Cyclofarnesoids and methylhexanoids produced from ß-carotene in Phycomyces blakesleeanus.

The oxidative cleavage of ß-carotene in the Mucorales produces three fragments of 18, 15, and 7 carbons, respective heads of three families of apocarotenoids: the methylhexanoids, the trisporoids, and the cyclofarnesoids (named after their 1,6-cyclofarnesane skeleton). The apocarotenoids are easily recognized because they are absent in white mutants unable to produce ß-carotene. In cultures of Phycomyces blakesleeanus we detected thirty-two apocarotenoids by LC, UV absorbance, and MS. With additional IR and NMR we identified two methylhexanoids and the eight most abundant cyclofarnesoids. Four of them were previously-unknown natural compounds, including 4-dihydrocyclofarnesine S, the most abundant cyclofarnesoid in young cultures. We arranged the apocarotenoids of the Mucorales in a scheme that helps classifying and naming them and suggests possible metabolites and biosynthetic pathways. We propose specific biosynthetic pathways for cyclofarnesoids and methylhexanoids based on structural comparisons, the time course of appearance of individual compounds, and the bioconversion of ß-apo-12-carotenol, an early precursor, to three more oxygenated cyclofarnesoids by the white mutants. Some of the reactions occur spontaneously in the increasingly acidic culture media. Mating increased the contents of methylhexanoids and cyclofarnesoids by ca. threefold in young cultures and ca. twelvefold in old ones (five days); cyclofarnesine S, the most abundant cyclofarnesoid in old cultures, increased over one hundredfold. We found no differences between the sexes and no activity as sexual pheromones, but we suggest that methylhexanoids and cyclofarnesoids could mediate species-specific physiology and behavior.

Expedient access to enantiopure cyclopentanic natural products: total synthesis of (-)-cyclonerodiol.

Following the principles of collective total synthesis, a number of natural products sharing an optically pure, multifunctional, cyclopentanic core were synthesized from a common precursor: plinol A (1). This intermediate was efficiently obtained in only four steps from (-)-linalool (2) using as the key step a Ti(III)-mediated diastereoselective radical cyclization. The feasibility of this approach was confirmed with the expedient enantiospecific synthesis of cyclonerodiol (3), and the formal synthesis of chocol G (4) and piperitone (5).

[Clinical practice variation in cataract surgery]. / Variabilidad clínica en la cirugía de la catarata.

PURPOSE: Cataract surgery rates have dramatically increased in the last two decades. However, clinical practice variation in cataract surgery has not been thoroughly studied. The aim of this review is to analyze clinical practice variation, including the causes and consequences of this phenomenon. Then, its role in health care planning and health care quality is focused, emphasizing the importance of reducing it and providing several practical strategies to accomplish it. RECENT FINDINGS: The latest researches are presented in this article. They identify the development and implementation of clinical practice guidelines as the best tool to standardize care processes. CONCLUSION: Managing unwarranted or unwanted variation would improve quality of care and may lead to a significant saving in health care spending.

First total synthesis of (+)-apotrisporin E and (+)-apotrientriols A-B: a cyclization approach to apocarotenoids.

The first total synthesis of the natural apocarotenoids (+)-apotrisporin E (1) and (+)-apotrientriols A and B (2-3) has been accomplished. The structure, relative stereochemistry and the assignation of the absolute configuration have been confirmed. This is a fast and easy access to this family of natural products whose key steps are a diastereoselective cyclization and a HWE olefination to attach the dienic side chain. This work also opens the door to the synthesis of other apocarotenoids such as trisporols and trisporic acids.

Structural diversity from the transannular cyclizations of natural germacrone and epoxy derivatives: a theoretical-experimental study.

Treatment of germacrone (1) with different electrophiles, and of its epoxy derivatives germacrone-4,5-epoxide (2), germacrone-1,10-epoxide (3) and isogermacrone-4,5-epoxide (4) with Brönsted/Lewis acids and Ti(III), gives rise to a great structural diversity. Thus, by using a maximum of two steps, the production of more than 40 compounds corresponding to 14 skeletons is described. Computational calculations rationalizing the structural divergence produced are also described. Finally, since some of the compounds generated are bioactive natural sesquiterpenes, the mechanisms of formation of these substances will provide new insights in their biosynthesis.

A comparison between radiochromic EBT2 film model and its predecessor EBT film model.

The manufacturer has introduced the new EBT2 film model so as to improve its predecessor, the EBT radiochromic film model. According to the manufacturer, some of its main advantages include a higher tolerance to light exposure and it can correct non-uniformity of the active layer thickness using a marker dye. However, the equivalence in uniformity between both models was questioned by some authors, and the asymmetrical configuration of layers of the EBT2 film model produces a new dependence on the film side being scanned (front and back orientation). In this study, the EBT2 radiochromic film model was compared with the EBT model and the new marker dye feature was assessed. We also compared this correction method with a pre-irradiated pixel value correction method. An Epson Expression 10000XL scanner in transmission mode was used to scan the films and the red channel response was analyzed. We confirmed the lower-measured signal dependence on the visible light exposure of the EBT2 film model. Differences in pixel values remained below 0.5% for a minimum of 15 days. In regard to the uniformity, similar results for EBT2 and EBT film models were obtained; in both cases inhomogeneity was found to be less than 1%, in relative pixel value from the mean. However, we found that the signal-to-noise ratio was reduced for low doses by 37% for old EBT2 batch and by 21% for new EBT2 batch compared to signal-to-noise ratio for EBT. The EBT2 film model's pixel value difference for the front and back orientation reached up to 1.0% in the red channel. Our results did not show a clear advantage between to use a pre-irradiated pixel value correction and to use the manufacturer's correction.

A minor dihydropyran apocarotenoid from mated cultures of Blakeslea trispora.

The heterocyclic C15 apocarotenoid 1 was isolated from mated cultures of the strains F986 (+) and F921 (−) of Blakeslea trispora. This new compound formed during sexual interaction is a minor constituent of the culture media and its structure was elucidated by spectroscopic data, including 2D-NMR. A plausible biosynthetic pathway involving a double degradation of β-carotene, followed by several oxidations of the resulting monocyclofarnesane C15 fragment is proposed.

Apocarotenoids in the sexual interaction of Phycomyces blakesleeanus.

A simple genetic test allowed us to carry out the first systematic study of the apocarotenoids in the Mucorales. We have identified 13 apocarotenoids in the culture media of the fungus Phycomyces blakesleeanus (Mucoromycota, Mucorales). Three of these compounds were novel apocarotenoids: (2S,8R,E)-8,14-epoxycyclofarnesa-4,6,9-triene-2,11-diol (6), (2S,6E,8E)-cyclofarnesa-4,6,8-triene-2,10,11-triol (7), and its 6Z isomer (8). Four of the remaining compounds have been reported previously from this fungus and six from other Mucorales. All of them belong to three families, the 18-carbon trisporoids, the 15-carbon cyclofarnesoids, and the 7-carbon methylhexanoids, derived from the three fragments that result when ß-carotene is cleaved at its 11',12' and 12,13 double bonds. The apocarotenoids were more varied and more abundant in mated cultures of strains of opposite sex than in single cultures. The presence of acetate in the medium blocked the production of many apocarotenoids while having little effect on the concentrations of the remaining ones.

Communic acids: occurrence, properties and use as chirons for the synthesis of bioactive compounds.

Communic acids are diterpenes with labdane skeletons found in many plant species, mainly conifers, predominating in the genus Juniperus (fam. Cupresaceae). In this review we briefly describe their distribution and different biological activities (anti- bacterial, antitumoral, hypolipidemic, relaxing smooth muscle, etc.). This paper also includes a detailed explanation of their use as chiral building blocks for the synthesis of bioactive natural products. Among other uses, communic acids have proven useful as chirons for the synthesis of quassinoids (formal), abietane antioxidants, ambrox and other perfume fixatives, podolactone herbicides, etc., featuring shorter and more efficient processes.

Characterization of microsatellite loci in Erysimum mediohispanicum (Brassicaceae) and cross-amplification in related species.

PREMISE OF THE STUDY: We have developed and optimized microsatellite loci from a genomic library of Erysimum mediohispanicum. Microsatellites were also tested for cross-amplification in 31 other Erysimum species. METHODS AND RESULTS: A total of 10 microsatellite loci were successfully amplified. They were polymorphic for 81 E. mediohispanicum individuals from two locations in Sierra Nevada (southeastern Spain), which showed similar patterns of genetic diversity. On average, microsatellites had 8.6 alleles per locus and an expected heterozygosity of 0.69. Only one locus significantly departed from Hardy-Weinberg equilibrium in both locations. Most of the markers successfully amplified in other Erysimum species. CONCLUSIONS: The genetic attributes of microsatellite loci will allow their application to population genetic studies in Erysimum, such as genetic differentiation and structure, gene flow, pollinator-mediated speciation, and hybridization studies.

New apocarotenoids and ß-carotene cleavage in Blakeslea trispora.

Mixed cultures of strains of opposite sex ("mated" cultures) of Blakeslea trispora contain trisporic acids and other apocarotenoids, some of which mediate the sexual responses of this fungus and other Mucorales. In mated cultures of the wild-type strains F986 and F921 we identified eleven apocarotenoids: two C(18) trisporoids, three C(15) compounds with a monocyclofarnesane skeleton, a C(13) compound, and five C(7) compounds with a 2-methylhexane skeleton. Six of them are new natural products and two others are new for Blakeslea. Their structures were established by NMR and mass spectra and those of the C(7) and C(13) compounds were confirmed by chemical synthesis. The finding of these compounds and the presence of approximately equimolecular amounts of the C(18), C(15), and C(7) families led to the conclusion that ß-carotene is initially split in three fragments by cleavage of its 13,14 and 11',12' double bonds.

Comparison of the essential oils of leaves and stem bark from two different populations of Drimys winteri a Chilean herbal medicine.

The chemical composition of the essential oils obtained by hydrodistillation of stem bark and leaves of Drimys winteri J.R. et G. Foster var. chilensis /DC A. Gray (Winteraceae) from Chiloe Island (ID) and Continental Chile (Santiago) (CD) were studied by GC and GC/MS. Sesquiterpene hydrocarbons constituted the main chemical groups in the stem bark oils, with alpha-santalene, trans-beta-bergamotene and curcumenes as the major components. Monoterpenes constituted the main chemical groups in the leaves of Island plants with alpha-pinene (23.1%) beta-pinene (43.6%) and linalool (10.5%) as the main components whereas sesquiterpenes (germacrene D 17.6%) and phenylpropanoids (safrole 20.8%) are the most abundant in the leaves of Continental plants.

Using complementary techniques to distinguish cryptic species: a new Erysimum (Brassicaceae) species from North Africa.

PREMISE OF THE STUDY: Cryptic species are superficially morphologically indistinguishable and therefore erroneously classified under one single name. The identification and delimitation of these species is usually a difficult task. The main aim of this study is to provide an inclusive methodology that combines standard and new tools to allow accurate identification of cryptic species. We used Erysimum nervosum s.l. as a model system. METHODS: Four populations belonging to E. nervosum s.l. were sampled at their two distribution ranges in Morocco (the Atlas Mountains and the Rif Mountains). Fifteen individuals per population were collected to assess standard taxonomic traits. Additionally, corolla color and shape were quantified in 30 individuals per population using spectrophotometry and geometric morphometrics, respectively. Finally, we collected tissue samples from each population per species to study the phylogenetic relationships among them. KEY RESULTS: Using the standard taxonomic traits, we could not distinguish the four populations. Nonetheless, there were differences in corolla color and shape between plants from the two mountain ranges. The population differentiation based on quantitative morphological differences were confirmed and supported by the phylogenetic relationships obtained for these populations and the rest of the Moroccan Erysimum species. CONCLUSIONS: The joint use of the results obtained from standard taxonomic traits, quantitative analyses of plant phenotype, and molecular data suggests the occurrence of two species within E. nervosum s.l. in Morocco, one located in the Atlas Mountains (E. nervosum s.s.) and the other in the Rif Mountains (E. riphaeanum sp. nov.). Consequently, we suggest that combining quantitative and molecular approaches with standard taxonomy greatly benefits the identification of cryptic species.

Use of the plant Bellardia trixago for the enantiospecific synthesis of odorant products.

An easy procedure to obtain extracts enriched in trixagol monomalonylesther (1) from aerial parts of the plant Belladia trixago chemotype Trix was developed. Preparation of (+)-dihydro-gamma-ionone (4) was carried out directly from the extracts with good yields by selective oxidation. Other interesting odorant products as alpha-ambrinol (5), ambraldehyde (6) and the tricyclic compound 7 were synthesized very efficiently using (4) as intermediate.

Protecting-group-free synthesis of chokols.

As a result of a combined theoretical and experimental study, we describe a two-step protocol for the preparation of an optically pure, multifunctional, cyclopentanic core shared by a number of natural products. This process is based on a hitherto unreported Ti(III)-mediated diastereoselective cyclization in which the hydroxy-directed template effect played by the Ti(III) species was found to be crucial for the stereoselective outcome of the reaction. The viability of this concept was confirmed with the first protecting-group free synthesis of three enantiopure chokols, namely, chokols K, E, and B.

Efficient synthesis of the anticancer ß-elemene and other bioactive elemanes from sustainable germacrone.

Highly efficient preparations of anticancer ß-elemene and other bioactive elemanes were carried out using the natural product germacrone as a renewable starting material. The syntheses were achieved in only 3-5 steps with excellent overall yields (43-54%). An enantioselective approach to these molecules is also described.

Differentiation between microcystin contaminated and uncontaminated fish by determination of unconjugated MCs using an ELISA anti-Adda test based on receiver-operating characteristic curves threshold values: application to Tinca tinca from natural ponds.

The aim of this study was to evaluate whether the enzyme-linked immunosorbent assay (ELISA) anti-Adda technique could be used to monitor free microcystins (MCs) in biological samples from fish naturally exposed to toxic cyanobacteria by using receiver operating characteristic (ROC) curve software to establish an optimal cut-off value for MCs. The cut-off value determined by ROC curve analysis in tench (Tinca tinca) exposed to MCs under laboratory conditions by ROC curve analysis was 5.90-µg MCs/kg tissue dry weight (d.w.) with a sensitivity of 93.3%. This value was applied in fish samples from natural ponds (Extremadura, Spain) in order to asses its potential MCs bioaccumulation by classifying samples as either true positive (TP), false positive (FP), true negative (TN), or false negative (FN). In this work, it has been demonstrated that toxic cyanobacteria, mainly Microcystis aeruginosa, Aphanizomenon issatchenkoi, and Anabaena spiroides, were present in two of these ponds, Barruecos de Abajo (BDown) and Barruecos de Arriba (BUp). The MCs levels were detected in waters from both ponds with an anti-MC-LR ELISA immunoassay and were of similar values (between 3.8-6.5-µg MC-LR equivalent/L in BDown pond and 4.8-6.0-µg MC-LR equivalent/L in BUp). The MCs cut-off values were applied in livers from fish collected from these two ponds using the ELISA anti-Adda technique. A total of 83% of samples from BDown pond and only 42% from BUp were TP with values of free MCs higher than 8.8-µg MCs/kg tissue (d.w.).

Intra-laboratory assessment of method accuracy (trueness and precision) by using validation standards.

Assessment of accuracy of analytical methods is a fundamental stage in method validation. The use of validation standards enables the assessment of both trueness and precision of analytical methods at the same time. Procedures of intra-laboratory testing of method accuracy using validation standards are outlined and discussed.

Splitting of beta-carotene in the sexual interaction of Phycomyces.

Two new 7-carbon compounds, 1 and 2, have been found in the culture medium of Phycomyces blakesleeanus. A genetic test showed that they derive from beta-carotene. These new molecules represent the missing link that proves that beta-carotene is split into fragments of 18, 15 and 7 carbon fragments, each head of a separate family of apocarotenoids.

Weakening C-O bonds: Ti(III), a new reagent for alcohol deoxygenation and carbonyl coupling olefination.

Investigations detailed herein, including density functional theory (DFT) calculations, demonstrate that the formation of either alkoxy- or hydroxy-Ti(III) complexes considerably decreases the energy of activation for C-O bond homolysis. As a consequence of this observation, we described two new synthetic applications of Nugent's reagent in organic chemistry. The first of these applications is an one-step methodology for deoxygenation-reduction of alcohols, including benzylic and allylic alcohols and 1,2-dihydroxy compounds. Additionally, we have also proved that Ti(III) is capable of mediating carbonyl coupling-olefination. In this sense, and despite the fact that for over 35 years it has been widely accepted that either Ti(II) or Ti(0) was the active species in the reductive process of the McMurry reaction, the mechanistic evidence presented proves the involvement of Ti(III) pinacolates in the deoxygenation step of the herein described Nugent's reagent-mediated McMurry olefination. This observation sheds some light on probably one of the mechanistically more complex transformations in organic chemistry. Finally, we have also proved that both of these processes can be performed catalytically in Cp(2)TiCl(2) by using trimethylsilyl chloride (TMSCl) as the final oxygen trap.