Phenolic compounds in peach (Prunus persica) cultivars at harvest and during fruit maturation

Six peach and six nectarine cultivars were evaluated for the phenolic content in their pulp and peel tissues. Chlorogenic acid, catechin, epicatechin, rutin and cyanidin-3-glucoside were detected as the main phenolic compounds of ripened fruits. The concentration was always higher in peel tissue, with average values ranging from 1 to 8 mg g⁻¹ dry weight (DW) depending on cultivar. Of the tested varieties, the white-flesh nectarine 'Silver Rome' emerged as the cultivar with the highest amount of total phenolics. Phenolic compounds were also profiled during fruit growth and ripening in the yellow nectarine cv. 'Stark Red Gold', which showed a decreasing concentration during fruit development in both peel and pulp tissues. Average amounts of total phenolics were approximately 25 mg g⁻¹ DW 60 days after full bloom and decreased to 3 mg g⁻¹ DW at ripening in pulp tissue. Differences among peel and pulp composition show the different dietetic and antioxidant potential of fruits consumed unpeeled and peeled.     

Detection of Leishmania DNA in phlebotomines captured in Campo Grande, Mato Grosso do Sul, Brazil

Over the past years, leishmaniases have become a public health issue in the Brazilian state of Mato Grosso do Sul, particularly in Campo Grande, the state capital. The purpose of this study was to detect the presence of Leishmania DNA in the population of phlebotomine sandflies using DNA amplification by PCR. Insect captures were carried out from 4 pm. to 7 am for 4 consecutive days each month from October 2005 to September 2006 in 16 neighborhoods located in 7 urban regions of Campo Grande. Traps were placed indoors and in the vicinity of households. As many as 971 males and 203 females were collected. One hundred and five naturally fed females were identified and grouped as 1- to 4-specimen pools. DNA extraction was carried out using whole insects. Lutzomyia longipalpis predominated, accounting for 99.15% of the phlebotomines captured. Also found was Nyssomyia whitmani, the vector of tegumentary leishmaniasis. Abundance was greatest in the vicinity of households (69.8% of the phlebotomines captured). As revealed by PCR, parasites were present in 1.9% of the Leishmania spp. specimens investigated and confirmed for visceral leishmaniasis.     

Ethical,legal and social aspects of the approach in Sudan

The global malaria situation, especially in Africa, and the problems frequently encountered in chemical control of vectors such as insecticide resistance, emphasize the urgency of research, development and implementation of new vector control technologies that are applicable at regional and local levels. The successful application of the sterile insect technique (SIT) for the control of the New World screwworm Cochliomyia hominivorax and several species of fruit flies has given impetus to the use of this method for suppression or elimination of malaria vectors in some areas of Africa including Northern State of Sudan. The research and development phase of the Northern State feasibility study has been started. Sudanese stakeholders are working side-by-side with the International Atomic Energy Agency in the activities of this important phase. Several ethical, legal and social issues associated with this approach arose during this phase of the project. They need to be seriously considered and handled with care. In this paper, these issues are described, and the current and proposed activities to overcome potential hurdles to ensure success of the project are listed.     

Xanthomonas maltophilia CBS 897.97 as a source of new 7beta- and 7alpha-hydroxysteroid dehydrogenases and cholylglycine hydrolase: improved biotransformations of bile acids

The paper reports the partial purification and characterization of the 7beta- and 7alpha-hydroxysteroid dehydrogenases (HSDH) and cholylglycine hydrolase (CGH), isolated from Xanthomonas maltophilia CBS 897.97. The activity of 7beta-HSDH and 7alpha-HSDH in the reduction of the 7-keto bile acids is determined. The affinity of 7beta-HSDH for bile acids is confirmed by the reduction, on analytical scale, to the corresponding 7beta-OH derivatives. A crude mixture of 7alpha- and 7beta-HSDH, in soluble or immobilized form, is employed in the synthesis, on preparative scale, of ursocholic and ursodeoxycholic acids starting from the corresponding 7alpha-derivatives. On the other hand, a partially purified 7beta-HSDH in a double enzyme system, where the couple formate/formate dehydrogenase allows the cofactor recycle, affords 6alpha-fluoro-3alpha, 7beta-dihydroxy-5beta-cholan-24-oic acid (6-FUDCA) by reduction of the corresponding 7-keto derivative. This compound is not obtainable by microbiological route. The efficient and mild hydrolysis of glycinates and taurinates of bile acids with CGH is also reported. Very promising results are also obtained with bile acid containing raw materials.     

Hydrolyses and transglycosylations performed by purified alpha-D-glucosidase of the marine mollusc Aplysia fasciata

The purification and characterisation of the alpha-glucosidase from the marine mollusc Aplysia fasciata are reported. Overall substrate specificity of the pure enzyme for both hydrolytic and transglycosylation reactions was studied. Remarkable characteristics of this enzyme are indicated by the results of the interesting survey of transglycosylation reactions reported: pyridoxine glucosylation, synthesis of chromophoric (pNP) di- and trisaccharides, glucosylation of cellobiose and sucrose. For these last two acceptors both the yields of reactions and the concentrations of products are comparable to those obtained using glycosyl transferases; in addition, synthesis of pyridoxine and chromophoric glycosides were still possible using a 1:1 ratio maltose:acceptor which is a very interesting characteristic from a synthetic point of view (effortless purification, productivity of each reaction batch, etc.).     

Effect of Cu2+ on the oxidative folding of synthetic maurotoxin in vitro

Maurotoxin (MTX) is a 34-mer scorpion toxin cross-linked by four disulphide bridges that acts on various K+ channel types. It folds according to an alpha/beta scaffold, i.e., a helix connected to a two stranded beta-sheet by two disulphide bridges. In a former study, various parameters that affect the oxidation and folding of the reduced form of synthetic MTX were investigated in vitro. It was found that MTX achieves its final 3-D structure by evolving over time through a series of oxidation intermediates, from the least to the most oxidized species. MTX oxidative intermediates can be studied by iodoacetamide alkylation of free cysteine residues followed by mass spectrometry analysis. Here, we have analysed the effect of Cu2+ (0.1 to 50 mM) on the kinetics of MTX oxidative folding and found that it dramatically speeds up the formation of the four-disulphide bridged, native-like, MTX (maximal production within 30 minutes instead of > 60 hours). This catalysing effect of Cu2+ was found to be concentration-dependent, reaching a plateau at 10 mM copper ions. Cu2+ was also found to prevent the slow transition of a three disulphide-bridged MTX intermediate towards the final four disulphide-bridged product (12% of total MTX). The data are discussed in light of the potential effects of Cu2+ on MTX secondary structure formation, disulphide bridging and peptidyl prolyl cis-trans isomerization.     

The glucose transport system of the hyperthermophilic anaerobic bacterium Thermotoga neapolitana

The glucose transport system of the extremely thermophilic anaerobic bacterium Thermotoga neapolitana was studied with the nonmetabolizable glucose analog 2-deoxy-D-glucose (2-DOG). T. neapolitana accumulated 2-DOG against a concentration gradient in an intracellular free sugar pool that was exchangeable with external source of energy, such as pyruvate, and was inhibited by arsenate and gramicidin D. There was no phosphoenolpyruvate-dependent phosphorylation of glucose, 2-DOG, or fructose by cell extracts or toluene-treated cells, indicating the absence of a phosphoenolpyruvate:sugar phosphotransferase system. These data indicate that D-glucose is taken up by T. neapolitana via an active transport system that is energized by an ion gradient generated by ATP, derived from substrate-level phosphorylation.     

The active site of Sulfolobus solfataricus aspartate aminotransferase

Aspartate aminotransferase from the archaebacterium Sulfolobus solfataricus binds pyridoxal 5' phosphate, via an aldimine bond, with Lys-241. This residue has been identified by reducing the enzyme in the pyridoxal form with sodium cyanoboro[3H]hydride and sequencing the specifically labeled peptic peptides. The amino acid sequence centered around the coenzyme binding site is highly conserved between thermophilic aspartate aminotransferases and differs from that found in mesophilic isoenzymes. An alignment of aspartate aminotransferase from Sulfolobus solfataricus with mesophilic isoenzymes, attempted in spite of the low degree of similarity, was confirmed by the correspondence between pyridoxal 5' phosphate binding residues. Using this alignment it was possible to insert the archaebacterial aspartate aminotransferase into a subclass, subclass I, of pyridoxal 5' phosphate binding enzymes comprising mesophilic aspartate aminotransferases, tyrosine aminotransferases and histidinol phosphate aminotransferases. These enzymes share 12 invariant amino acids most of which interact with the coenzyme or with the substrates. Some enzymes of subclass I and in particular aspartate aminotransferase from Sulfolobus solfataricus, lack a positively charged residue, corresponding to Arg-292, which in pig cytosolic aspartate aminotransferase interacts with the distal carboxylate of the substrates (and determines the specificity towards dicarboxylic acids). It was confirmed that aspartate aminotransferase from Sulfolobus solfataricus does not possess any arginine residue exposed to chemical modifications responsible for the binding of omega-carboxylate of the substrates. Furthermore, it has been found that aspartate aminotransferase from Sulfolobus solfataricus is fairly active when alanine is used as substrate and that this activity is not affected by the presence of formate. The KM value of the thermophilic aspartate aminotransferase towards alanine is at least one order of magnitude lower than that of the mesophilic analogue enzymes.     

Phorbol ester tumor promoter modulation of alloantigen-specific T lymphocyte responses

The phorbol diester tumor promoter PMA was studied for its effects on T lymphocyte activation in allogeneic human mixed lymphocyte cultures. Lymphocyte proliferation was significantly enhanced in cultures with PMA at concentrations greater than or equal to 12.5 ng/ml, and interferon-gamma production was synergistically increased in cultures with PMA at concentrations greater than or equal to 5 ng/ml. In contrast, the generation of HLA-specific cytotoxic T lymphocyte activity was completely inhibited in cultures with PMA at concentrations greater than or equal to 5 ng/ml. Fluorescence-activated cell sorter analysis with OKT mouse monoclonal antibodies indicated these PMA effects on lymphocyte activation are associated with an increase in the number of activated OKT4-positive T lymphocytes.