Synthesis of some racemicγ-fluoro-α-amino acids

Summary Versatile three-step procedures for syntheses of seven racemi-fluoro-a-amino acids are described. Alkylation oftert-butyl N-(diphenylmethylene) glycinate with 1-bromo-2-fluoroalkanes gave N-protected aminoacid esters both in anhydrous medium using lithium-diisopropylamide as base at low temperature or in a two phase system of 50% aqueous sodium hydroxide and methylene chloride with triethylbenzylammonium chloride as the phase transfer catalyst at room temperature. Subsequent two-step deprotection with citric acid and hydrochloric acid gave the title compounds in 13–33% overall yields.Dedicated to Professor Dr.mult., Dr.h.c. Alois Haas on the occasion of his 65th birthday     

Syntheses ofγ-fluoro-α-amino acids

Summary Methods for the synthesis of racemic and optically active title compounds are presented. Key step of these four-step procedures is the alkylation with 1-bromo-2-fluoroalkanes of glycine-ester-derived imines in anhydrous medium using lithium diisopropylamide as a base at low temperature or phase transfer catalyzed alkylation with 50% NaOH and triethylbenzylammoniumchloride as the phase transfer catalyst, respectively. Subsequent three-step deprotection gave the free acids in 13–33% overall yield. Deracemization of-fluoro--aminobutyric acid methyl and ethyl esters with-chymotrypsin was shown to give the (–)-enantiomers of the esters and (+)--fluoro--aminobutyric acid in >98% ee, while from thetert-butylester the opposite stereochemical result was observed giving the (–)-acid with 88% ee. Optically active-fluoro--amino acids were synthesized alternatively by phase transfer catalysis with N-benzyl-cinchonium chloride or using an auxiliary-directed asymmetric alkylation of the imine derived from (R)-(+)-camphor or (R)-(+)-2-hydroxypinan-3-one. These processes gave different enantiomers of-fluoro--aminobutyric acid via a monomeric lithium enolate in the first or a dimeric lithium enolate in the second case, respectively. The enantiomeric excess can be improved by lithium/magnesium exchange.     

Book reviews

Ohne Zusammenfassung

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Book reviews

     

Genetic dissection of Al tolerance QTLs in the maize genome by high density SNP scan

Background

Aluminum (Al) toxicity is an important limitation to food security in tropical and subtropical regions. High Al saturation on acid soils limits root development, reducing water and nutrient uptake. In addition to naturally occurring acid soils, agricultural practices may decrease soil pH, leading to yield losses due to Al toxicity. Elucidating the genetic and molecular mechanisms underlying maize Al tolerance is expected to accelerate the development of Al-tolerant cultivars.

Results

Five genomic regions were significantly associated with Al tolerance, using 54,455 SNP markers in a recombinant inbred line population derived from Cateto Al237. Candidate genes co-localized with Al tolerance QTLs were further investigated. Near-isogenic lines (NILs) developed for ZmMATE2 were as Al-sensitive as the recurrent line, indicating that this candidate gene was not responsible for the Al tolerance QTL on chromosome 5, qALT5. However, ZmNrat1, a maize homolog to OsNrat1, which encodes an Al³⁺ specific transporter previously implicated in rice Al tolerance, was mapped at ~40 Mbp from qALT5. We demonstrate for the first time that ZmNrat1 is preferentially expressed in maize root tips and is up-regulated by Al, similarly to OsNrat1 in rice, suggesting a role of this gene in maize Al tolerance. The strongest-effect QTL was mapped on chromosome 6 (qALT6), within a 0.5 Mbp region where three copies of the Al tolerance gene, ZmMATE1, were found in tandem configuration. qALT6 was shown to increase Al tolerance in maize; the qALT6-NILs carrying three copies of ZmMATE1 exhibited a two-fold increase in Al tolerance, and higher expression of ZmMATE1 compared to the Al sensitive recurrent parent. Interestingly, a new source of Al tolerance via ZmMATE1 was identified in a Brazilian elite line that showed high expression of ZmMATE1 but carries a single copy of ZmMATE1.

Conclusions

High ZmMATE1 expression, controlled either by three copies of the target gene or by an unknown molecular mechanism, is responsible for Al tolerance mediated by qALT6. As Al tolerant alleles at qALT6 are rare in maize, marker-assisted introgression of this QTL is an important strategy to improve maize adaptation to acid soils worldwide.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-153) contains supplementary material, which is available to authorized users.     

Aspidosperma (Apocynaceae) plant cytotoxicity and activity towards malaria parasites. Part II: experimental studies with Aspidosperma ramiflorum in vivo and in vitro

Several species of Aspidosperma plants are used to treat diseases in the tropics, including Aspidosperma ramiflorum, which acts against leishmaniasis, an activity that is experimentally confirmed. The species, known as guatambu-yellow, yellow peroba, coffee-peroba andmatiambu, grows in the Atlantic Forest of Brazil in the South to the Southeast regions. Through a guided biofractionation of A. ramiflorum extracts, the plant activity against Plasmodium falciparum was evaluated in vitro for toxicity towards human hepatoma G2 cells, normal monkey kidney cells and nonimmortalised human monocytes isolated from peripheral blood. Six of the seven extracts tested were active at low doses (half-maximal drug inhibitory concentration < 3.8 µg/mL); the aqueous extract was inactive. Overall, the plant extracts and the purified compounds displayed low toxicity in vitro. A nonsoluble extract fraction and one purified alkaloid isositsirikine (compound 5) displayed high selectivity indexes (SI) (= 56 and 113, respectively), whereas compounds 2 and 3 were toxic (SI < 10). The structure, activity and low toxicity of isositsirikine in vitro are described here for the first time in A. ramiflorum, but only the neutral and precipitate plant fractions were tested for activity, which caused up to 53% parasitaemia inhibition of Plasmodium berghei in mice with blood-induced malaria. This plant species is likely to be useful in the further development of an antimalarial drug, but its pharmacological evaluation is still required.     

Biodeterioration of Mayan buildings at uxmal and tulum,Mexico

Uxmal and Tulum are two important Mayan sites in the Yucatan peninsula. The buildings are mainly composed of limestone and grey/black discoloration is seen on exposed walls and copious greenish biofilms on inner walls. The principal microorganisms detected on interior walls at both Uxmal and Tulum were cyanobacteria; heterotrophic bacteria and filamentous fungi were also present. A dark‐pigmented mitosporic fungus and Bacillus cereus, both isolated from Uxmal, were shown to be acidogenic in laboratory cultures. Cyanobacteria belonging to rock‐degrading genera Synechocystis and Gloeocapsa were identified at both sites. Surface analysis previously showed that calcium ions were present in the biofilms on buildings at Uxmal and Tulum, suggesting the deposition of biosolubilized stone. Apart from their potential to degrade the substrate, the coccoid cyanobacteria supply organic nutrients for bacteria and fungi, which can produce organic acids, further increasing stone degradation.     

Quantitative measurements of activity ofAedes Aegypti (L.) (Culicidæ: Diptera) in response to changes in the hygrothermal environment

Reactions of A. AEGYPTI females to hygrothermal changes in the atmosphere were measured quantitatively by calculating the time between quantal cumulative response curves for take-off and settling. Reactions were compared among experimental groups derived from a stabilized colony and conditioned to a standard dark environment of 23 C and 100 per cent relative humidity.Atmospheric moisture was the limiting factor for duration of activity and for the initiation of the take-off or settling responses. Activity in response to hygrothermal conditions was mediated internally in relation to starvation and increased after a blood meal. Light operated interdependently with temperature and humidity to stimulate take-off after a period of rest.Activity appeared to be governed by the ability of mosquitoes to adapt to changes in the internal-external water balance.When the environmental water-vapor pressure remained near the adaptation level activity increased or decreased directly with temperature. Starving mosquitoes did not tolerate sudden decreases in water-vapor pressure, but did so within an hour following a blood meal. Large increases in watervapor pressure induced the resting state, which, if prolonged, took the form of prostrate immobilization. Maximum activity was related to the maximum hygrothermal change to which the mosquito could adapt without injury or without severe physiological consequences in water balance.

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Zusammenfassung Die Reaktionen von A. AEGYPTI Weibchen bei Änderungen der Temperatur und Feuchte in der AtmosphÄre wurden quantitativ durch die Reaktionskurven für Abflug und Aufsetzen gemessen. Die Reaktionen wurden mit denen von Mücken aus einer an Dunkelheit, 23 C und 100% R.F. adaptierten Standardkolonie verglichen. Der limitierende Faktor für die Dauer der AktivitÄt und für die Einleitung der Abflug und Landereaktion war die Feuchte. Bei der Änderung der Temperatur-Feuchte Bedingungen wurde die AktivitÄt durch Hunger beeinflusst und war nach einer Blutmahlzeit grosser. Licht stimulierte unabhÄngig von Temperatur und Feuchte den Abflug nach einer Ruheperiode. Die AktivitÄt der Mücken schien von der FÄhigkeit bestimmt zu werden, sich an Anderungen im intern-externen Wassergleichgewicht anzupassen.Wenn die Feuchte in der Höhe des Adaptationswertes lag, fiel oder stieg die AktivitÄt proportional zur Temperatur. Im Hunger konnten die Mücken einen plötzlichen Feuchteabfall nicht vertragen, wohl aber innerhalb einer Stunde nach einer Blutmahlzeit. Starker Anstieg der Feuchte leitete eine Ruhephase ein, die bei lÄngerem Bestehen zur EntkrÄftung führte. Maximale AktivitÄt war mit maximalen Temperatur-Feuchtewechsel verbundenen den die Mücken sich ohne ernste Folgen im Wasserhaushalt anpassen konnten.

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Resume La réaction de femelles d'A. AEGYPTI aux changements d'humidité et de température de l'atmosphère a été déterminée quantitativement au moyen des courbes de réaction fixant les intervalles de temps qui correspondent à l'envol et au retour. Les réactions ont été comparées à celles de moustiques appartenant à une colonie standard à l'obscurité, à 23 C de température et 100% d'humidité relative.L'humidité atmosphérique fut le facteur limitatif quant à la durée de l'activité et le déclenchement des réactions de repos et d'envol. L'activité en fonction des conditions d'humidité et de température fut influenÇable par la faim et montra un accroissement dans le cas de moustiques nourris de sang. Après une période de repos, une stimulation de l'envol eut lieu sous l'influence de la lumière, quelles que fussent les conditions de température et d'humidité.On constata que l'activité des moustiques fut dominée per leur faculté d'adaptation aux changements internes-externes de l'équilibre d'eau. Au cas où la pression de la vapeur d'eau environnante fut maintenue à proximité du niveau d'adaptation, l'activité augments ou décrût proportionellement à la température. A l'état affamé, les moustiques ne supportèrent pas les chutes rapides de la pression de la vapeur d'eau, mais, après qu'ils se fussent nourris de sang, le contraire eut lieu pendant une heure. Une forte augmentation de la pression de la vapeur d'eau provoqua le retour à l'état de repos, qui finit par prendre la forme d'un état d'épuisement. On constata que l'activité maximum correspond au changement maximum des conditions d'humidité et de température auquel les moustiques puissent s'adapter sans que soit porté préjudice à leur équilibre d'eau.

     

Further characterization of the binding of human recombinant interleukin 2 to heparin and identification of putative binding sites

We have previously provided compelling evidence that human recombinant interleukin 2 (IL-2) binds to the sulfated polysaccharides heparin, highly sulfated heparan sulfate and fucoidan. Here we show that IL-2 binding is dependent on heparin chain length, but with fragments as small as 15-mers retaining binding activity. The addition of exogenous heparin has no effect on the in vitro biological activity of IL-2. In addition soluble IL-2 receptor alpha and beta polypeptides do not compete with heparin for the binding of IL-2. IL-2 bound by heparin is still recognized by two IL-2 specific monoclonal antibodies, 3H9 and H2- 8, whose epitopes lie in the amino terminal region. Murine IL-2 unlike its human counterpart fails to bind to heparin. Human IL-2 analogs with single amino acid substitutions at positions Lys43, Thr51, and Gln126 analogs no longer bind to heparin. By contrast the Arg38Ala analog retains heparin full heparin binding activity. These experimental findings together with molecular modeling studies suggest two putative heparin binding sites on human IL-2, one involving four basic residues, Lys48, Lys49, Lys54, and His55, and the other being a discontinuous site comprising Lys43, Lys64, Arg81, and Arg83. Neither of these two clusters is completely conserved in murine IL-2. Overall our data suggest that the binding of human IL-2 to heparin and heparan sulfate does not interfere with IL-2/IL-2 receptor interactions. Therefore, binding to glycosaminoglycan may be a mechanism for retaining the cytokine in an active form close to its site of secretion in the tissue, thus favoring a paracrine role for IL-2.      

Brazilian Angiostrongylus cantonensis haplotypes,ac8 and ac9, have two different biological and morphological profiles

Angiostrongylus cantonensis is the etiologic agent of eosinophilic meningoencephalitis in humans. Cases have been recorded in many parts of the world, including Brazil. The aim of this study was to compare the differences in the biology and morphology of two different Brazilian haplotypes of A. : ac8 and ac9. A significantly larger number of L1 larvae eliminated in the faeces of rodents at the beginning of the patent period was observed for ac9 haplotype and compared to the total of L1 larvae eliminated, there was a significant difference between the two haplotypes. The ac9 haplotype showed a significant difference in the proportion of female and male specimens (0.6:1), but the same was not observed for ac8 (1.2:1). The morphometric analysis showed that male and female specimens isolated from ac8 haplotype were significantly larger with respect to body length, oesophagus length, spicule length (male) and distance from the anus to the rear end (female) compared to specimens from ac9. The morphological analysis by light microscopy showed little variation in the level of bifurcations at the lateral rays in the right lobe of the copulatory bursa between the two haplotypes. The biological, morphological and morphometric variations observed between the two haplotypes agree with the observed variation at the molecular level using the cytochrome oxidase subunit I marker and reinforce the possible influence of geographical isolation on the development of these haplotypes.