Depsidomycins B and C: New Cyclic Peptides from a Ginseng Farm Soil-Derived Actinomycete.

LC/MS-based chemical profiling of a ginseng farm soil-derived actinomycete strain, Streptomyces sp. BYK1371, enabled the discovery of two new cyclic heptapeptides, depsidomycins B and C (1 and 2), each containing two piperazic acid units and a formyl group at their N-terminus. The structures of 1 and 2 were elucidated by a combination of spectroscopic and chemical analyses. These new compounds were determined to possess d-leucine, d-threonine, d-valine, and S-piperazic acid based on the advanced Marfey's method and a GITC (2,3,4,6-tetra-O-acetyl-ß-d-glucopyranosyl isothiocyanate) derivatization of their hydrolysates, followed by LC/MS analysis. Depsidomycins B and C displayed significant antimetastatic activities against metastatic breast cancer cells (MDA-MB-231).

Screening of an FDA-approved compound library identifies levosimendan as a novel anti-HIV-1 agent that inhibits viral transcription.

Combination antiretroviral therapy (cART) has been proven to efficiently inhibit ongoing replication of human immunodeficiency virus type 1 (HIV-1), and significantly improve the health outcome in patients of acquired immune deficiency syndrome (AIDS). However, cART is unable to cure HIV-1/AIDS. Even in presence of cART there exists a residual viremia, contributed from the viral reservoirs of latently infected HIV-1 proviruses; this constitutes a major hurdle. Currently, there are multiple strategies aimed at eliminating or permanently silence these HIV-1 latent reservoirs being intensely explored. One such strategy, a recently emerged "block and lock" approach is appealing. For this approach, so-called HIV-1 latency-promoting agents (LPAs) are used to reinforce viral latency and to prevent the low-level or sporadic transcription of integrated HIV-1 proviruses. Although several LPAs have been reported, there is still a question of their suitability to be further developed as a safe and valid therapeutic agent for the clinical use. In this study, we aimed to identify new potential LPAs through the screening an FDA-approved compound library. A new and promising anti-HIV-1 inhibitor, levosimendan, was identified from these screens. Levosimendan is currently used to treat heart failure in clinics, but it demonstrates strong inhibition of TNFα-induced HIV-1 reactivation in multiple cell lines of HIV-1 latency through affecting the HIV-1 Tat-LTR transcriptional axis. Furthermore, we confirmed that in primary CD4+ T cells levosimendan inhibits both the acute HIV-1 replication and the reactivation of latent HIV-1 proviruses. As a summary, our studies successfully identify levosimendan as a novel and promising anti-HIV-1 inhibitor, which should be immediately investigated in vivo given that it is already an FDA-approved drug.

Svetamycins A-G, Unusual Piperazic Acid-Containing Peptides from Streptomyces sp.

Seven new halogenated peptides termed svetamycins A-G (1-7) have been isolated from laboratory cultures of a Streptomyces sp. Svetamycins A-D, F, and G are cyclic depsipeptides, whereas svetamycin E is a linear analogue of svetamycin C. Their structures were determined using extensive spectroscopic analysis, and their stereochemical configuration was established by a combination of NMR data, quantum mechanical calculations, and chemical derivatizations. Svetamycins are characterized by the presence of a hydroxyl acetic acid and five amino acids including a rare 4,5-dihydroxy-2,3,4,5-tetrahydropyridazine-3-carboxylic acid, a γ-halogenated piperazic acid, and a novel δ-methylated piperazic acid in svetamycins B-C, E, and G. Moreover, isotope-labeled substrate feeding experiments demonstrated ornithine as the precursor of piperazic acid and that methylation at the δ position of the piperazyl scaffold is S-adenosyl-l-methionine (SAM)-dependent. Svetamycin G, the most potent antimicrobial of this suite of compounds, inhibited the growth of Mycobacterium smegmatis with an MIC80 value of 2 µg/mL.

Worker illness related to newly marketed pesticides--Douglas County, Washington, 2014.

On April 10, 2014 the Washington State Department of Agriculture (WSDA) was notified by a local newspaper of a suspected pesticide poisoning incident in Douglas County involving pesticides not previously reported in the published literature to be associated with human illness. On that same day, WSDA notified the Washington State Department of Health, which investigated this incident by conducting a site visit, reviewing medical and applicator records, and interviewing affected farmworkers, pesticide applicators, and the farmworkers' employer. In addition, on April 11, WSDA collected swab, foliage, and clothing samples and tested them for residues of pyridaben, novaluron, and triflumizole. In this incident, all 20 farmworkers working in a cherry orchard became ill from off-target drift of a pesticide mixture that was being applied to a neighboring pear orchard. Sixteen sought medical treatment for neurologic, gastrointestinal, ocular, and respiratory symptoms. This event highlights the need for greater efforts to prevent off-target drift exposures and promote awareness about the toxicity of some recently marketed pesticides. Incidents such as this could be prevented if farm managers planning pesticide applications notify their neighbors of their plans.

High-performance liquid chromatography tandem mass spectrometry method for quantification of endothelin receptor antagonist drug, ambrisentan, in human plasma and its application in a pharmacokinetic study.

A simple and sensitive liquid chromatography tandem mass spectrometry method has been developed for the quantification of ambrisentan (AMB) in human plasma using midazolam (MID) as an internal standard (IS). Chromatographic separation was performed using a Beta Basic-8 (50 × 4.6 mm, 5 µm) column with an isocratic mobile phase. AMB and MID were detected with proton adducts at m/z 379.09 → 303.12 and 326.15 → 291.14 in multiple reaction monitoring-positive mode, respectively. A solid-phase extraction method was used for extraction of the analyte and IS from the plasma samples. The method was shown to be reproducible and reliable with within-run precision <11%, between-run precision <14% and linear concentration range from 10.0 to 2000.2 ng/mL, with a correlation coefficient (r(2) ) of >0.995. The method was successfully applied to a pharmacokinetic study of oral administration of AMB (10 mg) in 24 healthy Indian male human volunteers under fasting conditions.

Synthesis, HPLC enantioresolution, and X-ray analysis of a new series of C5-methyl pyridazines as N-formyl peptide receptor (FPR) agonists.

The synthesis of three racemates and the corresponding non-chiral analogues of a C5-methyl pyridazine series is described here, as well as the isolation of pure enantiomers and their absolute configuration assignment. In order to obtain optically active compounds, direct chromatographic methods of separation by HPLC-UV were investigated using four chiral stationary phases (CSPs: Lux Amylose-2, Lux Cellulose-1, Lux Cellulose-2 and Lux Cellulose-3). The best resolution was achieved using amylose tris(5-chloro-2-methylphenylcarbamate) (Lux Amylose-2), and single enantiomers were isolated on a semipreparative scale with high enantiomeric excess, suitable for biological assays. The absolute configuration of optically active compounds was unequivocally established by X-ray crystallographic analysis and comparative chiral HPLC-UV profile. All compounds of the series were tested for formyl peptide receptor (FPR) agonist activity, and four were found to be active, with EC50 values in the micromolar range.

Adsorption of diuron, fluridone and norflurazon on single-walled and multi-walled carbon nanotubes.

The sorption behaviors of diuron (DIU), fluridone (FLU) and norflurazon (NOR) by a single-walled carbon nanotube (SWCNT) and three multi-walled carbon nanotubes (MWCNT) samples including MWCNT10 (<10nm, outer diameter), MWCNT20 (10-20 nm), and MWCNT40 (20-40 nm) were investigated. All adsorption isotherms were nonlinear and were well fitted with the Freundlich model and Dubinin Ashtakhov (DA) model. The linear relationships between the organic carbon (OC)-normalized saturated adsorption capacity (Q(0)(OC)) and surface area (SA) suggest that SA is presumably responsible for the adsorption of DIU and NOR on CNTs. While FLU, DIU, and NOR OC-normalized distribution coefficients (logK(OC)) of CNTs increased with increasing their hydrophobicity (logK(OW)) and the positive relationships between the logK(OW)-normalized logK(OC) (i.e., logK(OC)/logK(OW)) of FLU, DIU, and NOR and their hydrogen bonding ability indicate that the adsorption of FLU, DIU and NOR was mainly controlled by the hydrophobic interaction and hydrogen bonding. The higher logK(OC) or Q(0)(OC) values of MWCNT10 and SWCNT relative to other large MWCNTs and carbonaceous adsorbents suggest that MWCNT10 has the potential to serve as an adsorbent used to reduce the mobility of herbicides in agricultural and environmental applications.

Sorption of fluorinated herbicides to plant biomass-derived biochars as a function of molecular structure.

Biochars produced at different heat treatment temperatures (HTT) are molecularly distinct and thus expected to show variable sorbent characteristics. We investigated the difference in sorption behavior of norflurazon (NORO) and fluridone (FLUN) to biochars from wood and grass feedstocks produced at different HTT. Amorphous biochars (HTT=400°C) exhibited the highest sorption parameter (K(OC)) for the two herbicides, emphasizing the importance of amorphous structural arrangement of aromatic moieties in these chars. Negative correlation between biochar aromaticity and isotherm nonlinearity (n) suggests that the n values were related mainly to total aromatic C content, not to that in the individual phases. Sorption of FLUN and NORO to low-temperature biochars (HTT=400°C) was about 1100 times and 6400 times greater, respectively, than a sediment sample, confirming that applications of low-temperature biochars to arable soils may reduce the mobility of FLUN and NORO, thus preventing unwanted herbicide leaching and subsequent contamination of sensitive water bodies.

Analytical and semi-preparative HPLC enantioseparation of novel pyridazin-3(2H)-one derivatives with α-aminophosphonate moiety using immobilized polysaccharide chiral stationary phases.

The direct HPLC enantioseparation of a novel series of chiral pyridazin-3(2H)-one derivatives with α-aminophosphonate moiety was performed on two immobilized polysaccharide chiral stationary phases (Chiralpak IA, Chiralpak IC) using n-hexane (n-Hex)/dichloromethane (DCM) mobile phase with 5% alcohol additive. Good baseline separation of the enantiomers was achieved using amylose tris-(3,5-dimethylphenylcarbamate) chiral stationary phases (Chiralpak IA) on analytical scale. The analytical method was further scaled up to semi-preparative loading to obtain small amounts of both the enantiomers of pyridazin-3(2H)-one derivative. The semi-preparative resolution of all compounds was successfully achieved with n-hexane/dichloromethane/ethanol (EtOH) as mobile phase using a semi-preparative Chiralpak IA column. The first fractions were isolated with purities of >99.9% (enantiomeric excess (e.e.), and the second fractions were obtained with purities of >98.2% (enantiomeric excess). The assignment of the absolute configuration was established for the F1 fraction of compound a-2 by single-crystal X-ray diffraction method.

Adsorption of chloridazon from aqueous solution on modified kerolite-rich materials.

The adsorption of chloridazon (5-amine-4-chloro-2-phenylpyridazin-3(2H)-one) on kerolite samples heated at 110 degrees C (K-110), 200 degrees C (K-200), 400 degrees C (K-400), 600 degrees C (K-600) and acid-treated with H(2)SO(4) solutions of two different concentrations (0.25 and 0.5 M) (K-0.25 and K-0.5, respectively) from pure water at 25 degrees C has been studied by using batch and column experiments. The adsorption experimental data points were fitted to the Freundlich equation in order to calculate the adsorption capacities (K(f)) of the samples; K(f) values ranged from 184.7 mg kg(-1) (K-0.5) up to 2253 mg kg(-1) (K-600). This indicated that the heat treatment given to the kerolite greatly increases its adsorption capacity for the herbicide whereas the acid treatment produces a clear decrease in the amount of chloridazon adsorbed. The removal efficiency (R) was also calculated; R values ranging from 52.8% (K-0.5) up to 88.3% (K-600). Thus, the results showed that the 600 degrees C heat-treated kerolite was more effective in relation to adsorption of chloridazon and it might be reasonably used in removing this herbicide from water.

4849F, a new metabolite produced by the Streptomyces sp. 4849 as an inhibitor of IL-4 receptor.

A new compound named 4849F was isolated from the Streptomyces sp. 4849. The structure of 4849F was elucidated by spectroscopic analyses. The immobilized ligand-binding assay showed that 4849F can competitively inhibit the binding of IL-4 with IL-4 receptor in a dose dependent manner with an IC50 value of 6.7 microM.

Inclusion complexes of alpha- and gamma-cyclodextrins and the herbicide norflurazon: I. Preparation and characterisation. II. Enhanced solubilisation and removal from soils.

The interaction of norflurazon with alpha- and gamma-cyclodextrins (CDs) yielded the formation of inclusion complexes at a 1:1 stoichiometric ratio in solution and in the solid state. Apparent stability constants of 50.7+/-1.6 and 37+/-1.7 M(-1) and an increase in herbicide solubility by up to five and fourfold for alpha- and gamma-CD, respectively, were determined from the phase solubility diagrams at 25 degrees C in water. Three processing methods (kneading, spray-drying and vacuum evaporation) were used to prepare norflurazon-CD solid inclusion complexes, which were characterised by infrared spectroscopy, differential scanning calorimetry and scanning electron microscopy. A high increase in the norflurazon dissolution rate was obtained with all the solid complexes with gamma-CD, but when alpha-CD was used, only the solid system obtained after the vacuum evaporation process showed a higher dissolution rate. This finding is a first step in the development of new, environmentally sound formulations of norflurazon (NFL), due to the capacity for increasing its dissolution rate and hydrosolubility, and thus diminishing the use of organic solvents. On the other hand, the effect of alpha- and gamma-cyclodextrin on the solubility of norflurazon in solution was also considered as a way of modifying its behaviour in the soil environment. Desorption studies of NFL from soils in the presence of alpha- and gamma-cyclodextrin were carried out using a batch equilibration method. The results obtained showed that alpha- and gamma-cyclodextrin greatly increased the removal of norflurazon previously adsorbed, proving the potential use of these CDs for in situ remediation of pesticide-contaminated soils.

Adsorption of chloridazon from aqueous solution on heat and acid treated sepiolites.

The adsorption of chloridazon on heat treated sepiolite samples at 110 degrees C (S-110), 200 degrees C (S-200), 400 degrees C (S-400), 600 degrees C (S-600) and acid treated samples with H2SO4 solutions of two different concentrations (0.25 and 1.0M) (S-0.25 and S-1.0, respectively) from pure water at 25 degrees C has been studied by using batch experiments. In addition, column experiments were carried out with the natural (S-110) and 600 degrees C (S-600) heat treated samples, using a 10.30 mg l-1 aqueous solution of chloridazon. The adsorption experimental data points have been fitted to the Freundlich equation in order to calculate the adsorption capacities (Kf) of the samples; Kf values range from 2.89 mg kg-1 for the S-1.0 sample up to 164 mg kg-1 for the S-600 sample; so, the heat treatment given to the sepiolite greatly increases its adsorption capacity for the herbicide chloridazon whereas the acid treatment produces a clear decrease in the amount of chloridazon adsorbed. The removal efficiency (R) has also been calculated; R values ranging from 5.08% for S-1.0 up to 60.9% for S-600. The batch experiments showed that the strongest heat treatment is more effective than the natural and acid treated sepiolite in relation to adsorption of chloridazon. The column experiments also showed that 600 degrees C heat treated sepiolite might be reasonably used in removing chloridazon from water. Thus, as this type of clay is relatively plentiful, these activated samples might be reasonably used in order to remove chloridazon from water.

Chloptosin, an apoptosis-inducing dimeric cyclohexapeptide produced by Streptomyces.

In the course of screening for apoptosis-inducing agents, chloptosin (1) was isolated from the culture broth of Streptomyces. The dumbbell-type structure of the dimeric cyclohexapeptide consisting of D-valine, (3S)- and (3R)-piperazic acids, O-methyl-L-serine, D-threonine, and (2S,3aR,8aR)-6-chloro-3a-hydroxy-2,3,3a, 8a-hexahydropyrrolo[2,3-b]indole-2-carboxylic acid was elucidated by spectroscopic and chemical degradation studies. The amino acid components in each cyclohexapeptide domain were presented in alternating R and S configurations. Chloptosin (1) was found to induce apoptotic activity in apoptosis-resistant human pancreatic adenocarcinoma cell line AsPC-1 and showed a strong antimicrobial activity against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus.

Enantiomeric separation of optically active pyridazinone derivatives by chiral HPLC.

Several 4,5-disubstituted 3(2H)-pyridazinone derivatives containing 2-hydroxymethylpyrrolidino moiety as a chiral building block were synthetized. Separation of enantiomers was carried out by chiral HPLC on Chiralcel OJ and OF columns. Mobile phases consisted of hexane, ethanol and 2-propanol. Chiralcel OJ column was capable of separating most of the enantiomeric pairs. For one type of compound. Chiralcel OF column was used for separation.

Chiral resolution and absolute configuration of the enantiomers of 5-acetyl-2-methyl-4-methylsulfinyl-6-phenyl-3(2H)-pyridazinone and evaluation of their platelet aggregation inhibitory activity.

In a series of 5-acyl-6-phenyl-2,4-substituted-3(2H)-pyridaziones the derivative 1a, with a sulfur stereogenic center, had the most potent activity as human platelet aggregation inhibitor. The resolution of rac-1a was successfully performed by chiral chromatography on Chiralcel OD-R, OD-H, and Chiralpak AD columns and scaled up to a preparative level. The absolute configuration of (-)-(S)-1a was determined by X-ray crystallographic analysis. In vitro human platelet aggregation inhibitory activity was evaluated. Both the enantiomers showed IC50 values in the same micromolar range, but the (-)-(S) isomer was slightly more potent [(S)/(R) potency ratio was 4/1].

Piperastatin B: a new selective serine carboxypeptidase inhibitor from Streptomyces lavendofoliae MJ908-WF13.

Piperastatin B, a new inhibitor of serine carboxypeptidase was purified from a culture broth of Streptomyces lavendofoliae MJ908-WF13 as a minor component by monitoring its inhibitory activity against carboxypeptidase Y (CP-Y). Its structure was determined to be N-formyl-Val-Thr-Leu-Val-Pip-Leu-Pip (pip: piperazic acid, hexahydropyridadine-3-carboxylic acid). Piperastatin B is a highly specific competitive inhibitor of CP-Y (Ki = 55 nM) with little effect on related enzymes and resembles the major component, piperastatin A, in these respects.

Piperastatin A, a new selective serine carboxypeptidase inhibitor produced by actinomycete. I. Taxonomy, production, isolation and biological activities.

Piperastatin A (structure, N-formyl-allo Ile-Thr-Leu-Val-Pip-Leu-Pip, Pip = hexahydropyridadine-3-carboxylic acid; molecular weight, 809), a new inhibitor of serine carboxypeptidase was discovered in the fermentation broth of Streptomyces lavendofoliae MJ908-WF13. It was purified by activated charcoal chromatography, YMC gel ODS-A chromatography and centrifugal partition chromatography (CPC) by monitoring its inhibitory activity against carboxypeptidase Y (CP-Y), and finally obtained as colourless needles. Piperastatin A is a competitive inhibitor of the enzyme with Ki = 52 +/- 6.2 nM. Piperastatin A is a highly specific inhibitor of the serine carboxypeptidases, CP-Y and platelet deamidase with little effect on related enzymes, has no antimicrobial activity and has low toxicity.

Zarzissine, a new cytotoxic guanidine alkaloid from the Mediterranean sponge Anchinoe paupertas.

From a CH2Cl2 extract of the Mediterranean sponge Anchinoe paupertas were isolated and characterized zarzissine [2], a new 4,5-guanidino-pyridazine compound and the known p-hydroxybenzaldehyde [1]. The structure of zarzissine [2] was elucidated by spectroscopic methods, including the application of a number of 2D nmr techniques. Biological activities of compounds 1 and 2 were also determined, with zarzissine exhibiting cytotoxicity against three human and murine tumor cell lines.

Matlystatins, new inhibitors of typeIV collagenases from Actinomadura atramentaria. I. Taxonomy, fermentation, isolation, and physico-chemical properties of matlystatin-group compounds.

During the course of a screening for inhibitors of typeIV collagenases, new metabolites, designated matlystatins, have been isolated from an actinomycete strain, which was identified as a strain of Actinomadura atramentaria. Matlystatins were composed of five congeners, which were separated and purified by n-butanol extraction and chromatography.