Potent Synergist Research Articles

Synergistic interactions between three insecticides and Beauveria bassiana (Bals.-Criv.) Vuill. (Ascomycota: Hypocreales) in lesser mealworm, Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae), larvae

Topical applications of insecticides β-cyfluthrin, imidacloprid, and spinosad in combination with Beauveria bassiana (topical and contact filter paper application) induced synergistic interactions in lesser mealworm larvae, increasing mortality and in some cases numbers of larval cadavers exhibiting conidiogenesis. Reduced concentrations (LC10, LC20, LC30) of the insecticides induced sublethal effects in lesser mealworm larvae, inhibiting development (mass, head-capsule width, moulting) after eight days’ exposure and movement behaviour (area explored and distance travelled) after 3 h’ exposure. The most potent synergist was ß-cyfluthrin, it strongly inhibited larval development and movement while significantly increasing mortality and conidiogenesis in B. bassiana-infected larvae. Imidacloprid also strongly inhibited larval development and movement, but only produced weak short-lived synergistic increases in mortality, with no increase in conidiogenesis. Spinosad induced no effect on development and limited effect on movement, but still induced moderate short-lived synergistic increases in mortality and conidiogenesis. Intoxicated larvae exposed to B. bassiana on filter paper for 3 h showed no synergistic interactions, except when intoxicated by spinosad.

Discovery of Novel Thiosemicarbazides Containing 1,3,5-Triazines Derivatives as Potential Synergists against Fluconazole-Resistant Candida albicans.

The clinical prevalence of antifungal drug resistance has been increasing over recent years, resulting in the failure of treatments. In an attempt to overcome this critical problem, we sought novel synergistic enhancers to restore the effectiveness of fluconazole against resistant Candida albicans. Based on the structural optimization of hit compound 8 from our in-house library, a series of novel 1,3,5-triazines derivatives was designed, synthesized, and biologically evaluated for synergistic activity in combination with fluconazole. Among them, compounds 10a-o, which contain thiosemicarbazides side chains, exhibited excellent in vitro synergistic antifungal potency (MIC80 = 0.125-2.0 μg/mL, FICI range from 0.127 to 0.25). Interestingly, compound 10l exhibited moderate C. albicans activity as monotherapy with an MIC80 value of 4.0 μg/mL, and also on several Cryptococcus strains (MIC80 ranging from ≤ 0.125-0.5 μg/mL) and C. glabrata (MIC80 ≤ 0.125 μg/mL). These effects were fungal-selective, with much lower levels of cytotoxicity towards human umbilical vein endothelial cells. Here, we report a series of thiosemicarbazides containing 1,3,5-triazines derivatives as potent synergists with fluconazole, and have preliminarily validated compound 10l as a promising antifungal lead for further investigation.

Structure‐Activity Relationship of Anionic and Cationic Polyamidoamine (PAMAM) Dendrimers against Staphylococcus aureus

Dendrimers are potent synergists, carriers, and delivery molecules for natural biological products and pharmaceuticals. Staphylococcus aureus (S. aureus) infection is causing serious diseases in humans and animals. Given the recorded antibacterial and antiviral activity of terminal‐charged PAMAM dendrimers, the relation between dendrimer charge type and generation is to be established against S. aureus. Three types of polyanionic dendrimers comprising terminal groups sodium carboxylate (generations 1.5, 2.5, 3.5, and 4.5), hydroxyl (generations 2, 3, 4, and 5), and succinamic acid (generations 2, 3, 4, and 5) and polycationic dendrimers containing primary amine (generations 2, 3, 4, and 5) were in antibacterial assays to determine their zone of inhibition and antibacterial activity. Cationic dendrimers were more potent than anionic dendrimers. The largest inhibition was shown by G(5)‐128NH2 followed by G(4)‐64NH2 primary amine dendrimers. Carboxylate, hydroxyl, and succinamic acid dendrimers showed weaker effects. Owing to their antibacterial actions, the addition of dendrimers to antibiotic preparations may increase their efficacy by their intrinsic and bacterial action by damaging the bacterial membranes as well as their usage in drug delivery.

Thrombin-Derived Peptides Potentiate the Activity of Gram-Positive-Specific Antibiotics against Gram-Negative Bacteria.

The continued rise of antibiotic resistance threatens to undermine the utility of the world’s current antibiotic arsenal. This problem is particularly troubling when it comes to Gram-negative pathogens for which there are inherently fewer antibiotics available. To address this challenge, recent attention has been focused on finding compounds capable of disrupting the Gram-negative outer membrane as a means of potentiating otherwise Gram-positive-specific antibiotics. In this regard, agents capable of binding to the lipopolysaccharide (LPS) present in the Gram-negative outer membrane are of particular interest as synergists. Recently, thrombin-derived C-terminal peptides (TCPs) were reported to exhibit unique LPS-binding properties. We here describe investigations establishing the capacity of TCPs to act as synergists with the antibiotics erythromycin, rifampicin, novobiocin, and vancomycin against multiple Gram-negative strains including polymyxin-resistant clinical isolates. We further assessed the structural features most important for the observed synergy and characterized the outer membrane permeabilizing activity of the most potent synergists. Our investigations highlight the potential for such peptides in expanding the therapeutic range of antibiotics typically only used to treat Gram-positive infections.

Comparison of kinetic inhibition of ethylene and methane hydrate formation when PEGs with low and high molecular weights meet common KHIs

In the current study, the focus was placed on the understanding of kinetic behavior of two common KHIs and PEGs with different molecular weight in ethylene-water system compared with methane-water system. First, the effect of PVP, Luvicap EG, PEG300, and PEG20000 was tested on the induction time and the growth rate of hydrate in ethylene-water system. The experimental results indicated that PVP and Luvicap EG could slow down the growth rate of ethylene hydrate, while PEG300 and PEG20000 were more effective for retarding the nucleation of ethylene hydrate. In addition, for understanding the performance of the KHIs in different systems, their potential as inhibitors in ethylene-water and methane-water systems were compared. The synergistic effects of PEGs were also evaluated in the above-mentioned systems. The results revealed that they are effective synergists, so that PEG20000 can be introduced as a new potent synergist to PVP and Luvicap EG in ethylene-water and methane-water systems.

Synergistic Effects of Thiosemicarbazides with Clinical Drugs against S. aureus.

Antimicrobial resistance spurred by the overuse and misuse of antibiotics is a major global health concern, and of the Gram positive bacteria, S. aureus is a leading cause of mortality and morbidity. Alternative strategies to treat S. aureus infections, such as combination therapy, are urgently needed. In this study, a checkerboard method was used to evaluate synergistic interactions between nine thiosemicarbazides (4-benzoyl-1-(2,3-dichloro-benzoyl)thiosemicarbazides 1–5 and 4-aryl-1-(2-fluorobenzoyl)thiosemicarbazides 6–9) and conventional antibiotics against S. aureus ATCC 25923, which were determined as the fractional inhibitory concentration indices (FICIs). For these experiments, amoxicillin, gentamicin, levofloxacin, linezolid, and vancomycin were selected to represent the five antimicrobial classes most commonly used in clinical practice. With one exception of 7-vancomycin combination, none of the forty-five thiosemicarbazide-antibiotic combinations tested had an antagonistic effect, showing promising results with respect to a combination therapy. The synergic effect was observed for the 2-linezolid, 4-levofloxacin, 5-linezolid, 6-gentamicin, 6-linezolid, and 7-levofloxacin combinations. No interactions were seen in combination of the thiosemicarbazide with gentamicin or vancomycin, whereas all combinations with linezolid acted in additive or synergism, except for 6-gentamicin and 7-linezolid. The 4-(4-chlorophenyl)-1-(2-fluorobenzoyl)thiosemicarbazide 6 showed a clear preference for the potency; it affected synergistically in combinations with gentamicin or linezolid and additively in combinations with amoxicillin, levofloxacin, or vancomycin. In further studies, the inhibitory potency of the thiosemicarbazides against S. aureus DNA gyrase and topoisomerase IV was examined to clarify the molecular mechanism involved in their synergistic effect in combination with levofloxacin. The most potent synergist 6 at concentration of 100 µM was able to inhibit ~50% activity of S. aureus DNA gyrase, thereby suggesting that its anti-gyrase activity, although weak, may be a possible factor contributing to its synergism effect in combination with linezolid or gentamycin.

Design, synthesis, and SAR study of 3-(benzo[d][1,3]dioxol-5-yl)-N-benzylpropanamide as novel potent synergists against fluconazole-resistant Candida albicans

Based on our previous discovery and SAR study on the lead compounds 7d, 5 and berberine which can significantly enhance the susceptibility of fluconazole against fluconazole-resistant Candida albicans, a series of 3-(benzo[d][1,3]dioxol-5-yl)-N-(substituted benzyl)propanamides were designed, synthesized, and evaluated for their in vitro synergistic activity in combination with fluconazole. The series 2a–f were designed by replacing the amide moiety of the lead compound 7d with retro-amide moiety, and compounds 2a and 2b showed more activity than the lead 7d. Furthermore, introducing biphenyl moiety into series 2d–f afforded series 3a–r, most of which exhibited significantly superior activity to the series 2d–f. Especially, compound 3e, at a concentration of 1.0µg/ml, can enhance the susceptibility of fluconazole against fluconazole-resistant Candida albicans from 128.0µg/ml to 0.125–0.25µg/ml. A clear SAR of the compounds is discussed.

Abstract B27: Melatonin synergistically enhances cisplatin-induced apoptosis via the dephosphorylation of ERK/p90 ribosomal S6 kinase/heat shock protein 27 in SK-OV-3 cells

Abstract To evaluate melatonin's ability to enhance ovarian cancer cells to cisplatin treatment for ovarian cancer, this study was performed. Melatonin by itself had no significant cytotoxicity against SK-OV-3 cells, while cisplatin suppressed the cell viability in a dose-dependent manner. Combined treatment with cisplatin and melatonin synergistically inhibited the viability of SK-OV-3 cells with the synergism between two drugs (1 > combination index). In contrast, melatonin revealed the protective effect against cisplatin-induced cytotoxicity in OSEN normal ovarian epithelial cells. Cotreatment with cisplatin and melatonin increased the sub-G1 DNA contents and TdT-mediated dUTP nick end-labeling (TUNEL)-positive cells compared with cisplatin control in SK-OV-3 cells, suggesting that melatonin augments cisplatin-induced apoptosis. Consistently, combined treatment of cisplatin and melatonin increased the cleavage of caspase-3 and poly-(ADP-ribose) polymerase (PARP). Importantly, melatonin synergistically inhibited the phosphorylation of extracellular signal-regulated kinase (ERK) along with dephosphorylation of 90-kDa ribosomal S6 kinase (p90RSK) and heat shock protein 27 (HSP27) induced by cisplatin. Furthermore, melatonin remarkably blocked the expression and colocalization of p90RSK and HSP27 by combination treatment with cisplatin. Taken together, our findings demonstrate that melatonin enhances cisplatin-induced apoptosis via the inactivation of ERK/p90RSK/HSP27 cascade in SK-OV-3 cells as a potent synergist to cisplatin treatment. Citation Format: Sunghoon Kim, Jihyun Kim, bonglee kim, Junghyo Kim. Melatonin synergistically enhances cisplatin-induced apoptosis via the dephosphorylation of ERK/p90 ribosomal S6 kinase/heat shock protein 27 in SK-OV-3 cells. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B27.

Melatonin synergistically enhances cisplatin‐induced apoptosis via the dephosphorylation of ERK/p90 ribosomal S6 kinase/heat shock protein 27 in SK‐OV‐3 cells

To evaluate melatonin's ability to enhance ovarian cancer cells to cisplatin treatment for ovarian cancer, this study was performed. Melatonin by itself had no significant cytotoxicity against SK-OV-3 cells, while cisplatin suppressed the cell viability in a dose-dependent manner. Combined treatment with cisplatin and melatonin synergistically inhibited the viability of SK-OV-3 cells with the synergism between two drugs (1 > combination index). In contrast, melatonin revealed the protective effect against cisplatin-induced cytotoxicity in OSEN normal ovarian epithelial cells. Cotreatment with cisplatin and melatonin increased the sub-G1 DNA contents and TdT-mediated dUTP nick end-labeling (TUNEL)-positive cells compared with cisplatin control in SK-OV-3 cells, suggesting that melatonin augments cisplatin-induced apoptosis. Consistently, combined treatment of cisplatin and melatonin increased the cleavage of caspase-3 and poly-(ADP-ribose) polymerase (PARP). Importantly, melatonin synergistically inhibited the phosphorylation of extracellular signal-regulated kinase (ERK) along with dephosphorylation of 90-kDa ribosomal S6 kinase (p90RSK) and heat shock protein 27 (HSP27) induced by cisplatin. Furthermore, melatonin remarkably blocked the expression and colocalization of p90RSK and HSP27 by combination treatment with cisplatin. Taken together, our findings demonstrate that melatonin enhances cisplatin-induced apoptosis via the inactivation of ERK/p90RSK/HSP27 cascade in SK-OV-3 cells as a potent synergist to cisplatin treatment.

Acetylbritannilactone is a potent synergist of celecoxib to inhibit cyclooxygenase‐2 expression and to induce apoptosis in human breast cancer cells

The use of cyclooxygenase‐2 (COX‐2) inhibitors is associated with a significant decrease in breast cancer risk. However, recent studies show that the long‐term use might be limited due to undesired side effects. Our previous study has demonstrated that acetylbritannilactone (ABL), isolated from Chinese herb Inula Britannica L, inhibit COX‐2 expression via blockade of nuclear factor‐kappa B (NF‐κB) activation. Here, the effects of ABL and celecoxib as single and their combination were examined in breast cancer cells, and CalcuSyn software was used to assess synergistic inhibition. Celecoxib (5 μM) and ABL (25–50 μM) synergistically inhibited cell growth and induced cell apoptosis. The results showed that ABL augmented celecoxib inhibition of COX‐2 expression and prostaglandin E2 synthesis. To confirm the mechanism by which ABL and celecoxib synergistically attenuate COX‐2 expression. Furthermore, the promoter activity of COX‐2 gene was analyzed by chromatin immunoprecipitation and DNA affinity protein‐binding assays. We found that the combination of ABL with celecoxib at lower concertration resulted in reduction of binding of CREB‐1, ATF‐2 and c‐fos to the COX‐2 promoter, and then decreased activity of promoter reporter gene. In addition, inhibition of Akt and p38 signaling pathways was required for the synergism, as a constitutively active Akt and p38 abrogated the reduction of COX‐2 transcription and protected cells against apoptosis induced by the combination. Taken together, the combination of ABL with celecoxib elicited significantly greater inhibition of cell growth and more apoptosis, compared with either drug alone. Thus, ABL is expected to be a potent synergist of celecoxib for treatment of breast cancer.

Evidence that (+)-endo-Brevicomin is a Male-Produced Component of the Southern Pine Beetle Aggregation Pheromone

Previous research indicated that the aggregation pheromone of the southern pine beetle, Dendroctonus frontalis, is produced only by females, the sex that initiates attacks. We provide evidence indicating that secondarily arriving males augment mass aggregation by releasing the attractive synergist (+)-endo-brevicomin. Healthy pines artificially infested with both sexes of D. frontalis were significantly more attractive to conspecifics than trees infested solely with females. Coupled gas chromatography-electroantennographic detection (GC-EAD) analyses of volatiles isolated from male beetles revealed substantially greater olfactory sensitivity by D. frontalis to endo-brevicomin than to any other component. The threshold of detection of both sexes for (+)-endo-brevicomin was four orders of magnitude lower than for its antipode and at least one order of magnitude lower than for either enantiomer of frontalin, the major female-produced aggregation pheromone component. Pairing with a female in a gallery stimulated individual male beetles to produce hundreds of nanograms of (+)-endo-brevicomin. (+)-endo-Brevicomin was detected in a small percentage of female D. frontalis, whereas (-)-endo-brevicomin was never detected in either sex. In field trapping bioassays, we confirmed that (+)-endo-brevicomin is a potent synergist for attractive combinations of frontalin and pine turpentine. However, (+)-endo-brevicomin failed to attract D. frontalis either when presented alone or in combination with turpentine. We postulate that mass colonization of host trees by D. frontalis is mediated by distinct semiochemicals from both sexes rather than females alone. Our discovery of a key aggregation pheromone component in such an apparently well-studied species implies that the pheromone models of other bark beetles could benefit from systematic reexamination using newer technologies. Additionally, baits fortified with (+)-endo-brevicomin may enhance pest management strategies that exploit attractants for D. frontalis.

Is prochloraz a potent synergist across aquatic species? A study on bacteria, daphnia, algae and higher plants

Fungicides inhibiting the biosynthesis of ergostrol, such as the triazoles and imidazoles, have been shown to enhance the effect of insecticides on birds, mammals and invertebrates in the terrestrial environment. The synergy is proposed to be due to an effect on P450 monooxygenase enzymes active in pesticide metabolism in these organisms. Fungicides often enter the aquatic environment jointly with other pesticides. It is therefore possible that they could act as synergists also in the aquatic environment. In this study we tested the joint effect of the imidazole fungicide prochloraz together with the herbicides acifluorfen, diquat and terbuthylazine, the fungicide azoxystrobin and the insecticides chlorfenvinphos, dimethoate, and pirimicarb on the bacteria Vibrio fischeri (six binary mixtures) , the crustacean Daphnia magna (four binary mixtures), the algae Pseudokirchneriella subcapitata (four binary mixtures) and the floating plant Lemna minor (three binary mixtures). All the binary mixtures were evaluated both in relation to the model of concentration addition (CA) and independent action (IA) using isobolograms. The study showed strong synergy in relation to CA between prochloraz and azoxystrobin, diquat and esfenvalerat on D. magna with sums of toxic units for the 50:50% effect mixture (∑TU 50:50) as low as 0.25. The mixture with dimethoate was however antagonistic with ∑TU 50:50 of 2.04. Four out of the six mixtures testes on V. fisheri showed synergy in relation to CA, but for three of the mixtures the response could be explained by IA. Only the mixture with diquat showed synergy in relation to both IA and CA with ∑TU 50:50 around 0.50. There was no significant synergy for any of the combinations tested on the plant and the algae species in relation to CA and only for diquat in the algae-test in relation to IA. Hence, prochloraz does synergise the effect of some pesticides in the aquatic environment, but not consistently across species. The organism most susceptible to synergy by prochloraz in this study was D. magna. Especially the combination with insecticides such as esfenvalerate, where the concentration needed to immobilize 50% of the daphnia was reduced from >3 μg L −1 to less than 0.5 μg L −1 when prochloraz was added, could be problematic as these concentrations are environmentally realistic. Furthermore, insecticides and ergostrol-biosynthesis-inhibitors (EBI-fungicides) are often applied together, and are therefore likely to co-exist in surface waters, enhancing the problem of the already very potent insecticides.

Male‐produced aggregation pheromone of Carpophilus sayi, a nitidulid vector of oak wilt disease, and pheromonal comparison with Carpophilus lugubris

Abstract 1 Carpophilus sayi, a nitidulid beetle vector of the oak wilt fungus, Ceratocystis fagacearum, was shown to have a male‐produced aggregation pheromone.2 Six male‐specific chemicals were identified from collections of volatiles. The two major compounds were (2E,4E,6E,8E)‐3,5‐dimethyl‐7‐ethyl‐2,4,6,8‐undecatetraene and (2E,4E,6E,8E)‐3,5,7‐trimethyl‐2,4,6,8‐undecatetraene, in a ratio of 100 : 18. These compounds, in a similar ratio, were previously reported to be the pheromone of Carpophilus lugubris, a closely related species. The four minor C. sayi compounds (less than 4% as abundant as the first) were also alkyl‐branched hydrocarbons and consisted of two additional tetraenes and two trienes.3 The pheromone of C. lugubris was re‐examined to refine the comparison with C. sayi, and C. lugubris was found to have the same additional, minor tetraenes as C. sayi, but not the trienes.4 A synthetic mixture of the two major compounds was behaviourally active for both sexes of C. sayi in oak woodlands in Minnesota. The pheromone was tested in combination with fermenting whole wheat bread dough (a potent synergist of nitidulid pheromones). The combination of the 500‐µg pheromone dose and dough attracted at least 30‐fold more C. sayi than either pheromone or dough by itself. The synergized pheromone has potential as a tool for monitoring insect vector activity in an integrated management program for oak wilt.5 Although C. lugubris was not present at the Minnesota test sites, two other Carpophilus species, Carpophilus brachypterus and Carpophilus corticinus, were clearly cross‐attracted to the synergized pheromone of C. sayi.

Effective pheromone lures for monitoring the rice leaffolder moth, Cnaphalocrocis medinalis (Lepidoptera: Crambidae)

Several factors affecting the attractiveness of traps baited with synthetic sex pheromone components of Cnaphalocrocis medinalis were examined in the laboratory and field. In the laboratory, among the four pheromone components Z11-18:Ald, Z13-18:Ald, Z11-18:OH and Z13-18:OH, the major component, Z13-18:Ald, was shown to be essential for attraction and Z11-18:Ald was a potent synergist for the major component. The alcohols also showed an apparent synergistic effect in the laboratory bioassay when their mixture was combined with an aldehyde mixture. The addition of the geometric isomers of the four components at levels greater than 5% of the purified 4-component blend showed significant inhibitory effects on the pheromonal activity in the laboratory. In contrast, no synergistic effect of the alcohols on the aldehydes was shown in the field. A range of loadings of the pheromone rubber septa (0.01–3.42 mg) was tested and all remained attractive for at least 1 month with optimum attractiveness of 0.86 mg/septum. No significant effects were obtained when the ratio of Z11-18:Ald to Z13-18:Ald was changed. Rubber septa containing 0.86 mg of the four components ( E-isomers <5%) at the natural ratio are being effectively used for monitoring C. medinalis.

Dicarba-closo-dodecaboranes as a pharmacophore. Retinoidal antagonists and potential agonists.

Synthesis and biological evaluation of the first dicarba-closo-dodecaborane (carborane) derivatives of retinoids are described. Their retinoidal activity were examined in terms of the differentiation-inducing ability toward human promyelocytic leukemia HL-60 cells. High retinoidal activity (agonist or antagonist for retinoic acid receptor (RAR) requires a carboxylic acid moiety and an appropriate hydrophobic group located at a suitable position on the molecule. The 4-carboranyl-substituted compounds (7, 11) showed antagonistic activity but no agonistic activity even in the presence of the potent synergist HX630. On the other hand, the 3-carboranyl-substituted compounds (8, 12) showed potential agonistic activity, but no antagonistic activity. The results indicates that carboranes are applicable as the hydrophobic moiety of biologically active molecules.

The helical domain of a G protein alpha subunit is a regulator of its effector.

The alpha subunit (Galpha) of heterotrimeric G proteins is a major determinant of signaling selectivity. The Galpha structure essentially comprises a GTPase "Ras-like" domain (RasD) and a unique alpha-helical domain (HD). We used the vertebrate phototransduction model to test for potential functions of HD and found that the HD of the retinal transducin Galpha (Galphat) and the closely related gustducin (Galphag), but not Galphai1, Galphas, or Galphaq synergistically enhance guanosine 5'-gamma[-thio]triphosphate bound Galphat (GalphatGTPgammaS) activation of bovine rod cGMP phosphodiesterase (PDE). In addition, both HDt and HDg, but not HDi1, HDs, or HDq attenuate the trypsin-activated PDE. GalphatGDP and HDt attenuation of trypsin-activated PDE saturate with similar affinities and to an identical 38% of initial activity. These data suggest that interaction of intact Galphat with the PDE catalytic core may be caused by the HD moiety, and they indicate an independent site(s) for the HD moiety of Galphat within the PDE catalytic core in addition to the sites for the inhibitory Pgamma subunits. The HD moiety of GalphatGDP is an attenuator of the activated catalytic core, whereas in the presence of activated GalphatGTPgammaS the independently expressed HDt is a potent synergist. Rhodopsin catalysis of Galphat activation enhances the PDE activation produced by subsaturating levels of Galphat, suggesting a HD-moiety synergism from a transient conformation of Galphat. These results establish HD-selective regulations of vertebrate retinal PDE, and they provide evidence demonstrating that the HD is a modulatory domain. We suggest that the HD works in concert with the RasD, enhancing the efficiency of G protein signaling.

Potent retinoid synergists with a diphenylamine skeleton.

4-[N-(5,6,7,8-Tetrahydro-5,5,8,8-tetramethyl-2-naphthyl)amino]benzoic acid (5) exhibited weak retinoidal and retinoid synergistic activities in HL-60 cell differentiation assay. N-Alkylation of 5 caused decrease or loss of differentiation-inducing activity, but enhanced the synergistic activity with a synthetic retinoid Am80 (2), as reflected in the potent synergistic EC50 (SEC50) values of DA023 (11, 1.6 x 10(-10) M) and DA113 (14, 1.4 x 10(-10) M) in the presence of 1.0 x 10(-10) M Am80 (2).

Antioxidant and pro-oxidant activities of the brown algae, Laminaria digitata, Himanthalia elongata, Fucus vesiculosus, Fucus serratus and Ascophyllum nodosum

The ability of Laminaria digitata, Himanthalia elongata, Fucus vesiculosus, Fucus serratus and Ascophyllum nodosum to scavenge peroxyl radicals was investigated by kinetic studies in a model system. The thermal initiated oxidation of methyl linoleate was performed at 60°C in heptanol, with or without antioxidants. When they reached 1% of the substrate, seaweed extracts exhibited antioxidant activities by extending the induction period, but they did not suppress the rate of oxygen uptake as did vitamin E. A synergistic effect occurred when both algal extracts (1.5 g L-1) and vitamin E (0.4 mmol L-1) were present, and the effectiveness of the combined antioxidants during the whole induction period was vitamin E effectiveness. The synergistic effect of L. digitata, however, was subject to seasonal variations: samples collected in summer were effective synergists, whereas samples collected in winter displayed a marked negative synergism. The phospholipid fractions of F. vesiculosus, F. serratus and A. nodosum, including pigments, accounted for only 6% of the total lipid fraction, and did not exhibit a large synergistic effect. The main phospholipid was not phosphatidyl ethanolamine as usually related, but phosphatidyl inositol. Fucoxanthin had some antioxidant activity per se under our experimental conditions, but did not act as a synergist of vitamin E. The most potent synergists were recognized as chlorophyll a and related compounds by the application of liquid-liquid partition and chromatography for the identification of active components.

Adding Methyl-Substituted Ammonia Derivatives to a Food-Based Synthetic Attractant on Capture of the Mediterranean and Mexican Fruit Flies (Diptera: Tephritidae)

Field trials were conducted in Guatemala to determine the effect of addition of methyl-substituted ammonia derivatives (methylamine, dimethylamine or trimethylamine) to a food-based synthetic attractant for the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), and the Mexican fruit fly, Anastrepha ludens (Loew). Addition of trimethylamine to traps baited with ammonium acetate and putrescine increased capture of C. capitata , but not A. ludens , over traps baited with ammonium acetate and putrescine alone, in all tests. Addition of methylamine or dimethylamine had no effect on either species. More C. capitata were captured in adhesive paper cylindrical traps baited with tested combinations of synthetic lures than in McPhail traps baited with liquid protein bait (torula yeast solution). In tests conducted during the dry season in Guatemala, more female A. ludens were captured in liquid protein-baited McPhail traps than in adhesive paper cylindrical traps baited with ammonium acetate, putrescine, and trimethylamine. However, during the rainy season, adhesive paper cylindrical traps baited with ammonium acetate, putrescine, and trimethylamine captured significantly more female A. ludens than liquid protein-baited McPhail traps. No C. capitata were captured in traps baited with trimethylamine alone, indicating that trimethylamine is a potent synergist to ammonium acetate and putrescine for the capture of C. capitata .

Synergistic effects of hepatocyte growth factor on human cord blood CD34+ progenitor cells are the result of c-met receptor expression.

Hepatocyte growth factor (HGF) is a pleiotropic growth factor which, in addition to its mitogenic potency for primary hepatocytes, also has a role in the regulation of cell motility, cell growth and morphogenesis. In the present study, we show that c-met, the high-affinity receptor for HGF, is expressed on human cord blood (CB) CD34+ progenitor cells and CD34+Thy-1+ Lin-(lin-) cells. We have investigated the capacity of HGF to synergize with other growth factors to induce colony formation by CB CD34+ progenitor cells. CD34+ cells were cultured in semisolid medium containing serum with increasing concentrations of GM-CSF, G-CSF, macrophage colony-stimulating factor (M-CSF), stem cell factor (SCF), interleukin 3 (IL-3) and IL-11 alone or in combination with HGF. HGF acted as a potent synergist and enhanced, up to fourfold, colony formation induced by GM-CSF, G-CSF or M-CSF. HGF in combination with SCF, IL-3 or IL-11 did not induce proliferation of colony forming units-granulocyte macrophage (CFU-GM) above control levels. In serum-deprived cultures, HGF was only detectably synergistic with IL-11, and all other culture combinations showed no proliferation. To determine whether the stimulatory effect of IL-11 and the synergistic effect of HGF in the absence of serum could be attributed to the effect of these two cytokines on stem cells, IL-11-stimulated and unstimulated lin- cells were analyzed for expression of c-met. CD34+Thy-1+Lin- cells were positive for c-met, both in the presence and absence of IL-11 stimulation, and Northern analysis indicated that c-met RNA expression was upregulated in response to IL-11 compared to unstimulated controls. These results provide strong evidence for upregulation of the HGF receptor on primitive hematopoietic cells by IL-11, and for the synergistic role of HGF in colony formation by hematopoietic stem cells.