Mechanisms of karyotype differentiation in Cassidinae sensu lato (Coleoptera,Polyphaga, Chrysomelidae) based on seven species of the Brazilian fauna and an overview of the cytogenetic data

Among the subfamilies of Chrysomelidae, Cassidinae sensu lato (s.l.) includes 6000 species distributed in 43 tribes. Approximately 100 of these species were cytogenetically analyzed and most of them presented 2n = 18 = 16 + Xy_p, which was smaller than 2n = 20 = 18 + Xy_p considered basal for Polyphaga. However, some groups of species presented maintenance of the basal diploid number and others showed increase in this number. Certain species of the latter group also exhibited variation in the type of sex chromosome system (SCS). Considering the recent taxonomic revision accomplished for the Cassidinae s.l. species, the existence of phylogenetic relationship for some species of this subfamily, the high diversity of species of this group in the Neotropical region, and the low number of Cassidinae s.l. species karyotyped so far, the aim of the present work was to establish the main mechanisms involved in the karyotype evolution of this subfamily through the study of seven species of the Brazilian fauna and overview of the cytogenetic data. The individuals were collected in southeast and south of Brazil. The chromosomal preparations obtained from embryo and testes of adult males were stained with Giemsa solution. The species Agroiconota inedita (2n = 42 = 40 + Xy_p), Charidotella (s.str.) immaculata (2n = 22 = 20 + Xy_p), Charidotella (s.str.) sexpunctata (2n = 22 = 20 + Xy_p), and Stolas chalybaea (2n = 24 = 22 + Xy_p) revealed diploid number higher than that established as basal for Polyphaga and biarmed chromosomes. The karyotype of Cteisella confusa, Deloyala cruciata, and Metriona elatior showed the chromosomal formulae 2n = 18 = 16 + Xy_p considered modal for Cassidinae s.l. and biarmed chromosomes. The seven species exhibited easily identified sex chromosomes due to their size and/or morphology. The analysis of meiotic cells of all the species showed pachytenes with a positively heteropycnotic block probably corresponding to the sex chromosomes; diplotenes with a high number of bivalents with two chiasmata and sex chromosomes in a parachute configuration, and metaphases II that confirmed the chromosomal morphology, the type of SCS, and the regular segregation of all chromosomes. The data regarding to the number and morphology of the chromosomes, their behaviour during meiosis, and type of SCS were inedit for the majority of these species. In relation to the all Cassidinae s.l. species that presented SCS of the Xy_p type, A. inedita was that with the highest diploid number. Furthermore, this work reported for the first time the cytogenetic information of representatives of the genera Cteisella and Metriona. Taking into account the phylogenetic and cytogenetic data of Cassidinae s.l. species, the karyotype differentiation of this group seems to have occurred from the basal karyotype of Polyphaga by decrease in the chromosome number and subsequent increase in this number. Pericentric inversion, centric fusion and fission seem to have been the main mechanisms that promoted the evolution of the autosomes. However, in the sex chromosome evolution, the mechanisms involved were centric fission and/or chromosomal translocation.     

Influence of nutrient medium composition on in vitro growth, polyphenolic content and antioxidant activity of Alchemilla mollis

Alchemilla mollis (Buser) Rothm. (Rosaceae) is a high-mountain medicinal plant growing in the Balkan Peninsula, with only one population in Bulgaria. Alchemilla plants (Lady's mantle) are commonly used in traditional medicine for treatment of many gynecological diseases. The commercial drugs "Herba Alchemillae" induce a rapid regeneration of skin epithelium and have styptic and anti-inflammatory actions. Because of the high content of phenolic compounds (tannins and flavonoids) and the ecological plasticity of the species, field cultivation or in vitro biomass production of A. mollis are possible alternatives to its collection from nature. Four MS based nutrient media differing in the concentration of the minerals and supplemented with alpha-naphthaleneacetic acid (NAA) and benzylaminopurine (BAP) were tested in order to examine their influence on the shoot multiplication effectiveness and the antioxidant activity of A. mollis, and also the possible relation between these parameters under the conditions of in vitro culture. The cultures grown for two months on these media differed significantly in their multiplication rates (p < 0.001), as well as in their morphological features--height, leaf color and root development. Methanol extracts of in vitro cultivated and ex vitro adapted and acclimated on Vitosha Mt. (1500 m a. s. l.) plants were analyzed for tannin and flavonoid content and for free radical scavenging activity. The contents of flavonoids and tannins in the in vitro cultures of A. mollis cultivated on the four tested media differed significantly (p < 0.05). The highest flavonoid content was found in the shoots cultivated on the control MS medium, as well as in the ex vitro adapted plants. The antioxidant activity of the in vitro cultures correlated positively with the concentrations of the PGRs in the respective media, and the ex vitro adapted plants had the highest antioxidant activity (IC50 13.1 +/- 1.9 microg/mL) commensurable with that of the commercial antioxidant butylated hydroxytoluene (BHT) used as a positive control, with an IC50 of 12.65 microg/mL.     

Supercritical fluid extraction of cynaropicrin and 20-hydroxyecdysone from Leuzea carthamoides DC

Leuzea carthamoides is an adaptogenic plant containing biologically active compounds as ecdysteroids and guaianolide-type sesquiterpene lactones, conventionally extracted from the plant with ethanol. It may be a potential source of the mentioned natural compounds. Ethanol-modified near-critical CO(2) was used as selective solvent with the aim to increase the level of 20-hydroxyecdysone in the extract from L. carthamoides roots and to remove selectively cynaropicrin, a sesquiterpene lactone of bitter taste, from the leaves. The extraction conditions were varied (pressure 20-28 MPa, temperature 40-60 degrees C, ethanol concentration in the solvent 0-7.1%) and the extraction yield and extract composition were compared with the results of ethanolic extraction. The supercritical fluid extraction (SFE) from finely powdered plant was controlled by phase equilibrium. Cynaropicrin was quantitatively removed from the leaves where 89% of 20-hydroxyecdysone was retained. The extraction yield of 20-hydroxyecdysone from roots with ethanol-modified CO(2 )was lower by 30% than with ethanol but its concentration in the extract was higher by 67%.     

One-Dimensional Lorentz Gas with Rotating Scatterers: Exact Solutions

The simplest solutions (orbits) to the recently introduced Lorentz gas with rotating scatterers are found by considering its one-dimensional one-particle reduction. This model has only one parameter which can be viewed as the amount of energy transfer between the scatterers and the particle during a collision. Exact solutions of the system are found for several values of this parameter. For some of these values, the dynamics is shown to be in many respects similar to the dynamics of the deterministic Lorentz lattice gases.     

Inverses depending holomorphically on a parameter in a Banach space

We show that the existence of a right inverse at each point for a holomorphic mapping from a pseudo-convex domain in a Banach space with an unconditional basis into a unital Banach algebra implies the existence of a holomorphic right inverse. Variations of this result are given.     

Validation of ELISA methods for search and quantification of β-n-methylamino-l-alanine in water and fish tissue

Water basins with low hydrodynamic activities can promote the growth and increase in algal biomass due to eutrophication, and toxic cyanobacteria species might then produce metabolites hazardous to human health. Over the last decade, a neurotoxic non-protein amino acid, (2S)-2-amino-3-(methylamino) propanoic acid, known as β-N-methylamino-L-alanine (BMAA), has become of particular interest because it has been hypothesised to be involved in progressive human neurodegenerative pathologies. This toxin can be found both in algal cells and free in water, as well as in some foods of aquatic and terrestrial origin. Analytical methods used for BMAA are often based on chromatography coupled with mass spectrometry, although these techniques involve long and expensive analysis. As the availability of a faster and cheaper screening method would be useful, we tested the only available Enzyme-Linked Immunosorbent Assay kit for BMAA evaluation and validated methods to verify their reliability for the analysis of water and fish muscle. For both matrices, we determined adequate selectivity and repeatability (relative standard deviation < 6%), with recoveries from 70% to 83% at the tested spiking levels; the methods were also robust. These data appear in contrast to a previous evaluation carried out on the same kit in 2013, although this might depend on an improvement to the kit performance. We can conclude that a preliminary determination of BMAA in water, and also in fish tissue after an adequate extraction procedure, can be performed efficiently with the tested kit, which provides for easier monitoring of this dangerous toxin.     

Oxidation of a P−C Bond under Mild Conditions

The reactivity of phosphenium dication [(Ph₃P)₂C‐P‐NiPr₂]²⁺, 1²⁺ , towards pyridine N‐oxide (O‐py) has been investigated. The resulting oxophosphonium dication [(Ph₃P)₂C(NiPr₂)P(O)(O‐py)]²⁺, 2²⁺ , was surprisingly stabilized by a less nucleophilic O‐py ligand instead of pyridine (py). This compound was then identified as an analogue of the elusive Criegee intermediate as it underwent oxygen insertion into the P?C bond through a mechanism usually observed for Baeyer–Villiger oxidations. This oxygen insertion appears to be the first example of a Baeyer–Villiger oxidation involving O‐py.     

SYNTHESIS AND ANTICANCER SCREENING OF DIASTEREOISOMERIC 4,5-DIPHENYL- AND 3-METHYL-4,5-DIPHENYLCYCLOPHOSPHAMIDES

Abstract

The four possible diastereoisomers of 4,5-diphenylcyclophosphamides (2–5) and their N-methyl analogues (6–9) have been synthesized via condensation of aminopropanols 10 and 11 (threo- and erythro) with bis-(2-chloroethyl)-phosphoramidic dichloride 12. Anticancer screening tests against L 1210 lymphoid leukaemia in mice did not show any therapeutic activity.