Synthetic dibenzylideneketones as promising anti-herpes simplex virus type 1 agents.

New antiviral agents for the treatment of herpes simplex virus type 1 (HSV-1) infection, which causes a highly prevalent and incurable disease, are needed. Here, we report for the first time the in vitro anti-HSV-1 activity of two dibenzylideneketone compounds: DBK1 and DBK2. DBK1 demonstrated virucidal activity, and high-resolution scanning electron microscopy showed that it caused morphological changes in the HSV-1 envelope. DBK2 was able to reduce HSV-1 plaque size in vitro. The DBKs are promising anti-HSV-1 candidates, as they exhibit low toxicity and exert an antiviral effect by acting at the early stages of HSV-1-host cell interaction.

Endophytic, extremophilic and entomophilic fungi strains biodegrade anthracene showing potential for bioremediation.

Anthropogenic activities have been increasing Polycyclic Aromatic Hydrocarbons (PAHs) release, promoting an urgent need for decontamination methods. Therefore, anthracene biodegradation by endophytic, extremophilic, and entomophilic fungi was studied. Moreover, a salting-out extraction methodology with the renewable solvent ethanol and the innocuous salt K2HPO4 was employed. Nine of the ten employed strains biodegraded anthracene in liquid medium (19-56% biodegradation) after 14 days at 30 °C, 130 rpm, and 100 mg L-1. The most efficient strain Didymellaceae sp. LaBioMMi 155, an entomophilic strain, was employed for optimized biodegradation, aiming at a better understanding of how factors like pollutant initial concentration, pH, and temperature affected this process. Biodegradation reached 90 ± 11% at 22 °C, pH 9.0, and 50 mg L-1. Futhermore, 8 different PAHs were biodegraded and metabolites were identified. Then, experiments with anthracene in soil ex situ were performed and bioaugmentation with Didymellaceae sp. LaBioMMi 155 presented better results than natural attenuation by the native microbiome and biostimulation by the addition of liquid nutrient medium into soil. Therefore, an expanded knowledge about PAHs biodegradation processes was achieved with emphasis to the action of Didymellaceae sp. LaBioMMi 155, which can be further employed for in situ biodegradation (after strain security test), or for enzyme identification and isolation aiming at oxygenases with optimal activity under alkaline conditions.

Acid catalyzed one-pot approach towards the synthesis of curcuminoid systems: unsymmetrical diarylidene cycloalkanones, exploration of their single crystals, optical and nonlinear optical properties.

In the present study crystalline unsymmetrical diarylidene ketone derivatives BNTP and BDBC have been prepared by two sequential acid catalyzed aldol condensation reactions in a one pot manner. The crystal structures of both compounds were confirmed by single crystal X-ray diffraction analysis which revealed the presence of H-bonding interactions of type C-H⋯O, along with weak C-H⋯π and weak π⋯π stacking interactions that are involved in the crystal stabilization of both organic compounds. Hirshfeld surface analysis is carried out for the broad investigation of the intermolecular interactions in both compounds. The quantum chemical investigation was performed on the optimized molecular geometries of BNTP and BDBC to calculate optical and nonlinear optical (NLO) properties. The density functional theory (DFT) study showed that the third-order NLO polarizabilities of compounds BNTP and BDBC are found to be 226.45 × 10-36 esu and 238.72 × 10-36 esu, respectively, which indicates noticeable good NLO response properties. Additionally, the BNTP and BDBC molecules also showed the HOMO-LUMO orbital gaps of 5.96 eV and 6.06 eV, respectively. Furthermore, the computation of UV-visible spectra of the titled compounds indicated a limited and/or no absorption above the 400 nm region, directing a good transparency and NLO property trade-off for both synthesized compounds that may play a significant contribution in the future for optoelectronic technologies.

Dibenzylideneacetone Induces Apoptosis in Cervical Cancer Cells through Ros-Mediated Mitochondrial Damage.

Cervical cancer is a health problem among women worldwide. Considering the limitations of prevention and antineoplastic chemotherapy against cervical cancer, research is needed to discover new, more effective, and safe antitumor agents. In the present study, we investigated the in vitro cytotoxicity of a new synthetic dibenzylideneacetone derived from 1,5-diaryl-3-oxo-1,4-pentadienyl (A3K2A3) against cervical cancer cells immortalized by HPV 16 (SiHa), and 18 (HeLa) by MTT assay. Furthermore, we performed spectrofluorimetry, flow cytometry, and Western blot analyzes to explore the inhibitory mechanism of A3K2A3 in cervical cancer cells. A3K2A3 showed cytotoxic activity against both cell lines. Mitochondrial depolarization and reduction in intracellular ATP levels were observed, which may be dependent on the redox imbalance between increased ROS and reduced levels of the antioxidant defense. In addition, damage to the cell membrane and DNA, and effective blocking of cell division in the G2/M phase were detected, which possibly led to the induction of apoptosis. This result was further confirmed by the upregulation of apoptosis-related proteins Bax, cytochrome C, and caspases 9 and 3. Our results provided the first evidence that A3K2A3 contributes to the suppression of cervical cancer in vitro, showing promise as a possible alternative for the treatment of this cancer.

MALDI-TOF Mass Spectrometry for the Diagnosis of Citrus Canker Caused by Xanthomonas citri subsp. citri.

Citrus canker, caused by the bacterium Xanthomonas citri subsp. citri (Xcc), is a disease that causes serious problems to the global citrus industry. Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-TOF) Mass Spectrometry (MS) has been used in human medicine to diagnose various diseases caused by both fungi and bacteria. In agriculture, this technique has potential for the diagnosis of diseases due to the low cost of large-scale analysis and quickness. This study showed that MALDI-TOF MS combined with chemometric analysis was effective for differentiating the macromolecule profile of orange leaves with canker lesions, healthy leaves, and leaves with phytotoxicity symptoms, proving that this technique may be used for the rapid diagnosis of citrus canker.

Synthesis, Structural, and Intriguing Electronic Properties of Symmetrical Bis-Aryl-α,ß-Unsaturated Ketone Derivatives.

Three symmetrical bis-aryl-α,ß-unsaturated ketone derivatives, 2,6-di((E)-benzylidene)-cyclohexan-1-one (DBC), 2,6-bis((E)-4-chlorobenzylidene)cyclohexan-1-one (BCC), and (1E,1'E,4E,4'E)-5,5'-(1,4-phenylene)bis(2-methyl-1-phenylpenta-1,4-dien-3-one) (PBMP), have been prepared using the aldol condensation approach toward ketones having two enolizable sites. The structures of DBC, BCC, and PBMP have been resolved via spectrometric methods. Moreover, the crystal structure of PBMP is determined by the single-crystal X-ray diffraction (SC-XRD) technique, which revealed that the PBMP molecular assembly is stabilized by the intermolecular C-H···O bonding and C-O···π and weak T-shaped offset π···π stacking interactions. The Hirshfeld surface analysis (HSA) of the PBMP crystal structure was performed as well, and the results were compared with the results of DBC and BCC. The density functional theory (DFT) study results revealed that the longer conjugated molecule of PBMP has smaller but still quite significant HOMO-LUMO gaps compared to the smaller molecules of BCC and DBC. The natural population analysis (NPA) and natural bonding orbital (NBO) analysis were performed. Accordingly, the hydrogen bonding and dipole-dipole interactions stabilize the crystal structures of these compounds. Additionally, the NBO analysis showed numerous high-energy stabilizing interactions for the PBMP compound due to the presence of numerous delocalized and relatively easily polarizable π-electrons, thus implying its significant thermodynamic stability. According to the global reactivity parameter (GRP) analysis, the compounds BCC and DBC are relatively stable in redox processes and have high thermodynamic stability and relatively lower reactivity in general. The molecular electrostatic potential (MEP) analysis results imply potential formation of the intermolecular hydrogen bonding and dispersion interactions, which stabilizes the crystal structures of these compounds.

Biodegradation of the pyrethroid cypermethrin by bacterial consortia collected from orange crops.

Pyrethroids, such as cypermethrin (CYP), are widely employed in agriculture, promoting environmental pollution and the need for efficient decontamination methods. In this study, bacteria from orange crops were explored for CYP biodegradation. Among 40 tested bacterial strains, 20 grew in the presence of CYP and 19 performed statistically significant CYP biodegradation in 5 days (20.5%-97.8%). In addition, 3-phenoxybenzoic acid, the main metabolite from CYP, was quantified ranging from 1.1 mg.L-1 to 32.1 mg.L-1. The five most efficient strains, and consortia composed of 5, 10 and 20 bacteria biodegraded the CYP formulation as sole carbon source in phosphate buffer and in minimum mineral medium. Under optimized conditions determined employing Response Surface Methodology, Bacillus sp. CSA-1 and the consortium composed of 10 strains biodegraded 71.0% and 71.6% CYP in 24 h, respectively. Moreover, metabolite identification enabled the proposal of an extended biodegradation pathway with 29 identified compounds, including different new amide and amine derivatives that expanded the knowledge about the fate of this compound in the environment. Experiments of bioaugmentation in soil using Bacillus sp. CSA-1 and the consortium of 10 bacterial strains resulted in faster CYP biodegradation than natural attenuation, showing that the selection of efficient strains for composing a consortium is an interesting approach for bioremediation of pyrethroids.

Development, validation, and application of an HPLC-MS/MS method for quantification of oxidized fatty acids in plants.

Oxylipins constitute a huge class of compounds produced by oxidation of long-chain unsaturated fatty acids either chemically (by radicals such as reactive oxygen species, ROS) or enzymatically (by lipoxygenases, LOX; cyclooxygenases, COX; or cytochrome P450 pathways). This process generates fatty acids peroxides, which can then be further modified in a broad range to epoxy, hydroxy, keto, ether fatty acids, and also hydrolyzed to generate small aldehydes and alcohols. In general, oxylipins are present in almost all living organisms and have a wide range of signaling, metabolic, physiological, and ecological roles depending on the particular organism and on their structure. In plants, oxylipins have been extensively studied over the past 35 years. However, these studies have focused mainly on the jasmonates and so-called green leaves volatiles. The function of early LOX products (like keto and hydroxy fatty acids) is yet not well understood in plants, where they are mainly analyzed by indirect methods or by GC-MS what requires a laborious sample preparation. Here, we developed and validated a straightforward, precise, accurate, and sensitive method for quantifying oxylipins in plant tissues using HPLC-MS/MS, with a one-step extraction procedure using low amount of plant tissues. We successfully applied this method to quantify the oxylipins in different plant species and Arabidopsis thaliana plants treated with various biotic and abiotic stress conditions.

Purification and characterization of two new antimicrobial molecules produced by an endophytic strain of Paenibacillus polymyxa.

An endophytic bacterium inhibiting pathogenic bacteria was isolated and the strain was genetically identified as Paenibacillus polymyxa. Biochemical characterization of fermentation broth indicated the presence of peptidic antimicrobial molecules. Liquid-liquid partition resulted in an organic fraction (OF) and an aqueous fraction (AF). OF presented a broad spectrum of activity against a panel of pathogenic bacteria and a fungus whereas the AF was active only against Gram-negative bacteria. AF was sequentially submitted to ion-exchange, desalting and reverse phase (RP) chromatography. A molecule with an RT of 2.45 min exhibited activity against all Gram-negative pathogenic strains tested beside P. mirabilis. The primary structure of the molecule, named AMP-Pp, was determined as Gly-Glu-Hyp-Gly-Ala by N-terminal sequencing. The molecular mass and amino acid sequence were confirmed by MS/MS. With a molecular mass of 463 Da, AMP-Pp is one of the smallest active natural peptides reported, yet. RP chromatography of OF resulted in four peaks. The first three peaks corresponded to known antimicrobials. MS analysis of peak 4 revealed the presence of an ion with m/z 3,376.4 Da, whose proposed molecular formula is C182H321N29O29. The compound, named polycerradin, showed a spectrum of activity against Gram-positive bacteria, Gram-negative bacteria (beside P. mirabilis) and a fungus.

New Role for a Commercially Available Bioinsecticide: Bacillus thuringiensis Berliner Biodegrades the Pyrethroid Cypermethrin.

The microbial diversity of several environments has been explored by researchers for the biodegradation of pyrethroids. In this study, a new approach was employed aiming at the use of Bacillus thuringiensis Berliner, a strain commercially available as bioinsecticide, for Cypermethrin (Cyp) biodegradation. This bacterial strain grew in the presence of Cyp and biodegraded this xenobiotic in a liquid medium. A central composite design for surface response methodology was employed for biodegradation. Under optimized conditions (50 mg·L-1 of Cyp, pH 8.5, 37 °C), 83.5% biodegradation was determined with the production of 12.0 ± 0.6 mg·L-1 3-phenoxybenzoic acid after 5 days. Moreover, a biodegradation pathway with the 18 compounds identified by GC-MS and LC-MS/MS was proposed. Experiments in soil for 28 days at 30 °C were performed, and 16.7% Cyp degradation was determined under abiotic conditions, whereas 36.6 ± 1.9% biodegradation was observed for B. thuringiensis Berliner with the native microbiome, indicating that bioaugmentation with this strain promoted a significant increase in the Cyp decontamination. Therefore, B. thuringiensis Berliner can act as biodegrader agent and insecticide at the same time, promoting decontamination of chemicals as Cyp while maintaining the protection of crops against insects. Moreover, B. thuringiensis species can produce bacteriocins with antifungal activity, which may increase agricultural productivity.

New Δ8,9-pregnene steroids isolated from the extremophile fungus Exophiala oligosperma.

Three pregnene steroids, two of them new, were isolated from ethyl-acetate partition of liquid-cultivation of the extremophyle fungus Exophiala oligosperma, found in a pH 1.5 hydrochloric acid aqueous solution. Spectroscopic studies using NMR and HRMS, allowed the identification of the molecular structures of both Δ8,9-pregnenes, still not described in the literature.

Biodegradation of the fungicide Pyraclostrobin by bacteria from orange cultivation plots.

The pesticides belonging the strobilurin group are among the most common contaminants in the environment. In this work, biodegradation studies of the strobilurin fungicide Pyraclostrobin by bacteria from orange cultivation plots were performed aiming to contribute with the development of a bioremediation method. Experiments were performed in triplicate with validated methods, and optimization was performed by Central Composite Design and Response Surface Methodology. The strains were evaluated in liquid nutrient medium containing 100 mg L-1 of Pyraclostrobin, and decreased concentrations of 61.5 to 100.5 mg L-1 were determined after 5 days at 37 °C and 130 rpm, showing the importance of strain selection. When the five most efficient strains (Bacillus sp. CSA-13, Paenibacillus alvei CBMAI2221, Bacillus sp. CBMAI2222, Bacillus safensis CBMAI2220 and Bacillus aryabhattai CBMAI2223) were used in consortia, synergistic and antagonistic effects were observed accordingly to the employed combination of bacteria, resulting in 64.2 ± 3.9 to 95.4 ± 4.9 mg L-1 residual Pyraclostrobin. In addition, the formation of 1-(4-chlorophenyl)-1H-pyrazol-3-ol was quantified (0.59-0.01 mg L-1), and a new biodegradation pathway was proposed with 15 identified metabolites. Experiments were also performed in soil under controlled conditions (30 °C, 0-28 days, 100 mg kg-1 pesticide), and the native microbiome reduced the pesticide concentration to 70.4 ± 2.3 mg L-1, whereas the inoculation of an efficient bacterial consortium promoted clearly better results, 57.2 ± 3.9 mg L-1 residual Pyraclostrobin. This suggests that the introduction of these strains in soil in a bioaugmentation process increases decontamination. However, the native microbiome is important for a more efficient bioremediation.

Antiproliferative activity of the dibenzylideneacetone derivate (E)-3-ethyl-4-(4-nitrophenyl)but­3-en-2-one in Trypanosoma cruzi.

Chagas disease is one of the most prevalent neglected diseases in the world. The illness is caused by Trypanosoma cruzi, a protozoan parasite with a complex life cycle and three morphologically distinct developmental stages. Nowadays, the only treatment is based on two nitro-derivative drugs, benznidazole and nifurtimox, which cause serious side effects. Since the treatment is limited, the search for new treatment options for patients with Chagas disease is highly necessary. In this study we analyzed the substance A11K3, a dibenzylideneacetone (DBA). DBAs have an acyclic dienone attached to aryl groups in both ß-positions and studies have shown that they have biological activity against tumors cells, bacteria, and protozoa such as T. cruzi and Leishmania spp. Here we show that A11K3 is active against all three T. cruzi evolutionary forms: the epimastigote (IC50 = 3.3 ± 0.8), the trypomastigote (EC50 = 24 ± 4.3) and the intracellular amastigote (IC50 = 9.3 ± 0.5 µM). A cytotoxicity assay in LLCMK2 cells showed a CC50 of 239.2 ± 15.7 µM giving a selectivity index (CC50/IC50) of 72.7 for epimastigotes, 9.9 for trypomastigotes and 25.9 for intracellular amastigotes. Morphological and ultrastructural analysis of the parasites treated with A11K3 by TEM and SEM revealed alterations in the Golgi complex, mitochondria, plasma membrane and cell body, with an increase of autophagic vacuoles and lipid bodies. Biochemical assays of A11K3-treated T. cruzi showed an increase of ROS, plasma membrane ruptures, lipid peroxidation, mitochondrial membrane depolarization with a decrease in ATP and accumulation of autophagic vacuoles. The results lead to the hypothesis that A11K3 causes death of the protozoan through events such as plasma membrane and mitochondrial alterations and autophagy, characteristic of cell collapse.

Digestion of Intact Gluten Proteins by Bifidobacterium Species: Reduction of Cytotoxicity and Proinflammatory Responses.

Celiac disease (CD) is a chronic illness characterized by an inflammatory process triggered by gluten protein intake. Recent evidence has suggested that the lower relative abundance of bifidobacteria in the intestinal lumen may be associated with CD. Herein, we assessed the effect of the Bifidobacterium species Bifidobacterium bifidum, Bifidobacterium longum, Bembidion breve, Bifidobacterium animalis alone, and also a Bifidobacterium consortium on the digestion of intact gluten proteins (gliadins and glutenins) and the associated immunomodulatory responses elicited by the resulting peptides. The cytotoxicity and proinflammatory responses were evaluated through the activation of NF-kB p65 and the expression of cytokines TNF-α and IL-1ß in Caco-2 cell cultures exposed to gluten-derived peptides. The peptides induced a clear reduction in cytotoxic responses and proinflammatory marker levels compared to the gluten fragments generated during noninoculated gastrointestinal digestion. These results highlight the possible use of probiotics based on bifidobacteria as a prospective treatment for CD.

Rapid differentiation of graft Citrus sinensis with and without Xylella fastidiosa infection by mass spectrometry.

RATIONALE: Xylella fastidiosa causes citrus variegated chlorosis (CVC) in sweet orange trees. A diagnostic method for detecting CVC before the symptoms appear, which would inform citrus producers in advance about when the plant should be removed from the orchard, is essential for reducing pesticide application costs. METHODS: Chemometrics was applied to high-performance liquid chromatography diode array detector (HPLC-DAD) data to evaluate the similarities and differences between the chromatographic profiles. A liquid chromatography/atmospheric pressure chemical ionization mass spectrometry selected reaction monitoring (LC/APCI-MS-SRM) method was developed to identify the major compounds and to determine their amounts in all samples. RESULTS: We evaluated the effect of this bacterium on the variation in the chemical profile in citrus plants. The organs of C. sinensis grafted on C. limonia were analyzed. Chemometrics was applied to the obtained data, and two major groups were differentiated. Flavonoids were observed in one group (leaves) and coumarins in the second (roots), both at higher concentrations in the plants with CVC symptoms than in those without the symptoms and those in the negative control. The rootstocks also interfered in the metabolism of the scion. CONCLUSIONS: The developed LC/APCI-MS-SRM method for detecting CVC before the symptoms appear is simple and accurate. It is inexpensive, and many samples can be screened per hour using 1 mg of leaves. Knowledge of the influence of the rootstock on the chemical profile of the graft is limited. This study demonstrates the effect of the rootstock in synthesizing flavonoids and increasing its content in all parts of the graft.

Activity and Cell-Death Pathway in Leishmania infantum Induced by Sugiol: Vectorization Using Yeast Cell Wall Particles Obtained From Saccharomyces cerevisiae.

Visceral leishmaniasis, caused by Leishmania infantum, is a neglected tropical disease, to which efforts in the innovation of effective and affordable treatments remain limited, despite the rising incidence in several regions of the world. In this work, the antileishmanial effects of sugiol were investigated in vitro. This compound was isolated from the bark of Cupressus lusitanica and showed promising activity against L. infantum. In spite of the positive results, it is known that the compound is a poorly water-soluble diterpene molecule, which hinders further investigation, especially in preclinical animal studies. Thus, in an alternative delivery method, sugiol was entrapped in glucan-rich particles obtained from Saccharomyces cerevisiae yeast cell walls (YCWPs). To evaluate the activity of sugiol, the experiments were divided into two parts: (i) the in vitro investigation of antileishmanial activity of free sugiol against L. infantum promastigotes after 24, 48, and 72 h of treatment and (ii) the evaluation of antileishmanial activity of sugiol entrapped in glucan-rich particles against intracellular L. infantum amastigotes. Free sugiol induced the cell-death process in promastigotes, which was triggered by enhancing cytosolic calcium level and promoting the autophagy up to the first 24 h. Over time, the presence of autophagic vacuoles became rarer, especially after treatment with lower concentrations of sugiol, but other cellular events intensified, like ROS production, cell shrinkage, and phosphatidylserine exposure. Hyperpolarization of mitochondrial membrane potential was found at 72 h, induced by the mitochondria calcium uptake, causing an increase in ROS production and lipid peroxidation as a consequence. These events resulted in the cell death of promastigotes by secondary necrosis. Sugiol entrapped in glucan-rich particles was specifically recognized by dectin-1 receptor on the plasma membrane of macrophages, the main host cell of Leishmania spp. Electron micrographs revealed particles containing sugiol within the infected macrophages and these particles were active against the intracellular L. infantum amastigotes without affecting the host cell. Therefore, the YCWPs act like a Trojan horse to successfully deliver sugiol into the macrophage, presenting an interesting strategy to deliver water-insoluble drugs to parasitized cells.

Synthesis, characterization, molecular docking evaluation, antidepressant, and anti-Alzheimer effects of dibenzylidene ketone derivatives.

Novel bioactive compounds as synthetic analogs of the potent herbal medicines can be optimized as potential drug candidates for various neurologic disorders. This study was performed to investigate the newly synthesized dibenzylidene ketone derivatives: (2E,6E)-2,6-dibenzylidene cyclohexanone (A1K1) and (1E,4E)-5-(2,3-dichlorophenyl)-1-(4-methoxyphenyl)-2-methylpenta-1,4-diene-3-one (A2K2) and evaluate its potential anti-Alzheimer's and anti-depressant properties. Both the derivatives are chemically characterized by using HNMR and CNMR techniques. Auto Dock Vina program was used to investigate ligand-protein affinity. Forced swim test, tail suspension test, open field test, Y-maze test, and Morris water maze test (MWM) models were employed to evaluate anti-depressant and anti-Alzheimer's activity of dibenzylidene ketone derivatives in mice. Both A1K1 and A2K2 showed high binding affinities against various proteins involved in depression and Alzheimer's mechanisms like monoamine oxidase B, acetylcholinesterase, norepinephrine transporter 2, serotonin transporter, dopamine receptor, serotonin receptor modulator, and beta-amyloid targets. A1K1 and A2K2 dose-dependently (0.1-1 mg/kg) decreased immobility time, while increased swimming and climbing time of mice in forced swim test (FST). A1K1 and A2K2 decreased animal immobility time in TST. In the open field test, both A1K1 and A2K2 increased the number of ambulations and rearings. A1K1 and A2K2 dose-dependently (0.5-1.0 mg/kg) increased spontaneous alternation behavior (%) and the number of entries of mice in Y-maze test. In the MWM test, A1K1 and A2K2 decreased escape latency time. Overall, both in-silico and in-vivo investigations of A1K1 and A2K2, report their therapeutic potential for antidepressant and anti-Alzheimer properties. Hence, these compounds possess potent neuroprotective properties and may be further evaluated for their therapeutic potential in various neurological disorders.

Draft Genome Sequence of the Fungus Penicillium brasilianum (Strain LaBioMMi 136), a Plant Endophyte from Melia azedarach.

Penicillium brasilianum (strain LaBioMMi 136) has been reported to be a great producer of secondary metabolites and a source of enzymes of biotechnological interest. Here, we report the draft genome sequence of P. brasilianum (strain LaBioMMi 136), isolated as an endophyte from the plant Melia azedarach (family Meliaceae).

Conjugation of antifungal benzoic acid derivatives as a path for detoxification in Penicillium brasilianum, an endophyte from Melia azedarach.

In this study, the consumption of 4-bromobenzoic acid and 4-chlorobenzoic acid by the fungus Penicillium brasilianum, an endophyte from Melia azedarach is evaluated. This fungus metabolizes these halobenzoic acids to produce three new brominated compounds, which have been isolated and characterized, and three new chlorinated derivatives identified by HRMS. Among these products, (4-bromobenzoyl)proline has been also chemically synthesized and employed in biological assays, thus providing insights for the elucidation of the defense mechanism of P. brasilianum towards these halobenzoic acids.

Photolysis of parabens using medium-pressure mercury lamps: Toxicity effects in MCF7, Balb/c 3T3 cells and Ceriodaphnia dubia.

Degradation studies of the propylparaben (PrP), butylparaben (BuP) and of the propylparaben-butylparaben mixture (PrP-BuP) in deionized water and surface river water was investigated as a function of pH and initial concentration of the reactants using a medium-pressure mercury lamp. The photolysis of parabens (concentration ranging from 5 to 30 mg L-1) followed apparent pseudo-first-order kinetics, with rate constants (k) in deionized water and surface river water changed from 1.80 × 10-1 to 3.68 × 10-2 min-1 and 1.43 × 10-1 to 1.45 × 10-2 min-1, respectively. Degradation reaction was faster at pH 5 in comparison with pH 7 or 11. The photolysis of parabens was greater than 91%, with low mineralization (26.15%) observed in acidic medium after 95 min. Analysis by chromatography coupled to mass spectrometry (LC-MS/MS) showed that only one product was generated during the degradation reaction and has UV bands similar to 3,4-dihydroxybenzoic acid. Estrogenic activity tests showed that non-degraded parabens stimulated the growth of breast adenocarcinoma (MCF-7) cells and this effect was evaluated after the photolysis. Cytotoxicity assays using fibroblasts cells (Balb/C 3T3 clone A31) indicated that the parental compounds and degradation products were not cytotoxic. On the contrary, non-degraded parabens were toxic to Ceriodaphnia dubia, but the product of photolysis was not. Overall, the photolytic method presented was able to degrade these parabens providing safe and non-estrogenic reaction product.