economically important pathogen that is mainly controlled by synthetic fungicides. Synergistic fungicide mixtures can help reduce fungicide residues in the environment and mitigate the development of fungicide-resistant strains. In this study, we screened microbial culture extracts on Botrytis cinerea to identify an antifungal synergist for tebuconazole. Among the 4,006 microbial extracts screened in this study, the culture extract from Schizophyllum commune displayed the most enhanced activity with a sub-lethal dosage of tebuconazole, and the active ingredient was identified as schizostatin. In combination with 5 μg/ml tebuconazole, schizostatin (1 μg/ml) showed disease control efficacy against gray mold on tomato leaf similar to that achieved with 20 μg/ml tebuconazole treatment alone. Interestingly, schizostatin showed demethylation inhibitor (DMI)-specific synergistic interactions in the crossed-paper strip assay using commercial fungicides. In a checkerboard assay with schizostatin and DMIs, the fractional inhibitory concentration values were 0.0938-0.375. To assess the molecular mechanisms underlying this synergism, the transcription levels of the ergosterol biosynthetic genes were observed in response to DMIs, schizostatin, and their mixtures. Treatment with DMIs increased the erg11 (the target gene of DMI fungicides) expression level 15.4-56.6-fold. However, treatment with a mixture of schizostatin and DMIs evidently reverted erg11 transcription levels to the pre-DMI treatment levels. These results show the potential of schizostatin as a natural antifungal synergist that can reduce the dose of DMIs applied in the field without compromising the disease control efficacy of the fungicides.

Encapsulation in alginate beads has always been limited by the leakage due to the too wide distribution of pore sizes. Mixing alginate with other polymers have sometimes reduced the problem. Hydrocolloids from seven tropical vegetal species (barks of Triumfetta cordifolia and Bridelia thermifolia, seeds of Irvingia gabonensis and Beilschmiedia obscura, and leaves of Ceratotheca sesamoides, Adansonia digitata and Corchorus olitorius) were screened for synergistic interactions with alginate in dilute aqueous solution. Mixtures with alginate were made at different volume proportions and deviations from the initial viscosity set at 1 were evaluated. In distilled water, the gums from T. cordifolia, B. obscura, C. sesamoides and C. olitorius presented synergies with alginate. In 2 mM calcium chloride, the seven gums showed positive synergy. Interactions are favored by gum flexibility and the presence of charges, although high charges reduced the interactions. Alginate fraction of maximum viscosity enhancement depends on the ability to conformational order of the gum. The measure by laser diffraction of alginate-gum particles sizes at different fractions showed that the cooperative interactions did not always involve the largest complexes formed in gums associations. The occurrence of these interactions predicts the formation of homogeneous mixed gels at higher polymer and calcium concentrations., Food Science; Food Technology; Physical Chemistry; Alginate; Calcium Chloride; Synergistic interactions; Tropical vegetal gums.

Genetically engineered crops simultaneously produce defensive allelochemicals and Bacillus thuringiensis (Bt) toxin proteins to kill some of the world's most devastating insect pests. How the two types of toxins, when ingested sequentially or simultaneously, interact at both lethal and sublethal doses in these pests remains underexplored. Here, we examined the toxicological interactions between the Bt toxin Cry1Ac and the flavonoid allelochemical flavone in Helicoverpa armigera. Simultaneous exposure of H. armigera neonates to lethal doses (LC25 ) of Cry1Ac and flavone caused a mortality significantly higher than that of either toxin alone and their expected additive mortality. Preexposure for 24 h to a sublethal dose (LC10 ) of Cry1Ac followed by 6-d simultaneous exposure to the same dose of Cry1Ac plus a lethal dose (1.6 mg/g diets, LC50 ) of flavone resulted in a mortality significantly higher than that of the LC50 dose of flavone alone and the expected additive mortality of the LC50 dose of flavone plus the LC10 dose of Cry1Ac. One-day preexposure to the sublethal dose (LC10 ) of flavone followed by 6-d simultaneous exposure to the LC50 dose (6 ng/cm2 ) of Cry1Ac plus the LC10 dose of flavone yielded a mortality significantly higher than that of the LC50 dose of Cry1Ac but similar to the expected additive mortality of the LC50 dose of Cry1Ac plus the LC10 dose of flavone. The results suggest that Cry1Ac induces and synergizes the toxicity of flavone against H. armigera larvae.

Cajal bodies (CBs) are subnuclear domains involved in the formation of ribonucleoproteins (RNPs) including small nuclear RNPs (snRNPs). CBs associate with specific gene loci, which impacts expression and provides a platform for the biogenesis of the nascent transcripts emanating from these genes. Here we report that CBs can associate with the C19MC microRNA (miRNA) gene cluster, which suggests a role for CBs in the biogenesis of animal miRNAs. The machinery involved in the formation of miRNAs includes the Drosha/DGCR8 complex, which processes primary-miRNA to precursor miRNA. Further processing of precursor miRNA by Dicer and other components generates mature miRNA. To test if CBs influence the expression and formation of miRNAs, we examined two representative miRNAs (miR-520 h and let-7a) in conditions that disrupt CBs. CB disruption correlates with alterations in the level of primary and mature miRNA and the let-7a mRNA target, HMGA2. We have also found that the processing of some small CB-specific RNAs (scaRNAs) is directly mediated by the Drosha/DGCR8 complex. ScaRNAs form scaRNPs, which play an important role in snRNP formation. Collectively, our results demonstrate that CBs and the miRNA processing machinery functionally interact and together contribute to the biogenesis of miRNAs and snRNPs.

Cel48A of Thermobifida fusca (TfCel48A) is a processive exocellulase that contains an active site tunnel and digests lignocellulosic biomass via synergistic interactions between different cellulases. Cel48A possesses a number of aromatic amino acids lining the tunnel entrance, which are highly conserved across a diverse number of microbial species and appear to play a role in the selection and threading of individual strands of cellulose from highly recalcitrant substrates. In this study, we sought to further elucidate the roles of these tunnel entrance aromatic amino acids by creating a series of double mutants and examining their effect on TfCel48A activity, processivity, and synergistic interactions with the well-studied processive endocellulase TfCel9A. Our results provide further insight concerning the mechanism of Cel48A kinetics with soluble and insoluble substrates and could play an influential role in the application of Cel48A and other exocellulases for industrial purposes.

Panicum mosaic virus (PMV) (genus Panicovirus, family Tombusviridae) and its molecular parasite, Satellite panicum mosaic virus (SPMV), synergistically interact in coinfected proso and pearl millet (Panicum miliaceum L.) plants resulting in a severe symptom phenotype. In this study, we examined synergistic interactions between the isolates of PMV and SPMV by using PMV-NE, PMV85, SPMV-KS, and SPMV-Type as interacting partner viruses in different combinations. Coinfection of proso millet plants by PMV-NE and SPMV-KS elicited severe mosaic, chlorosis, stunting, and eventual plant death compared with moderate mosaic, chlorotic streaks, and stunting by PMV85 and SPMV-Type. In reciprocal combinations, coinfection of proso millet by either isolate of PMV with SPMV-KS but not with SPMV-Type elicited severe disease synergism, suggesting that SPMV-KS was the main contributor for efficient synergistic interaction with PMV isolates. Coinfection of proso millet plants by either isolate of PMV and SPMV-KS or SPMV-Type caused increased accumulation of coat protein (CP) and genomic RNA copies of PMV, compared with infections by individual PMV isolates. Additionally, CP and genomic RNA copies of SPMV-KS accumulated at substantially higher levels, compared with SMPV-Type in coinfected proso millet plants with either isolate of PMV. Hybrid viruses between SPMV-KS and SPMV-Type revealed that SPMV isolates harboring a CP fragment with four differing amino acids at positions 18, 35, 59, and 98 were responsible for differential synergistic interactions with PMV in proso millet plants. Mutation of amino acid residues at these positions in different combinations in SPMV-KS, similar to those as in SPMV-Type or vice-versa, revealed that A35 and R98 in SPMV-KS CP play critical roles in enhanced synergistic interactions with PMV isolates. Taken together, these data suggest that the two distinct amino acids at positions 35 and 98 in the CP of SPMV-KS and SPMV-Type are involved in the differential synergistic interactions with the helper viruses.

Cannabidiol (CBD), a nonpsychoactive phytocannabinoid, has recently emerged as a potential cytotoxic agent in addition to its ameliorative activity in chemotherapy-associated side effects. In this work, the potential interactions of CBD with docetaxel (DOC), doxorubicin (DOX), paclitaxel (PTX), vinorelbine (VIN), and 7-ethyl-10-hydroxycamptothecin (SN−38) were explored in MCF7 breast adenocarcinoma cells using different synergy quantification models. The apoptotic profiles of MCF7 cells after the treatments were assessed via flow cytometry. The molecular mechanisms of CBD and the most promising combinations were investigated via label-free quantification proteomics. A strong synergy was observed across all synergy models at different molar ratios of CBD in combination with SN−38 and VIN. Intriguingly, synergy was observed for CBD with all chemotherapeutic drugs at a molar ratio of 636:1 in almost all synergy models. However, discording synergy trends warranted the validation of the selected combinations against different models. Enhanced apoptosis was observed for all synergistic CBD combinations compared to monotherapies or negative controls. A shotgun proteomics study highlighted 121 dysregulated proteins in CBD-treated MCF7 cells compared to the negative controls. We reported the inhibition of topoisomerase II β and α, cullin 1, V-type proton ATPase, and CDK-6 in CBD-treated MCF7 cells for the first time as additional cytotoxic mechanisms of CBD, alongside sabotaged energy production and reduced mitochondrial translation. We observed 91 significantly dysregulated proteins in MCF7 cells treated with the synergistic combination of CBD with SN−38 (CSN−38), compared to the monotherapies. Regulation of telomerase, cell cycle, topoisomerase I, EGFR1, protein metabolism, TP53 regulation of DNA repair, death receptor signalling, and RHO GTPase signalling pathways contributed to the proteome-wide synergistic molecular mechanisms of CSN−38. In conclusion, we identified significant synergistic interactions between CBD and the five important chemotherapeutic drugs and the key molecular pathways of CBD and its synergistic combination with SN−38 in MCF7 cells. Further in vivo and clinical studies are warranted to evaluate the implementation of CBD-based synergistic adjuvant therapies for breast cancer.

The emergence of multidrug resistant (MDR) pathogens is a global threat and has created problems in providing adequate treatment of many infectious diseases. Although the conventional antimicrobial agents are quite effective against several pathogens, yet there is a need for more effective antimicrobial agents against MDR pathogens. Herbal drugs and phytochemicals have been used for their effective antimicrobial activity from ancient times and there is an increasing trend for development of plant based natural products for the prevention and treatment of pathogenic diseases. One of the strategies for effective resistance modification is the use of antimicrobial agent-phytochemical combinations that will neutralize the resistance mechanism, enabling the drug to still be effective against resistant microbes. These phytochemicals can work by several strategies, such as inhibition of target modifying and drug degrading enzymes or as efflux pumps inhibitors. A plethora of herbal extracts, essential oils and isolated pure compounds have been reported to act synergistically with existing antibiotics, antifungals and chemotherapeutics and augment the activity of these drugs. Considerable increases in the susceptibility pattern of several microbes towards the natural antimicrobials and their combinations were observed as indicated by significant decline in minimum inhibitory concentrations. This review paper summarizes the current developments regarding synergistic interactions of plant extracts and isolated pure compounds in combination with existing antibacterial, antifungal agents and chemotherapeutics. The effect of these agents on the susceptibility patterns of these pathogens and possible mechanisms of action are described in detail. In conclusion, many phytochemicals in combination with existing drugs were found to act as resistance modifying agents and proper combinations may rescue the efficacy of important lifesaving antimicrobial agents.

Bacterial species in the human gut predominantly exist in the form of mixed-species biofilms on mucosal surfaces. In this study, the biofilm-forming ability of many human gut bacterial strains (133 strains recovered from human faeces) on mucin-coated and non-coated polystyrene surfaces was determined. A significant variation (P < 0.05) in the biofilm-forming ability of many bacterial species on both surfaces was noticed. Based on some preliminary trials, four bacterial species were selected (Bifidobacterium bifidum, Bifidobacterium longum subsp. infantis, Parabacteroides distasonis and Bacteroides ovatus), which could not form any abundant biofilm individually under the in vitro conditions investigated, but produced abundant biofilms when co-cultured in different combinations of two, three and four species, giving an evidence of synergistic interactions in multispecies biofilm formation. There was a 4.74-fold increase in the biofilm mass when all strains developed a biofilm together. Strain-specific qPCR analysis showed that B. bifidum was the most dominant species (56%) in the four-species biofilm after 24 h, followed by B. longum subsp. infantis (36.2%). Study involving cell free supernatant of the cooperating strains showed that cell viability as well as physical presence of cooperating cells were prerequisites for the observed synergy in biofilms. The molecular mechanism behind these interactions and subsequent effects on the functionality of the strains involved were not determined in our study but merit further work.

A series of HPW/Zn-Ti layered double hydroxides (LDHs) were prepared by immobilizing phosphotungstic acid (HPW) on the Zn-Ti LDHs. Systematically characterizations were performed to elucidate their structure. These HPW/Zn-Ti LDHs hybrids were used as heterogeneous catalysts in the epoxidation of fatty acid methyl ester (FAMEs) with H2O2. The HPW/Zn-Ti LDHs showed higher activity than bulk HPW and Zn-Ti LDHs. The conversion of FAMEs and the selectivity to epoxy fatty acid methyl ester (EFAME) were up to 90.6 and 86.0%, respectively. (PO4[WO2(O2)]4)3- was presumed to be the active site for the epoxidation in this work. The formed oxygen-donating intermediate Ti-OOH transferred oxygen free radicals to (PO4[WO2(O2)]4)3- in the catalytic process, accelerating the transfer of oxygen free radicals. Therefore, high catalytic activity of HPW/Zn-Ti LDHs is related to the potential synergistic interactions of Ti-OOH and PW anions.

Background Mortality rate of invasive Candida infections is raising mainly amongst immunocompromised patients. These infections are hard-to-treat mainly due to the increasing incidence of resistance. The overexpression of ATP-binding cassette and major facilitator superfamily transporters is the main responsible for the failure of antifungal therapies. In a Saccharomyces cerevisiae model, β-lapachone inhibited Pdr5p, a transporter homologous to those found in Candida albicans. Aims To determine whether β-lapachone reverses the resistance phenotype mediated by efflux transporters in C. albicans clinical isolates. Methods The antifungal activity of β-lapachone combined with fluconazole was measured by agarose chemosensitization and microdilution assays. CaCdr2p and CaMdr1p activities were evaluated through fluorescent dyes accumulation. ATPase activity was assessed using transporter-enriched plasma membranes. Results β-lapachone reverted antifungal resistance of S. cerevisiae and C. albicans strains overexpressing CaCdr2p and CaMdr1p transporters by inhibiting these proteins activities. CaCdr2p ATPase activity was not impaired by the compound. Conclusions β-lapachone is a promising drug candidate to be used as an adjuvant in the treatment of candidiasis caused by fluconazole-resistant C. albicans strains.

Plants and fungi are closely associated through parasitic or symbiotic relationships in which bidirectional exchanges of cellular contents occur. Recently, a plant virus was shown to be transmitted from a plant to a fungus, but it is unknown whether fungal viruses can also cross host barriers and spread to plants. In this study, we investigated the infectivity of Cryphonectria hypovirus 1 (CHV1, family Hypoviridae), a capsidless, positive-sense (+), single-stranded RNA (ssRNA) fungal virus in a model plant, Nicotiana tabacum. CHV1 replicated in mechanically inoculated leaves but did not spread systemically, but coinoculation with an unrelated plant (+)ssRNA virus, tobacco mosaic virus (TMV, family Virgaviridae), or other plant RNA viruses, enabled CHV1 to systemically infect the plant. Likewise, CHV1 systemically infected transgenic plants expressing the TMV movement protein, and coinfection with TMV further enhanced CHV1 accumulation in these plants. Conversely, CHV1 infection increased TMV accumulation when TMV was introduced into a plant pathogenic fungus, Fusarium graminearum. In the in planta F. graminearum inoculation experiment, we demonstrated that TMV infection of either the plant or the fungus enabled the horizontal transfer of CHV1 from the fungus to the plant, whereas CHV1 infection enhanced fungal acquisition of TMV. Our results demonstrate two-way facilitative interactions between the plant and fungal viruses that promote cross-kingdom virus infections and suggest the presence of plant–fungal-mediated routes for dissemination of fungal and plant viruses in nature.

Farnaz Minooei,1 Joel R Fried,1 Joshua L Fuqua,2– 4 Kenneth E Palmer,3– 5 Jill M Steinbach-Rankins2– 5 1Department of Chemical Engineering, University of Louisville Speed School of Engineering, Louisville, KY, USA; 2Department of Bioengineering, University of Louisville Speed School of Engineering, Louisville, KY, USA; 3Department of Pharmacology and Toxicology, University of Louisville School of Medicine, Louisville, KY, USA; 4Center for Predictive Medicine, University of Louisville, Louisville, KY, USA; 5Department of Microbiology and Immunology, University of Louisville School of Medicine, Louisville, KY, USACorrespondence: Jill M Steinbach-RankinsDepartment of Bioengineering, University of Louisville Speed School of Engineering, 505 S. Hancock St., Room 623, Louisville, KY, 40202, USATel +1 502-852-5486Email jmstei01@louisville.eduIntroduction: Human immunodeficiency virus (HIV) remains a persistent global challenge, impacting 38 million people worldwide. Antiretrovirals (ARVs) including tenofovir (TFV), raltegravir (RAL), and dapivirine (DAP) have been developed to prevent and treat HIV-1 via different mechanisms of action. In parallel, a promising biological candidate, griffithsin (GRFT), has demonstrated outstanding preclinical safety and potency against HIV-1. While ARV co-administration has been shown to enhance virus inhibition, synergistic interactions between ARVs and the oxidation-resistant variant of GRFT (Q-GRFT) have not yet been explored. Here, we co-administered Q-GRFT with TFV, RAL, and DAP, in free and encapsulated forms, to identify unique protein-drug synergies.Methods: Nanoparticles (NPs) were synthesized using a single or double-emulsion technique and release from each formulation was assessed in simulated vaginal fluid. Next, each ARV, in free and encapsulated forms, was co-administered with Q-GRFT or Q-GRFT NPs to evaluate the impact of co-administration in HIV-1 pseudovirus assays, and the combination indices were calculated to identify synergistic interactions. Using the most synergistic formulations, we investigated the effect of agent incorporation in NP-fiber composites on release properties. Finally, NP safety was assessed in vitro using MTT assay.Results: All active agents were encapsulated in NPs with desirable encapsulation efficiency (15– 100%), providing ∼ 20% release over 2 weeks. The co-administration of free Q-GRFT with each free ARV resulted in strong synergistic interactions, relative to each agent alone. Similarly, Q-GRFT NP and ARV NP co-administration resulted in synergy across all formulations, with the most potent interactions between encapsulated Q-GRFT and DAP. Furthermore, the incorporation of Q-GRFT and DAP in NP-fiber composites resulted in burst release of DAP and Q-GRFT with a second phase of Q-GRFT release. Finally, all NP formulations exhibited safety in vitro.Conclusions: This work suggests that Q-GRFT and ARV co-administration in free or encapsulated forms may improve efficacy in achieving prophylaxis.Keywords: griffithsin, nanoparticles, electrospun fibers, antiretrovirals, synergy, HIV-1 prevention, microbicide

Christia vespertilionis locally known as ‘rerama’ has gained much attention as herbal preparation to treat various ailments and diseases although there is no conclusive evidence of its therapeutic values. Self-medication is commonly practiced by patients, underpinned by health believes that affect their adherence to medication regimens, and impacting on treatment outcomes. Therefore, in this study the phytochemical profile and potential interaction of this plant extracts with conventional chemotherapy drug cyclophosphamide were investigated. Water decoction and maceration method with 70% ethanol were used to extract the plant leaves. Phytochemical profiling of the extracts and compound identification was done using HPLC. Isoorientin was identified as the major phytochemical component of the extracts. MTT assay was used to screen cytotoxicity of the extract towards CRL2522, HaCaT, HepG2, MCF-7 and WRL68 cell lines. Combination index (CI) and isobologram were derived from combination treatment of extract and cyclophosphamide. Ethanolic extract has low cytotoxicity against all cell lines (IC50>1mg/mL). However, combination treatment of ethanolic extract and cyclophosphamide at IC15 and IC25 concentration resulted in synergism (CI

The interactions between six commonly occurring fungal species in damp or water-damaged houses in southern California were studied. These fungal species were Alternaria alternata, Aspergillus niger, Chaetomium globosum, Cladosporium herbarum, Penicillium chrysogenum and Stachybotrys chartarum. In the damp building materials, S. chartarum was found to be associated with A. niger, C. globosum, and P. chrysogenum but not with A. alternata and C. herbarum. Stachybotrys chartarum showed strong antagonistic effect against A. alternata and C. herbarum and significantly inhibited in vitro growth of A. alternata and C. herbarum but had no effect on A. niger, C. globosum, and P. chrysogenum. Two trichothecenes, produced by S. chartarum, trichodermin and trichodermol, significantly inhibited spore germination and in vitro growth of A. alternata and C. herbarum but had no effect on A. niger, C. globosum, P. chrysogenum and S. chartarum. In the damp building materials (drywall, ceiling tile, and oak woods), S. chartarum significantly inhibited the growth of A. alternata and C. herbarum and had no effect on the growth and colonization of A. niger, C. globosum, P. chrysogenum in these substrata.

Soil saltiness is a noteworthy issue as it results in loss of profitability and development of agrarian harvests and decline in soil health. Microorganisms associated with plants contribute to their growth promotion and salinity tolerance by employing a multitude of macromolecules and pathways. Plant growth promoting rhizobacteria (PGPR) have an immediate impact on improving profitability based on higher crop yield. Some PGPR produce 1-aminocyclopropane-1-carboxylic (ACC) deaminase (EC 4.1.99.4), which controls ethylene production by diverting ACC into α-ketobutyrate and ammonia. ACC deaminase enhances germination rate and growth parameters of root and shoot in different harvests with and without salt stress. Arbuscular mycorrhizal fungi (AMF) show a symbiotic relationship with plants, which helps in efficient uptake of mineral nutrients and water by the plants and also provide protection to the plants against pathogens and various abiotic stresses. The dual inoculation of PGPR and AMF enhances nutrient uptake and productivity of several crops compared to a single inoculation in both normal and stressed environments. Positively interacting PGPR + AMF combination is an efficient and cost-effective recipe for improving plant tolerance against salinity stress, which can be an extremely useful approach for sustainable agriculture.

Candida auris is an emerging multidrug-resistant pathogen implicated in numerous outbreaks worldwide, with a notably high mortality rate (ca. 60%). A significant challenge with treatment of these infections is the resistance of C. auris to most antifungal drugs used clinically. Thus, finding co-drugs capable of overcoming resistance to frontline antifungals is of prime clinical importance. In this study, the ability of the combination of different sulfa drugs with azole antifungals to inhibit the growth of azole-resistant C. auris isolates was evaluated. Among the active sulfa drugs, sulfamethoxazole exhibited the most potent in vitro synergistic interactions with voriconazole and itraconazole. The sulfamethoxazole-voriconazole combination restored voriconazole's fungistatic activity against three of eight voriconazole-resistant clinical isolates. Similarly, the sulfamethoxazole-itraconazole combination restored itraconazole's fungistatic activity against three of four itraconazole-resistant clinical isolates. This activity was further confirmed in vivo in a Caenorhabditis elegans model of C. auris infection. The sulfamethoxazole-voriconazole combination enhanced survival of nematodes infected with C. auris by nearly 70%. Notably, these data indicate that the efficacy of this novel combination is dependent on the underlying mechanism of azole resistance. Mutant strains demonstrating azole resistance by either overproduction of or decreased affinity for the azole target (ERG11p) were found highly to be susceptible to the sulfamethoxazole-azole combination. However, this synergistic interaction was ineffective against mutant strains that demonstrated azole resistance via efflux pump hyperactivity. In conclusion, sulfamethoxazole represents a promising co-drug that can restore the efficacy of certain azole antifungal drugs against some azole-resistant isolates of C. auris.

Simple Summary Endophytic bacteria are plant-associated bacteria that live in the internal tissues of the plant without harming the host plant. They have an important role in plant growth promotion, as they directly or indirectly promote plant growth. They do it by inhibiting the growth of plant pathogens, and/or by producing various secondary metabolites. They are used in the agricultural sector as an eco-friendly alternative tool that helps to improve crop yield. Detection of plant defense response and identification of compounds synthesized by root endophytes are an effective means for their utilization in the agriculture sector as biofertilizers. Therefore, it is important to study the diversity of root endophytic microbial community, endophyte-host plant interactions and their colonization, and their activity for successful application in agricultural lands. Here, in this review, the potential of the root endophytic microbial community, colonization, and role in the improvement of plant growth has been explained. This could mark the potential use of endophytes for the benefit of plant growth and enhanced yield. Abstract The plant root is the primary site of interaction between plants and associated microorganisms and constitutes the main components of plant microbiomes that impact crop production. The endophytic bacteria in the root zone have an important role in plant growth promotion. Diverse microbial communities inhabit plant root tissues, and they directly or indirectly promote plant growth by inhibiting the growth of plant pathogens, producing various secondary metabolites. Mechanisms of plant growth promotion and response of root endophytic microorganisms for their survival and colonization in the host plants are the result of complex plant-microbe interactions. Endophytic microorganisms also assist the host to sustain different biotic and abiotic stresses. Better insights are emerging for the endophyte, such as host plant interactions due to advancements in ‘omic’ technologies, which facilitate the exploration of genes that are responsible for plant tissue colonization. Consequently, this is informative to envisage putative functions and metabolic processes crucial for endophytic adaptations. Detection of cell signaling molecules between host plants and identification of compounds synthesized by root endophytes are effective means for their utilization in the agriculture sector as biofertilizers. In addition, it is interesting that the endophytic microorganism colonization impacts the relative abundance of indigenous microbial communities and suppresses the deleterious microorganisms in plant tissues. Natural products released by endophytes act as biocontrol agents and inhibit pathogen growth. The symbiosis of endophytic bacteria and arbuscular mycorrhizal fungi (AMF) affects plant symbiotic signaling pathways and root colonization patterns and phytohormone synthesis. In this review, the potential of the root endophytic community, colonization, and role in the improvement of plant growth has been explained in the light of intricate plant-microbe interactions.

The main reasons for the discontinuation of contact lens wear are ocular dryness and discomfort. Proteoglycan 4 (PRG4), a mucinous glycoprotein, and hyaluronic acid (HA), a nonsulfated linear glycosaminoglycan, are naturally present in the eye and contribute to ocular hydration and lubrication. This study aimed to investigate the impact of the structure of the recombinant human PRG4 (rhPRG4)/HA complex on contact lens properties, when one agent is grafted and the counterpart is physisorbed on the surface of model conventional or silicone contact lens materials. Investigation of the wettability, water retention, antifouling, and boundary lubricant properties of the prepared hydrogels showed that the rhPRG4/HA interactions varied with the rhPRG/HA configuration on the hydrogel surface as well as the composition of the underlying substrate used. The rhPRG4-physisorbed/HA-grafted sample was characterized by better antifouling and boundary lubricant properties on the model conventional hydrogels, while the HA-physisorbed/rhPRG4-grafted sample exhibited improved surface wettability, antifouling, and water-retentive properties on the model silicone hydrogels. The results of this study contribute to the design of biomimetic contact lens surfaces that work synergistically with ocular fluid-phase biological agents to enhance compatibility between the contact lens and the ocular environment, alleviating dry eye symptoms and improving comfort.

Candida auris is an emergent fungal pathogen that causes severe infectious outbreaks globally. The public health concern when dealing with this pathogen is mainly due to reduced susceptibility to current antifungal drugs. A valuable alternative to overcome this problem is to investigate the efficacy of combination therapy. The aim of this study was to determine the in vitro interactions of isavuconazole with echinocandins against C. auris. Interactions were determined using a checkerboard method, and absorbance data were analyzed with different approaches: the fractional inhibitory concentration index (FICI), Greco universal response surface approach, and Bliss interaction model. All models were in accordance and showed that combinations of isavuconazole with echinocandins resulted in an overall synergistic interaction. A wide range of concentrations within the therapeutic range were selected to perform time-kill curves. These confirmed that isavuconazole–echinocandin combinations were more effective than monotherapy regimens. Synergism and fungistatic activity were achieved with combinations that included isavuconazole in low concentrations (≥0.125 mg/L) and ≥1 mg/L of echinocandin. Time-kill curves revealed that once synergy was achieved, combinations of higher drug concentrations did not improve the antifungal activity. This work launches promising results regarding the combination of isavuconazole with echinocandins for the treatment of C. auris infections. This research was funded by Consejería de Educación, Universidades e Investigación of Gobierno Vasco-Eusko Jaurlaritza, GIC15/78 IT-990-16 and by FIS, Spain, PI17/01538. UC was funded by a Ph.D. grant from the University of the Basque Country UPV/EHU, PIF 17/266.

Microbial sulfate-reduction coupling polycyclic aromatic hydrocarbon (PAH) degradation is an important process for the remediation of contaminated sediments. However, little is known about core players and their mechanisms in this process due to the complexity of PAH degradation and the large number of microorganisms involved. Here we analyzed potential core players in a black-odorous sediment using gradient-dilution culturing, isolation and genomic/metagenomic approaches. Along the dilution gradient, microbial PAH degradation and sulfate consumption were not decreased, and even a significant (p = 0.003) increase was observed in the degradation of phenanthrene although the microbial diversity declined. Two species, affiliated with Desulfovibrio and Petrimonas, were commonly present in all of the gradients as keystone taxa and showed as the dominant microorganisms in the single colony (SB8) isolated from the highest dilution culture with 93.49% and 4.73% of the microbial community, respectively. Desulfovibrio sp. SB8 and Petrimonas sp. SB8 could serve together as core players for sulfate-reduction coupling PAH degradation, in which Desulfovibrio sp. SB8 could degrade PAHs to hexahydro-2-naphthoyl through the carboxylation pathway while Petrimonas sp. SB8 might degrade intermediate metabolites of PAHs. This study provides new insights into the microbial sulfate-reduction coupling PAH degradation in black-odorous sediments.

Submerged macrophyte Vallisneria spiralis and sediment microbial fuel cell (SMFC) systems are cost-effective methods for the remediation of polycyclic aromatic hydrocarbon (PAH)–polluted sediments. This study evaluates whether the combination of these two ecological approaches could further improve the removal efficiency of PAHs from sediments and investigates the possible mechanisms of removal. Sediments, macrophytes, electrodes, and plexiglass columns were used to construct an experimental microcosm. A 65-day comparative study was performed with six treatments as follows: SMFC without PAH (SMFC); PAH only (PAH); SMFC with PAH (SMFC-PAH); V. spiralis without PAH (macrophyte); V. spiralis with PAH (macrophyte-PAH); V. spiralis with SMFC and PAH (M-SMFC-PAH). Pyrene and phenanthrene were added to raw sediments to obtain initial PAH concentrations of 10 mg kg−1 dry sediment. The monitored parameters were sediment oxidation–reduction potential (ORP); low molecular weight organic acids (LMWOAs) and Fe (II) concentrations in pore water; electron acceptor (sulfate and Fe (III)), humic acid (HA), and PAH concentrations in sediments; and plant morphology and root physiology. High-throughput 16S rRNA gene sequencing was also performed to assist mechanistic understanding. The M-SMFC-PAH treatment obtained the highest sediment ORP and PAH removal efficiency. The average ORP level in M-SMFC-PAH was increased by 57.2, 59.1, and 168.4 mV, compared with the SMFC-PAH, macrophyte-PAH, and PAH-only treatments, respectively, with a mean value of 121.7 mV observed during the whole experimental period. The pyrene (phenanthrene) dissipation ratios at the end of the experimental period were 29.1% (35.4%), 45.5% (56.3%), 59.8% (67.3%), and 79.4% (88.2%) for PAH only, SMFC-PAH, macrophyte-PAH, and M-SMFC-PAH treatments, respectively. The highest correlation was observed between PAH concentration and sediment ORP value, in the coupled M-SMFC-PAH system. Results suggest that the interactions between the anode and rhizosphere of V. spiralis were synergistic during PAH removal. The coexistence of anodic and rhizospheric oxygen loss in sediments had a synergistic effect on PAH degradation. Plant presence facilitated the electrogenic degradation of PAHs. The inhibited growth of V. spiralis due to PAH toxicity was reduced by electrogenesis, thus facilitating the removal of vegetable PAHs from sediments. Coordinated growth of anaerobic and aerobic PAH degrading bacteria on the anode was a key factor in the optimal removal of PAHs in coupled systems.

Objective Rehmanniae Radix has been traditionally used to treat diabetes. Catalpol (CAT) and stachyose (STA) are two of the main bioactive compounds in Rehmannia Radix and found to have similar therapeutic effects on diabetes and its complications. In this paper, we aimed to investigate whether there were synergistic therapeutic effects of CAT and STA on diabetes. Methods Streptozotocin (STZ) with the feeding of high-sugar-high-fat diet (HFD) was applied to induce diabetic C57BL/6 mice. STZ-HFD induced diabetic mice were then divided into model and six medical-treated groups: metformin (MET), STA, CAT, and three combinations of CAT:STA (1:1, 1:2, 2:1). Blood, liver, and kidney samples were isolated after six-week oral administration for biochemical assays of serum lipids, the indicators of kidney and liver functions and HE staining for liver tissues. Results It turned out that CAT, STA and their three combinations (1:1, 1:2, 2:1) could effectively control body weight, blood glucose, kidney weight and liver weight index, and well regulate levels of TC, HDL-c, TG, ALT, and TBA. In addition, CAT and its combination with STA at the ratio of 2:1 could significantly improve albumin content, compared to that in model group. STA and CAT and their combinations showed the improvements on kidney function in terms of urinary creatinine (Ucr). However, there were no such consistent observations on serum creatinine (Scr) and creatinine clearance rate (Ccr). The combination of CAT and STA at the ratio of 1:1 exhibited the better adjusting effects on kidney weight and liver weight indexes and the levels of ALT, Ucr, Scr, and Ccr. Our results demonstrated that the combinations of CAT and STA especially 1:1 showed similar or better improvements on diabetes-associated complications, compared to the sole CAT or STA treatment. Conclusion Thus, we concluded that there were synergistic therapeutic effects between CAT and STA on STZ/HFD-induced type 2 diabetes. This project provided insights and technical supports for the innovation of discovering bioactive constituents in Rehmannia Radix and studying its integrative mechanism in curing diabetes.

An understanding of the inter-species relationships, especially their metabolic network in a mixed-culture system, is crucial to design an effective inoculum for enhancing the power generation of wastewater fed microbial fuel cell (MFC). In the present study, the influence of microbial mutualistic interactions on the power generation of palm oil mill effluent fed MFCs has been widely investigated by designing several co-culture and mixed culture inoculums. Among the different inoculum compositions, the highest power density of 14.8 W/m3 was achieved by Pseudomonas aeruginosa and Klebsiella variicola co-culture inoculum due to their synergistic relationships which were inter-linked via fermentation-based metabolites. Besides, the interaction of K. variicola and Bacillus cereus positively influenced the power generation resulting in a maximum power density of 11.8 W/m3 whereas the antagonistic relationship between B. cereus and P. aeruginosa resulted in a lower power generation of 1.9 W/m3. The microbial mutualistic interactions were investigated with polarization, cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), as well as by using metabolite and biofilm analysis. It was observed that the synergism between bacteria enhanced power generation through the production of higher electron shuttling mediators and efficient biofilm formation as evidenced by polarization, CV and EIS analysis. In contrast, the antagonistic relationship resulted in production of cell inhibiting metabolites leading to the formation of ineffective biofilm. These findings demonstrate that the synergistic interaction between or within microorganisms is emergent in designing co-culture or mixed-culture inoculum for achieving maximum power generation in MFCs.

Type 1 diabetes mellitus (T1DM) is an autoimmune endocrine disease, in which both gene-gene and gene-environmental factors play a key role in the development and progression of the disease. The main aim of the present study was to determine the association of Angiotensin Converting Enzyme (ACE) and Apolipoprotein E (APOE) gene polymorphisms in T1DM patients. A cohort of 196 T1DM patients (n = 196) and age/sex matched controls (n = 196) were enrolled for typing of ACE and APOE by PCR-SSP and PCR-RFLP methods. We observed an increased frequencies of ‘DD’ (P: OR = 2.40; M: OR = 2.08 and F: OR = 2.90), ‘ID’ (P: OR = 1.94; M: OR = 2.01; F: OR = 1.84) genotypes and ‘D’ allele (P: OR = 2.43; M: OR = 2.21; F: OR = 2.74) in patients confirmed their susceptible association. Whereas, the ‘II’ genotype (P: OR = 0.25; M: OR = 0.28; F: OR = 0.21) and ‘I’ allele frequencies (P: OR = 0.41; M: OR = 0.45; F: OR = 0.36) were decreased in patients suggested their protective association. Multivariate analysis revealed that the ‘GAD65+’, ‘IA2+’ and ‘GAD65+ + IA2+’ autoantibodies were significantly associated with patients. Genetic model of inheritance analysis showed that, the additive (P: OR = 5.15; M: OR = 5.52; F: OR = 4.66), recessive (P: OR = 2.40; M: OR = 2.41; F: OR = 2.41), dominant (P: OR = 3.99; M: OR = 4.79; F: OR = 3.21) and co-dominant (P: OR = 1.94; M: OR = 2.60) models were significantly associated in patients. No significant association was observed for the APOE genotypes with T1DM. However, ‘e3’ allele (M: OR = 0.191; F: OR = 0.128) was decreased in patients confirming its moderate protection. Analysis on synergistic association have revealed that: ‘DD + e3/e4’ (P: OR = 11.59) and ‘ID + e3/e3’ (P: OR = 1.66) were susceptible; and ‘II + e3/e3’ (P: OR = 0.25) as the protective combinations. Thus, the present study conclude the stronger susceptible risks for the combinations ‘DD’ + ‘e3/e4’ and ‘ID’ + ‘e3/e3’ towards the development of T1D in south India.

BACKGROUND: Eosinophilic esophagitis (EoE) is an esophageal inflammatory disease associated with atopic diseases. Thymic stromal lymphopoietin (TSLP) and calpain 14 (CAPN14) genetic variations contribute to EoE, but how this relates to atopy is unclear. OBJECTIVE: The purpose of this study was to explore the relationship between EoE, atopy, and genetic risk. METHODS: EoE-atopy enrichment was tested by using 700 patients with EoE and 801 community control subjects. Probing 372 single nucleotide polymorphisms (SNPs) in 63 atopy genes, we evaluated EoE associations using 412 nonatopic and 868 atopic disease control subjects. Interaction and stratified analyses of EoE-specific and atopy-related SNPs were performed. RESULTS: Atopic disease was enriched in patients with EoE (P < .0001). Comparing patients with EoE and nonatopic control subjects, EoE associated strongly with IL-4/kinesin family member 3A (IL4/KIF3A) (P = 2.8 × 10(−6); odds ratio [OR], 1.87), moderately with TSLP (P = 1.5 × 10(−4); OR, 1.43), and nominally with CAPN14 (P = .029; OR, 1.35). Comparing patients with EoE with atopic disease control subjects, EoE associated strongly with ST2 (P = 3.5 × 10(−6); OR, 1.77) and nominally with IL4/KIF3A (P = .019; OR, 1.25); TSLP’s association persisted (P = 4.7 × 10(−5); OR, 1.37), and CAPN14’s association strengthened (P = .0001; OR, 1.71). Notably, there was gene-gene interaction between TSLP and IL4 SNPs (P = .0074). Children with risk alleles for both genes were at higher risk for EoE (P = 2.0 × 10(−10); OR, 3.67). CONCLUSIONS: EoE genetic susceptibility is mediated by EoE-specific and general atopic disease loci, which can have synergistic effects. These results might aid in identifying potential therapeutics and predicting EoE susceptibility.

Carbapenem-resistant Klebsiella pneumoniae (CR-Kp) can cause biofilm-related bloodstream infections associated with significant morbidity and mortality worldwide. We investigated the bactericidal activities of colistin (CST), rifampin (RIF), meropenem (MEM), gentamicin (GEN), and tigecycline (TGC) alone and that of CST in combination with RIF, MEM, GEN, or TGC against CR-Kp mature biofilms. Twenty CR-Kp blood isolates were derived from an equal number of bloodstream infections in adult patients. Biofilm formation was assessed by staining with 0.4% crystal violet and measuring the optical density spectrophotometrically at 545 nm. Biofilm damage was measured as the percent reduction of metabolic activity by an XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt] assay. The MIC(50) for biofilms was determined as the minimum concentration that caused ≥50% bacterial damage compared to that for untreated controls. Antibacterial drug interactions were analyzed by the Bliss independence model. Four of the 20 CR-Kp isolates were biofilm producers. Biofilm MIC(50)s of CST, RIF, MEM, GEN, and TGC for these isolates were 64, 8, >256, 128, and 8 mg/liter, respectively. Synergistic interactions were observed at 32 to 64 mg/liter of CST combined with 0.25 to 4 mg/liter of RIF, at 32 mg/liter of CST combined with 0.007 to 0.25 mg/liter of MEM, and at 16 to 32 mg/liter of CST combined with 16 to 64 mg/liter of TGC. The synergy was highest for CST plus RIF, with a mean ΔE ± standard error (SE) of 49.87% ± 9.22%, compared to 29.52% ± 4.97% for CST plus MEM (P < 0.001) and 32.44% ± 6.49% for CST plus TGC (P < 0.001). Indifferent results were exhibited by CST plus GEN. None of the combinations exhibited antagonism. These drug interaction findings, especially those for CST with RIF, may be of importance in the treatment of biofilm-related CR-Kp infections.

BACKGROUND In recent years, concern has been raised over honey bee colony losses, and also among wild bees there is evidence for extinctions and range contractions in Europe and North America. Pesticides have been proposed as a potential cause of this decline. Bees are exposed simultaneously to a variety of agrochemicals, which may cause synergistically detrimental impacts, which are incompletely understood. We investigated the toxicity of the fungicide imazalil in mixture with four common insecticides: fipronil (phenylpyrazoid), cypermethrin (pyrethroid), thiamethoxam, and imidacloprid (neonicotinoids). Ergosterol biosynthesis inhibitor (EBI) fungicides like imazalil can inhibit P450 detoxification systems in insects and therefore fungicide - insecticide co-occurrence might produce synergistic toxicity in bees. We assessed the impact of dietary fungicide - insecticide mixtures on the mortality and feeding rates of laboratory bumble bees (Bombus terrestris L.). RESULTS Regarding mortality, imazalil synergised the toxicity of fipronil, cypermethrin and thiamethoxam, but not imidacloprid. We found no synergistic effects on feeding rates. CONCLUSION Our findings suggest that P450-based detoxification processes are differentially important in mitigating the toxicity of certain insecticides, even those of the same chemical class. Our evidence that cocktail effects can arise in bumble bees should extend concern about the potential impacts of agrochemical mixtures to include wild bee species in farmland. © 2017 Society of Chemical Industry.

The combined effect of four abiotic factors on Microcystis aeruginosa growth and toxin production was assessed by culturing the cyanobacterium under different light intensities (10190 mol photons·m 2·s 1), CO2 concentrations (010% (v/v)), temperatures (1540 °C), and pH values (6.59.5). Results indicate a significant influence caused by the synergistic effect of environmental factors over growth-related parameters and cyanobacteria toxicity. The combined use of low to medium light intensities (50120 mol photons·m 2·s 1) and CO2 concentration (16% v/v) led to higher cell concentrations, while specific growth rate and biomass productivity were favoured by medium to high light intensities (110190 mol photons·m 2·s 1), CO2 concentrations (49.5% v/v) and temperatures (2939 °C). Regarding microcystin (MC) production, higher concentrations were obtained at low light intensities and low CO2 concentrations while approximately 2000-fold lower MC concentrations were achieved by simultaneous use of high values of light intensity, CO2 concentration and temperature., info:eu-repo/semantics/publishedVersion

Background The herbal medicinal product Myrrhinil-Intest®, a combination of myrrh, chamomile flower extract and coffee charcoal is used for the maintenance therapy of inflammatory bowel disease. In vitro studies revealed that the herbal components influence chemokine signaling of human macrophages as part of an anti-inflammatory mechanism. However, the occurrence of synergistic effects remains unexplored. Aim The present study aims to investigate the synergistic effect of dual combinations of the plant extracts on the pro-inflammatory chemokine (CXCL13) release from activated human macrophages. Methods The single effect of myrrh, chamomile flower and coffee charcoal extract on CXCL13 release from lipopolysaccharide stimulated human macrophages was investigated using ELISA and IC50 values were determined. Budesonide served as positive control. To characterize the combined effect, IC50 values were used to prepare combinations of two plant extracts in different proportions to each other (3:5; 1:1; 5:3). Interpretation of the data was based on isobologram analysis and calculation of a combination index (CI). Results LPS-induced CXCL13 release from human macrophages was inhibited after treatment with myrrh (IC50 = 19 μg/ml), chamomile flower (IC50 = 82 μg/ml) and coffee charcoal (IC50 = 106 μg/ml) whereby the extent of inhibition was comparable to budesonide. All combinations of two plant extracts resulted in synergistic effects with varying magnitude (CI from 0.82 to 0.42). The combination of myrrh and coffee charcoal (ratio of 3:5) exhibited the strongest synergistic effect (CI = 0.42). Increasing amounts of coffee charcoal resulted in increased synergistic activity. Conclusion Synergistic effects between all plant components contribute to the anti-inflammatory activity of all dual combinations of the plant components and support the composition of the herbal combination.

New drugs or therapeutic combinations are urgently needed against Mycobacterium abscessus . Previously, we demonstrated the potent activity of indole-2-carboxamides 6 and 12 against M. abscessus . We show here that these compounds act synergistically with imipenem and cefoxitin in vitro and increase the bactericidal activity of the β-lactams against M. abscessus . In addition, compound 12 also displays synergism with imipenem and cefoxitin within infected macrophages.

In this work, the essential oil (EO) and supercritical CO2 fluid extract (SF extract) of four Valerianaceae plants (Valeriana officinalis L., Valeriana officinalis L. var. latifolia Miq., Valeriana jatamansi Jones and Nardostachys chinensis Bat.) were chemically characterized. GC-MS analysis identified 74 compounds, representing 35.2%–82.4% of the total EOs and SF extracts. The EO was dominated by low-molecular-weight components while the SF extract was rich in fatty acids. Bornyl acetate and camphene were the characteristic compounds in EO and SF extracts. The efficacy of six extracts against three stored-product insects was investigated. In contact assays, V. officinalis exhibited strongest toxicity to red flour beetle (LD50 = 10.0 μg/adult), and V. jatamansi EO was the most active one against the cigarette beetle (LD50 = 17.6 μg/adult), while V. officinalis var. latifolia EO showed outstanding efficacy against the booklouse (LD50 = 40.2 μg/cm2). Binary mixtures of two major compounds (camphene and bornyl acetate) were assessed for the contact toxicity to the red flour beetle. Additive effect existed in the natural proportion of V. officinalis, and synergism was observed in that of V. officinalis var. latifolia. This work confirmed the insecticidal efficacy of the species of the Valerianaceae family, and it would offer some information for the development of botanical insecticide.

The in vitro activities of two antifungal drugs in combination with four nonantifungal ophthalmic agents were evaluated using a broth microdilution method and a collection of eight Fusarium ocular isolates that exhibited resistance to both natamycin (MICs, 14 to 32 μg/ml) and voriconazole (MICs, 4 to >128 μg/ml). Synergistic and indifferent interactions were observed for natamycin and 5-fluorouracil and natamycin with timolol dependent on the Fusarium isolate tested. Isolate-dependent synergistic and indifferent interactions were also observed for natamycin with EDTA and natamycin with dorzolamide. Synergistic or indifferent interactions were observed for voriconazole with timolol and voriconazole with 5-fluorouracil depending on Fusarium isolate. Taken together, these data suggest that commonly used ophthalmic agents enhance the in vitro activity of antifungal drugs against drug-recalcitrant ocular fungal pathogens.

Monotonous cucumber double-cropping systems under plastic greenhouse vegetable cultivation (PGVC) previously intensified by long-term anthropogenic activities and manipulative treatments leads to a crop productivity reduction and soil biota disturbances. In this study, the role of the indigenous arbuscular mycorrhizal strain (AM: Glomus versiforme L.) and organic substrate (GS: Garlic stalk) application were assessed for plant microbe interaction and crop productivity feedback in a greenhouse (2016&ndash, 2018) under a cultivated Anthrosol characterized as a replanted degraded soil. We found that repetitively adding AM inocula with organic substrates (GS) improved the cucumber growth and physiology. The useful trait of AM symbiosis with C-amended organic substrates preferentially manifested as increased root colonization, hyphal density proliferation, AM sporulation, root activity, and suppressed Fusarium incidence. The post AM development further prevailed the synergistic interaction, and the co-inoculation effect resulted in an increase in fruit nutrition uptake, seasonal cucumber yield and fruit quality attributes. Illumina MiSeq analysis of the 18S rRNA gene amplicons revealed that the dominant AM genera that are particularly enriched with the Glomus taxon may be important ecological drivers associated with plant productivity and fruit quality characteristics. These results suggest that the AM-organic substrate association might be a pragmatic option for use as an economic and efficient biological resource and as a newly-sustainable plant microbe mediator to enhance the regional ecosystem services and plant productivity of the anthropogenic PGVC of this region.

Since plant essential oils are composed with vast numbers of constituents, they often show complex interactions among the components. Although the research interest on the insecticidal activity of plant essential oils has been gaining more attention recently, most of the studies still focus on simple screening of active plant source or identification of active compounds. In the present study, insecticidal activity and synergistic interactions among the four major constituents of lemongrass (Cymbopogon citratus) essential oil were examined via topical application against the third instar larvae of the cabbage looper, Trichoplusia ni. Two synergistic binary combinations—citral + limonene and citral + geranyl acetate—were identified at the equivalent mixing ratios. The former was especially synergistic in larvae and additionally with respect to cytotoxicity in the ovarian cell line of the cabbage looper. Morphological observations indicated different cytotoxic modes of action of citral and limonene. GC–MS analysis of larval extracts in vivo revealed several metabolites of citral and limonene, with geranic acid and neric acid (from citral), and limonene-1,2-diol (from limonene) as the major ones. The insecticidal activity of geranic acid was very similar to that of the parent compound, citral, but limonene-1,2-diol failed to show any toxicity, indicating that inhibition of the metabolism could be a good strategy to enhance toxicity. Further, larval extracts following topical administration of a binary mixture of the two compounds revealed higher internal concentrations of both compared to their individual application, suggesting the possibility of enhanced cuticular penetration as the mechanism of synergy.

Made available in DSpace on 2018-12-11T16:58:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-03-01 Locust bean gum synergistic interactions with whey proteins are widely described in terms of functional properties. The aim of this work is to assess how these interactions affect whey protein-based film properties. Blended films were manufactured using whey protein isolate (WPI), four different concentrations levels of locust bean gum (LBG) and two different thermal treatments. A rheological study was performed to assess interaction between WPI and LBG. The influence of glycerol on WPI/LBG interactions was also verified. Barrier, mechanical, and optical properties, as well as microstructure, solubility and moisture sorption behavior of films were evaluated. The results show that interaction between WPI and LBG and more severe heat treatments provide stronger, more flexible and less soluble films with lower permeability to carbon dioxide and oxygen and lower transparency. These findings suggest that the addition of locust bean gum to WPI can be used to tune the properties of WPI-based edible films to meet specific food packaging and edible coating needs. REQUIMTE, LAQV, Departamento de Engenharia Química, Faculdade de Engenharia, Universidade do Porto, Rua Dr. Roberto Frias Department of Food Engineering and Technology, Institute of Biosciences, Language and Physical Sciences (IBILCE), UNESP - São Paulo State University, Rua Cristóvão Colombo 2265 Department of Food Engineering and Technology, Institute of Biosciences, Language and Physical Sciences (IBILCE), UNESP - São Paulo State University, Rua Cristóvão Colombo 2265

Staphylococcus aureus (S. aureus) is the leading cause of a broad range of human infections and associated with food poisoning worldwide. Nisin can be considered as a natural preservative and is widely used by the food industry. However, resistance of nisin has been reported in several species of bacteria. Identification of antibacterial agents that are synergistic with nisin is a method of minimising resistance to nisin. Cinnamaldehyde (CA), a well-studied essential oil isolated from cinnamon bark, has been reported to exhibit a broad spectrum of antimicrobial activity across a wide range of microorganisms. Previous studies have demonstrated that CA could be used in the food industry due to its active antibacterial activity and its safety. In this study, the synergistic antibacterial activity in vitro of nisin and CA against 13 food-borne isolates of S. aureus and S. aureus ATCC 29213 were evaluated by a checkerboard assay. The FICI values of nisin combined with CA ranged from 0.1875 to 0.375, which demonstrated a synergistic effect. The positive interactions were verified using the challenge tests in pasteurized milk and agar diffusion assays. The LIVE/DEAD BacLight experiment and scanning electron microscope (SEM) assay were also performed to distinguish the intact and damaged both of cell walls and membranes with the compounds, the combination group displayed stronger damage both of cell wall and cell membrane than when nisin or CA was used alone. These findings indicated that the combination of nisin and CA could not only be used as a promising naturally sourced food preservative but also to reduce the problem of bacterial resistance.

Leaf litters commonly interact during decomposition in ways that can synergistically increases rates of decay. These interactions have been linked to moisture availability, suggesting that drought could slow decomposition rates by disrupting litter interactions. Slowed decomposition may reduce competitive ability of exotic species that exploit rapid decomposition rates as part of niche construction mechanisms. Here, we evaluated the impacts of drought on interactions between native and exotic species' litter decomposition. We considered litter mixtures of Lupinus polyphyllus (exotic N-fixing forb), Trifolium pratense (native N-fixing forb), Senecio inaequidens (exotic non-N-fixing forb), and Senecio jacobaea (native non-N-fixing forb) with the native grass Alopecurus pratensis and evaluated the difference between the observed rate of decay and the one expected based on species decomposing in monocultures. Litters were deployed in Belgium and Germany and exposed to a 56 day drought, which resembled local millennium drought (statistical recurrence of duration in local precipitation series > 1000 years). Litter interactions reduced mass remaining by 81% in Belgium and 15% in Germany, averaged across mixtures. Similarly, litter interactions reduced N remaining by 93% in Belgium and 14% in Germany. Drought consistently removed these interactions and resulted in additive litter decay. Litters of native and exotic species did not differ in their response to drought. These findings support moisture availability as a key regulator of interactions between litters during decomposition. Thus, increasing frequency of drought may slow nutrient cycling to a greater extent than previously thought.

Algal biofilms were grown to investigate the interaction effects of bulk medium CO2 concentration and photon flux density (PFD) on biomass productivities. When increasing the CO2 concentration from 0.04% to 2%, while maintaining a PFD of 100μmol/m(2)/s, biomass productivities increased from ∼0.5 to 2.0g/m(2)/d; however, the productivities plateaued when CO2 concentrations were incrementally increased above 2-12%. Statistical analysis demonstrates that there is a significant interaction between PFD and CO2 concentrations on biomass productivities. By simultaneously increasing PFD and CO2 concentrations, biomass productivities were significantly increased to 4.0 and 4.1g/m(2)/d in the experimental and modeled data, respectively. The second order model predicted increases in biomass productivities as both PFD and CO2 simultaneously increased yielding an optimum at 440μmol/m(2)/s and 7.1%; however, when conditions were extended to the highest end of their respective ranges, the conditions were detrimental to growth and productivities decreased.

Legumes can host rhizobia and mycorrhizal fungi, and this triple symbiosis might be exploited to improve saline soil fertility. Therefore, a greater understanding of the interaction of rhizobia and arbuscular mycorrhizal fungus during legume growth in saline soil is required. We investigated the efficiency of salt tolerance conferred by rhizobia in mycorrhizal Sesbania cannabina. Greenhouse experiments were conducted in which S. cannabina plants inoculated with Glomus mosseae BGC NM03D (GM), and two rhizobia strains Agrobacterium pusense YIC4105 (4105) and Neorhizobium huautlense YIC4083 (4083), were exposed to 100 and 200 mM NaCl. Under 200 mM NaCl stress, plants inoculated with 4105, rather than 4083, showed significant increases in shoot and root dry mass compared with non-inoculated plants. Simultaneously, a significant increase over GM-inoculated plants in mycorrhizal colonization and dependency was recorded for 4105 + GM-inoculated plants compared with 4083 + GM-inoculated plants. In addition, under NaCl stress, significant increases in the number and mass of nodules, nitrogenase activity, and leghemoglobin content of nodules occurred in 4105 + GM-inoculated plants compared with 4083 + GM-inoculated plants. Furthermore, the activities of antioxidant enzymes in rhizobia-inoculated plants were significantly higher in the GM + 4105 group than the 4083 + GM group. The malondialdehyde content of plants from the 4105 + GM group was significantly lower than in the 4083 + GM group. Thus, the results revealed a synergistic relationship among the 4105 and GM in alleviating salt stress in S. cannabina. Salt-tolerant rhizobia might improve the salinity tolerance of S. cannabina by enhancing the antioxidant system.

The interactions of four natural compounds including apigenin, naringin, emodin and quercetin were investigated on inhibiting 3T3-L1 preadipocyte differentiation and pancreas lipase activity. Oil Red O staining was conducted to visualise and quantify lipid accumulation. The difference between experimental and calculated results was utilised for determining the interaction types. Interestingly, emodin synergistically interacted with the other three compounds, and the combination of emodin and apigenin exhibited the strongest synergistic effect in both differentiation and pancreas lipase assays. Results implied that the combination of apigenin and emodin may be regarded as a promising complementary therapy for management of overweight or obesity.

Accumulation of heavy metals in soil causes loss of cover vegetation and increases the production of reactive oxygen species (ROS). ROS accumulation induces the expression of genes encoding antioxidant enzymes and other proteins involved in redox homeostasis. This study aimed to evaluate the interaction between a saprophytic fungal consortium and mycorrhizal Rhizophagus irregularis with regard to the oxidative stress and molecular responses of Solanum lycopersicum L. grown in a soil contaminated with heavy metals. We determined the effects of the saprophytic fungal consortium (Bjerkandera adusta and Mortierella sp) and the mycorrhizal fungus Rhizophagus irregularis on the plant antioxidant response and the expression levels of genes encoding metallothioneins (MT), phytochelatins (PC), the NRAMP transporter and heat shock protein (HSP) in Solanum lycopersicum cultivated in a heavy metal-contaminated soil. The fungal consortium increased plant growth, and the co-inoculation with R. irregularis synergistically improved soil biochemical activities. Superoxide dismutase activity decreased in all treatments. Peroxidase activity (ascorbate and guaiacol) increased in plants inoculated with R. irregularis and the fungal consortium. Dual inoculation decreased the malondialdehyde content in the leaves and increased transcription of the NRAMP, GR, MT2b, PCS and HSP90 genes. Our results demonstrate that co-inoculation contributes to reduced plant stress by improving defence mechanisms and homeostasis

This study aims at performing a chemical analysis of the essential oils of Laurus nobilis (LNEO), Lavandula stoechas (LSEO) and Mentha pulegium (MPEO) by GC-MS. The possible synergistic effects of their combinations on anticholinesterase and antioxidant activities were investigated. LNEO was dominated by 1,8-Cineole. Pulegone was found to be the major constituent of MPEO, while fenchone was the major component present, along with camphor in LSEO. LSEO was found to have the highest AChE inhibitor (17.73 ± 2.23 μg/mL) that matched the capacity of galantamine and the highest BChE inhibitor (1.61 ± 0.34 μg/mL) which is more than galantamine. A total of 13 combinations were obtained by mixing the EOs in binary and ternary mixtures at different ratios. CompuSyn software was used to allow automated simulation of synergism and/or antagonism. Of these, LSEO in very strong synergy with MPEO at a 2:1 ratio displayed 0.08 combination index (CI) on BChE. Even, the combination of LNEO with LSEO at different ratios showed synergistic effect against both enzymes. A significant antioxidant activity was displayed by LNEO. Of interest, LNEO is considered to interact very strong synergistically with either LSEO or MPEO in a 2:1 ratio in CUPRAC. The same combinations exhibited strong synergistic action in DPPH and ABTS. Summing up, the present work highlights the importance of the synergistic interaction of EOs as promising and ideal alternatives to the food and pharmaceutical industries as it leads to diminishing their effective doses consequently improving the safety profile with lower side effects while increasing their efficiency.